Anatomical Society Summer Meeting in Galway

S1 Chemotherapy‐induced neuropathy in human survivors of cancer

Topp K. ,¹ Mastick J.² and Miaskowski C.¹

¹School of Medicine, University of California, San Francisco, CA, USA and ²School of Nursing, University of California, San Francisco, CA, USA

In 2016, there were 1,685,210 new diagnoses of cancer in the United States, and more than 14 million cases worldwide. With improvements in diagnosis and treatment, approximately 15 million people are now living as survivors of cancer in the U.S. One of the most life‐impactful consequences of treatment is chemotherapy‐induced neuropathy (CIN), experienced by 10–60% of people with a cancer diagnosis treated with taxane or platinum chemotherapy. Although multiple compounds have been tested to prevent or treat persistent CIN, currently only one medication is recommended for use in clinical practice. To improve patient care and outcomes, we completed a systematic investigation of the sensory and motor symptoms, functional status, and quality of life of cancer survivors who did and did not develop CIN following treatment with a platinum and/or taxane compound. The study was approved by the UCSF Committee on Human Subjects. Of 1450 survivors screened, 754 were enrolled, and 623 completed all aspects of the study. Survivors with CIN (68%) had completed chemotherapy ≥3 months before enrollment, had changes in sensation and/or pain in their feet and/or hands and had a rating of ≥ 3 on a 0–10 numeric rating scale for one or more sensations from the Pain Qualities Assessment Scale. Survivors without CIN (32%) had received chemotherapy and did not have persistent changes in sensation and/or pain. Numbness, unpleasant, and tingling in the hands and/or feet were the most common pain qualities described by survivors with CIN. A significantly higher percentage of survivors with CIN had impaired light touch sensations, as measured with Semmes‐Weinstein monofilament testing, and higher vibration thresholds, as measured with a biothesiometer. Additionally, significantly more survivors with CIN demonstrated weakness in grip strength and plantarflexion, and poorer balance, assessed with the Timed Up and Go test and the Fullerton Advanced Balance test. Our findings are in agreement with those of other investigators and demonstrate the significant burden of persistent CIN in a large sample of cancer survivors. This work was supported by grants (Miaskowski) from the National Cancer Institute, the American Cancer Society, and the UCSF‐CTSI.

S2 From neurons to neuroblastoma: the developmental biology of a childhood cancer

O'Keeffe G. , Al Shangiti A. , Hegarty S. and Sullivan A.

Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland

During development the sympathetic branch of the nervous system is assembled by the differentiation of immature neurons (neuroblasts) into mature sympathetic neurons. In rare cases this process does not happen correctly and the cells become cancerous. This type of cancer is called a Neuroblastoma that occurs almost exclusively in infants and children. Neuroblastoma is the most common solid tumor that occurs outside the brain in infants and children and it is responsible for almost 15% of cancer deaths in children. Although the survival rates are good for some patients, those children diagnosed with high‐risk neuroblastoma have survival rates as low as 35%. Moreover relapses are common and these relapses are often treatment resistant. For this reason neuroblastoma remains a significant clinical challenge and the development of novel treatment strategies is essential.

Microarray data suggests that neuroblastoma may arise as a result of a primary failure of neuritogenesis. Therefore we examined distinct anatomical regions of the developing sympathetic nervous system to identify signalling molecules that were physiological relevant signals that promoted neuritogenesis in sympathetic neurons during development. All experiments were carried out with ethical approval. We identified one member of the transforming growth factor‐beta superfamily known as growth differentiation factor‐5 (GDF5) that promoted neuritogenesis in sympathetic mouse explant cultures in vitro and was responsible was directing normal sympathetic innervation in vivo. We next tested whether GDF5 signalling could promote neuritogenesis in human neuroblastoma cell lines with and without amplification of the MYCN gene which is an important prognostic marker of poor survival. Using a modified line intercept we found that GDF5 exerted a dose dependent increase in neuritogensis in neuroblastoma cells that required bone morphogenetic protein receptor signalling. This study suggests that analysing the mechanism through which GDF5 promotes neuritogenesis in neuroblastoma may help identify new therapies or therapeutic targets for the treatment of neuroblastoma in infants and children.

S3 Endometrial and cervical cancers – how do they affect the functional morphology of the human uterine tube?

Hughes B. and Kölle S.

School of Medicine, University College Dublin, Dublin, Ireland

The human fallopian tube is known to be the site of gamete transport, fertilization and early embryogenesis. It is a dynamic structure which in recent years has become associated with the early stages of ovarian cancer. Due to a lack of imaging technologies, most studies on the fallopian tube to date have been performed under in vitro conditions. Therefore the aim of our study was to investigate alterations in tubal morphology and function in response to endometrial and cervical cancer using newly established Live Cell Imaging technology and scanning electron microscopy. Experiments were performed on the ampulla of 18 women undergoing hysterectomy. The study included 10 patients with endometrial carcinoma, three patients with cervical carcinoma and five control patients. Ethical approval was granted for this study by UCD (No LS‐14‐23) and the Mater Hospital (No: 1/378/1645). All samples were obtained with informed patient consent. The fallopian tubes of all patients were investigated immediately after surgery. Ampulla with and without incubation of frozen/thawed sperm was examined qualitatively and quantitatively using a digital video microscopic system allowing for real‐time observations. 1 cm samples of fallopian tube were fixed in 1% glutaraldehyde and analysed using scanning electron microscopy (SEM). Our studies revealed that endometrial and cervical cancer resulted in the fusion of ampullar folds protruding into the tubal lumen. The tubal epithelial cells showed increased numbers of apoptotic and necrotic cells with cellular debris accumulating in the lumen. However, ciliary beat frequency was not altered in cancer patients as compared to controls. Even in stages of early cancer the tubal cells revealed high variations in form and size which was in strong contrast to control patients. Live cell imaging showed the presence of T helper cells attacking cancer cells. The formation of the sperm reservoir was seen, however spermatozoa only bound to healthy uniform cells.

Our results imply that endometrial and cervical cancers are concomitant with severe alterations in form and function of tubal cells. Thus, in vivo imaging of the salpinx might be a valuable tool for the early diagnosis of cancer, for developing new therapeutic concepts or for establishing efficient individual prognoses.

S4 Harnessing light control of kinase activity to regulate neuronal function in rodent neurons

Coffey E.

Turku Centre for Biotechnology, Åbo Akademi University, Biocity, Turku, Finland

Protein kinase regulation of brain function is recognised as important. However global inhibition of kinase activity in the complex cellular landscape that makes up brain tissue reveals only crude information on mechanism. Moreover, individual neurons themselves are highly compartmentalised into molecularly distinct regions: axons, dendrites and dendritic spines, among others. Real understanding of kinase function in neurons requires spatial control of activity. We have previously shown that compartment‐specific targeting of a c‐Jun kinase (JNK) inhibitor to the nucleus or the cytosol, reveals separable functions in death and cytoskeletal regulation. This presentation will describe recent work where we have developed optogenetic and viral approaches that have helped shed new light on subcellular JNK function in the context of structural plasticity of synapses and at the cellular level using viruses, to identify a specific population of neurons from where JNK controls mood.

All work was performed by licensed personnel, with ethical approval from the Animal Experimentation Board in Finland.

S5 Non‐canonical hippocampal circuit organization and function in the mouse

Xu X.

Department of Anatomy and Neurobiology, University of California, Irvine, CA, USA

The hippocampal formation plays essential roles in learning and memory, and spatial navigation. It is traditionally viewed as having a feed‐forward, unidirectional circuit organization which promotes propagation of excitatory processes. While the substantial forward projection from hippocampal CA1 to the subiculum has been very well established, we recently discovered a non‐canonical but significant set of back‐projections comprised of both excitatory and inhibitory elements from the subiculum to CA1 in the mouse. Our quantitative analysis indicates that non‐canonical back projections from the subiculum to CA1 have connectivity strengths similar to the well‐studied medial septal projections to CA1. This discovery suggests previously unconsidered functional roles for direct subicular modulation of hippocampal CA1 circuit activity. In this talk, I will present our recent progress in studying the synaptic circuit organization and function of the subiculum‐CA1 back‐projection pathway in the mouse using recent technological advancements. We hypothesize that subicular‐CA1 back projections are topographically organized and originate from a distinct group of subicular neurons. These neurons integrate strong CA1 and local subicular inputs to exert direct feedback regulation of CA1 circuit activity, thus supporting hippocampus‐associated learning and memory behaviors. To test the hypotheses, we map the topographical relationship of anatomical connections between the subiculum and CA1, using monosynaptic rabies tracing from different CA1 sub‐regions in vivo, and using fast voltage dye imaging in slice preparations, respectively. We also map and compare global circuit input and output connections of CA1‐projecting and other excitatory subicular neurons using retrograde monosynaptic rabies tracing and anterograde directed herpes simplex virus (H129 strain) in intact brains, respectively. In conjunction with behavior analysis and genetically targeted neuronal activation and inactivation, in vivo GCaMP6‐based calcium imaging of CA1 at single‐cell resolution in freely moving animals is used to resolve how CA1‐projecting subicular neurons modulate CA1 place cell activity and how they contribute to location dependent learning and memory. Together, these studies provide new insights into this subicular‐CA1 pathway, and improve our understanding of how the subiculum interacts with CA1 to regulate hippocampal circuit activity and learning and memory behaviors. All animal experiment procedures and protocols including anaesthetic and surgical procedures were approved by the Institutional Animal Care and Use Committee of the University of California, Irvine.

S6 Alkylating histone deacetylase inhibitor treatment in rodent models of MPO‐ANCA vasculitis

Dooley D. ,¹ van Timmeren M. ,² O'Reilly V. ,¹ O'Brien E. ,¹ Fazekas B. ,¹ Hickey F. ,¹ Pusey C. ,³ Tam F. ,³ Mehrling T. ,⁴ Heeringa P.² and Little M.¹

¹Trinity Health Kidney Centre, Trinity Translational Medicine Institute, Trinity College Dublin, Dublin, Ireland; ²Department of Pathology and Medical Biology, University of Groningen, Groningen, Netherlands; ³Renal and Vascular Inflammation Section, Department of Medicine, Imperial College London, UK and ⁴Mundipharma EDO GmbH

Anti‐neutrophil cytoplasmic autoantibody (ANCA)‐associated vasculitis (AAV) is a systemic inflammatory, autoimmune condition that affects the microvasculature. The lungs and kidneys are most frequently affected, leading to lung haemorrhage and glomerulonephritis (GN). When left untreated, AAV has a mortality rate of around 80% in year one and a 25% five‐year mortality rate with current conventional treatments consisting of cyclophosphamide and steroids. However, these therapies do not prevent disease relapse and patients often require long‐term treatment which is associated with severe morbidity. Recently, histone deacetylase inhibitors (HDACi) were shown to have beneficial effects in inflammatory rodent models and have been found to act synergistically with a diverse range of pharmacological agents including cyclophosphamide. EDO‐S101, a small molecule compound developed by Mundipharma EDO GmbH, is an alkylating HDACi fusion molecule which combines the strong DNA damaging effect of bendamustine, with a fully functional pan‐HDACi, vorinostat. In this study, we investigated the effects of EDO‐S101 in two well established rodent models of AAV. These consisted of a passive mouse model of anti‐myeloperoxidase (MPO) IgG‐induced GN and an active rat model of MPO‐ANCA microscopic polyangitis: experimental autoimmune vasculitis (EAV). All experiments were approved by the local ethical committees and performed according to the guidelines described on the protection of animals used for scientific purposes at Trinity College Dublin and University Medical Centre Groningen. Our data indicate that although pre‐treatment with EDO‐S101 reduced circulating leukocyte populations, it did not affect development of anti‐MPO IgG‐induced GN in mice. On the other hand, EDO‐S101 in EAV significantly reduced the degree of lung haemorrhage, severe GN and almost completely abolished crescent formation. EDO‐S101 treatment in EAV also significantly depleted B and T cells compared with vehicle‐treated controls, suggesting a selective effect on the adaptive immune response. Taken together, we have demonstrated that EDO‐S101 is a promising novel therapy for treatment of AAV and operates primarily through its effects on the adaptive immune response to the autoantigen MPO. T. Mehrling is an employee of Mundipharma‐EDO GmbH.

S7 Inner ear mitochondria exhibit structural features that differ between hair cells, afferents and efferents: a rodent study

Lysakowski A.

University of Illinois at Chicago, Chicago, IL, USA

In other organs in the body, the structure of mitochondria and their role in apoptosis and cell death is a topic of intense research interest. Such a framework of structural studies is missing in the inner ear. This study is an attempt to fill that gap by studying hair cell mitochondria for the purpose of addressing hair cell damage and death in mitochondrial‐associated forms of deafness (both non‐syndromic and antibiotic‐induced). We wanted to know if variations in the physical structure and corresponding molecular composition in various sub‐populations of hair‐cell mitochondria could cause mitochondria to be affected differently by ototoxic insults, such as aminoglycoside antibiotics or chemotherapeutics.

Using electron microscope (EM) tomography and 3D‐reconstructions, we have found that mitochondria in rat vestibular endorgans come in different sizes (large, medium and small) and exhibit different internal structures (lamellar vs. tubular cristae). Mitochondria found in hair cells are mostly medium‐sized with lamellar cristae; those found in afferents (both calyces and boutons) are also medium‐sized but they have tubular cristae. Efferent bouton mitochondria are small with tubular cristae. Finally, the largest mitochondria, which have lamellar cristae, are found in the subcuticular plate region in central type I hair cells.

We have analyzed the internal structure of these various types of mitochondria and find differences in crista surface areas and volumes, which affect their ATPase‐carrying capacity. In addition, they appear to be not only tethered to various hair cell organelles and to each other, but are also arranged in particular orientations in relation to these organelles. As previously shown in auditory brainstem mitochondria, the cristae have openings to the inner mitochondrial membrane termed “crista junctions” and there are 1.5–2× more crista junctions on the side of the mitochondria facing toward structures relevant to hair cell function than on the side away from these structures. These crista junctions may be a way of directing ATP and Ca2+ toward energy‐requiring organelles, such as ribbon synapses, stereociliar rootlets, striated organelles, etc. Several short animations will illustrate these structural differences.

All experiments were performed in an ethical manner and approved by the UIC IACUC.

Supported by NIH R21 DC‐013181‐01.

S8 Mesentery; structure, function and role in disease

Coffey J.C.

Centre for Interventions in Infection, Inflammation and Immunity, University of Limerick, Limerick, Ireland

The classic anatomic model of the mesentery described separate mesenteries and absence of a right and left mesocolon. Recent advancements have demonstrated the universal presence of a right and left mesocolon. In turn, the mesentery distal to the duodenojejunal (DJ) flexure becomes a single, continuous structure. It fans out to span the entire intestine and in tandem with the intestine, reaches up to two metres in length. Small intestinal, transverse and sigmoidal regions of mesentery are mobile whilst right and left mesocolic regions are flattened against the posterior abdominal wall.

Reappraisal of mesenteric anatomy prompts a review of the macroscopic anatomy of related structures including the fascia and surrounding peritoneum. Where mesentery flattens against the posterior abdominal wall it does not “insert” into the abdominal wall. Instead, Toldt's connective tissue fascia is interposed between mesentery and true retroperitoneum. The peritoneum is reflected onto and is continuous with the intestinal serosa. At the mesenteric pole of the intestine, the serosa continues medially as mesenteric mesothelium. At medial border of the mesentery, the mesothelium extends as the peritoneal reflection to the abdominal wall.

The intact mesentery can only be fully detached from the posterior abdominal wall by sharp division of the peritoneal reflection, and separation from the underlying fascia. These activities are the cornerstone of mesenteric based colorectal surgery. This aims to detach an intact mesenteric organ with a view to its formal disconnection.

The key properties of the anatomic model are mesenteric, intestinal, fascial and peritoneal continuity and contiguity. These properties hold at all levels distal to the DJ flexure. They explain all flexural anatomy. They hold in all forms of mesenteric congenital anomaly. They occur both across and within species. This anatomic model is thus universally applicable.

Given the mesentery is a substantive and distinct anatomical structure with several essential functions it merits redesignation as an organ. This has considerable clinical utility as it as it improves our understanding of the pathobiology of numerous abdominal and non‐abdominal diseases. It greatly aids in the radiological interpretation of abdominal diseases and in the development of mesenteric‐based treatment strategies for these.

For all living human work ethical approval was obtained from the University Hospital Limerick Ethics Committee. For cadaveric work, all material used was bequeathed to the Medical School, National University of Ireland, Galway, Ireland, for the advancement of medical knowledge. Associated legislation include the Anatomy Act 1832, the Anatomy Act 1871, Health order 1949, Medical Practitioners Act 2007, Ireland.

S9 Using motion capture, virtual reality, and 3D fluoroscopy for clinical reasoning and rehabilitation in humans

Foreman K.B.^1,2,3

¹Department of Physical Therapy and Athletic Training (Primary), University of Utah, Salt Lake City, UT, USA; ²Department of Orthopaedics, University of Utah, Salt Lake City, UT, USA and ³Department of Veteran Affairs, Salt Lake City, UT, USA

Kinetic and kinematic data from human movement can provide insight into clinical decision‐making to augment treatment and rehabilitation for patients with various pathologies and condition. To demonstrate this, several research projects and associated techniques performed at the University of Utah Motion Capture Core Facility will be highlighted. For the presented projects, participants consented and procedures were approved by the Institutional Review Board at the University of Utah. The first project investigated transhumeral loading during advance upper extremity activities of daily living to calculate the axial force, bending, and torsional moments at three virtual amputation levels for the purpose of direct skeletal attachment of upper extremity prosthetics through percutaneous osseointegrated implants. This was accomplished using motion capture techniques that tracked the position of reflective markers adhered to the skin over bony landmarks to track body segments. Once collected, this information was used to develop biomechanical models to estimate kinematics and kinetics of the upper limb. The second project was to quantify in‐vivo joint kinematics, specifically to examine hip articulations/impingements and to determine the roles of the tibiotalar and subtalar kinematics in patients with chronic ankle instability. To accomplish this, we utilized high‐speed dual fluoroscopy. Dual fluoroscopy is an imaging modality that accurately measures three‐dimensional in‐vivo bone movement and can be used to identify the independent roles of specific joints with sub‐millimeter accuracy. The third project is the investigation of using virtual reality as a tool for training individuals with Parkinson disease in an immersive environment that provides modifiable challenges in a safe and controlled environment for the purpose of increasing mobility and improving balance. Currently we are finalizing the methods that utilize a Cave Automatic Virtual Environment (CAVE), motion capture data to track body segments, and a Unity game engine to deploy varying environments and assets to create these immersive virtual three‐dimensional environments. Clinical studies will begin within the next 6 months. In summary, the use of presented technologies to study anatomical form and function have the potential to augment current clinical decision‐making processes and rehabilitation methods with the overall goal to improve function and quality of life for patients. For the presented projects, participants consented and procedures were approved by the Institutional Review Board at the University of Utah

S10 A cadaveric study of aortic arch branching pattern in an Irish population

O'Malley A. , Debebe H. , Burukan A. , El Kininy W. and Davy S.

Department of Anatomy, Trinity College, Dublin, Dublin, Ireland

The aim of this study was to establish the incidence of aortic arch (AA) branching variation in an Irish population.

Over a period of 2 years, cadaveric dissection of 24 subjects was conducted in the dissection theatre (DT) at Trinity College, Dublin (TCD). Using gross dissection, AA's and their branches were identified, recorded, photographed and the following measurements made; the angle of the AA to the coronal plane, the distance from each branch of the AA to the midline, length of the brachiocephalic trunk (BCT) and distance from AA to each vertebral artery (VA).

The ‘normal’ branching pattern was observed in 19 subjects (79%). Three subjects (13%) displayed a two‐branched AA (bovine variant) where the BCT gave rise to both common carotids arteries (CCA) and right subclavian artery (RSCA). Two subjects (8%) displayed a four‐branched pattern where an aberrant left VA arose directly from the AA. The mean distance from the midline to the BCT, left CCA and left subclavian artery (LSCA) was 9.05 mm, 10.76 mm and 21.44 mm respectively. The mean length of the BCT was 34.09 mm. The mean distance from LSCA to left VA was 39.79 mm and was greater than its counterpart on the right side (23.38 mm). The mean angle of the AA to the coronal plane was 59.02 degrees.

Anatomical variation in the branching pattern of the AA has significant radiological and surgical significance. Patients with these variants may be at higher risk of stroke or thoracic aortic aneurysm (TAA). In addition, pre‐operative diagnosis of an AA variation would inform surgeons of the need to use alternative endovascular stents with corresponding branches in TAA surgery. Furthermore, knowledge of these variants is relevant to interventional radiologists who access VA's and CCA's in cases of clot retrieval in acute stroke and in cases of intracranial aneurysm coiling.

All donors in this study had previously consented for their bodies to be donated for the purpose of education and research, no ethical approval was required for the study.

S11 Analysis of dermal composition of the breast and donor sites to determine suitability for autologous breast reconstruction

Gorman N. ,¹ Beecher S. ,² Chua D. ,² Kelly J.² and Stanley A.¹

¹Anatomy, NUI Galway, Galway, Ireland and ²Department of Plastic and Reconstructive Surgery, NUI Galway, Ireland

Breast cancer is the most frequent cancer diagnosis and the leading cause of cancer related death in women. Mastectomy, the partial or complete removal of the breast is a well‐established procedure for the treatment of breast cancer. In addition, the increased screening and awareness of genetic predisposition to breast cancer increases the rate of elective mastectomy. However, mastectomy can provoke an array of emotions, including feelings of mutilation, low self‐esteem and a lack of sense of femininity.

Breast reconstruction aims to restore the appearance of the breast, providing aesthetic satisfaction and restoring quality of life for the patient. Autologous breast reconstruction with a chosen skin flap is often a preferred method. However, which is the most suitable skin flap for breast reconstruction is still debated. The composition of the dermis is a factor that should be considered in this debate as this would have a major impact on providing a more natural feeling breast for the patient. Therefore this study compared dermal composition of breast to that of Anterolateral Thigh (ALT), Deep Inferior Epigastric Perforator (DIEP), Latissimus Dorsi (LD) and Transverse Upper Gracilis (TUG) flaps. Samples were taken from female cadavers (n = 5), processed for light microscopy. 5 μm sections were stained with Masson's Trichrome with Gomori's Aldehyde Fuchsin. Slides were scanned using the Olympus VS120 digital slide scanning system. 10 non‐overlapping areas for quantification were selected in superficial, middle and deep regions (i.e. 30 images per section). A stereological square grid was superimposed on the images using the grid function in ImageJ and percentage of dermis occupied by collagen, elastin, space, blood vessel, other (nerve endings, hair follicles and glands) was calculated.

Statistically significant differences were found in the dermal composition of breast tissue compared to DIEP, LD and TUG samples. There were no statistically significant differences found between the dermal composition of the breast and ALT samples. The results of this study indicate that the ALT flap is the most suitable for autologous breast reconstruction based on dermal composition.

This study was carried out with full ethical approval from NUI Galway Research Ethics Committee.

S12 Turning to the “Cloud” in histology education

Felszeghy S. ,^1,2 Seppänen S.¹ and Mahonen A.¹

¹Institute of Biomedicine, University of Eastern Finland, Kuopio, Finland and ²Institute of Dentistry, University of Eastern Finland, Kuopio, Finland

Here we report the findings of a 390 student‐scale study that documented the advantages of the use of digital technology for learning histology at the Institute of Biomedicine, University of Eastern Finland (UEF). Although we have a long tradition providing classical and didactic teaching impetus, instruction and guidance regarding to the anatomy, biochemistry and physiology of the human body, there is a strong tendency at UEF, where traditional lectures‐lessons share less and less proportion of contact hours in our medical and dental curriculum. In 2016, a new digital system powered by Webmicroscope^® was introduced in histology education in UEF. It has the full advantage of current state‐of‐the‐art digital imaging technology designed for education: 1, students gain access to new digital tools analyzing histological specimens. 2, it combines independent out of class preparation with in class small group discussion. 3, Web stored histological images are available for review from any location, even remotely. 4, Webmicroscope^® allows students to “consume” histological images, to develop their ideas during class‐independent web‐based discussions. All these facts are educators’ dream to improve the basic knowledge on the subject, therefore the aim of the study was to analyze the core histology examination performances of medical and dental students. Assessment of histological knowledge was based on final written, subject examinations. Moreover Google Analytics has been applied to understand how students behaved on Webmicroscopy^® sites and apps after they clicked to drive their better performance in histology. Our analysis indicates that the average test score of the digital histology exams was different when compared to the conventional histology score. A comparison between the performance of Webmicroscopy^® users and non‐users was found to be that they were significantly higher scores on final course assessments and less student failed on written examination. The Google Analytics data indicate that the students appear to profit from Web‐system‐based modalities using intensively it outside the UEF network, as well. A shift away from the traditional teacher‐based approach towards integrated, Web‐based and multimodal teaching pattern might require further research to evaluate the suitability and student perceptions of, to meet learning outcomes at UEF.

No ethical approval was required for this study.

S13 Mapping medical school anatomy learning outcomes against the core anatomical syllabus to gauge anatomical knowledge: medical students’ perspective

Evans B. ,¹ Love A. ,¹* Burch A. , Mowatt A. ,¹ Matthan J. ,² and Phillips A.^2,3

¹Newcastle Medical School, Newcastle, UK; ²Faculty of Medical Sciences, School of Medical Education, Anatomy and Clinical Skills Department, Newcastle Medical School, Newcastle, UK and ³The Newcastle upon Tyne Hospitals Foundation Trust, Newcastle, UK *Presenting author

The Anatomical Society's core anatomy syllabus was originally published in 2003 and revised in 2007. The latest revision, 2016, used a modified Delphi technique to identify 156 learning outcomes (LO) applicable to medical curricula; this can be considered the gold standard for anatomy education in medicine, the General Medical Council's anatomy LOs being merely recommendations. The syllabus is of particular value to medical schools and aspiring surgeons wanting to ensure their anatomy knowledge is adequate. The LOs of an integrated undergraduate MBBS curriculum were mapped against the core syllabus to gauge students’ depth of knowledge and identify gaps when preparing for speciality training programmes.

All pre‐clinical LOs across the curriculum – clinical and communication skills, anatomy and physiology – were identified to ensure that no anatomical LOs were missed. LOs were allocated an anatomical region (Head and Neck, Vertebral Column, Thorax, Upper Limb, Lower Limb, Abdomen, Pelvis and Perineum) by two independent researchers. Discrepancies were assessed by a team of four researchers, assessed for merit and accuracy of LO expression. 140 of 156 of the anatomy LOs mapped against the core syllabus. 100% of abdominal, 95% upper limb, 91% lower limb, 88% thoracic and vertebral column, 86% head and neck and 84% of pelvic and perineal anatomy LOs were identified. While 90% of LOs are present, three areas for improvement were identified: clinical LOs, overarching LOs and imaging‐related LOs. In comparison to the core syllabus, where LOs are specific and clear anatomical learning objectives and clinical relevance evident, broader LOs were used that were unhelpful in understanding relevance. Few imaging‐related LOs exist, whereas the core syllabus identified these as key for personal development. Of note was an insufficiently broad LO on the lymphatic system, where the core syllabus named lymphatic drainage and nodes in multiple LOs in different anatomical areas.

The core anatomy syllabus should be compared to medical school curricula and improvement areas identified. Currently, further mapping of medical school curricula is ongoing against this criteria to identify where changes can be suggested and realised by medical schools, with the ultimate aim of ensuring consistency in anatomy teaching nationwide.

S14 Social media use within medical education: a systematic review to develop a pilot questionnaire on how social media can be best used at BSMS

Whyte W. and Henessy C.

Brighton and Sussex Medical School, Brighton, UK

Since the early 2000s social media has become a major part of our daily lives, and over the past decade it has found its way into the medical profession. Despite its ubiquity, only 5 systematic reviews exist on the subject of social medial use within medical education. The reviews conclude that there are positive correlations linked to social media use however the studies are restricted by the same limitations: a lack of quantitative data and the fact that social media research fast becomes outdated. This review will therefore examine the latest studies in order to identify which questions remain to be answered and what areas need further development in order for social media to become a credible resource within medical education. The information gained from this process will be amalgamated to create a valid questionnaire which will produce quantitative data.

A systematic review of Pubmed, Cochrane, PsychINFO, ERIC & Scopus was conducted following the Preferred Reporting Items for Systematic Reviews and Meta‐Analyses (PRISMA) guidelines. The search was from 1st January 2014 to the 12th January 2017 and included keywords linked with social media and medical education. 27 papers were identified: 12 qualitative and 15 quantitative. From this data a questionnaire was drafted and put to a focus group in order for it to be validated. Research Governance and Ethics Committee at BSMS confirmed that ethical approval was not needed for piloting a questionnaire.

Six major themes were identified and analysed: community & interactivity, communication & feedback, learning theories, social media vs traditional didactic lectures, role of faculty and professionalism. Quantitative data was limited but highlighted the efficiency of social media use especially when Facebook and Twitter were used. After the analysis a validated questionnaire was produced.

Social media can be a useful tool within the medical curriculum if implemented correctly. The final questionnaire can be used to generate quantitative data on the following questions: which platforms are most effective and for what purposes? How beneficial is social media to teaching? and What do students understand the benefits/disadvantages of academic social media platforms to be?

S15 Medical students favoring active learning methods seem to prefer to integrate ultrasound anatomy in to the preclinical curriculum

Viranta‐kovanen S. ,¹ Ilkka V. ,¹ Vehmas T.^1,2 and Luoma K.^1,3

¹University Of Helsinki, Helsinki, Finland; ²Finnish Institute of Occupational Health, Finland and ³Helsinki and Uusima District University Hospital, Finland

Diverse learning methods and teaching preclinical subjects in a clinical context may improve medical students’ motivation to learn anatomy. Radiological anatomy can be integrated in the basic anatomy curriculum. Ultrasound (US) may help to comprehend anatomy as it provides a manual, self‐performed method to observe both the spatial relationships between organs and their functions and movements.

We integrated, as collaboration between Department of Radiology and Department of Anatomy (University of Helsinki) a guided US exercise into the musculoskeletal surface anatomy session of the first year students in the Faculty of Medicine. In order to evaluate the intervention we asked the students (n = 172) to fill a questionnaire. We asked their background information before the US session and their feedback and experience after the US session. Descriptive, bivariate and multivariate statistical methods were used to analyze the results.

Most students (94%) found ultrasound anatomy (UA) beneficial and 91% thought that more radiology should be integrated in their basic anatomy studies. Students favoring active learning methods (PBL, surface anatomy, dissection) were more likely to feel that UA supported their learning. 92% announced that they were going to train themselves independently in the future and 86% would recommend the use of US to their fellow students. After the first exercise, 89% of the students felt that they are able identify bones and blood vessels in the US picture.

The positive feedback of the integrated US teaching has led to a permanent change in anatomy teaching in our faculty. Currently, every first year student receives hands‐on US‐guidance and they are then encouraged for independent practicing. Ultrasound devices are available for all the students in a special learning center.

No ethical committee approval was needed for this survey.

S16 The use of a Padlet^® for the distribution and sharing of anatomy mnemonics among medical students

Sweeney E.^1,2

¹Queen's University Belfast, Belfast, Northern Ireland and ²Ross University School of Medicine, Portsmouth, Dominica

Mnemonics are learning strategies to aid memorisation, they involve encoding information into a less abstract form by linking it with familiar objects or concepts already held in long‐term memory. Mnemonic use by medical students is widespread and in some cases has been successfully commercialised (Picmonic^®, Anatomonics^®). Some consider mnemonics to be representative of a surface learning strategy, but their use has been shown to improve scores on not only memorisation tests but also higher‐order thinking and comprehension tests. Many medical students dedicate time to sourcing or creating mnemonics, this is a time costly approach in an already time constrained curriculum. The aim of this project was to provide a platform for distribution and peer‐sharing of anatomy mnemonics among medical students.

A free virtual bulletin board (Padlet^®) was embedded in the online learning management software (eCollege^®) of first year medical students at Ross University School of Medicine, Dominica. This platform allows upload of text, image, audio or video‐based mnemonics sourced or generated by academic staff and additionally enabled students to add (moderated) mnemonics to share with their peers. The upload of relevant posts was paced to the curriculum progression. Feedback was collected anonymously using a separate Padlet^®.

This poster describes in detail the creation and embedding of Padlet^® in a virtual learning environment. Also presented are numerous gross anatomy examples from the two major subtypes of mnemonics defined by Bellezza, these are encoding (visual, phonetic, semantic, digit‐consonant encoding) and organisational (rhymes, links, stories, peg‐words, loci, first‐letter encoding).

Feedback received from students was entirely positive and indicates that the use of a Padlet^® for sharing anatomy mnemonics is an easily implemented, worthwhile endeavour.

S17 Do postgraduate trauma and orthopaedic surgery learning outcomes compare with those in the undergraduate core anatomy syllabus? A review of the two syllabi

Haynes E. ,¹ Love A. ,¹* McMahon D. ,¹ Creissen A.¹ and Matthan J.²

¹Newcastle University Medical School, Newcastle, UK and ²Anatomy and Clinical Skills Department, Newcastle University Medical School, Newcastle, UK *Presenting author

Regulatory bodies, primarily the General Medical Council (GMC), now advocate an outcomes‐based approach for teaching medicine, with a greater emphasis on clinical transferability of knowledge. This has reduced the time available to deliver core knowledge, such as the anatomy teaching previously considered fundamental to medical graduates, and has led to debate over the amount of anatomical knowledge required to ensure patient safety. The Anatomical Society first produced a syllabus for regional anatomy teaching in undergraduate medicine in 2003 using a modified Delphi technique. The revised version from 2015 contains 156 learning outcomes (LO) considered fundamental to medical graduates. We compared these LOs to the intercollegiate surgical curriculum programme (ISCP) for Trauma and Orthopaedics’ (T&O) knowledge requirements to ascertain their utility and value for T&O trainees.

LOs for upper limb, lower limb and the vertebral column were selected from the anatomical syllabus. These were compared to the ISCP knowledge requirements found in the Applied Clinical Knowledge Syllabus section of the T&O programme document. All outcomes that contained the terms ‘anatomy’ or ‘physiology’ were selected. The remainder – relating to clinical skills, pathology, investigations, treatments and complications – were excluded.

The core anatomy syllabus details 20 LOs for the upper limb, 21 for the lower limb and 7 for the vertebral column. This compares to 38 ISCP outcomes, once duplications were removed from sub‐specialties. 24 ISCP LOs are evident in the core anatomy syllabus. The remaining 14 ISCP LOs relate to surgical approaches and are not deemed relevant for undergraduate teaching.

A striking contrast between the two syllabi is the level of detail evident in the core anatomy syllabus; the ISCP LOs are markedly vague and a single LO broad enough to cover multiple core anatomy syllabus outcomes, making comparison difficult. In an era of outcomes‐driven learning, this could potentially mean that postgraduate trainees lack direction as to the level of anatomy knowledge required of them for safe practice in T&O. The T&O ISCP should be directed by the undergraduate core syllabus and should model its anatomy syllabus on it to ensure breadth and clarity for trainees.

No ethical approval was required.

S18 Implantation and early pregnancy: from laboratory models to basic mechanism to clinical translation

Aplin J.

Division of Developmental Biology & Medicine, University of Manchester, Manchester, UK

Low rates of implantation of embryos after fertilisation in vitro is a long‐standing problem in assisted reproduction. We have been modelling the early phase using various in vitro models. The results suggest how embryos might be helped to attach more efficiently to the endometrial surface. Attachment releases signals from maternal cells that trigger profound responses in the embryo, leading to the early developmental decisions that initiate placental morphogenesis. This will be the topic of the first part of the presentation.

Growth of the embryo in pre‐ and post‐implantation phases affects the eventual size of the offspring. In the second part, I will describe a novel approach to treatment of fetal growth restriction, using peptide targeting to deliver two different drugs to the early placenta. I will show results from our efforts to correct FGR in mice.

All mouse procedures were performed in accordance with the UK Animals (Scientific Procedures) Act of 1986 under a Home Office project licence. There were no surgical procedures with live animals. Dams were euthanased 2 days after mating or at E18.5, by cervical dislocation.

Work with anonymised human embryos was carried out with approval of Central Manchester Local Research Ethics Committee (03/CM/684) under a license granted by the Human Fertilisation and Embryology Authority (HFEA). Informed consent was obtained from couples.

S19 Engaging student learning at School, University and internationally using microscopy

Anderson S.I.

University of Nottingham School of Medicine, Nottingham, UK

This talk aims to give an overview of the activities in support of learning carried out at University, using a virtual microscope to develop and assess histology and pathology and at School, in support of science learning and engagement using microscopy.

The University of Nottingham Virtual Microscope (VM) has steadily increasing numbers of student and staff users. Evaluation of the resource indicated that students used a combination of guidance from staff and student generated e‐workbooks to optimise histology and pathology learning. Further development of this technology promoted independent study, in groups by engaging students across our UK and international campuses. Staff allocated mixed student groups and provided a didactic lecture and resources detailing appropriate digital slides and guidance. Students led the process of deciding how to interact as a group, allocate tasks and decided on structure and style of coursework. The standard and detail of coursework was very high and students reported that they understood tissue structure better as they had engaged actively and conducted a careful analysis of virtual slides. They generally enjoyed the experience of working with their international counterparts. Preliminary analysis of assessment data reveals that exam performance was substantially improved over previous cohorts. We aim to continue and further develop this technology to enhance student experience.

As part of leading the Education and Outreach activities of the Royal Microscopical Society, there have been major developments in (1) promoting engagement of young people in science education through the medium of microscopy (2) supporting continued professional development of professional microscopists and (3) public awareness and engagement concerning the value and applications of microscopy. One key way of achieving these aims have been to develop a Microscope Activity Kit (MAK), containing curriculum mapped activities, along with samples and microscopes to engage primary school children and their teachers. These are loaned free to Schools for a term and have now reached almost 50 000 children with excellent feedback. We also partner with the Primary Science Teaching Trust. Projects in Ireland have also engaged several thousand young people and the scheme is now in operation in Malaysia. A second strand has been the development of a diploma qualification for microscopists. This is based on a portfolio including a major project, technical essay, course attendance and presentation/outreach/writing skills development. Public engagement is achieved via contributing to a variety of science festivals and exhibitions. Our current focus is on accreditation of the DipRMS, internationalising the MAK, working in partnership with other Societies and Universities as well as measuring and evidencing the educational impact of these activities for young people.

S20 E‐learning resources for the anatomical sciences–what does it take to make them work?

Hortsch M. ,¹ Zureick A.H. ,¹ Burk‐Rafel J. ,¹ Purkiss J. ,² Goldman H.³ and Lee L.⁴

¹University of Michigan Medical School, Ann Arbor, MI, USA; ²Baylor College of Medicine, Houston, TX, USA; ³Drexel University College of Medicine, Philadelphia, PA, USA and ⁴University of Colorado School of Medicine, Aurora, CO, USA

New instructional technologies have been increasingly incorporated into the learning environment, including lecture video recordings as a substitute for live lecture attendance. The literature presents varying conclusions regarding how this impacts students’ academic success. Previously, a multi‐year study of the M1 histology component at the University of Michigan Medical School revealed that live lecture attendance was positively and lecture video use negatively correlated with learning success. Here, we surveyed three cohorts of University of Michigan M1 students in greater detail regarding lecture attendance and video usage, focusing on study behaviors that may influence learning outcomes in histology, anatomy, biochemistry, and physiology. Students who always attended lectures scored the highest average, while choosing an inconsistent strategy is associated with poorer performance. Several behaviors were negatively associated with histology performance. Students who attended lectures and engaged in “non‐lecture activities” and students who watched lecture videos and reported being interrupted, feeling sleepy/losing focus, or engaging in “non‐lecture activities” had lower scores. Watching the lecture video at up to 2×‐accelerated speed had no influence. These results suggest that interruptions and distractions during learning activities—whether live or recorded—can have an important impact on learning outcomes.

Electronic resources not only have changed the instructional landscape for students, but also for educators. The use of virtual microscopy has become the norm for teaching microanatomy. Unfortunately, the technological parameters and expenses that are required to create virtual microscopy file collections often pose a barrier for the use of this powerful approach, especially for new universities or in developing countries. To help educators and researchers worldwide, we created the Virtual Microscopy Database (VMD) as a resource‐sharing website. It allows registered users to upload and download virtual microscopy image files to be used in their local environment. Copyright of the images remains with the donor and use of VMD images is governed by a Creative Commons License. The VMD's purpose is to offer its users access to a wide selection of virtual images. New users and donors can register at http://www.virtualmicroscopydatabase.org This study is IRB‐exempt. The VMD is supported by a grant from the American Association of Anatomists.

S21 The impact of the anatomical society core syllabus in gross anatomy

Smith C. ,¹ Finn G.² and McHanwell S.³

¹University of Sussex, Brighton, UK; ²University of York, York, UK and ³Newcastle University, Newcastle, UK

The questions of what students need to know and when they need to know it are important for curriculum planners, anatomy educators and their students, as ever‐expanding knowledge changes of emphasis in medical treatment. The new syllabus published 2016 contains 156 learning outcomes grouped by body region.

The new syllabus has defined the ‘what do I need to know question’, however the ‘how’ and ‘with what’ resources require further exploration. This project set out to explore how has the new core syllabus impacted on undergraduate anatomy? The study used mixed methodology and utilized A, Questionnaire, B, focus groups of anatomy educators and C, Metrics such as download and Social Media impact. Ethical approval was granted for this study.

The questionnaire, anatomists and clinicians (N = 24) revealed 100% had knowledge of the new syllabus and that the syllabus had been well received. Through the focus groups seven key themes were identified: standardisation, validation, leverage, need, praise, limitations, feelings and perception. Overwhelming the discussions were positive with praise for the new syllabus. There was concern over the uptake in clinical years. Additionally, some further needs were identified in the form of other core syllabi. The syllabus has been downloaded 1235 times with 38 click links being through Twitter. The altmetric score is 16.

One consideration under the theme standardization was the feeling of being comparable, teaching to the right level and the new syllabus provided reassurance to teachers. It also helped with leverage to different groups when teaching content was threatened especially as it is referenced by the GMC. Measuring the impact the new syllabus has made to doctors and in turn patients is much harder to judge. It could be inferred that increasing awareness about the level of expected knowledge has improved students’ knowledge and ensured greater consistency between medical schools which will be of benefit to patients in the future.

The new syllabus has been accepted within the anatomical community in the UK as the gold standard and further work is needed on para‐syllabuses including embryology and neuroanatomy and core syllabi for different health care professions.

S22 Learning through dissection: a student perspective on the transition from formalin to Thiel‐embalmed cadaveric material

Sanders K. and Bazira P.

Hull York Medical School, York, UK

Dissection has been a corner stone of medical education for centuries, and in that time preservation techniques have evolved to suit medical school demands. Currently, formalin embalming methods are prevalent, as they provide stable tissues long‐term which are ideal for prosection preservation. More recently, the option of Thiel embalming has become available. The main benefit of Thiel over formalin‐based fixing methods is that tissue remains more lifelike and flexible – a profound advantage for studying clinical anatomy.

In 2016, the first cohort of MSc in Clinical Anatomy students at Hull York Medical School (HYMS) began musculoskeletal dissection on formalin‐embalmed cadaveric specimens. In term two, they then transitioned to Thiel‐embalmed cadavers for dissection of the trunk. This study explores student perspectives of the transition from the use of formalin‐embalmed specimens to Thiel‐embalmed specimens.

Submission of a reflective portfolio formed part of the summative assessment for both Clinical Anatomy of the Limbs and Back (formalin‐embalmed) and Clinical Anatomy of the Trunk (Thiel‐embalmed). All submitted student portfolios (n = 15 per module) underwent thematic analysis to identify recurrent themes, areas of corroboration and contradiction, and points of interest regarding student perspectives of musculoskeletal and visceral dissection of formalin and Thiel specimens.

Portfolios were submitted anonymously and are now part of the programme's domain; ethical approval to bypass student consent has been granted by the HYMS Human Research Ethics Committee. Furthermore, student reflections are based on using cadaveric material sourced from HYMS’ body donation programme, in which informed, written consent is already provided by donors for use in teaching and research.

The context of student statements will contribute to the literature on the appropriateness of formalin and Thiel‐embalmed tissues for dissection of different anatomical regions. The outcomes of this study will inform the future planning of dissection sessions, and the introduction of further preservation techniques.

S23 A survey of the anatomical knowledge of the general public; a tool for public engagement and student education

Taylor A. ,¹ Diggle P.¹ and Wessels Q.²

¹Lancaster Medical School, Lancaster University, Lancaster, UK and ²Department of Anatomy, School of Medicine, University of Namibia, Windhoek, Namibia

Public knowledge of the anatomical “self” is lacking and evidence points towards a growing interest and need for anatomy education to the public. Limited information exists on how much anatomy the general public knows. This should be factored into current pedagogical strategies in order for medical students to appreciate the importance of health literacy as a bridge between healthcare professionals and patients. The aim of this study was to provide an interactive platform to assess the anatomical knowledge of the general public

We offered the public of Lancaster an opportunity to learn human anatomy and complete an anatomical knowledge survey afterwards. Sixty‐three participants volunteered following obtaining informed consent to attempt to place 20 anatomical structures on a human body outline. Study approval by the Faculty of Health and Medicine Research Ethics Committee, Lancaster University. Responses were scored independently by 2 anatomists and analysed; associations with age group, gender, education, previous healthcare visit were assessed by one‐way ANOVA and Kruskal‐Wallis post‐test, we examined via a mixed effects logistic model associations with gender and organ type as factors and subject (structures).

There was a significant increase in the number of correct answers in the age group 18–29 and 40–49 compared to the under 18′s (P < 0.05). There were no significant differences between the total correct answers compared to academic qualification, working in healthcare or not, visiting a healthcare professional in the last 7 days or not and comparing males versus females. Logistical regression showed males were better at answering muscles than females. The current study demonstrates the public's eagerness to learn anatomy despite their limited knowledge of the human body. Furthermore, it raises awareness of the anatomical literacy needs of the public, especially in school children and young adults and emphasises the value of health literacy and competent knowledge base as a focus in undergraduate medical education. Anatomy literacy appears to be neglected and our experience provides an example of a possible mode of public engagement in anatomy coupled with a resource to highlight the necessary knowledge needed for medical practitioners.

S24 The importance of comparative anatomy in orthopaedic research

McNulty M.^1,2 and Richbourg H.²

¹Indiana University School of Medicine, IN, USA and ²Louisiana State University School of Veterinary Medicine, Louisiana, USA

Translational research relies on choosing the appropriate animal model of human disease. However, many variables must be considered when choosing a suitable species, including, but not limited to, availability, cost, and analogy of the species to the human. The normal anatomy in quadrupedal species, especially musculoskeletal anatomy, differs from humans in many ways, and it is not always clear whether or how this will impact research. Understanding the normal anatomy of the experimental species is critical in orthopaedic research, which relies on a full understanding of the musculoskeletal anatomy and the implications with regards to biomechanics and other functional and pathological outcomes. Prompted by work evaluating bone and joint lesions secondary to infection by the arthritogenic chikungunya virus in the mouse, we have characterized the normal anatomy of the tarsus in common United States inbred laboratory murine strains (M. muscus). The anatomy of this species was compared to the skeletal anatomy of relative outgroups, including the white‐footed mouse (P. leucopus), common laboratory rat (R. norvegicus), eastern mole (S. aquaticus), and nine‐banded armadillo (D. novemcinctus). Through this work, we have identified a normal fusion between tarsal bones in the inbred laboratory strains of mice that is not present in the aforementioned related species. This anatomical characteristic has been previously identified in the inbred laboratory mouse and published in the early 1970's as a pathological lesion, thereby raising questions as to the origin, nature, and apparent wide presence of this fusion in inbred strains today. Similarly, this fusion has implications for ongoing work evaluating pathological bone and joint lesions secondary to other diseases, such as arthritis and inflammatory lesions of bone, and raises questions as to whether variations in related anatomy in this region, such as vasculature and musculature, would influence study results using the mouse as an animal model. Related work has examined the anatomy of the distal hind limb of the nine‐banded armadillo, with this species in mind as a potential model for human disease. Overall, the work presented will underscore the need for continued characterization of the anatomy of those quadrupedal species being utilized for translational orthopaedic research. No ethical approval was required for this work.

S25 Digital anatomy and computed morphometrics: old questions and new problems

Bruner E.

Centro Nacional de Investigación sobre la Evolución Humana, Paseo Sierra de Atapuerca 3, Burgos, Spain

The advent of digital tools and multivariate morphometrics has represented a major advance in anatomy, most of all during the last 15 years. Biomedical imaging and shape analysis have introduced new professional frameworks, and many anatomists are nowadays “specialists” in pixels and numerical models, integrating biology with electronics, informatics, and statistics. The improvement was noticeable, in terms of resolution (the details available), statistical power (the efficiency of the quantitative approaches and the sample sizes), and biological reliability (the possibility to investigate in vivo anatomical systems). These tools have introduced new questions and new challenges, but they have also rescued old problems left unresolved since decades, because of the limitations associated with physical dissections and small samples. Nonetheless, the complexity of these tools also increases the possibility of methodological and technical biases and pitfalls, mostly when taking into account that programs, devices and databases are available to a wide range of users, with different background and expertise. Collaboration between biologists and medical doctors is probably the best way to design teams and projects based on fruitful and competent multidisciplinary perspectives, integrating distinct skills and different scopes to approach shared problems and common interests.

web: http://paleoneurology.wordpress.com/

S26 Comparative anatomy of the intertrabecular angle in mammalian femora

Felder A. ,¹ Domander R. ,¹ Reznikov N.² and Doube M.¹

¹The Royal Veterinary College, London, UK and ²Imperial College London, London, UK

The intertrabecular angle (ITA) has been recently proposed as a new parameter to describe the topology of trabecular bone in three dimensions. It has been measured in the human femoral head, showing maximally spanning properties, possibly relating to the impact dampening function of trabecular bone. However, we do not know whether these properties are similar in other species. In this study, we measured ITA distributions in X‐ray microtomography images of trabeculae in the femoral head and condyle from 10 different mammalian species. We used a novel, open‐source implementation of ITA in the development version of BoneJ, a collection of libraries for ImageJ. Like in humans, the mean angles are close to 120° for trabecular junctions with 3 branches, and close to 109° for trabecular junctions with 4 branches, suggesting that the maximal spanning organization of trabeculae may be conserved throughout mammal species.

No ethical approval was required for this study.

S27 Fantastic beasts and how to image them

Metscher B.

Department of Theoretical Biology, University of Vienna, Vienna, Austria

Driven by the fundamental necessity of detailed and realistic visualisations for the study of anatomy in development, function, evolution, and diversity, technologies for making 3D images have progressed from the meticulously reconstructed wax‐plate models of the late 19th century to high‐resolution direct 3D microscopy, while increasingly sophisticated tools for rendering and analysing volume images have allowed us to move from depicting organs and tissues as simple coloured blobs to viewing anatomical structures with their full internal complexity.

X‐ray microtomography (microCT) was the first direct non‐destructive 3D imaging method and has now come of age as a general‐purpose tool for anatomy, especially as a non‐destructive 3D complement to other important imaging modalities, particularly EM, conventional histology, and block‐face imaging methods. With soft‐tissue contrast enhancement, microCT offers 3D in statu imaging of complete embryos and other intact specimens at histological scales. Digital 3D models of embryos can serve as virtual specimens in teaching and research, and we are currently generating high‐resolution 3D embryo images for atlases of rat and chick developmental anatomy. Related projects include characterising a teratogenic rat model of oesophageal atresia; a study of the relationship between the vestigial lung and pulmonary vasculature in snakes; research on horse early renal development, combining microCT and TEM to maximize the information obtained from rare specimens; and demonstrating the role of gut endoderm in the development of cement organs and other extreme anterior structures in basal fishes. MicroCT is remarkably effective for imaging insects and other arthropods, and internal micro‐morphological characters can be essential for distinguishing and describing species as well as for analysing function. We have recently published the first new millipede species description to be based partly on its cybertype – a set of virtual specimens made from microCT images of the physical type material for the species.

Our other current efforts include refinement of molecular probe imaging with microCT, developing X‐ray‐dense tissue‐selective stains, and imaging of melanocytes in development and in tumours. These methods will work effectively with dual‐energy (spectrally‐sensitive, “two‐colour”) microCT, which allows simultaneous high‐resolution imaging of different tissues or contrast agents using lab‐based systems.

S28 Testing long‐term neurodevelopment following preterm birth and prolonged respiratory suppor, using preterm lambs

Albertine K.H.

Departments of Pediatrics, Medicine, and Neurobiology & Anatomy, University of Utah School of Medicine, Utah, USA

The brain of chronically ventilated preterm infants is vulnerable to injury during the days, weeks, or months of ventilation support with oxygen‐rich gas that are necessary to keep them alive. Severe lesions are germinal matrix hemorrhage‐intraventricular hemorrhage, posthemorrhagic hydrocephalus, or periventricular leukomalacia, particularly with parenchymal cysts. However, awareness is growing about more subtle diffuse lesions that also lead to abnormal neural function and subsequent suboptimal neurodevelopmental outcome. Two persisting questions that await answers are (1) What are the long‐term consequences of diffuse brain injury on neurodevelopmental delays and/or neurological deficits? and (2) What is the molecular basis of the brain injury? For this presentation, focus will be on the first question, using our unique preterm lamb model of BPD and its co‐morbidities, including diffuse brain injury. An advantage of the chronically ventilated preterm lamb model is its setting of preterm birth uncomplicated by antenatal conditions, such as intrauterine infection or asphyxia that may potentiate brain injury. On the other hand, the less complicated setting for the large‐animal models may be viewed as a disadvantage because the models lack antenatal conditions that may potentiate injury to the lung and brain. Nonetheless, new mechanistic insights are being discovered as the model is developed for long‐term structural and functional outcomes. Evidence will be provided that invasive mechanical ventilation leads to electroencephalographic evidence of seizure‐like activity, loss of white and gray matter, loss of neurons and glial subtypes, and delay in development of gray matter. These effects are associated with neurobehavioral impairments in learning and memory of former preterm lambs. Our mechanistic studies are revealing that preterm birth and sequelae following prolonged usual clinical care for respiratory failure lead to epigenetic changes in the brain, changes that are common to the epigenetic profile in the lung.

Our studies are approved by the University of Utah IACUC.

S29 Qualitative studies of the calcified tissues in the lumbar vertebral body of the ageing human

Boyde A. and Mills D.

Queen Mary University London, London, UK

We studied the degree of mineralisation of trabecular and cortical bone and cartilage and thickness of the cortex in lumbar vertebral bodies. We report correlated 3D microscopic imaging methods concerning osteoporotic changes.

We examined 34 archival macerated para‐sagittal slices of L4 (17 male, 17 female) by 3D BSE‐SEM and 69 L2 (38 male, 31 female, 70 ± 15 years) obtained through European Union Concerted Action Biomed 1 ‘Assessment of bone quality in osteoporosis’, which were embedded in PMMA: plane block surfaces were carbon coated for quantitative 20 kV backscattered electron scanning electron microscopy (qBSE‐SEM); and later polished and stained with iodine vapour to study uncalcified matrix, providing very high resolution imaging of entire vertical sections. High contrast x‐ray microtomography (48 h per scan) was conducted at 30 μm voxel resolution. Thin sections from block faces were prepared by laser‐ablation for correlative light microscopy.

Central compression failure modes encompass calcified end‐plate cartilage and implosion of disc content into the cancellous domain, where cartilage growth and calcification continue. Trabecular and cortical fractures repair with initially low density woven bone, but instigation may involve high density acellular mineralised infill. The anterior cortex, especially, is frequently very thin or absent. In wedge compression and collapse fractures it thickens with inclusion of extra‐bony tissues during drift and collapse.

QBSE‐SEM showed 50% of all ‘bone’ to lie in bin 5 of 8 of the normal qBSE‐SEM range (corresponding to 1.99 g/ml peak found density gradient centrifugation): 29.10% in males and 27.44% in females is very highly mineralised (bins 6–8). Much of this is not bone, but calcified cartilage in the end‐plates and Sharpey fibre bone and calcified ligament and fibrous periosteum, which become incorporated into the cortices as they drift outwards and anteriorly: spaces originally outside the cortex become included within the bone organ and develop marrow histology.

The abundance of calcified ligament and periosteum should be factored into thinking about failure properties of the anterior cortex. More highly mineralised phases may not be ‘bone’: they will be artefactually represented as ‘thicker’ in clinical CT. They have a different ‘grain’ than ‘bone’ and will have different tendencies to fracture.

No ethical approval was required for this study.

S30 Alcohol use is associated with reduced anterior cingulate cortex thickness in bipolar disorder and healthy participants’

Martyn F. , McPhilemy G. , Nabulsi L. , Akudjedu T. , Natj P. , Ambati S. , McLoughlin J. , Hallahan B. , McDonald C. and Cannon D.

The Centre for Neuroimaging & Cognitive Genomics (NICOG), Clinical Neuroimaging Lab, NCBES Galway Neuroscience Centre, University Of Ireland Galway, Ireland

Cortical thinning, in particular of the anterior cingulate cortex (ACC), is associated with both bipolar disorder and alcohol‐use. Despite this potential confound the effects of moderate alcohol‐use on cortical thickness in bipolar disorder remain unclear. We hypothesized that ACC thinning would be negatively related to alcohol consumption levels in participants with bipolar disorder to a greater extent than control participants.

Thirty participants meeting DSM‐V diagnostic criteria for bipolar disorder (50% female, mean age ± SD 43 ± 12 years), and 32 healthy controls (56% female, 38 ± 14) underwent structural MRI (3Tesla‐MPRAGE) and the AUDIT‐C, a validated psychometric instrument to assess alcohol use. ACC thickness was determined using Freesurfer (v.5.3.0) and examined in relation to alcohol‐use using a non‐parametric correlation (Spearman's rho). Ethics approval was granted by The Galway University Hospital Research Ethics Committee.

There was no diagnostic‐group difference in age (U = 393, P = 0.22), alcohol‐use (U = 355, P = 0.08) or ACC thickness (right U = 368, P = 0.115; left U = 388, P = 0.197). A significant negative correlation between alcohol‐use (AUDIT‐C score) and left (r = −0.352, P = 0.004) but not right (r = 0.120, P = 0.357) ACC thickness was detected, that was not explained by age or diagnosis. This left ACC effect was driven by the bipolar group (r = −0.347, P = 0.061), and potentially by the control group (r = −0.206, P = 0.259), the latter only after removal of two outliers (r = −0.356, P = 0.054, defined as three standard deviations above mean).

The ACC mediates emotional regulation, and subsequent behaviour, it is thinner with greater alcohol‐use within a range of moderate to harmful consumption, this may be more pronounced in bipolar disorder. This work emphasizes the importance of controlling for and reporting moderate and harmful alcohol‐use when examining diagnosis based cortical thickness rather than excluding for dependence alone.

We gratefully acknowledge the participants and the support of the Wellcome‐Trust HRB Clinical Research Facility and the Centre for Advanced Medical Imaging, St. James Hospital for their support of this research.

This research was funded by the Health Research Board (HRA‐POR‐324).

S31 In vivo optical imaging of structure and function

Leahy M.

Physics, National University of Ireland, Galway, Ireland

Since Malpighi used an early microscope to prove Harvey's theory of the circulation of the blood through the capillaries, imaging has been a key resource for discovery related to the structure and function of tissues. Modern microscopy can produce exquisite images of the tissue with subcellular resolution, but is typically limited to ˜100 μm, which is approximately the mean free path for light before scattering in typical tissues. Recently several methods have been developed to image structure and function deeper in the body. In particular Optical Coherence Tomography uses low coherence interferometry to select unscattered photons has to image up to about 1 mm into scattering tissues which is sufficient to reveal biology in its native state e.g. in the skin, retina, mouse brain… after about 1 mm there no ballistic photons so diffuse optics is necessary. Photoacoustic imaging takes advantage of diffuse photons, which penetrate several centimetres. This paper will discuss the advantages of using optical methods to elucidate structure and function beyond the depth traditionally accessible to microscopy.

S32 In vivo imaging in the human male and female genital tract

Kölle S.

School of Medicine, University College Dublin, Belfield, Dublin 4, Ireland

Successful treatment of patients requires early diagnosis of the disease and constant monitoring of the effects of treatment. The use of novel in vivo imaging technologies will allow innovative diagnostic and therapeutic strategies in the daily routine of clinicians in the future. In the present, these technologies are impressive tools for researchers to get new and stunning insights into physiology and pathology of the male and female reproductive tract.

The lecture will cover three in vivo imaging technologies: digital videomicroscopy, probe‐based confocal laser endomicroscopy (pCLE) and optical coherence tomography (OCT). Ethical approval was granted for these studies by the University of Munich (No161‐12, No 371‐12), by UCD (No LS‐14‐23) and by the Mater Hospital (No: 1/378/1645). Using digital videomicroscopy the physiology of gamete transport, fertilization and early embryogenesis under near in vivo conditions is illustrated. Unique movies within the tube documenting the interaction between sperm and oviduct, the communication between cumulus‐oocyte‐complex and tube, as well as the process of fertilization and early embryogenesis are presented. This includes the first videos revealing the secret life of sperm in the human oviduct. Furthermore the first establishment of probe‐based confocal laser endomicroscopy (pCLE) as diagnostic tool in andrology and gynaecology is shown. This technology allows a three‐dimensional analysis of the oviduct, uterus, cervix, testis and epididymis on a cellular level and in‐real time. The probe for this technology is around 1 mm in size so that it can easily be used in the working channel of the endoscope. Finally optical coherence tomography (OCT) which measures reflexion of light at interfaces in a contactless way is covered. This technology allows a 3D monitoring of pathologies in tube, in the uterus and in the testis on a cellular level and in real‐time.

This lecture aims to provide new insights in imaging technologies for researchers and physicians and to stimulate the further development of imaging technologies in the field. The lecture should also motivate interested researchers to apply these technologies to find answers to the questions they are dealing with in their special field of interest.

S33 Simultaneous en‐face imaging of a human finger tip using multiple reference optical coherence tomography

Neuhaus K. ,¹ Alexandrov S. ,¹ O'Gorman S. ,¹ McNamara P. ,^1,2 Hogan J. ,² Wilson C.² and Leahy M.^1,3

¹Tissue Optics and Microcirculation Imaging facility, National University of Ireland, Galway, Ireland; ²Compact Imaging, Inc. Mountain View, CA, USA and ³Royal College of Surgeons (RCSI), Dublin, Ireland

We propose a novel imaging method, based on multiple reference optical coherence tomography (MR‐OCT). The MR‐OCT technology is based on the principle of a Michelson interferometer using a single scanning path delay and a partial mirror generating multiple reference beams relative to the reflected beam to form multiple interference signals at different depths. The method has been validated by imaging a reference sample and a fingerprint in vivo. The principle of scanning multiple selected layers is shown by imaging a partial fingerprint with 200 × 200 × 200 voxels of 3 × 3 × 0.5 mm size and obtaining an arbitrary amount of layers merely by digital processing. A mirror scan frequency for the path delay of 1 kHz was achieved using a custom voice coil actuator which allows scanning A‐lines at a rate of 2 kHz.

The layer‐scanning method for MR‐OCT (lsMR‐OCT) enables higher A‐line rates compared to previous MR‐OCT systems that could be increased beyond 2 kHz. MR‐OCT offers a low‐cost OCT platform comparable to simple technologies known from CD/DVD‐ROM pick‐up heads. The lsMR‐OCT technique provides fit for purpose OCT quality and is considered a complementary and mobile platform. The image quality can provide for several applications in the area of ophthalmology, biometrics, and others.

The author has performed the scans of the partial fingerprints on his own fingers using an irradiation power of about 6 mW, wavelength 1330 nm with about 300 ms exposure time, and a spot size of about 28 μm for each point scan, which poses no danger according to Health Physics guidelines. The size of the fingerprint pattern does not allow for identification or poses any risks to privacy.

The acquisition of the in‐vivo samples do not affect any ethical, health or privacy concerns and where performed according to the ethical regulations and safety standards of the NUI Galway.

S34 Sensing the morphome and the future of nanoscale morphomics

Lucocq J.M.

School of Medicine, University of St. Andrews, Fife, UK

Morphological analysis is fundamental to understanding cell and tissue function. Cells and organelles comprise rich arrays of 3D data (morphomes) that can be mined across the imaging scales of light and electron microscopy using morphomics methods that are characterized by systematic and quantitative 3D analysis. At the nanometer scale morphomics could yield petabytes of data and a big question is how to estimate high quality data easily and rapidly. I will discuss solutions to this “large data” problem and strategies for removing or negotiating technological roadblocks to higher‐throughput. I will also address issues such as phenotypic screening, automated recognition, the integration of EM data with readouts from other imaging modes and how the morphome integrates into the ‘omics framework.

S35 Peri‐vascular and glia limitans GFAP+ astrocytes are upregulated in guinea pig inferior colliculus at hearing loss onset and downregulated in late stage presbycusis

Orton L.^1,2

¹Manchester Metropolitan University, Manchester, UK and ²Newcastle University, Newcastle, UK

Age‐related hearing loss, presbycusis, is an increasingly prevalent condition associated with the ageing population. Presbycusis is associated with decreased speech comprehension and concomitant deterioration of emotional, social and cognitive processing. We know little of the mechanisms that mediate changes in the brain following hearing loss. This study sought first to identify the onset of sound threshold elevation in guinea pig auditory midbrain, and second, to investigate whether this is accompanied by a concomitant change in the number of GFAP+ astrocytes. Single units were recorded in the inferior colliculi (IC) of guinea pigs, anaesthetised with a cocktail of urethane (0.6–1 g/kg, 20% solution, i.p) and Hypnorm (0.4–1 mL, i.m.). Responses to sound, at time points during the first year of life, identified reductions in auditory driven spiking responses and raised thresholds at 1 year of age, suggestive of hearing loss onset. Following recordings, animals were maintained under deep anaesthesia via sodium pentobarbitone administration (200 mg/kg, i.p.) and transcardially perfused with phosphate buffered saline and paraformaldehyde. Immunohistochemistry revealed increased numbers of GFAP+ astrocytes (+22%) in the IC at 1 year of age compared to 6 months. Peri‐vascular GFAP+ processes were clearly identifiable and adjacent astrocyte somata were numerous and densely packed. To identify changes in IC associated with advanced presbycusis, aged animals were also stained for GFAP. Thirty‐seven month old animals showed reduced numbers of GFAP+ astrocytes (−90%) compared to 6 months, with few identifiable peri‐vascular GFAP+ processes. Furthermore, the cross‐sectional area of the IC increased at 37‐months, suggestive of neuroinflammation. These data suggest that GFAP+ astrocytes may be involved in changes associated with onset of sound threshold elevation. Furthermore, in advanced stages of presbycusis, dysregulation of peri‐vascular astrocytes may be involved in degenerative changes associated with reduced auditory function. The procedures undertaken in the study were approved by the Local Ethical Review Committee of Newcastle University and complied with the Animals (Scientific Procedures) Act, 1986.

S36 From video atlas to virtual reality: anatomy in every dimension

McGilvary I.

Department of Surgery, University of Toronto, Toronto, Canada

It has become increasingly difficult to teach complex points of anatomy during surgery of the liver and pancreas. Anatomical variations are common, important, and often obscured by adjacent structures; moreover, lines of sight are difficult around a crowded operating table. An additional issue is one of perception – experience is critical to pattern recognition but difficult to acquire, particularly as changes to the education of surgical trainees has resulted in fewer operations being seen and done during their training. The Toronto Video Atlas of Surgery (http://www.TVASurg.ca) was created as a way to teach difficult anatomy and surgical procedures by combining story‐telling, anatomical modeling, 3D computer animation, and real‐time surgical footage. Over the last year we have also developed an interest in virtual reality. This immersive modality is ideal for teaching anatomy and may well represent the future of surgical, medical and anatomical instruction. In this talk we present examples of how anatomical‐based modeling and animation lends itself to creating virtual reality modules that can be used to teach surgical anatomy and principles.

S37 The skeletons in our closet: E‐learning tools and what happens when one side doesn't fit all

Van Nuland S. and Rogers K.

Department of Anatomy and Cell Biology, The University of Western Ontario, ON, Canada

In anatomical sciences, computer‐aided instruction and online learning tools have become a critical component of teaching when physical space and cadaveric resources are limited. Our previous research (n = 70) compared a simple 2‐dimensional e‐learning tool (A.D.A.M. Interactive Anatomy) to a more complex tool that allows for a more 3‐dimensional perspective (Netter's 3D Interactive Anatomy). Despite the differences in how these e‐learning tools present information, student ability to learn anatomical material, and their mental effort while doing so, known as cognitive load, were identical between e‐learning tools. However, the results showed that e‐learning tool software design can differentially influence students based on their spatial ability, but it remains to be determined if traditional kinesthetic‐tactile learning approaches, such as manipulating a skeleton, are also impacted by a student's spatial ability. Using a novel dual‐task methodology with a cross over design, undergraduate anatomy students from The University of Western Ontario, Canada (n = 77) were evaluated as they studied a bony joint using a physical skeleton as well as a simple commercial software program (A.D.A.M. Interactive Anatomy). We hypothesized that the acquisition of anatomical knowledge by students, regardless of their spatial ability, would be superior when learning is associated with a real model, rather than currently available e‐learning tools. Students were assessed using a baseline knowledge test, Stroop task response times (a measure of cognitive load), MRT scores (a measure of spatial ability), and an anatomy post‐test. Our results showed that knowledge recall was significantly higher when students studied using a physical skeleton than with a simple 2D e‐learning tool (P < 0.001, large practical effect). Furthermore our results demonstrated that students with low spatial ability are at a significant disadvantage when they learn the bony anatomy of a joint and are tested on images of the contralateral joint (P = 0.046, R = 0.326). This study highlights a major weakness in the strategy to move traditional anatomical education online, and suggests that we should be teaching both anatomy and surgical procedures on both sides of the human body, to ensure that all students, regardless of spatial ability, can take their anatomical knowledge into the clinic and perform successfully. Research was carried out in compliance with The University of Western Ontario Research Ethics Board (protocol #104961).

S38 Usage and preferential features of e‐resources for anatomy learning: the student‐perspective

Ahmed W. ,¹ Jones C. ,^1,2 Tarr J.¹ and Chinnah T.¹

¹University of Exeter Medical School, Exeter, UK and ²Peninsula College of Medicine and Dentistry, Plymouth, UK

Anatomy e‐resources (AERs) have become increasingly important in undergraduate medical education. However, student perceptions and usage of such resources remains poorly understood. We aimed to determine how students use AERs at a single UK medical school.

The study employed a mixed‐method triangulation approach. An exploratory focus group informed the design of a questionnaire survey which consisted of Likert scale questions (where 1 = strongly disagree, 5 = strongly agree), multiple‐choice questions and free‐text options. The questionnaire was circulated at a single medical school to students studying various disciplines.

The questionnaire was completed by 129 students (48 male, 80 female, 1 prefer not to say). Medicine and medical science BSc were the most common programmes of study (59.7% and 24.8%). The majority of respondents were in their first (44.2%), second (34.9%) and third (17.1%) years of study. AERs had been used by 81.4% and were used regularly by 51.2% of students. The majority of students reported using them for <1 h (40.3%) or for 1 h (26.4%) on each occasion. 57.4% reported that AERs were useful or very useful. AERs were used in preference and as an adjunct to books by 24% and 57% of students, respectively. AERs such as YouTube Videos (69.0%), Acland's Video Atlas of Human Anatomy (20.2%) and Anatomy TV (18.6%) were reported to have been regularly used. Easy accessibility (95.3%), interactivity (90.7%), an attractive interface (83.6%), life‐like graphics (82.0%), consolidation of learning quizzes (80.4%), and audio narration (58.6%) were felt to be important features of AERs.

Respondents reported that increasing the accessibility of AERs, the provision of training in their use and greater integration and signposting of AERs within other aspects of the undergraduate medical curriculum delivery would likely promote their uptake.

Students reported that e‐resources are a helpful adjunct for anatomy learning. Accessibility and the ability to interact were the most important features of AERs. Students suggested that educators should aim to improve accessibility by integrating the AERs into other modalities of teaching.

This project was deemed to be a service evaluation project and was, therefore, exempt from the need for ethical approval.

S39 Transition from educational to healthcare environment: Emphasizing non‐traditional discipline independent skills in the anatomy course

Pawlina W. and Lachman N.

Mayo Clinic, Rochester, Minnesota, USA

Patient safety and high quality of healthcare delivery are essential indicators of health care outcomes. Studies in surgical environment show that sub‐optimal cognitive and interpersonal skills (non‐technical skills also called non‐traditional discipline independent skills) contribute to adverse surgical events. Importance of reinforcing these skills has received increasing attention in current literature. Promoting early development of non‐traditional discipline independent skills (NTDIS) and emphasizing their relevance for safe clinical practice is a responsibility for both clinical and basic science educators. In current basic science medical curriculum there is little emphasis on teaching and assessment of NTDIS. Anatomy courses delivered in a team‐based learning environment provide unique opportunity to teach and evaluate these skills. As students work together within the laboratory and classroom environments, they are able to evaluate and self‐ reflect on team interaction, communication, leadership, and professionalism. A variety of modalities are used to assess skills through self‐, peer‐, near‐peer and faculty evaluations. Additionally, quality of dissection and team based formative assessments add to the learning outcome that simulates the surgical team environment. Feedback regarding NTDIS assessment is shared with students during the course. Teaching faculty facilitate feedback sessions by guiding students through their interpretation of both self‐ and peer reflections and evaluations. Strong mentorship is a key to helping students internalize feedback and plan future goals for improvement. In the final course written report, the assessment of NTDIS are presented as competency alongside that of anatomical knowledge. Incorporation of NTDIS into the basic science curriculum provides a bridge between medical education and healthcare environments. Awareness of NTDIS assessment in anatomy curriculum has shown positive effect on team cohesiveness and interaction and had no adverse effects on academic performance as measured by the National Board of Medical Examiners (NBME) subject examination and United States Medical Licensing Examination (USMLE) Part 1 Anatomy scores.

S40 Internationalisation of medical education: fostering early student collaborations during the anatomy courses between medical students in the USA, Germany and Japan

Wu A. ,¹ Goelkel C. ,² Sakurai T. ,³ Kielstein H.² and Bernd P.⁴

¹Department of Medicine and Pathology and Cell Biology, Columbia University, New York, USA' ²Department of Anatomy and Cell Biology, Martin Luther University Halle‐Wittenberg, Germany; ³Department of Drug Discovery Medicine, Kyoto University Graduate School of Medicine, Japan and ⁴Department of Pathology and Cell Biology, Columbia University, New York, USA

Global health is important at a time of global interconnectedness. Internationalisation of medical education (IoM) can help with global health education and prepare future physicians to practice with a broader frame of reference and better understanding of cultural differences. The purpose of this pilot study was to introduce IoM early in medical school – during the Gross Anatomy course. We connected preclinical medical students internationally via structured videoconferencing during the Gross Anatomy course and via early student exchanges. We determined if this approach was feasible and desirable.

Medical students from the USA (Columbia University), Germany (Martin Luther University) and Japan (Kyoto University) were partnered to participate in a series of videoconferences during a 6‐week period. All students were in their first 2 years of medical school, either enrolled in or just completed their respective Anatomy courses. First, small groups of students participated in exploratory conferences – including discussions about Anatomy and Anatomy course related questions. In subsequent conferences students chose a project in which they compared their medical training, healthcare coverage or health policies in the different countries. Students from all three continents presented the results of their findings to each other during the final large group conference that included all students. Furthermore, students are continuing to communicate via a common Facebook page, while several students chose to visit their partner school to do research during their summer vacation. Questionnaires were sent after completion of the video conference. This pilot study indicated that group videoconferencing between students in different countries was liked by all participants and is feasible, although challenging, given the time differences and some technical issues. The students liked the interaction which proved informative because most students had limited knowledge of medical issues in each other's countries. Anchoring this interaction in the Gross Anatomy course was appreciated by the students and gave them a commonality that they all shared. Results from the student exchanges are still pending. Early international exposure is liked by students and may lead to a new role for Clinical Gross Anatomy courses in initiating internationalization of preclinical medical education.

S41 Flipped classroom at the university of Utah school of medicine: hammers, trumpets, basketball, Socrates and guacamole

Morton D.

University of Utah School Of Medicine, Utah, USA

A goal of higher education is to promote the synthesis of information to solve problems. However, many classroom activities require little to no preparation on the part of the student and often are the first exposure to the information. If a goal of higher education is to promote synthesis of information to solve problems, how much practice are these students getting in the classroom? The flipped classroom (FC) has emerged as one solution by exposing students to course material prior to classroom sessions via tutorials and workbooks. As a result, classroom time revolves around activities synthesizing the learning objectives. However, some studies measuring student performance in FC relative to lecture classroom (LC) have shown mixed results. What factors could play a part in the success or failure FC implementation? The University of Utah School of Medicine (UUSOM) implemented the FC for all anatomy lecture content in 2014. This presentation focuses on the lessons learned over 3 years of FC implementation: 1) The FC is one of many tools in education and like any tool is successful when used properly. (2) The FC enables students to prepare for the classroom activities and then practice solving problems with the professor's assistance to reinforce good reasoning and correct errors. 3) The rigor of the problems sets used in the classroom determines the level of discussion, learning and retention. (4) The skills and attributes of the professor guiding the classroom discussion are essential to cultivating a successful active learning environment. (5). Each element of the FC (learning objectives, pre‐class work, in‐class problem sets, student discussion, homework, and summative assessment) is required for a successful educational experience. This oral presentation details the lessons learned and provides examples of each.

This study was considered exempt by the Institutional Review Board at the UUSOM.

S42 Superficial learning is associated with decreased performance in a professional veterinary anatomy curriculum

McNulty M.^1,2 and Richbourg H.²

¹Indiana University School of Medicine, Indianapolis, USA and ²Louisiana State University School of Veterinary Medicine, Louisiana, USA

The importance of methods to promote deep learning of course material within professional curricula is widely accepted. However, to our knowledge, no studies have directly correlated assessments of superficial vs. deep learning strategies with performance within professional veterinary curricula. Thusly, the purpose of this study was to correlate these learning styles of professional veterinary students with performance within a veterinary anatomy precourse and subsequent professional curriculum. Professional veterinary students participated in a previously published, dissection‐based anatomy precourse prior to the start of the professional curriculum. The precourse included a final written and practical examination, and the subsequent professional anatomy curriculum included four examinations with both a written and practical component. At the start of the precourse, students took the published and validated Approaches and Study Skills Inventory for Students (ASSIST) to describe, in a systematic way, how students learn or study, specifically evaluating whether they are a superficial or deep learner. Their scores in each ASSIST category and subcategory were correlated to performance on each examination within the veterinary anatomy precourse and the professional veterinary anatomy curriculum. Results confirm previous studies by the authors that show precourse performance significantly predicts performance in the professional curriculum. Several significant correlations were identified between ASSIST scores and exam performance. A high score in the “Total Surface Apathetic Approach” category indicates a student who utilizes more superficial methods of learning. A significantly negative correlation was found between this score and students’ performance on both precourse examinations, as well as three examinations within the professional curriculum, indicating those students who employed more superficial learning strategies performed worse than their peers. Subcategories, including “Unrelated Memorizing” and “Fear of Failure” significantly and negatively correlated with precourse and professional curriculum exams, indicating students who utilized rote memorization of material and had a higher fear of failure performed worse than their peers. Taken with other data, these results provide direct evidence of a negative correlation between superficial learning styles and performance within a professional‐level veterinary anatomy course. Further work will be conducted regarding how students perceived their experience in the precourse and whether their experiences differed by learning styles.

This study was approved by the Louisiana State University Institutional Review Board

S43 Bone implant fixation: novel approaches for improving and imaging in the rat

Sumner R.

Rush University Medical Center, Chicago, USA

The principles for establishing and maintaining fixation of orthopaedic and dental implants are now generally agreed upon, including immediate mechanical stability and close initial contact with the host skeleton. All studies referred to in this review were approved by the author's Institutional Animal Care and Use Committee (IACUC). The material composition and surface characteristics of the implant must also be appropriate. Under these ideal conditions, most efforts at enhancing implant fixation focused on modifications to the implant itself, including various surface treatments. Locally delivered biologics such as bone morphogenetic protein or transforming growth factor‐beta and systemically administered drugs such as bisphosphonates, intermittent PTH and sclerostin antibody can enhance implant fixation in this situation. There is now interest in enhancing implant fixation when the microenvironment is not ideal, including bone loss due to the presence of particulate debris or osteoporosis. In these situations, most of the effort is on the systemic agents with the treatment tested as a preventive strategy, a logical place to start for proof‐of‐concept studies. A bigger challenge is the treatment of established disease. For instance, we found that sclerostin antibody is effective at preventing particle‐induced peri‐implant osteolysis in a rat model when the treatment is initiated early, but we have not yet determined if sclerostin antibody is effective at rescuing implant fixation in the presence of established particle‐induced bone loss. In the rat model of ovx‐induced bone loss, we found that sclerostin antibody enhances implant fixation but not nearly as well as when there is normal bone volume at the time of implant placement. Since sclerostin antibody works by stimulating bone formation on existing bone surfaces, the reduction in surface area associated with ovx presents a significant challenge. In this low bone volume microenvironment, future work should focus on the use of bone inducing agents followed by anabolic agents. Our lab has recently developed a microcomputed tomography method for assessing bone implant contact in the rat model. Using this method, we are now able to predict the fixation strength of the implant to the host bone quite accurately using a statistical model. Thus, in future studies mechanical testing may be avoidable, permitting other types of analyses to be performed.

S44 Tuning alginate bioinks to modulate their printability and stiffness in order to spatially direct MSC fate within bioprinted tissues

Freeman F. ^1,2 and Kelly D.^1,2,3

¹Trinity Centre For Bioengineering, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland; ²Department of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, Dublin, Ireland and ³Advanced Materials and Bioengineering Research Centre (AMBER), Royal College of Surgeons in Ireland and Trinity College Dublin, Dublin, Ireland

A common bioink selected for use in 3D bioprinting is alginate. It is a highly biocompatible hydrogel whose physical properties can potentially be tailored to direct 3D cell growth and differentiation both in vitro and in vivo. Molecular weight (MW) of alginate, choice of crosslinker and gelling conditions have all been shown to influence the mechanical properties of such hydrogels. In turn, the stiffness of such alginate hydrogels has been shown to be a key determinant of Mesenchymal Stem Cell (MSC) differentiation. This suggests that modulation of alginate bioink stiffness represents a promising strategy to regulate MSC fate within bioprinted tissues. The first objective of this study was to investigate the effect of MW, gelling conditions and the choice of ionic crosslinker (Calcium Carbonate (CaCO3), Calcium Chloride (CaCl2) or Calcium Sulphate (CaSO4)) has on the bioprintability and mechanical properties of alginate bioinks. A printability window was identified for alginate of differing MW by systematically varying the ratio of alginate to ionic crosslinker within the bioink. The optimal crosslinking ratio for printing depended on the MW of the alginate. MSC viability remained high (˜80%) within 3D printed alginate constructs regardless of the printing conditions. The MW of alginate bioinks and the choice of ionic crosslinker were also found to affect the mechanical properties (Young's Modulus, Degradation Rate, and Growth Factor Release) of a 3D printed alginate construct. We next explored if spatially modulating the stiffness of 3D bioprinted hydrogels could be used to direct different MSC fate decisions inside the printed construct. A construct with a soft core of 5 mm diameter and a stiff annulus of 10 mm diameter was bioprinted in an attempt to spatially direct MSC differentiation. By using the same alginate and crosslinker, but varying the crosslinking ratio, it is possible to bioprint constructs with spatially varying mechanical microenvironments. Moreover, these varying microenvironments were found to have a significant effect on the fate of MSCs encapsulated within the alginate bioinks, with softer regions of the bioprinted construct preferentially supporting adipogenesis and stiffer regions supporting osteogenesis.

S45 An injectable hyaluronic acid hydrogel modified with RGD to enhance human stem cell delivery to the infarcted heart

Gallagher L. ,^1,2,3 Dolan E. ,^1,4 Cavanagh B. ,⁵ O'Sullivan J. ,^1,2,3 Farrell T. ,³ O'Brien F.^1,2,3,6 and Duffy G.^1,2,3,6,7

¹Tissue Engineering Research Group, Department of Anatomy, RCSI, Dublin, Ireland; ²Trinity Centre for Bioengineering, Dublin, Ireland; ³Department of Anatomy, RCSI, Dublin, Ireland; ⁴Advanced Materials and Bioengineering Research (AMBER) Centre, Dublin, Ireland; ⁵Cellular and Molecular Imaging Core, RCSI, Ireland; ⁶School of Pharmacy, Trinity College Dublin, Dublin, Ireland and ⁷Anatomy, School of Medicine, RCSI, Dublin, Ireland

Myocardial infarction (MI) is a leading cause of mortality and morbidity worldwide. A typical MI results in the death of up to 1 billion cardiomyocytes, which leads to additional stress on the remaining cardiac muscle and compromises heart function. Current treatments may prolong survival but are unable to address the underlying damage to cardiomyocytes. Cell therapy has the potential to achieve the elusive goal of cardiac regeneration; however, transplanted cells display low survival and retention, a problem that affects the success of cardiac tissue engineering.

Our research aims to alleviate this problem through the combination of cells with biomaterial carriers. We focus on an in situ‐forming injectable hydrogel system, functionalized hyaluronic acid, to enhance the longevity of cells under conditions of hypoxia and reduced nutrients, as is present in ischemic injury sites. We hypothesize that the functionalization of our hydrogels with the adhesion tri‐peptide sequence, arg‐gly‐asp or “RGD” will increase encapsulated cell survival by inhibiting cell death by anoikis.

Human bone marrow‐derived mesenchymal stem cells (hMSCs) were encapsulated in HyA and HyA‐RGD hydrogels and maintained under standard culture conditions. hMSC viability was confirmed over a period of 11 days, with significant cell spreading observed in the RGD‐modified hydrogel from day 4. Having confirmed the biocompatibility of our hydrogels, encapsulated hMSCs were subjected to conditions frequently found at sites of tissue ischemia, namely hypoxia (1% O2) and reduced nutrients and the effect of the RGD modification on cell survival and function was assessed. While RGD significantly enhanced encapsulated hMSC spreading, hMSC survival in HyA‐RGD hydrogels was not increased compared to the control.

Following these results, we hypothesised that the inclusion of a 4 day preconditioning step under standard culture conditions was necessary to allow encapsulated hMSCs to adhere to the RGD peptide before transferral to adverse culture conditions. Our data demonstrates that the inclusion of a precondition step leads to an increase in the percentage of live cells within the HyA‐RGD construct along with a corresponding decrease in the number of dead cells. These results show progress towards establishing a hydrogel system to enhance delivery of cells already under clinical investigation. No ethical approval was required for this study.

S46 Close to the bone: engineering research into the biology of osteoporosis

McNamara L.

Biomedical Engineering, College of Engineering and Informatics, National University of Ireland, Galway, Ireland

While the phenomena of bone adaption to mechanical loading has been long observed, the biological mechanisms by which bone cells sense mechanical stimulus and produce a corresponding biochemical response are not fully understood. Osteoporosis renders bones susceptible to fracture, and represents a significant healthcare and economic burden worldwide. Existing drug treatments only reduce fracture susceptibility by 50%, possibly because the mechanisms initiating the disease are poorly understood. Our research has identified complex changes in bone tissue mechanics and mineral composition during osteoporosis, which are more prevalent at specific anatomical regions. However, as estrogen depletion is systemic, such changes might be related to the mechanical environment, which varies considerably with anatomical location. Thus we use multidisciplinary research techniques to enhance understanding of the mechanisms by which bone cells can sense changes in their local mechanical environment and to determine the mechanical environment of bone cells in vivo. Specifically, we have identified and quantified αvβ3 integrins and primary cilia within bone cells, which might play an important role in osteocyte mechanotransduction. We have developed a custom‐designed micromechanical loading device compatible with confocal microscopy imaging, which has been implemented a with digital image correlation (DIC) technique to characterise the local mechanical environment of osteocytes and osteoblasts. We have developed multiscale (organ, tissue, cellular), multi‐physics (fluid, solid, thermal) and anatomically representative models of bone cells both in vivo and in vitro. We have discovered that (1) the expression of mechanosensors is altered in bone cells from osteoporotic animals compared to controls and (2) the mechanical environment of osteocytes is altered at the onset of osteoporosis, but a mechanobiological response restores the homeostatic mechanical environment of the cells in the longer term. Our multidisciplinary experimental and computational research has predicted that changes in local matrix mineralisation could occur in response to such altered osteocyte stimulation during osteoporosis. Our recent in vitro studies have revealed that estrogen withdrawal alters calcium signalling, mineralisation, biochemical responses and osteogenic gene expression when bone cells are exposed to an applied fluid shear stress. Together these studies are enhancing understanding of the role of mechanobiology in osteoporosis aetiology. All animal procedures (ovariectomy) were carried out using appropriate anaesthetic and surgical procedures, following institutional ethical approval and under an animal license No. B100/4424 granted by the Irish Department of Health.

PS1. 1 Engineering the enthesis in vitro using graded mineral hydrogels

Graute M. ,¹ Pietryga K. ,² Pamula E.² and Paxton J.¹

¹Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK and ²AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Krakow, Poland

In the human body, the transition from bone to tendon (the enthesis) is gradual through distinct areas with decreasing mineral density. This allows the transition of force from hard to soft tissue through the concurrent decreasing compressive and increasing tensile strengths. This study investigates the use of novel graded mineral bone anchors in combination with a tissue‐engineered tendon and aims to replicate the native graded mineral content observed in vivo in an in vitro model.

Graded mineral bone anchors were manufactured using 0.7% gellan gum hydrogels with 5 mg/ml alkaline phosphatase addition and mineralised from one side by incubation in 0.1 m calcium glycerophosphate for 24 h. Experimental bone‐tendon constructs were manufactured using graded and non‐graded bone anchors consisting of 0.7% gellan alone. Both anchor types were pinned into custom‐designed Sylgard‐walled culture wells in 6‐well plates, to improve the formation of the bone‐tendon construct and increase similarly to the normal anatomical arrangement. Tissue‐engineered tendons were manufactured using 350 μl of fibrin hydrogel and 50 000 chick tendon fibroblasts. Bone‐tendon constructs were incubated at 37°C, 5% CO2 for the duration of the experiments. Constructs were removed from culture at day 7 and 14, fixed in 4% formaldehyde and processed for histological analysis.

The presence of a mineralisation gradient in the graded anchors was confirmed through gross digital imaging. Digital imaging was also employed to observe the formation of bone‐tendon constructs in vitro and assess the suitability of the novel graded anchors in the existing culture system. Thin sections (10 μm) of graded anchors and bone‐tendon constructs were stained with von Kossa, Alizarin Red or H&E stains to observe mineral deposition and/or gross tissue morphology. The graded anchors appeared to perform well in relation to the engineered tendon and this indicates them for use in forming an artificial enthesis.

This study has demonstrated the potential use of combining graded mineral anchors with an existing tissue‐engineered tendon model to recapitulate the unique anatomical arrangement at the enthesis. Further work is required to assess the similarity of the in‐vitro anatomy with the native structure in order to create an engineered enthesis. No ethical approval was required for this study.

PS1. 2 Dermal fibroblasts in morphologic monitoring of biodegradable materials: methodological basis of poiential application evaluation in dog dentistry

Kapustin R. ,¹ Krasnikov A. ,² Annikov V. ,² Krasnikova E.² and Starchenko N.³

¹Belgorod State Agrarian University Named After V.J. Gorin, Belgorod, Russia; ²Saratov State Agrarian University named after N.I. Vavilov Saratov, Russia and ³Belgorod Interregional Veterinary Laboratory, Belgorod, Russia

To evaluate functional state of dermal fibroblasts cultivated on titanium pieces for implants with different concentrations of a polymer film and propolis put on their surface; to determine polymer and propolis concentrations which may provide biodegradable coating nontoxicity and the required adhesive and proliferative cell culture potential.

As a material for our research, human dermal fibroblasts extracted from healthy donor skin were used. Culture viability and proliferative activity were evaluated in 1 mm of medium by means of an autocellcounter. Before the experiment was started the study titanium samples had been treated in the dry air sterilizer (“Across”, Russia), then placed in 24‐well plate (“Costar”, USA) with the follow‐up cell culture seeding (concentration – 1 × 100000 cells per a sample in 2 ml of the medium). The research was conducted with adhering to all existing ethical practices.

In the course of the experiment the optimal doses of the composite components which did not exert an inhibitory effect on fibroblasts were determined. The research demonstrated good cell adhesion (the first 5 types). Close to the presented samples high proliferative activity was also being observed. Then inhibition of cell growth followed by cell death was being observed, as evidenced by shape change (rounding) and growth cessation (types 6 and 7). The concentration of the polymer was consistently decreasing, the cell culture adhesive and proliferative potential were being improved (types 8, 10), as evidenced by the cells of the distinctive shape close to the sample. The best results were received for type 10. In 24 h after the control microscopy it was indicated that the fibroblast culture was in good condition, the cells were mainly fusiform, the processes are prominent, the nucleuses are clearly contoured.

For creating a biodegradable coating on the implant material surface the following concentrations are to be used: propolis (as a an active agent) – 1.25 mg per 1 ml, polymer – maximum of 0.0001%. Absence of culture cell damage (type 10) with a possibility of the required adhesion and proliferation on the study substances is an evidence of the biodegradable coating nontoxicity.

PS1. 3 Enhancing the efficacy of DNA‐origami delivery to cells using an on‐demand drug delivery system

Kearney C. ,^1,2 Gollapalli S.¹ and Sitram R.¹

¹Tissue Engineering Research Group, Dept. of Anatomy, Royal College Of Surgeons In Ireland, Dublin, Ireland and ²Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin, Ireland

DNA origami is the combination of a long strand of DNA with multiple short complimentary staple strands that fold the long strand into specific predesigned nanoscale shapes. These 2‐ or 3‐dimensional structures can be loaded with drugs and other molecules and then can act as therapeutics or biosensors. The precise nanoscale shapes have been shown to improve drug efficacy by changing drug‐cell interactions.

Controlled drug delivery systems position drugs locally at the target site, typically inside a polymeric depot, which controls the release rate and also has the advantage of protecting sensitive drugs from breakdown (e.g. DNA breakdown by endogenous nucleases). Temporal specificity in drug delivery can benefit a wide variety of treatments including the delivery of chemotherapy in breast cancer and tissue engineering strategies by delivering the correct cue at the appropriate time to enhance healing. DNA origami is a novel therapeutic that could show enhanced efficacy by being delivered in a spatiotemporally‐controlled fashion but this has not previously been tested.

In this work, we incorporated DNA origami into a previously designed on‐demand delivery system that incorporates drugs into an alginate hydrogel and stimulates their release using ultrasound [1]. The stability of the DNA origami in various media and time‐points was first observed using gel electrophoresis. Subsequently, the origami was incorporated in ultrasound‐responsive alginate and stimulated with ultrasound. The results demonstrated the stability of the DNA in the buffers used for release experiments and the successful release of DNA origami from the on‐demand system. This confirms the potential of the system as a delivery tool for DNA‐origami nanotechnologies. Future work will incorporate bioactive DNA origami in the delivery system and examine its therapeutic potential.

No ethical approval required for this study.

PS1. 4 Histologic evaluation of chicken thymus as one of the components for immunosupressive effect evaluation

Kapustin R.¹ and Tarasov M.²

¹Belgorod State Agrarian University Named After V.J. Gorin, Belgorod, Russia and ²Ltd. SPE NanoVetProm, Belgorod, Russia

Morphologic monitoring of immune organs enables to determine qualified dosing policy and strategy for the investigational drugs. The objective of the present research is to give thymus structural evaluation upon testing the immunosuppressive effect of the Sky‐Forse drug which is a synergistically effective nanostructural comprehensive medicine.

The experiment was initiated and conducted by Ltd. SPE NanoVetProm. It used broiler chickens divided into five groups, 10 in each group. The first group received the Sky‐Forse in the dose of 150 mg/kg of a live weight; the second one – 225 mg/kg of a live weight; the third one – 300 mg/kg of a live weight; the fourth one – 375 mg/kg of a live weight and the fifth one is controlling. The experiment duration was 23 days (from the age of 14–36 days old). The research was conducted with adhering to all existing ethical practices.

At the age of 19 days old there was no evidence of the pathologic processes in the thymus, from the age of 19–24 days old the increase of a specific content of the thymus lobule cortical areas and the lobule size was identified. There were not any differences between the groups, including the control group. At the age of 29 days old cases of Hassal's corpuscles vacuolation and focal capillary hyperemia were registered. By the age of 36 days old changes in the brain area and the Hassal's corpuscles were increasing likely accompanied by accelerated thymus involution by means of the brain area size reduction, degeneration of a part of the Hassal's corpuscles and amyloid deposits instead of the degraded corpuscles.

As the process is mostly localized i. e. there are invaded and not invaded lobules – and is not related: to the experiment, it may surely be stated that the Sky‐Forse has no immunosuppressive effect. More significant changes in the control group may even be regarded as favorable effect of the investigative drug, as functional load relief of the immune organs is surely able to reduce negative effects of possible decrease of thymus functional activity.

PS1. 5 Potential for polycaprolactone polymers in neural stem cell transplantation

O'Malley A. ,¹ Rajendiran M.² and Barry D.¹

¹Discipline of Anatomy, Trinity College Dublin, Dublin, Ireland and ²CRANN, Trinity College Dublin, Dublin, Ireland

The benefits of pluripotent stem cells in neurodegenerative disorders are well recognized. These pluripotent stem cells are intended to proliferate and differentiate in response to the local environment and produce functional myelin, thereby optimising their assimilation into diseased CNS regions. Direct application of stem cells into the injury site can result in low survival rates and early differentiation. Technological developments including the seeding of neuronal stem cells in nanofibers generated from biopolymers or hydrogel matrixes and their subsequent transplantation have shown promising results in overcoming these obstacles. Polycaprolactone (PCL) is a synthetic polyester, with strong mechanical and outstanding electrospinning properties. Electrospinning provides a 3‐dimensional scaffold with nanoscale fibre diameters and heavily interconnected pores. PCL is hydrolytically and enzymatically degraded in a physiological environment into non‐toxic products, making it an outstanding candidate as a scaffold material in tissue engineering. In this study, we demonstrate that electrospun PCL 3D scaffolds have potential in supporting the growth and transplantation of CNS progenitors. The viability, phenotype and morphology of dissociated E15 cortical and spinal cord cells seeded in these scaffolds was assessed. Electrospun PCL 3D scaffolds show great potential as a transplant conduit for neural precursor cells to the site of an injury following CNS trauma.

No ethical approval required for this study.

PS1. 6 Elucidating the role of porcine extracellular matrices from various tissue sources to modulate human mesenchymal stromal cell differentiation

Browe D. ,^1,2,3 Buckley C.^1,2 and Kelly D.^1,2,3

¹Trinity Centre for Bioengineering, Dublin, Ireland; ²Department of Mechanical and Manufacturing Engineering, Dublin, Ireland and ³Advanced Materials and Bioengineering Research Centre (AMBER), Dublin, Ireland

The interactions between cells and extracellular matrix (ECM) plays a key role in several integral processes in vivo such as cell proliferation, differentiation and the regeneration of damaged tissues. Xenogeneic ECM‐derived biological scaffolds have been used for various tissue engineering applications in clinical settings due to their ability to both provide both structural support and biological cues to aid tissue repair. The key objectives of this study are to evaluate the potential use of ECM‐derived biological scaffolds fabricated from three tissue sources for musculoskeletal tissue repair applications and to elucidate the role of these matrices in promoting and modulating mesenchymal stromal cell (MSC) differentiation.

ECM was isolated from porcine ligament (LIG), articular cartilage (AC) and growth plate cartilage (GP). The tissue was manually diced and subsequently solubilised using pepsin. Salt precipitation was performed to preferentially extract the collagen from the solubilised tissues. Following dialysis, the solubilised collagen ECM was lyophilised. To examine the paracrine role of ECM in 2D, tissue culture plastic (TCP) was coated with varying concentrations of AC, GP or LIG ECM onto which human infrapatellar fat pad (IFP) derived MSCs were seeded for 48 or 72 h. MSC proliferation, growth factor secretion and regulation of key differentiation genes was examined. To determine the optimal biomaterial based scaffold for both cartilage and bone repair applications; porous scaffolds were fabricated from lyophilised LIG, AC and GP ECM and then seeded with bone marrow or IFP derived MSCs and allowed to differentiate for 28 days.

In 2D, AC ECM was found to upregulate genes that promote chondrogenic differentiation of MSCs. MSCs cultured on AC and GP ECM were found to secrete higher levels of vascular endothelial growth factor (VEGF) when compared to TCP and LIG ECM. Under 3D conditions; scaffolds fabricated from AC ECM were found to significantly promote chondrogenesis to a greater extent that LIG or GP scaffolds. Conversely, scaffolds fabricated from GP ECM promoted osteogenesis to a greater extent that LIG or AC scaffolds. In conclusion, the choice of tissue source is a critical parameter to consider when developing ECM‐derived biomaterials for musculoskeletal tissue repair applications. Human tissue was obtained following ethical approval by the research ethics committee of the Sports Surgery Clinic Dublin (SSC). Informed consent was obtained from all patients prior to tissue donation.

PS1. 7 Engineering collagen scaffolds for advanced ex vivo culture of breast tumour

Monahan D. ,^1,2,3 McBryan J. ,⁴ O'Brien F.^5,6,7 and Levingstone T.^5,6,7,8,9,

¹Department of Anatomy, National University of Ireland Galway, Ireland; ²The Regenerative Medicine Institute (REMEDI), NUIG, Ireland; ³CÚRAM, NUIG, Ireland; ⁴Department of Molecular Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland; ⁵Tissue Engineering Research Group (TERG), Dept. of Anatomy, RCSI; ⁶Trinity Centre for Bioengineering, TCD, Dublin, Ireland; ⁷Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, Dublin, Ireland; ⁸School of Mechanical and Manufacturing Engineering, Dublin City University, Dublin, Ireland and ⁹Centre for Medical Engineering Research, Dublin City University, Dublin, Ireland

Current 2D culture methods involve growing cells in plastic flasks which does not accurately resemble cell‐cell and cell‐matrix interactions in vivo. This has led to the development of 3D culture methods which provide a more accurate model by allowing these interactions. Recently a gelatin based 3D method has been developed for the ex vivo culture of breast tumour; however gelatin poorly resembles the biological microenvironment. The overall aim of this research is to develop a collagen based scaffold to more accurately resemble the biological microenvironment for the advanced ex vivo culture of breast tumour with a future goal of adding hydroxyapetite to mimic bone metastases. This was achieved by fabricating (i) 0.5% collagen (ii) 0.5% collagen + 0.1% gelatin (iii) 0.5% collagen + 0.25% gelatin scaffolds using freeze drying methods. These scaffolds along with a commercial gelatin scaffold (Spongostan) and a collagen scaffold that received dehydrothemal (DHT) treatment were characterised by their structural properties using porosity measurements and scanning electron microscopy (SEM) followed by degradation and mechanical properties. Frozen patient tumours were grown on the scaffolds which exhibited the best characteristics. Fresh xenograft tumours were grown on selected scaffolds and scaffolds with a fibronectin coating to tailor a more specific environment. Cultured tumours and tumours before culture were stained with haematoxylin and eosin (H&E) for morphology and Ki67 to measure proliferation. It was found that all fabricated scaffolds exhibited better structural, mechanical and degradation properties than Spongostan. H&E staining revealed frozen tumours did not survive in culture and that fresh tumours maintained their morphology after culture. Ki67 staining revealed the optimal scaffold for advanced ex vivo culture of breast tumour was the 0.5% collagen + 0.1% gelatin group as it provided a 71% proliferation rate. It was also found the addition of fibronectin has the potential to offer a more specific environment in some scaffold groups. Patient tumour samples were obtained under ethical approval from the Beaumont Hospital Ethics Committee (Protocol 07/74). Mouse experiments were performed in accordance with the European Communities Council Directive (86/609/EEC) and approved by Research Ethics Committee under license from the Department of Health.

PS1. 8 Design, development and pre‐clinical assessment using a porcine model of a minimally invasive endocardial catheter for delivery of stem cells in fast‐gelling hyaluronic acid hydrogels

Dolan E. ,^1,2,3,4 Kohutova L. ,⁵ O'Neill H. ,⁴ Daro D. ,⁶ Cooney G. ,^2,3 Bellavia G. ,⁷ Straino S. ,⁷ Cavanagh B. ,⁸ Flanagan A. ,⁹ Kelly H. ,^1,4 Duffy G. ^4,10,11 and Murphy B.^2,3,10

¹School of Pharmacy, The Royal College Of Surgeons Ireland, Dublin; ²Trinity Centre for Bioengineering, Trinity College Dublin, Ireland; ³Department of Mechanical and Manufacturing Engineering, Trinity College Dublin, Ireland; ⁴Tissue Engineering Research Group, Department of Anatomy, the Royal College of Surgeons in Ireland; ⁵Tissue Engineering Group, Contipro, Czech Republic; ⁶Celyad SA, Mont‐Saint‐Guibert, Belgium; ⁷Explora Biotech Srl, G. Peroni, Rome, Italy; ⁸Cellular and Molecular Imaging Core, Royal College of Surgeons in Ireland, Dublin; ⁹Boston Scientific, Ballybrit Business Park, Ballybrit, Galway, Ireland; ¹⁰Advanced Materials and BioEngineering Research Centre (AMBER), Trinity College Dublin & the Royal College of Surgeons Ireland and ¹¹Department of Anatomy, National University of Ireland Galway, Ireland

Stem cells have shown promising preliminary data in cardiac regenerative therapy, but studies have shown poor retention/survival. Stem cell therapy could be optimised by using hydrogels to enhance cell viability/survival. However, achieving an injectable hydrogel for minimally invasive delivery is difficult due to their viscous nature, the length and small profile of the delivery device and the “blind” nature of endocardial delivery. The aim of this study is to develop and assess the pre‐clinical feasibility of a minimally invasive endocardial catheter for delivery of cell‐loaded hyaluronic acid hydrogels to the myocardium.

To determine the effect of delivery of the hydrogel through the developed catheter, mechanical and viscoelastic characterisation of the hydrogels and viability of encapsulated Adipose Derived Stem cells (ADSCs) was performed pre‐ and post‐injection. The pre‐clinical feasibility of the catheter was tested in vivo in a porcine model (Ethics approval No.904/2015‐PR). Under fluoroscopic guidance the catheter was advanced through the abdominal aorta, the aortic arch and into the left ventricle. Once the desired site of injection was attained the hydrogel precursor solutions were injected through the catheter at 200 μl/12 s. Three 200 μl injections were made 20 min before the animal was euthanised by IV injection of Tanax (0.3 ml/kg BW). Post‐mortem MRI was used to confirm hydrogel presence within the left ventricle wall.

In this study, we have developed a catheter which enables the delivery of a fast‐gelling hyaluronic acid hydrogel containing 20 million ADSCs/mL. The pre‐clinical feasibility and efficacy of the catheter was demonstrated by accessing the left ventricle of a pig by abdominal aortic access and demonstrating that the hydrogel was retained in the myocardium after injection. The mechanical properties of the hydrogels were reduced by passage through the catheter, however, the viability of the ADSCs encapsulated within the hydrogel were not compromised with 80% of the cells remaining viable 48 h after injection through the catheter. We have demonstrated pre‐clinical feasibility of the endocardial catheter for the delivery of a high density of cells embedded in a fast‐gelling hyaluronic acid hydrogel.

Conflict of Interest: Authors affiliations 5, 6, 7 and 9 are companies involved in the AMCARE consortium (grant agreement no.604531).

PS1. 9 The trabecular architecture of human perinatal lumbar vertebrae: a genetically predetermined structure?

Goodchild S. , Cunningham C. and Black S.

Centre for Anatomy and Human Identification, University Of Dundee, Dundee, UK

The lumbar vertebral column plays a vital role in the transfer of load to the pelvic complex. As trabecular bone architecture is organised to facilitate optimal load transfer, the architecture of human lumbar vertebrae has been studied extensively in the adult. Thick, well‐spaced, vertically oriented trabeculae have been observed in the central portion of the vertebral body, reinforced by dense horizontally oriented trabeculae at the superior and inferior end plates, enabling the lumbar vertebrae to withstand significant compressive forces. More recently, there has been a focus on documenting the development of trabecular architecture through the use of non‐invasive imaging techniques. Understanding the development of trabecular architecture aids in understanding the possible genetic, epigenetic and external factors that lead to the attainment of optimal trabecular patterns observed in the adult.

In this study, micro‐computed tomography (μCT) was utilised to visualise the trabecular bone structure in eleven perinatal lumbar vertebrae from the Scheuer collection, University of Dundee. Skyscan CTAnalyser was used to identify and analyse 27 volumes of interest (VOIs) within each lumbar vertebra. These volumes differentiated between superior, inferior and central areas of the perinatal centrum, as well as anterior, lateral and posterior aspects. A number of histomorphometric parameters were computed including trabecular number, trabecular separation, trabecular thickness, degree of anisotropy, structural model index and bone volume fraction. Preliminary findings suggest that the perinatal trabecular architecture mirrors that of the adult, with increased trabecular thickness and separation in centrally positioned VOIs and increased trabecular number with decreased trabecular thickness and separation in superior and inferior VOIs. While in the adult, this pattern has been attributed to weight bearing, the perinatal lumbar vertebra is a non‐weight bearing structure. This suggests that trabecular architecture during early development is predetermined and subsequently reinforced later in development. No ethical approval was required for this study.

PS1. 10 The arcuate fasciculus and verbal learning in psychosis: a constrained spherical deconvolution study in humans

McPhilemy G. ,¹ Kenney J. ,¹ McInerney S. ,^1,2,3 Najt P. ,¹ Scanlon C. ,¹ Arndt S. ,¹ Scherz E. ,¹ Byrne F. ,¹ Leemans A. ,⁴ Jeurissen B. ,⁵ Hallahan B. ,¹ McDonald C.¹ and Cannon D.¹

¹Neuroimaging, Cognition & Genomics Centre (NICOG), Clinical Neuroimaging Laboratory, NCBES Galway Neuroscience Centre, College of Medicine Nursing and Health Sciences, National University of Ireland Galway, Galway, Ireland; ²Department of Psychiatry, St Michaels Hospital, Toronto, Canada; ³Department of Psychiatry, University of Toronto, Canada; ⁴Images Sciences Institute, University Medical Centre Utrecht, Netherlands and ⁵iMinds‐Vision Lab, University of Antwerp, Belgium

Cognitive dysfunction is a prominent aspect of psychotic illness, with performance in verbal learning among the more severely affected cognitive domains. Schizophrenia is associated with disturbances in the anatomy of the arcuate fasciculus, which forms a fronto‐parieto‐temporal network associated with semantic aspects of working memory. The effect these white matter alterations have on verbal learning deficits in psychosis remains unclear.

We sought to investigate the relationship between microstructural properties of the arcuate fasciculus and verbal learning performance in psychosis and healthy control cohorts. Individuals with psychosis (schizophrenia, schizoaffective disorder, delusional disorder, mania, psychotic depression or psychosis not otherwise specified) who were clinically stable at the time of study participation, and healthy controls, provided written informed consent approved by the National University of Ireland Galway and Galway University Hospital Research Ethics Committees. Verbal learning was assessed using the Hopkins Verbal Learning Test‐Revised for 22 individuals with a diagnosis of psychotic disorder (14M/8F, mean age ± SD = 34 ± 9) and 27 controls (15M/12F, 38 ± 9). Long, anterior and posterior arcuate fasciculus segments were isolated following constrained spherical deconvolution‐based tractography of each participant's diffusion‐MRI data. Anisotropy measures and laterality indices for each segment were compared between diagnostic groups and correlated with verbal learning scores.

Individuals with psychosis showed significantly higher hindrance modulated orientational anisotropy (HMOA) in the direct pathway linking frontal and temporal cortices in the right hemisphere compared with healthy controls (F = 5.08, P = 0.03). Microstructural properties of the arcuate fasciculus did not correlate with verbal learning performance in either healthy control or psychosis groups. However, greater leftward asymmetry of the posterior segment was associated with increased verbal learning performance in controls (r = 0.50, P = 0.01).

Our results support previous findings of microstructural alteration of the arcuate fasciculus in psychosis. Greater leftward asymmetry of the posterior segment was associated with increased verbal learning performance in controls. The absence of this relationship in the psychosis group may be related to deficits in verbal learning. Future work should clarify the neural networks involved in this cognitive domain in healthy individuals so the effect of disturbances in these pathways in psychosis can be better understood.

PS1. 11 Functional morphology of the tongue and its lingual papillae in pygmy hippopotamus (Choeropsis liberiensis; Artiodactyla); LM, SEM study

Jackowiak H. , Skieresz‐Szewczyk K. and Godynicki S.

Department of Histology and Embryology, Poznań Univeristy of Life Sciences, Poznań, Poland

Anatomy of the tongue, arrangement of gustatory papillae, which are responsible for the perception of taste sensations and mechanical papillae, aiding in transfer of food over lingual mucosa, documented in mammals, constitute traits typical of taxonomic units. Important factor affecting the lingual structures is the manner of food ingestion and the method of food passage.

The study aim to describe the anatomy of the tongue and microstructure of the lingual papillae in the pseudo‐ruminant pygmy hippopotamus in light of fulfilled functions. The study was conducted on tongues of 32 and 34 years old pygmy hippopotamuses obtained from the Poznań Zoo Garden under guidelines of Ethic Committee. The samples of the tongue were prepared for LM and SEM observations.

The tongue in the pygmy hippopotamus is massive organ with movable broad apex with lateral wings and positioned ventrally by fibro‐muscular lyssa. The narrower body of the tongue with muscular torus are stable embedded at bottom of the mandible. The analysis of distribution of the gustatory papillae on the dorsal surface of the tongue reveals the presence of fungiform papillae on apex and anterior part of the body, and foliate papillae on posterolateral part of the tongue. The remarkable is lack of vallate papillae, which are constant features in mammals. Mechanical papillae are represented by short filiform papillae on the anterior part of the tongue, numerous giant filiform papillae forming a dense brush on the surface of the torus and conical papillae on the lingual root.

The obtained results in comparison with behavioural observations show the morphological similarity of the tongue of pygmy hippopotamus to the Equidae. The species specific trait is accumulation of gustatory papillae in anterior part of the tongue linked to nocturnal foraging and forage selection. The unique lyssa help in movement of fleshly lingual apex after gripping the herbage. The brush of keratinized papillae on the torus is similar to donkeys and allows keeping both aquatic and terrestrial plants during grinding with molars and helps in transport of bolus. Mentioned features can be considered as one of the many terrestrial adaptations of this semi‐aquatic mammal.

PS1. 12 The anatomy of the tongue and microstructure of the lingual mucosa in the Andean condor (Vultur gryphus, Cathartiformes)

Skieresz‐Szewczyk K. and Jackowiak H.

Departemt of Histology and Embryology, Poznan University of Life Sciences, Poznań, Poland

The tongue anatomy in birds reflects evolutionary adaptations shaped by the type of food and method of food intake. The special group among birds are species feeding mainly on carrion. Up to date the literature describe mainly the feeding behavior of scavenger birds, but there is no exactly informations about the morphology and microstructures of oral cavity organs.

The aim of the present study is to investigate the anatomy and microstucture of the lingual mucosa in the Andean condor.

The studies were conducted on the tongue of the adult Andean condor collected from the Poznań Zoo Garden by using LM, SEM and CT methods. The study was conducted in accordance with guidelines set out by the Ethics Commission.

The anatomical observations reveals the unique shape of the tongue, where the lateral part of the lingual body are raised and form kind of a gutter supported ventrally by strongly keratinized lingual nail. The edges of the raised lateral parts are serrated and form 37 to 39 teeth‐like, keratinized processes. The caudal part of the tongue is flat and covered by mucosa with openings of lingual glands. The internal skeleton of the tongue is constituted by the anterior part of hyoid apparatus, namely the paraglossal cartilage and basibranchial bone. The CT studies reveal that two separated wings of paraglossal cartilage form the skeleton of the gutter. The rectangle basibranchial bone is widening in the caudal part and its raised lateral edges form a depression for lingual glands. The microscopic observations show presence of two kind of the multilayered epithelium i.e. orthokeratinized epithelium locate in places undergoing mechanical pressure during food ingestion and nonkeratinized epithelium in place of occurrence of the lingual glands openings.

The unusual gutter‐shaped tongue with the teeth‐like processes on the edges in the Andean condor seems to be a specialized tool to collect and hold soft tissue from the carcass. The previous observation of the tongue in the Griffon vulture belonging to Accipitriformes reveals quite similar morphology, what could be a sign of structural convergence resulting of the same type of food and mechanism of food intake.

PS1. 13 Comparative ultrastructure and mineral composition of canine sesamoid bones

Baird A. , Kumar A. and Kilroy D.

University College Dublin, Dublin, Ireland

Sesamoid bones are small, flat bones that are embedded within tendons. To date, it has been thought that these bones develop within tendons in response to mechanical signals. This analysis of the ultrastructure and composition of canine sesamoid, patella and long bones is used to determine if the degree of mechanical modulation is related to chemical or to structural features. Sesamoid bone development is regulated by mechanical load; for example, patella separation is perturbed in mutant embryos that are devoid of contracting muscle.

The level of intrinsic control regulating the rate differential bone growth is significant. Differences between long bones and sesamoid bones reflect the variety of regulatory systems for each bone type. It is anticipated that all bones are influenced, to different degrees, by the same factors: genetic control, hormone levels, mechanical loading, bioavailability of solutes such as nutrients and signalling molecules, and autocrine and paracrine influences.

Sesamoid bones within the tendon of the common or long digital extensor muscle were removed from six greyhound cadavers. Along with six patellae and portions of tibia, the extracted sesamoids were subjected to Scanning Electron Microscopy in order to produce images and an atomic profile for each bone type. In addition, image analysis (using ImageJ software) was conducted to calculate bone density for each specimen.

Our data have shown that there is a difference between bone types both structurally and compositionally. Principal differences were in the Ca/P ratio and in the composition of trace elements. These factors support a variable hierarchy of influencing factors that affect growth of different bone types.

This project was exempt from full ethical review.

PS1. 14 Reptile development biology in Russia (1870–1900)

Kapustin R. , Zabolotnaja I. and Starchenko N.

¹Belgorod State Agrarian University Named After V.J. Gorin, Belgorod, Russia; ²Russian State University of Physical Education, Sport, Youth and Tourism, Moscow, Russia and ³Belgorod Interregional Veterinary Laboratory, Belgorod, Russia

There are several works on reptile development by Russian embryologists – A. Kovalevskii, A. Ostroumov, P. Mitrofanov, A. Severtsov, etc. For instance, A. Kovalevskii studied embryogenesis of the European tortoise (Emys europea). While working in Kazan’, A. Kovalevsky had collected rich embryologic material and in 1870 published an article about the tortoise development (Kiev) in which the only work available at that time in literature on embryology of these reptiles by Agassiz was subjected to Kovalevsky's critical scrutiny. Contrary to Agassiz's opinion that embryonic membranes of the tortoise consist of one neuroepidermal layer and participate in formation of nervous system, Kovalevsky clearly demonstrated that not only neuroepidermal layer, but also mesoblast participates in formation of embryonic membranes and that nervous system is formed in tortoises as well as in other vertebrate animals by means of upper primitive issue layer i.e. neuroepidermal layer. Another scientist who worked on reptile embryology was Kovalevsky's closest collaborator A. Ostroumov. He studied the development of Central Asian lizards (Phrynocephalus helioscopus Pall.) and in 1888 in Kazan’ issued a quite detailed research on history of lizard development filling in the gap in embryology of these reptiles. Before his research only embryogenesis of Lacerta genus had been described in literature. Having collected great embryologic material at the Caspian east seaside which was then thoroughly treated, A. Ostroumov greatly improved and made clear previous scientific understanding of embryogenesis of these reptiles. Initial stages of the reptile development were studied by P. Mitrofanov in the end of the nineteenth century (Warsaw, 1900). Except for A. Kovalevskii and A. Ostroumov, A. Severtsov contributed to reptile embryology as well. At the Villafrance Russian Zoological Station A. Severtsov collected great material on gecko embryology (about 400 samples). This rich embryologic material thoroughly treated by A. Severtsov formed the basis for his edition “To the history of Ascalabotes fascicularis development” (1900).

PS1. 15 Bioengineered urethra and its implication for urethral reconstruction

Danisovic L. , Galambosova M. , Varga I. and Ziaran S.

Faculty of Medicine, Comenius University in Bratislava, Bratislava, Slovakia

Urethra is the part the lower urinary tract and in males, is the part of the genital tract as well. It is responsible for transport of urine from urinary bladder to outside of the body. In males, as the part of the genital tract, is also involved in the ejaculation of semen. Many pathological processes during all developmental stages may affect urethra and thus negatively affect quality of life of the patients. In many cases the surgical treatment is the only option to resolve this problem. However, in many cases these approach use to fail. Moreover, it represent high burden to patient. Recently, a great hope is provided by tissue engineering (TE). Various TE approaches are used in order to replace missing or damaged part of urethra. The use of buccal mucosa graft is feasible, surgical technique is established, but has certain limitations, such as harvesting of buccal graft is painful, with the risk of future complications (scarring, bleeding, infection, etc.). To counteract these limitations, TE provides several possibilities in damaged urethral repair for clinical use, including the use of scaffolds, and cell seeded scaffolds. Cell culture techniques, using adipose‐derived stem cells, urine‐derived stem cells, human urothelial cells human umbilical cord mesenchymal stem cells and keratinocytes have been studied. To reduce invasivity, autologous urinary stem cells and autologous adipose‐derived stem cells show much promise, as they can be expanded in vitro. After choosing the cell source and cell culture technique, the choice of scaffold (biological, synthetic, composite) is another determinant of clinical outcome. Cell seeded grafts have encouraging clinical results, but optimal protocol‐based cell expansion and optimal scaffold for urethral replacement is still not established. Recently, 3D bioprinting with cells shows much promise. However, more the clinical experience is warranted to be fully established as a therapeutical option in a clinical setting.

Support: This work was supported by grant APVV No. 15‐0111.

PS1. 16 Anatomical evaluation of corpus callosum in patients with Alzheimer's disease in MRI image

Akin D. ,¹ Yilmaz M.T. ,¹ Aydin Kabakci A.D. ,¹ Guney F. ,² Kiresi D.³ and Guler I.⁴

¹Meram Medicine Faculty, Anatomy Department, Necmettin Erbakan University, Konya, Turkey; ²Neurology Department, Necmettin Erbakan University, Konya; ³Meram Medicine Faculty, Radiology Department, Necmettin Erbakan University, Konya and ⁴Medicine Faculty, Radiology Department, Selçuk University, Konya, Turkey

Alzheimer's disease is a neurological disease which appears by aging and cause severe damages on the brain during advanced stages. Corpus callosum (CC) which comprises of thick myelinated fibers is the largest and most important path connecting the relevant centers on right and left brain as well as enabling information exchange between the hemispheres. Therefore, the aim of the present study was to detect the modification of morphometric measurements of CC in AD when compared with healthy individuals and to reveal a regression formula of the modification appeared according to stages of AD.

Within the scope of the present study, cerebral MRI images of 100 patients who referred neurology polyclinic between 2010 and 2016 were evaluated. The patient group included 40 male and 40 female patients who were diagnosed with AD whereas 10 healthy males and 10 healthy females were included as the control group. Age, gender, stage of AD, the difference of CC measurements between first and last MRI images, CC circumference measurements, CC site measurements, CC angle measurements and parameters of the cerebral formation were evaluated in the MRI images obtained.

A regression formula was revealed from AD stage and CC measurements as corpus callosum width (CCW) and CCW = 6.974‐(0.599*AD stage); proportion of corpus callosum width to corpus callosum length (CCW/CCL) and CCW/CCL = 0.098‐(0.009*AD stage); genu width 1 (GW1) and GW1 = 11.211‐(0.748*AD stage) and maximum splenium width (SWMAX/CCL) and SWMAX/CCL = 0.172‐(0.008*AD stage). Proportion of maximum splenium width to cerebral length (SWMAX/FOU) was found through the formula SWMAX/FOU = 0.082‐(0.005*AD stage) and cerebral structure and CSF = 382.176 + (74.44*AD stage); proportion of parenchymal tissue quantity to total cerebral volume (P/T) through the formula P/T = 0.731‐(0.054*AD stage).

We believe that the data obtained in the present study may be used as reference values related to the age, gender and stages of AD. Use of regression formulas that we obtained during evaluation phase of the clinical findings of the patients may provide more precise information about stage of AD. *Approval for this study was received from the Instutional Review Board (Ref:2016/658).

PS1. 17‐ Immunohistochemical investigation of the effect of the green tea and vitamin E to streptozotocin induced diabetic rats’ vascular structure

Buru E. ,¹ Bahcelioglu M. ,² Kaplanoglu G.T. ,³ Gozil R.⁴ and Erdogan D.³

¹Faculty of Medicine, Department of Anatomy, Yuzuncu Yıl University, Ankara, Turkey; ²Faculty of Medicine, Department of Anatomy, Gazi University, Ankara, Turkey; ³Faculty of Medicine, Department of Histology and Embryology, Gazi University, Ankara, Turkey and ⁴Faculty of Medicine, Department of Anatomy, Yuksek Ihtisas University, Ankara, Turkey

Diabetes mellitus is a chronic metabolic disease due to an absolute or relative lack of insulin with tissue damage caused by the free radicals mainly superoxide anions and decreased antioxidant mechanism. Our aim was to investigate the possible effect of the green tea and vitamin E on cell proliferation and apoptosis in streptozotocin induced‐diabetic rats’ vascular structure.

The experimental protocol was approved by the local Ethical Committee for animal studies and conducted at Gazi University Faculty of Medicine. In this study, 54 Wistar albino rats were divided into nine different groups and 6 weeks after the streptozotocin (STZ) injection, some groups received orally green tea (300 mg/kg) and/or vitamin E (0.4 mg/kg) for 4 week. At the end of the tenth week, all the animals, after anesthesia, were sacrificed by cardiac puncture blood collection and then, the vessels were removed and fixed in neutral formalin for 72 h and processed for paraffin embedding. Immunohistochemical examination of anti‐AT1, anti‐PDGF and anti‐eNOS were done with Photo‐light microscope and Leica DFC280 plus camera.

Immunohistochemical investigations reveals that green tea has more protective effect than the vitamin E on angiotensine induced damage, while vitamin E is more effective and protective on PDGF expression related damage. Both antioxidant showed a maximum protective effect on nitric oxide sentetase mechanism when they are administered together.

Our results indicate that green tea has more protective effect than the vitamin E on angiotensine induced damage, PDGF expression related damage and nitric oxide sentetase mechanism. Also, the antioxidant effect of green tea is decreased with mutual usage of vitamin E indicating that the vitamin E has a controversial effect. So, the formation of antioxidant capsule with green tea should be prepared with other reagents. All our findings pointed out that several mechanisms are responsible for the oxidative stress induced tissue damage in diabetes and one antioxidant is not enough to prevent these changes. So, it is necessary to investigate the combination of several antioxidants with different effect in diabetic individuals.

PS1. 18‐ Anatomical evaluation of corpus callosum in MS patients’ MR images

Aydin Kabakci A.D. ,¹ Buyukmumcu M. ,¹ Akin D. ,¹ Poyraz N.² Uca A.U.³

¹Meram Medicine Faculty, Anatomy Department, Necmettin Erbakan University, Konya, Turkey; ²Meram Medicine Faculty, Radiology Department, Necmettin Erbakan University, Konya, Türkiye and ³Meram Medicine Faculty, Neurology Department, Necmettin Erbakan University, Konya, Türkiye

The corpus callosum (CC) is the largest interhemisferic commissural pathway which connecting cortial and subcortical areas of cerebral hemispheres. The brain structures especially CC have a tendency to be affected by morphological changes in the development and growth process. With the usage of tractography and functional MR; it's much more easy to understand neurological development process and possible local effection of the brain by neurodegenerative diseases. According to all these informations; this study had been planned to investigate morphological changes that can be occured on corpus callosum caused by MS.

This study was conducted on restrospective images of 50 patients (20 males and 30 females) whom applied to University of Necmettin Erbakan, Faculty of Meram Medicine, Neurology Clinic with Relapsing‐Remitting Multiple Sclerosis (RRMS) diagnosis, defined as RRMS or treated for RRMS. We measured many parameters related CC and distance between CC and surrounding structures on midsagittal MRI images. We assessed whether there was a relationship between the parameters CC was divided into 7 sub‐regions based on Witelson classification. The surface area of the each subregion was measured on midsagittal views.

In conclusion, morphometric changes and its statistical significant has been determined in both corpus callosum and its related surrounding structures (P < 0.05). The results of our study suggest that CC tend to have frontal localization depending on course of disease in male and female patients. Also, positive and negative correlation relation has been determined in the conclusion of the study betweens many parameters. There was a negative correlation between age and genu surface area measurement.

When we consider the localization and its function, under the many factors CC may have structural changes. Especially for the diagnosis of the diseases, structural changes of CC has Hint Point importance. We think that the obtained data from this study could be morphometric data sets for MS patients and a preliminary phase for future studies.

Key Words: Corpus callosum, ventricle, morphometry, magnetic resonance imaging (MRI), multiple sclerosis (MS).

*Approval for this study was received from the Instutional Review Board (Ref: 2015/288).

PS2. 1 3D printing in anatomy is it the future?

Smith C.

University of Sussex, Brighton, UK

Errors in medicine are on the rise. The expansion of knowledge means that junior doctors have much more to learn. Learning anatomy is essential and dissection of donors is the gold standard. However, donor levels do not meet current demand and digital learning lacks the 3D experience.

Brighton and Sussex Medical School has created a takeaway menu of 3D body parts printed on a 3D printer from a CT dataset of a donor that medical students can take away after their dissection sessions to continue their learning. Focus groups with medical students (n = 15) assessed students experiences and a RCT has shown a rise in student retention when 3D prints were used (P = 0.0006).To date over 300 body parts have been printed. An average price per print is £2.50. To assess the impact on medical student learning focus groups yielded six key themes: model properties, teaching integration, resource integration, assessment, clinical imaging, and pathology and anatomical variation. Learning anatomy is a multimodal subject and dissection of donors is a powerful and vital experience, the 3D prints really allowed students to continue their learning in a physical way after the timetabled sessions and helped students to apply their learning better to patients. 3D‐printed anatomical models can be successfully produced from the CT data set of a deceased donor. These models can be used in anatomy education as a teaching tool in their own right, as well as a method for augmenting the curriculum and complementing established learning modalities, such as dissection‐based teaching. 3D printing from donor cadavers offers an additional resource of high educational value to students, yet it remains to be proven to provide high quality education on its own. Ethical approval was obtained for this study.

PS2. 2 A modified Delphi to develop an anatomy core syllabus for undergraduate nurses

Connolly S.

University of Edinburgh, Edinburgh, UK

Purpose

The project aims to create an anatomical syllabus for undergraduate nurses.

Background

Biosciences required by nurses are anatomy, physiology, biochemistry, pathology, pathophysiology, genetics, microbiology, pharmacology and biophysics. Bioscience is an umbrella term used by nurse educators to describe these subjects as an amalgamate. Primary studies have identified a “bioscience problem” regarding a deficit in student nurses and registered nurses’ knowledge, understanding and application of the biosciences throughout the United Kingdom. Based on this evidence and little clarity on the specific biosciences that are problematic for students, we hypothesise that there is an existing anatomical deficit based on anatomy being incorporated and cited under the term bioscience. The bioscience deficit in nursing has been attributed, in part, to national widespread variability within the curriculum.

Methodology

Using the anatomical societies core anatomical syllabus for medical students as a template, participants have been invited to rewrite 181 learning outcomes for undergraduate nursing students. Ethical approval has been sought to use a modified delphi method to gain consensus. The template will be revised and refined by twenty‐four anonymised participants representing twenty‐three higher education institutes nationally. The study will be conducted in two phases. Participants will be asked to accept, modify, reject and free text comment for justification of each learning outcome. On completion, the survey results will go back to the research group. The second phase is to send out the revised syllabus from phase one to the original participants. This time, they may only accept or reject statements and make typographical modifications. Outcomes which were accepted with over 90% at each stage will automatically be included, all others will be removed, modified and amended as per the majority.

Conclusions

The literature has reached theoretical saturation regarding perceived deficits consistently reported and no conclusive performance markers identified. To date no standardised measurement has been formulated to nationally appraise if there is an actual knowledge deficit or a perceived one. Without national subject specific learning outcomes, we cannot assure quality of anatomical provision in undergraduate nursing programmes. Based on the anecdotal evidence, competency in this area may be compromised.

PS2. 3 Cadaveric surgical simulation – what is the best model?

Smith C. ,¹ Burton M.¹ and Harry L.²

¹University Of Sussex, Brighton, UK and ²Department of Plastic Surgery, Queen Victoria Hospital, East Grinstead, UK

Cadaveric dissection is now at the forefront of surgical education. Therefore, it is crucial that high quality surgical simulation is provided. To best prepare trainees for operating on patients a simulation model that most closely represents in vivo tissues should be used. This study developed a model surgical simulation programme using soft‐fix thiel and formalin cadavers to evaluate the utility of soft‐fix thiel in surgical simulation. Post‐foundation Year 2 doctors performed 3 basic surgical procedures on formalin embalmed and soft‐fix thiel cadavers. Participants completed an evaluation questionnaire after simulation on each tissue type. The results show that there are numerous differences between the tissue handling properties of formalin and soft‐fix thiel cadavers. A significant aspect of these differences are attributed to the moisture content of the tissues and there consequent pliability. Participants overall found that soft‐fix thiel cadavers were a more satisfying model for surgical simulation than formalin cadavers. This study demonstrates that soft‐fix thiel cadavers offer a more ‘realistic’ simulation model than formalin embalmed cadavers. They have a more accurate representation of in vivo tissues based on two characteristics; firstly, soft‐fix thiel cadavers have an appearance that is more lifelike, secondly, the tissue handling properties i.e. response to manual and instrumental manipulation is more reflective of tissues in surgery. This study adds to the literature that soft‐fix thiel cadaver's are a promising model for surgical simulation for the future. No ethical approval was required for this study.

PS2. 4 Twelve tips for utilising innovative artistic and creative learning and teaching methods in anatomy education

Keenan I. and Bell K.

School of Medical Education, Newcastle University, Newcastle, UK

It is essential within the modern environment and context of integrated anatomy, life sciences and medical education that innovative methods of delivering learning and teaching are developed in order to enhance learning and optimise student satisfaction while simultaneously maximising costs and resources. There are strong links between the utilisation of artistic learning methods and the enhancement of learning processes including both visual and haptic observation, as well as visualisation, reflection and visuospatial abilities. These skills are essential for students of anatomy and can also be important for the learning of medical students in other aspects of their training in the life sciences and clinical skills.

Based on recent research, educational theory and personal experiences surrounding and underpinning the use of innovative creative and artistic approaches in the delivery of anatomy and medical education, recommendations have been identified for educators seeking to incorporate such approaches into their own teaching practice. Our proposals have been produced collaboratively from both the academic educator and undergraduate student perspective. We describe twelve tips for educators for the utilisation of artistic methods as tools for learning, with a focus on a strong theoretical and research‐led basis; collaboration with artists and drawing instructors; student learning preferences; key pedagogical elements including flexible and active learning, time on task and multisensory techniques.

Practical recommendations are described as a guide for anatomy educators seeking to incorporate various learning modalities and processes with respect to innovative, creative and artistic learning methods into their teaching practice. The recommendations described here can be valuable to teachers of anatomy when incorporating artistic learning and teaching methods into medical curricula.

Ethical approval was not required for this work.

PS2. 5 Dual‐extrusion 3D printing of anatomical models for education

Smith M. and Jones J.

University College Dublin, Ireland

Two material 3D printing is becoming increasingly popular, cheap and accessible. In this paper, we describe how freely available printable files and dual extrusion fused deposition modeling can be combined to create a myriad of functional anatomical models. To represent muscle and bone we extruded Filaflex filament and polylactic acid filament respectively via a single 0.4 mm nozzle using a Big Builder printer. As an exemplar, we focused on the muscles, bones, and cartilage of upper airway and neck. The results show that it is possible to create articulations by combining flexible muscle and hard bone in a single print.

In terms of print resolution, a layer height of 0.1 mm with a shell thickness of 0.4 mm resulted in the highest quality print for both materials. Looking at accuracy, for the material PLA 5 cubes were analyzed for XYZ accuracy and these displayed the following % errors in their respective XY and Z directions 1.84 (1.28), 1.64 (1.49) and 0.24 (0.26). For Filaflex the % errors were a little larger 2.6 (2.03), 4.84 (3.26) and 2.2 (1.34). There was a statistically significant difference in the accuracy of the 2 materials for the X and Z directions (P = 0.01 and P = 0.02 respectively).

The models can be manufactured as dissectible, functional multi‐layered units and offer rich possibilities for sectional and/or reduced anatomy at relatively low cost. It has been calculated that these models are cheap to produce (ranging from $1.37–$13.20) and thus quite affordable for medical students. The approach described can liberate the anatomist from the constraints of inflexible hard models or plastinated specimens of the present and engage in the design of class‐specific models of the future. The combination of hard and elastic materials permits a much greater diversity in the manufacture of customized models which can illustrate Anatomy, biomechanics or principles of function.

PS2. 6 The role of 3D printing in anatomy education and medical training: a narrative review

Li K. ,^1,2 Kui C. ,¹ Lee E. ,¹ Chan J. ,³ Wong S. ,^2,4 Wu W. ,^4,5 Li G. ,⁷ Liu T. ,⁷ Yan B. ,^2,8 Tse G.^2,4 and Keenan I.¹

¹School of Medical Education, Newcastle University, UK; ²Department of Medicine and Therapeutics, Chinese University of Hong Kong; ³Department of Education, University of Oxford, UK; ⁴Li Ka Shing Institute of Health Sciences, Faculty of Medicine, Chinese University of Hong Kong; ⁵Department of Anaesthesia and Intensive Care, State Key Laboratory of Digestive Disease, LKS Institute of Health Sciences, The Chinese University of Hong Kong; ⁶School of Life Sciences, Chinese University of Hong Kong; ⁷Department of Epidemiology and Preventive Medicine, Monash University, Melbourne, Australia and ⁸Tianjin Key Laboratory of Ionic‐Molecular Function of Cardiovascular Disease, Department of Cardiology, Tianjin Institute of Cardiology, Second Hospital of Tianjin Medical University, China

The popularity of three‐dimensional printing (3Dp) has increased during the last decade in parallel with the development and availability of this beneficial and effective technology. 3Dp concerns the production of accurate 3D objects from a processed data set. This technology is therefore appropriate for the production of anatomical models from magnetic resonance imaging (MRI) and computed tomography (CT) data.

To understand the current relevance and importance of 3Dp, it is essential to determine the best evidence supporting the use of this technology. It is also necessary to establish the presence of any barriers preventing successful utilisation of this approach. We have conducted a narrative review to identify recent literature that describes primary research with respect to the evaluation of 3Dp in anatomy and medical education. We seek to establish the effectiveness and potential roles of 3Dp in terms of the theoretical and practical benefits of this technology in anatomy education as well as cost and resource implications. We have also investigated the utility and capability of 3Dp in medical practice and the influence of this technology on medical research in order to establish and illustrate the broad applications of 3Dp in medicine.

This narrative review was performed through utilising a student partnership approach whereby undergraduate students have worked collaboratively with academic educators. Terms related to 3Dp were agreed upon and PubMed searches identified primary studies published up to March 2017. Relevance of each study was determined against the four core principles: (i) Anatomy education; (ii) Surgical training; (iii) Medical research; (iv) Medical usage. We have identified several key areas where good evidence exists in support of the use of 3Dp in anatomy education and surgical training when compared to established and traditional approaches. We have also identified limitations with respect to the usage of 3Dp. While we conclude here that the current literature supports incorporation of anatomical 3Dp models into education and training, this narrative review is intended as the basis of a future systematic review, with or without meta‐analysis, of 3Dp in anatomy, surgery and medicine.

Ethical approval was not required for this work.

PS2. 7 Inclusion of foetal alcohol syndrome (FAS) in health profession education: a practical approach in the chick

McLaughlin D. , Keane A. and Tipping N.

Queens University Belfast, Belfast, Northern Ireland

Foetal alcohol spectrum disorder (FASD) is an umbrella term used to define a range of birth defects caused by ethanol. The most severe and clinically recognisable form of FASD, is foetal alcohol syndrome (FAS) In the United Kingdom there is no reliable evidence for the incidence of FASD, while specifically in Northern Ireland there is currently no data available. The British Medical Association reported that there was a lack of awareness of FASD within the general population and among medical healthcare professionals. They have highlighted a pressing need for guidance on the diagnosis and management of FASD in the United Kingdom. A more consistent use of diagnostic criteria by healthcare professionals in addition to thorough training and education, particularly in degree programmes, may increase, improve and enhance practitioner awareness and reporting, and thus impact on awareness in the general population.

The aim of this study was to develop a characteristic avian model of FAS, to potentially be used as a teaching aid in national medical curriculums across the United Kingdom.

Either 40 or 80 μl phosphate buffered saline (PBS) or equivalent volume of 33% ethanol was injected into the egg yolk sac of viable fertilised eggs at Day 2 of incubation. On Day 8 of incubation, treated viable embryos were fixed in 4% formalin/PBS, dissected out and eye measurements were obtained.

Treatment with ethanol in an avian model of FAS (n = 9–15) significantly reduced eye diameter in the vertical plane (P < 0.05).

The present study uses a chick model that exhibits some features of FAS, highlighting its potential use as a teaching aid. By incorporating direct practical experience of the acute effects of ethanol exposure on an avian model into medical curriculums, it is hoped that graduates will provide effective, considered and factual advice to pregnant patients.

Due to the time frame used in this investigation with regard to the development of chick embryos, no ethical approval or UK Home Office Animal License was needed to conduct these experiments.

PS2. 8 #anatomymcq – Using Twitter's survey tool for formative assessment – a pilot study

Bhattacharjee S. and Carty M.

University College Dublin (UCD), Dublin, Ireland

Abdominal and pelvic anatomy presents with layers of complexities including 3D relations of organs, and embryology along with extraordinary amount of vasculature and innervation. As coordinator of abdominal and pelvic anatomy education modules, a pilot study was initiated by the lead author to investigate the efficiency of Twitter to engage students during the second semester of 2016–2017 academic year in UCD.

The study design included posting one MCQ (multiple choice questions) from abdominal and pelvic anatomy each working day of the week (excluding public holidays) on lead author's Twitter profile (@souravdoc) with the hashtag #anatomymcq using Twitter's survey tool. The polls were open for 24 h for voting except the one posted on Friday where the polls were open until following Monday. The correct answer for #anatomymcq was tweeted the following day. The study was conducted from 23rd January‐3rd May 2017 (except two weeks of mid‐semester break in March) and altogether 67 MCQs were posted.

The MCQs received 351 votes in total with 5.24 votes in average/MCQ. The number of votes/MCQ ranged from 0 (lowest) to 31 (highest). The #anatomymcq hashtag received >10k impressions in Twitter over the total period. Few trends emerged: (i) more votes were casted on first three weekdays (Mon/Tue/Wed) than the last two (Thurs/Fri); (ii) voting rate dropped significantly after semester ended; (iii) MCQs from reproductive anatomy received less votes on average (˜1 vote/MCQ).

This presentation/poster will detail positive feedback from students as they liked the flexibility of interacting, fun of quizzing and the disciplined daily practice approach. It proved to be a good way for teachers to practice their skills on designing MCQs. Limitations encountered are: (i) having Twitter account is mandatory to be able to vote (ii) number of options/MCQ was limited to 4 only, and (iii) no figures could be embedded in MCQs.

The study provided interesting insights on engaging students using Twitter while deciphering certain trends which are helpful while teaching anatomy. The authors plan to continue the study next academic year and include other anatomy modules e.g. thoracic anatomy.

The study was conducted with approval from UCD Human Research Ethics Committee.

PS2. 9 Are anatomy learning outcomes taught at UK medical schools equitable and fit for purpose? A preliminary study

Creissen A. ,¹ McMahon D. ,¹ Cox M. ,¹ Haynes E. ,¹ Singh A. ,¹ Ttofa‐Roberts L. ,¹ Evans B. ,¹ Burch A. ,¹ Love A. ,¹ Mowatt A. ,¹ Philips A.¹ and Matthan J.²

¹Newcastle Medical School, Newcastle, UK and ²Anatomy and Clinical skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle, UK

Pedagogical content of UK medical curricula is guided largely by General Medical Council (GMC) recommendations. Nevertheless individual medical schools remain autonomous bodies and retain the ability to tailor learning outcomes (LOs) and delivery methods to their own curricula. Considerable variation in course structures across UK medical schools has resulted in lack of uniformity in the delivery and implementation of anatomical content on the different medical programmes, potentially adversely impacting on doctors’ diagnostic ability and/or limiting career choices. This study aimed to map the anatomy LOs of four UK medical schools against those defined by the Anatomical Society's 2015 core anatomy syllabus, some 156 anatomy LOs deemed foundation knowledge for undergraduate medical students, to gauge the extent of use of the anatomy syllabus in guiding basic science delivery and delivery of equitable anatomy knowledge across medical schools.

Anatomy LOs of four medical schools, with integrated course structures, were reviewed. The LOs were mapped against the suggested core anatomy syllabus by two independent assessors and disparities were identified. To ensure reliability of results and achieve consistency, assessor discrepancies were discussed.

Core anatomy outcomes present in the analysed medical schools’ LOs were at 61%, 63%, 85% and 90%, respectively. The variation of nearly 30% highlights the differences in anatomy education being received and delivered at different institutions, a finding potentially impacting on those wishing to pursue careers where anatomy knowledge is considered fundamental. Moreover, the LOs of many medical schools were overarching outcomes, often covering several LOs from the core syllabus. This lack of clarity potentially adversely impacts on knowledge acquisition for inexperienced students unable to decide on what should be considered key information.

This preliminary study suggests that anatomy education may require a nationwide overhaul to meet GMC standards, the anatomical aspects of which are informed by the core anatomy syllabus. A wider study mapping all UK medical schools’ anatomy LOs is imperative to raise awareness of the relevance of the core anatomy syllabus as the minimum requirement for all UK medical undergraduates and to ensure equity of learning across UK medical schools. No ethical approval was required for this preliminary study.

PS2. 10 Surgical approaches to human cadaveric dissection for the anatomical training of aspiring surgeons: medical student perspectives on dissection and the incidental discovery of pathology

Creissen A. ,¹ Haynes E. ,¹ McMahon D.¹ and Matthan J.²

¹Newcastle Medical School, Newcastle, UK and ²Anatomy and Clinical skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle Medical School, UK

Anatomical education in medical curricula has shifted from traditional whole‐body dissections to relying heavily on the use of prosected cadaveric specimens. This limits the opportunity for students to gain comprehensive knowledge of how anatomical structures relate to each other, whilst neglecting development of dissection skills. Student‐selected components offered during the clinical phase of medical degrees can act to remediate this, affording an opportunity for students to bolster both their surgical acumen and anatomical knowledge. Dissection projects performed by three medical students in their clinical years sought to simulate surgical approaches to joints commonly operated on during orthopaedic practice.

Two students performed a Hardinger (direct lateral) approach to a hip on the same cadaver whilst the third, on a separate cadaver, performed medial parapatellar and posterior approaches to the knee. The cadavers were of an 86‐year‐old male and an 88‐year‐old female respectively; preservation occurred using Vickers Embalming Fluid Number 4, administered via gravity‐feed, without drainage, into the common carotid artery.

Utilising a surgical approach, as opposed to the traditional tissue dissection employed when prosecting specimens for teaching, introduced methodologies of use in future surgical careers, whilst allowing the students to appreciate the importance of surrounding anatomy and the necessity of careful dissection when limited by the constraints of surgical, outcome‐led practice. Specifically, the Hardinger approach allowed for identification of the femoral and inferior gluteal nerves, both structures being at risk of iatrogenic injury with this approach. Furthermore, the presence of a previously undocumented left neck of femur fracture provided valuable insight into the soft‐tissue and anatomical changes associated with such an injury and how to adapt to these accordingly during dissection. The medial parapatellar approach to the knee helped develop a familiarity of the challenges associated with approaching the knee joint and a recognition of the structures within the joint itself; a posterior approach facilitated an improved understanding of the boundaries, content and importance of the popliteal fossa, in particular of the importance of careful dissection when operating in its environs.

No further ethical approval was required beyond the remit of the Human Tissue Act.

PS2. 11 Towards development of an online formative assessment tool to increase student engagement in anatomy dissection

Labberté M. and Bhattacharjee S.

School of Veterinary Medicine, University College of Dublin, Ireland

An existing challenge in anatomy education is to augment the learning experience of students during dissection classes where they have the opportunity to develop in‐depth understanding on location, relations and functionalities of various anatomical structures. A sound pre‐dissection preparation for students is pivotal in making their dissection sessions more rewarding, helpful and effective. Therefore, a series of formative assessments for students were developed within UCD School of Veterinary Medicine and executed through the online Blackboard suite which provided the students an opportunity to test their preparation before engaging in dissection on a weekly basis. The tests were termed as “online tickets” which – although not mandatory – conceptualized a “ticket” which students needed to obtain/earn before participation in dissection. Each of such six ‘tickets’ contained 10 to 25 single correct answer type MCQs tailor‐designed to test knowledge on the topics of focus in the dissection classes scheduled that week. The tests were made available to students through online blackboard interface for a 24 h window on the weekday before the dissection class. Two student cohorts – graduate‐ and undergraduate‐entry – were subjected to such formative assessments.

Despite being an optional component, the “online tickets” enjoyed high participation from students with positive feedback. For undergraduate class 99% of students engaged with the tests, with 73% taking all six tests. For the graduate entry class the uptake was even higher with 100% of students engaging and 93% taking all tests. Such exercise also exerted a positive impact on the time engagement with other online teaching resources. The pooled data obtained through blackboard provided with an excellent platform to plot, track and trace students’ engagement at both individual and group‐levels. Furthermore, it promises real‐time numerical read‐outs which will be able to distil out trends and correlations including that of student performance within definite time periods. Such data analysis investigating correlations of performances by students in formative assessments with their final summative grades for these modules are currently being undertaken in our group.

The study was performed with permission and in accordance with policies laid by UCD Human Research Ethics Committee – Sciences (HREC‐LS).

PS2. 12 Splendid learning: blended learning in anatomy

McLaughlin D.

Queens University Belfast, Belfast, Northern Ireland

In 2013, the Centre for Biomedical Sciences Education (CBMSE) introduced an MSc in Clinical Anatomy at Queen's Univeristy Belfast. Modern students have 21st century skills, yet anatomy in Queen's has for many years, still used a 19th century teaching approach. Anatomy lends itself well to online learning as the main subject content is universal, and does not change year‐on‐year. Along with colleagues from Information Services, staff members from CBMSE developed a series of ‘web apps’; small web pages hosted within the University. This approach to studying anatomy was thought to be attractive to clinical practitioners who were not able to commit full time to the MSc course offered in the University.

The implementation of blended learning in Clinical Anatomy aims to provide working healthcare professionals an opportunity to achieve a PgCert or PgDip in Clinical Anatomy. The course provides students with a greater understanding of advanced human anatomy including appropriate radiological anatomy, microscopic anatomy and embryology.

Students are introduced to the topic through short descriptive texts outlining the aims of each ‘app.’ Instructional narrated demonstration videos orientate the student with gross, functional and applied anatomy. A self‐directed learning section summarises the main learning outcomes of the topic area, as well as suggesting further areas for study and investigation, such as recent scientific journal articles.

Blended Learning is a relatively new concept within CBMSE. While there is a significant initial investment of staff time to develop and create content, it is a very time efficient model. Nonetheless, it is an extremely useful tool for teaching a highly visual subject, such as anatomy. It seems likely that technologies will become more embedded in the way anatomy is taught at Queens. However, given the many ways and resources in which anatomy is now supported in the University, the integration of ‘digital anatomy’ alongside ‘traditional anatomy’ should be a smooth process.

PS2. 13 Using an automated response system as a pedagogical strategy to enhance anatomy & physiology teaching in an undergraduate nursing curriculum

Meskell M. , Davies C. and Redmond C.

School of Nursing, Midwifery & Health Systems, University College Dublin, Dublin, Ireland

Anatomy & Physiology is a subject that students on pre‐registration nursing programmes find particularly challenging. Much of the emerging research has focussed on the promotion of active learning strategies to better engage students with anatomy & physiology content in curricula.

Using Audience Response Systems (ARS) such as clickers can enhance formative assessment and classroom room learning by promoting participation with lecturers and peers and active involvement in one's own learning. This study sought to investigate if the use of ARS technologies had a positive influence on student's perception of anatomy & physiology teaching & learning. Ethical exemption approval was granted by the UCD Research Ethics Committee. The study population was 1st year students on a Bachelor Nursing & Midwifery programme that had attended the Nervous System Unit as part of the Anatomy & Physiology module. The clicker technology system used the student's smartphone device, wireless system and a software package to allow active engagement in the lecture theatre. During lectures students answered anatomy and physiology MCQ's using the ARS.

Data was collected from students on the use of the ARS using a semi‐structured questionnaire (n = 91). Data measurement included a Likert scale and open ended questions. The questionnaire included 12 items and five scales related to formative assessment theory.

The majority (89%) of the students reported that the clicker system helped them engage more in content during class compared to traditional lectures. A more modest 64% agreed that clickers increased their concentration in class. The sharing of class answers were reported as significant for confidence building. 86% agreed that the clickers helped them to be more active in their learning, with 81% agreeing that they enhanced their learning during class.

The highest impact on learning related to the item on receiving feedback which is significant with 93% reporting that they felt the feedback from the lecturer helped clarify concepts and enhance learning. The students attributed greater engagement, active learning, goal setting and enhanced learning compared to traditional lectures. There was also a strong appreciation that the interaction of the lecturer and inclusion of feedback improved the student's experience.

PS2. 14 Assessment of a 3D tooth morphology quiz for dental students

Lone M. ,¹ Vagg T. ,² Theocharopoulos A. ,³ Cryan J. ,¹ McKenna J. ,³ Downer E.⁴ and Toulouse A.¹

¹Department of Anatomy and Neuroscience, University College Cork, Cork, Ireland; ²Department of Computer Science, University College Cork, Ireland; ³Cork University Dental School and Hospital, Ireland and ⁴Discipline of Physiology, Trinity College Dublin, Dublin, Ireland

The capacity to recognize the 3‐dimensional (3D) features of teeth (tooth morphology) has a central role in the dental curriculum and provides the foundation for the clinical years and dental practice. Teaching has traditionally relied on methods that include studying with extracted teeth and/or plastic models, drawings and wax/chalk carvings. In recent years, these have sometimes been supplemented with 3D computer generated models to further consolidate skills acquisition.

To supplement tooth morphology teaching a 3D quiz application (Tooth Morphology Quiz, TMQ) was developed. This 3D resource enables students to study tooth morphology actively by selecting teeth from the interactive quiz, modify their viewpoint and level of zoom of a particular tooth. Additionally, students are able to rotate the tooth to obtain a 3D spatial understanding of the different surfaces of the tooth. Research has shown that the use of quizzes (with feedback) promotes active learning, assists in examination preparation and keeps learners motivated.

Using a cross‐over study design, the efficacy of the newly developed quiz was compared to the traditional study of extracted teeth and plastic models. Students were assessed on their capacity to identify teeth and recognize specific features using extracted teeth and 3D images from the TMQ. Results showed no difference in performance between students using the TMQ or extracted teeth. However, in the early stages of learning tooth morphology, male participants performed better than females, possibly due to a better grasp of 3D complexity as measured by a mental rotation test. In addition, correlations show that student performance is positively correlated throughout the various phases of the assessment. Finally, a comparison of two student cohorts (2016 v. 2017) shows that students have significantly benefited from the implementation of the TMQ as class assessment was significantly higher for the 2017 group.

In conclusion, our results show that the 3D tooth morphology quiz is of benefit to students in developing their capacity to recognise the individual morphological features of teeth.

PS2. 15 How do student's achievements reflect curriculum in problem based learning?

Dabužinskienė A. ,¹ Burkauskiene A. ,¹ Udani N.² and Kleina R.³

¹Institute of Anatomy, Lithuanian University of Health Sciences, Kaunas, Lithuania; ²Faculty of Medicine, Lithuanian University of Health Sciences, Kaunas, Lithuania and ³Department of Pathology, Riga Stradins University, Latvia

One of the modules of Lithuanian University of Health Sciences in problem based learning studies is ‘‘Locomotion’’ module. It includes 13 subjects with curriculum 213 student's workload hours (h), 8 ECTS credits and they give correspondingly percentage for cumulative evaluation of students achievements – 100%: Anatomy (45 h – 20%), General surgery (25 h – 10%), Physiology (23 h – 10%), Essentials for Medical Diagnostics (19 h – 8%), Orthopaedics and Traumatology (16 h – 6%), Radiology (14 h – 6%), Physical therapy (13 h – 6%) Human Histology and Embryology (13 h – 6%), Biochemistry (13 h – 6%), Environment and occupational medicine (11 h – 6%), Pharmacology (8 h – 6%), Nursing (8 h – 6%), and Biology (5 h – 4%). Student's achievements in module are evaluated in 10‐score system. Goal of our study was to estimate correlation between curriculum and studying results. We particularly rate and compare subject's assessment and final evaluation of 762 students in second year for two academic years. We determine an average of student's results in Environmental and Occupational medicine – 88.3%, Physical therapy – 88.0%, Biology – 87.3%, Anatomy – 80.7%, Radiology – 81.2%, Nursing – 79.6%, Orthopaedics and Traumatology – 75.2%, Human Histology and Embryology – 72.7%, Biochemistry – 67.3%, Essentials for Medical Diagnostics – 66.3%, Physiology – 62.8%, Pharmacology – 61.9%, General surgery – 57.0%. The average grade of final exam module is 7.3. We find that, if the students started the module with previous basic knowledge in biology, anatomy, and histology, they were able to master knowledge and get higher assessment. The assay shows that student's lack knowledge in physiology, biochemistry, and pharmacology. We conclude that these results demonstrate that students achieve worst in clinical subjects. We suppose that basic knowledge is of great significance and students must acquire them before problem based learning modules.

PS2. 16 Faculty perspectives of embryology in North American medical education

Cassidy K.

University of Nebraska Medical Center, Omaha, USA

The anatomical sciences are experiencing a notable decrease in the time and resources devoted to embryology in North American medical education. With more changes assured, it is necessary to investigate the current trends in curriculum, pedagogy, and personal experiences of embryology educators. An original online mixed methods survey was distributed to current faculty who teach embryology in a medical school, and followed by voluntary interviews. This study was performed with the understanding and consent of the subjects, and ethical approval from the researcher's home institution. The researcher used grounded theory methodology to analyze the qualitative survey components and descriptive statistics to analyze the quantitative survey components. Of the 42 participants, 88% teach at allopathic medical schools, 31% with at least 16 years of experience teaching embryology and 31% with less than 5 years. The integration of embryology into other courses was present in 70% of the participants’ institutions while the other 30% were stand‐alone embryology courses. Sixty‐three percent of these individuals teach embryology because they volunteered, 25% were assigned, and 12% cited other reasons. When asked to rate their confidence level teaching embryology on a scale of 1 (low) to 10 (high), 79% reported at least a 7, with a mode of 7.5. Analysis of Likert items show that generally faculty find embryology interesting, moderately difficult, and both important and applicable to the education of future physicians. A combined grounded theory methodology and phenomenological approach was used to analyze the interviews with faculty. This generated a lived experience narrative of the phenomenon of teaching embryological content to medical students in the modern world, which led to a better understanding of the needs and challenges that face this subject matter and those who teach it. These include improved integration and curricular placement, enhanced educational resources, and a greater focus on clinical application. In this fluid era of medical education reform, the perspectives of faculty who teach embryology content are invaluable to assessing the status of embryology and formulating evidence‐based recommendations for the future.

PS2. 17 Deciphering the limitations of neuroanatomy teaching web‐resources – a perspective

Javaid M. , Cryan J. , Schellekens H. and Toulouse A.

University College Cork, Cork, Ireland

Neuroanatomy is perceived as a difficult topic and impairments in the understanding of neuroanatomy have been linked with a reduced confidence in managing neurology patients by health care trainees and professionals. Online web‐resources could facilitate learning intricate neuroanatomical concepts, especially in the context of diminishing teaching hours. However, despite the abundance of neuroanatomy web‐resources, students continue to report difficulties in learning the subject. Hence, we have sought students’ perception regarding the limitations of existing resources in the specific context of learning the neuroanatomical pathways. Deciphering such limitations could have clinical implications as facilitation of learning could improve confidence in clinical situations.

A systematic search for neuroanatomy web‐resources was performed using custom search strings. A stringent inclusion/exclusion criteria generated 159 web‐resources. This list was reduced to 24 resources using a scoping review criteria where exclusion was based on (1) histology‐only information, (2) lack of interactivity (3) partial information,. This was narrowed down to 13 and subjected to and “educators’ evaluation”, which was informed by educational/cognitive learning theories. Five UCC anatomy educators ranked these resources based on their usefulness in the context of student‐learning of spinal pathways. The top three web‐resources, were subsequently compared and ranked by undergraduate students (42 participants) based on how well various features were employed to aid in learning spinal pathways (Likert‐questions). Ethical approval was obtained from Social Research Ethics Committee, UCC.

Educators’ evaluation revealed that all the web‐resources analysed had limitations with regards to educational and cognitive learning outcomes. Students ranked the following pedagogical domains in terms of their usefulness in aiding neuroanatomy‐learning: (1) clarity of explanation, (2) explanation of key principles, (3) step‐by‐step drawing of pathways, (4) usage of cross‐sectional images, (5) online quizzes and feedback, (6) radiology images, (7) animations, (8) 3D computer models (9) linkage of neuroanatomy with disciplines such as neurophysiology, neuroradiology, (10) solving neurological cases and (11) various features helping in 3D visualization and orientation of brain structures.

Our study has identified a number of web‐resource features that are useful with regards to student learning experience while the educators’ perspective provides an optimal educational framework. This will inform instructional design of future resources to better achieve learning outcomes for undergraduate students and reduce the prevailing neurophobia.

PS2. 18 Does human anatomy teaching in clinical settings improve understanding of basic anatomical concepts in pre‐clinical medical students? A pilot study

Eltayeb M. ,¹ Altahir A.² and Matthan J.³

¹Newcastle Medical School, Newcastle University, Newcastle, UK; ²Newcastle Medical School, Newcastle University and ³Anatomy and Clinical Skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle Medical School, Newcastle University, Newcastle, UK

Medical students, as future clinicians, will be required to apply their anatomy knowledge in medical imaging. Numerous clinical and radiological resources are available for students to learn anatomy and apply it clinically. Few studies show early exposure to real‐life clinical settings in the teaching of anatomy. We compared head and neck anatomy teaching between two student groups, one taught in the Dissecting Room and the other additionally in a clinical setting, to gauge whether early exposure to anatomy teaching in a clinical context improves anatomy knowledge retention.

Teaching within a clinical setting was incorporated into head and neck anatomy practical sessions. 30 students participated in this pilot study. Students were divided into three groups. Initially, teaching in clinical environments was conducted with one student who felt comfortable with exposure to different imaging modalities on a neurosurgical ward. After exposure to imaging, the student was shown patients with different treatment devices and was encouraged to work out the use of these devices. Later, an entire group was taken to the ward after dissecting room teaching and the same order of teaching conducted. All sessions were conducted by two anatomy demonstrators and a post‐session quiz was conducted.

Preliminary results suggest that students felt more knowledgeable and confident about their anatomy knowledge after early clinical exposure. Students taught in a clinical setting in addition to the dissecting room were more confident with their anatomy knowledge when compared to the group of students only exposed to dissecting room teaching. They considered this adjunct to traditional teaching to have improved their understanding of anatomy considerably.

Our results suggest that early introduction to anatomy teaching within a clinical setting may increase students’ understanding and knowledge of basic anatomical concepts and 3D visualisation. Early exposure to clinical settings may prime students for later encounters with better understanding of applied anatomy. A larger‐scale study comparing the two cohorts would help elucidate any further benefits to anatomy teaching in a clinical setting in the pre‐clinical years.

Full ethical approval will be sought for a more extensive study. No ethical approval was required for this pilot study.

PS2. 19 The development and evaluation of a digital histological and pathological learning resource for medical education

Calladine K.

Division of Medical Sciences & Graduate Entry Medicine, School of Medicine, University of Nottingham, Royal Derby Hospital Centre, Nottingham, UK

Medical Education is a constantly changing environment, in which virtual microscopy (VM) and gamification are relatively new concepts. There are several established beneficial factors from using VM, rather than light microscopy for histology. Development of learning resources (LRs) which incorporate both virtual microscopy and gamification have not been used for histology. Therefore, this study aims to assess the value of both VM and gamification within medical education.

The study and aims were separated into three parts. The first study assessed the VM in the education of first year BSc students. The second study assessed VM in education of second year BSc students, followed by further assessment of a learning resource combining VM and gamification. Finally, the third study assessed the use of gamification as an engaging tool in education for first year graduate medical students and assessed performance before and after exposure to the LR.

Study one: n=48–49 students from a cohort of 54 students participated. There was a significant increase in the mean scores following the VM session (P‐value = 0.0148), demonstrating VM as an efficient educational tool. Study two: n=6–18 students from a cohort of 25 showed an increase in performance in mean scores following the VM session (P‐value = 0.0047). This increased further following the second intervention (LR). Study three: n=12–20 from a cohort of 86 showed a significant increase in the mean scores following the use of the LR (P‐value = 0.0291). Post‐evaluation questions demonstrated that the participants preferred the gamification aspects as opposed to traditional methods. Post‐evaluation questions demonstrated engagement and metacognition in all three studies.

The VM sessions within the University of Nottingham have shown to enhance the knowledge of students in both histological and pathological concepts. The use of LRs has proven to be useful in supplementing medical student understanding. Use of gamification has also increased student engagement, which demonstrated attention span and active learning. Qualitative data collected demonstrated metacognition in participants. This study supports the continued use of VM, its expansion into medical education and the potential for gamification to actively engage students in critical evaluation of histopathological material.

PS2. 20 Online anatomy assessment versus the traditional methods: a comparative study of student performance

Kilroy D. and Kumar A.

University College Dublin, Dublin, Ireland

As the veterinary curriculum prepares students for a competitive professional career, it is essential that the assessment of learning outcomes can stratify merit. Hence objective comparison of different assessment formats is essential. In this study, we aimed to determine whether an online multiple choice question examination was a suitable way of testing anatomy knowledge and if it provided a potential alternative to the traditional blended format of assessment (essay paper, practical exam and viva).

The final exam results of second year veterinary students (85±5 students/year) were compared over a three‐year period (2014–17) for the Neuroanatomy and Locomotion modules. These modules use different methods of assessment. In neuroanatomy, each student sits a 2‐hour essay paper and is required to answer four of five questions; this constitutes 40% of the final grade. The module examination for locomotion is a 90‐minute online MCQ which contributes 70% of the final mark.

In each year surveyed, the cohort performed better on the MCQ examination, with more students on average attaining an A grade (average 15 versus 2) and fewer failing students (average 9 versus 21). The distribution of grades B through D was broadly similar for both modules.

We concluded that the online format of examination has some obvious advantages (absence of choice, objective grading and reducing the marking load of examiners). Given the grade distributions, the MCQs should be standard set to prevent clustering of results around the upper end of the marking scale.

This project was exempt from full ethical review.

PS2. 21 Eye – one of the most complex organ in our body studied with help of clinical imaging resources

Dolaszynska B.

James Paget University Hospital, Great Yarmouth, UK

From 230,000 residents in the area, 50,000 are the patients seen in Eye Clinic yearly. Someone said that “Our eyes are made up of over 200 million working parts”. If we think about “how it is possible by such a small organ”, then well …. what do we know about our eye structure and what is the effective method of learning it?

Presented abstract emphasise importance of interesting way of teaching on the example of studying anatomy of the human eye and it correlation with disease.

Study shows that in a cohort group of 30 students of medical school, anatomy of the structure of the eye is not fully understandable. 100% of participant cannot name the layers of retina correctly and in order.

Those, who are thinking about career in ophthalmology are trying to memorise the sequence without understanding which is often temporary knowledge only.

Study was conducted between 2 groups each of 15 medical students. Results showed how important is introduction of pathological aspect while studying just eye anatomy. Making correlation between anatomy and disease of presented ophthalmological structure had very positive impact on the results of later on knowledge evaluation test. In the study, explanation of the anatomy of the eye based on the imaging system like OCT or autofluorescence, has 95% of success in reflection of knowledge checked by test. 80% of students successfully finish the MCQs regarding pathological finding presented on above mentioned investigations.

Among the diseases presented on the imaging were age‐related macular degeneration, central serous retinopathy, choroidal tumours and nevi, inflammatory diseases, optic nerve head drusen, pattern dystrophies, retinal toxicity and retinal detachments and others. Mean of correctly named disease presented on the image was 87%. No ethical approval was required for this study.

PS2. 22 The impact of spatial awareness training on the spatial ability of medical students learning anatomy

Gonzales R. , Smith C. and Ferns G.

Brighton & Sussex Medical School (BSMS), Brighton, UK

Spatial ability is the capacity to understand and remember the relationship between different parts of an object, and between the object and its surroundings. This ability is important for orientation and navigation in the human body and for decision making during surgery. The question of this study was to determine if this type of training has an impact on medical students learning anatomy. This project was approved by Brighton and Sussex Medical School's Research Governance and Ethics Committee (BSMS RGEC). Each participant gave written consent. A standard spatial ability test of rotation and a shortened two‐hour version of a training programme were adapted for this study. Of the 196 first and second year medical student taking the spatial ability test, 29 received training on spatial skills (intervention). These students were tested after training and their scores were compared to 75 classmates, who also took the second test but did not receive training (controls). Training comprised two workshops on spatial skills. The average of the first spatial ability score was 65.5% (SD = 18.8). There was a gender difference with female students scoring less than male students (Females Mdn = 60, Males Mdn = 75, P ≤ 0.05). Those re‐tested, whether they received the training or not, had an improved score on the second test (P ≤ 0.001). The intervention group scored higher than the controls (difference 3.8% P= 0.19); this small‐sized effect was larger in males. The improved scores in the second test suggests a combination of “learning effect”, despite one to three months of separation between tests, and the theoretical effect of learning anatomy, which is known to enhance this ability. The high scores in spatial ability support the hypothesis that medical students may represent a high‐achieving sample of the general population. Studies found that students who gain high scores in spatial ability tend to choose sciences and technology courses. The small effect of training could be related to the short duration of the intervention. Training could be extended in future trials or integrated in the anatomy curriculum to enhance content quality and ease the learning curve.

PS2.23 The effect of peripheral nerve injury and chronic inflammation on HCN channel subunit expression in rat dorsal root ganglion cells

Boyle K. , Robb A. and Hughes D.

University of Glasgow, Glasgow, UK

Hyperpolarisation‐activated cyclic nucleotide‐gated (HCN) channels are tetrameric non‐selective cation channels which perform several important functions in both central and peripheral neurons, including dendritic integration of synaptic input, setting resting membrane potentials and regulating spiking frequency. There are four HCN channel subunits (HCN1‐4), and each confers distinct properties on functional channel complexes. Little is known about the expression of HCN channel subunits in functional populations of sensory afferents, or how these change in chronic pain states. The aims of this study were to i) characterise the expression of HCN channels in neurochemically‐defined subpopulations of dorsal root ganglion neurons (DRGs), and ii) determine whether these HCN expression patterns change following the development of allodynia in animal models of chronic pain.

We looked at the pattern of HCN subunit expression in the cell bodies of cutaneous low threshold mechanoreceptive myelinated afferents that innervate allodynic skin territories in rats after either unilateral peripheral nerve injury (spared nerve injury model) or chronic inflammation (intraplantar co‐injection of CTb and complete Freund's adjuvant, CFA). All experiments were performed in accordance with UK Animals (Scientific Procedures) Act 1986. Surgical procedures were carried out under isoflurane anaesthesia, and animals were later terminated by overdose with pentobarbitone.

The proportions of DRGs expressing HCN1 in both nerve‐injured and inflamed animals (87.0 and 82.3%) were similar to naïve rats (89.1%), however, the proportions expressing HCN4 were higher in both pain models (42.8 and 68.7% in neuropathic and inflamed rats, respectively, compared to 19.4%). The proportions of lesioned DRGs expressing HCN2 were lower in neuropathic animals (26.4%) than in naïve (48.4%), but similar in the chronically‐inflamed group (43.3%).

Our results show that HCN channel subunit expression in myelinated afferents changes following the development of tactile allodynia in both neuropathic and chronic inflammatory models. Such plasticity is likely to result in aberrant afferent fibre activity, and may contribute to the development of tactile allodynia seen in these pain models. Targeting this apparent plasticity in HCN channel subunit expression may therefore offer novel avenues for developing new treatments to alleviate the chronic pain associated with peripheral nerve injury and chronic inflammation.

PS2. 24 A Qualitative radiographic analysis of human femoral development pertaining to gait acquisition and maturation

Fitton C. , Cunningham C. and MacLean S.

University Of Dundee, Dundee, UK

The premise of bone functional adaptation dictates that bone will respond to the mechanical forces exerted upon it. As, the loading conditions encountered throughout development are not consistent, a bone will experience differential biomechanical stresses throughout ontogeny. This study aims to investigate if the biomechanical demands on the femur, at advancing developmental milestones, are reflected in its gross internal structure.

Radiographs of seventy‐three femora from the Scheuer Collection, University of Dundee were selected for this study, representing individuals from the perinatal period to 7–9 years. A five‐colour gradient map was superimposed onto each radiograph. This produced two‐dimensional colour map representations of the internal structural intensities of the femur. The proximal and distal extremities were analysed separately with the age of specimens remaining blind to the author. Specimens were grouped according to external morphology and bone intensity patterns. Increased intensity was prevalent in the least mature specimens both proximally and distally, but declined up to approximately 6 months post‐partum.

In‐utero movements and the accumulation of a calcium reservoir offer the most likely explanations for this increased intensity although both genetic and vascular influences are also possible.

The decrease in calcium parallels the rapid growth characteristic of the first year of life, as the bones temporarily prioritise growth over strength. The distal femur exhibited increased lateral intensity, contrary to the medial load bearing of the adult. Differences in muscle activation and gait patterns could explain the discrepancy, however the paucity of literature regarding the distal femoral ontogeny, limits comparisons.

This research documents a previously unknown perspective of femoral bone development offering to expand current knowledge of normal bone development and functional adaptation of bone. This has the potential to be used in developing a standardised qualitative assessment used by clinicians to determine if a patient presents with normal bone density.

No ethical approval was required for this work.

PS2. 29 Sexual dimorphism of the human masseter muscle and endocranium

Humphreys C. ,^1,2 and Jeffery N.^1,2

¹University Of Liverpool and ²Institute of Ageing & Chronic Disease, Liverpool, UK

Considerable interest in skull ontogeny and phylogeny has focussed on the interplay among the functional units associated with it, in particular the brain and masticatory muscles. The present study aims to test for and investigate associations between the Masseter muscle (MV) and Endocranial volumes (ECV) among adult modern humans in order to map, amongst other things, sexual dimorphic trends between these functional units. .

Masseter and Endocranial volumes were measured from T1 and T2 weighted magnetic resonance images (IXI dataset – imperial college,), respectively. Datasets were excluded for obese persons and individuals with poor dentition. The resulting sample consisted of 58 male and 69 female individuals from 20 to 82 years of age. Quotients (MVQ & ECVQ) were also calculated with BMI as the denominator to mitigate against body size scaling effects. Bivariate analysis demonstrated a positive scaling trend between ECV and MV across males and females combined (r=0.51616, P= 5.26E‐10), but not within each gender (males: r=0.20818, P= 0.11684; females: r= 0.13605, P= 0.265). Further statistical analysis found that ECV is significantly different between males and females (P=1.04E‐14) as is the MV (P=1.06E‐16). This was also found to be the case when accommodating for body size in both MVQ (P=2.78E‐11) and ECVQ (P=5.33E‐04). There appears to be significant volumetric differences between male and females for both MV and ECV even when accounting for body size differences. This dimorphism is consistent with that reported in previous studies and implies that males follow a distinct growth pattern. The next challenge is to explore associated differences of skull morphology, especially in relation to differences of muscle volume as a proportion of endocranial size.

No ethical approval required as study carried out on ethically approved dataset using consenting patients from IXI database (http://brain-development.org/ixi-dataset/.

Keywords: muscle, masticatory, volume, masseter, endocranial, dimorphism, sexual

PS2. 30 A comparison of histomorphometric parameters in decalcified and non‐decalcified porcine and bovine bone

Sangchay N. , Felts P. and Cunningham C.

Centre for Anatomy and Human Identification, University Of Dundee, Dundee, UK

Skeletal histology has been demonstrated to play an important role in forensic investigations when severely damaged or fragmented remains are recovered. A number of histological methods have been used to generate tissue sections for such analysis, and it is necessary to ensure that morphological parameters are not altered by histological preparation, particularly as bone analysis becomes more detailed and sophisticated. While paraffin embedding and sectioning using a rotary microtome are readily utilized for soft tissues, the hardness of bone precludes the use of these histological techniques. For histological examination, bone must either be cut using a precision saw equipped with a blade designed for use with hard materials, or the tissue must be decalcified before being processed like any other soft tissue. The former preserves the morphological integrity of the osteon however it requires expensive specialized equipment not found in many histology laboratories. The latter can be achieved in any histology laboratory, however when morphological parameters must be examined precisely, for example when using morphometrics to differentiate human bone from non‐human bone, it is necessary to establish whether the decalcification process modifies the parameters being measured. Although many studies have examined the relative efficacy and speed of decalcifying agents in removing calcium, there is little information on the effect of decalcifying protocols on morphometric parameters which have been used in the analysis of bone. In this study we compared measurements of osteonic morphometric variables in sections of porcine and bovine femora prepared using a precision diamond saw, with sections from adjacent bone which had been decalcified using a commonly‐employed method utilizing 10% formic acid. We found no statistically significant differences in the osteon parameters measured (minimal and maximal diameter, perimeter, and osteon area) or Haversian canal variables measured (diameter, perimeter, and area) between the two methods. Our findings suggest that the decalcification of bone using 10% formic acid does not significantly alter bone morphometric parameters and thus can be used to simplify and reduce the cost of sample preparation for forensic examination. No ethical approval was required for these experiments.

PS2. 31 Cellular development is altered in the embryonic and postnatal rat spinal cord following maternal immune activation

Anderson R. ,¹ Foley T. ,² Radford J. ,² Allardyce J. ,¹ O'Hallaron S. ,¹ O'Keeffe G.² and McDermott K.¹

¹University Of Limerick, Limerick, Ireland and ²University College Cork, Cork, Ireland

Studies suggest that maternal immune activation (MIA) during gestation may be a factor in the development of neurological disorders such as schizophrenia and epilepsy. These studies suggest that the timing of the inflammatory insult and the region of the CNS examined may be crucial. This study investigates the temporal and spatial dependency of the effect of MIA on cell populations of the spinal cord (SC).

Immunohistochemical studies investigated the effect of MIA on oligodendrocyte populations (Olig2), microglial populations (Iba‐1) and reelin expression in the embryonic and postnatal SC. Animal procedures were carried out in accordance with Republic of Ireland Department of Health and Children licenses compliant with the Institutional Animal Care and Use Committee. Time‐mated dams received a single intraperitoneal injection of 100 μg/kg lipopolysaccharide (LPS) or saline on embryonic day (E)12 or 16. Dams were sacrificed 5 hr after injection to allow embryo harvest by laparotomy, or continued through pregnancy without further intervention. Offspring of these dams were sacrificed at postnatal day (P)14. Offspring were fixed with 4% paraformaldehyde, and embryos or dissected SCs frozen, and cryosectioned.

At E12 Olig2+ cell number was unchanged 5 hr after MIA but cell number had decreased in the ventral grey matter by P14. At E16 Olig2+ cell number was decreased in grey and white matter 5 hr after MIA. At E12 Iba‐1+ cell number remained unchanged in offspring 5 hr post‐MIA and at P14. At E16 Iba‐1+ cell number was decreased in grey and white matter regions 5 hr after MIA. Preliminary data suggests that reelin expression is unchanged in P14 offspring following MIA at E12. Reelin fluorescence intensity is decreased at E16 5 hr after MIA.

Collectively these data suggest that MIA affects cell development in the SC in a temporally and spatially dependent manner. The impact of MIA on developing cells may not always be evident acutely as here olig2+ cell number was unchanged at E12 but decreased by P14. In contrast MIA at E16 clearly does acutely impact olig2 and other cellular makers. Further work will elucidate how cell populations in the postnatal SC are affected by MIA at E16 and the mechanisms underlying such changes.

PS2. 32 Intrasutural (Wormian) bone in horse skulls: its frequency of occurrence and its effect on bone stiffness

Zambrano Martinez L. and Kilroy D.

University College Dublin, Dublin, Ireland

Intrasutural (Wormian) bone is well described in the medical literature but few studies have been done on domestic animals. This work aimed to determine the frequency of occurrence of intrasutural bone in the calvarium of the horse and, through bending tests, to determine if the presence of intrasutural bone altered the stiffness of the bone.

Sectioned pieces of equine parietal bone from ten healthy horse cadavers were subjected to microCT scanning and the resultant images were examined for the presence of intrasutural bone.

Stress tests were carried out on these bones to determine maximum load and whether intrasutural bone had an effect on bone stiffness.

Results were collated on the basis of age and sex and indicated that intrasutural bone did not affect the maximum load that could be applied to the parietal bones being studied. Bone stiffness was reduced by the presence of intrasutural bone, which was a more common finding in male horses than in female horses.

As the horse skulls were acquired from local abattoirs, ethical approval was not required for this work.

PS2. 33 An anatomical investigation into the flexibility of the human temporomandibular joint between formalin‐fixed, fresh‐frozen and Thiel fixed specimens

Quinn C. , Betts‐Dawson P. and Tierney C.

Craniofacial Lab, HARC, University Of Liverpool, Liverpool, UK

Formalin‐fixation [F] is often regarded as the traditional gold‐standard of cadaveric preservation and is used for teaching of anatomy via dissection whereas fresh‐frozen fixation [FF] is often prioritised for short‐term surgical training and biomedical research due to time limitations of use before decomposition of the material. Thiel's fixation [T] is a relatively new method of fixation that produces flexible, realistic anatomical specimens while remaining preserved and durable for extended periods of time. This study aims to investigate the flexibility of the temporomandibular joint (TMJ) between three types of fixation methods mentioned using mandibular goniometry and muscle volume of the masseter and temporalis masticatory apparatus. Three types of TMJ specimens sectioned at the cervico‐thoracic level were used: formalin‐fixed [F], 1 fresh‐frozen [FF] and Thiel‐fixed [T] (n=2). Mandibular depression (MD) was measured using an angle goniometer in open and closed positions while muscle geometrics and volume was measured in‐situ and via water‐displacement. Results show that there was a significant difference in the mobility of the Thiel and fresh‐frozen mandibular depression when compared to formalin specimens (P<0.001) with post‐hoc analysis revealing Thiel TMJ having the greatest mobility – 29.2° + 2.69° greater degree of movement compared to formalin TMJ. Muscle geometry and volume were also evaluated and it was found that while there was no difference in muscle geometrics due to the different fixative techniques measured in‐situ, there was a significant difference in muscle volume both for masseter (P<0.001) and temporalis (P<0.001) between the three different fixative techniques. Post‐hoc Tukey analysis revealed Thiel and fresh‐frozen‐fixed masticatory apparatus specimens had a significantly lower muscle volume when compared to formalin‐fixed specimens (P<0.017). It can be concluded that the mobility of the TMJ in Thiel‐fixed specimens is greatly increased when compared to formalin‐fixed TMJ's and the volume of the masticatory muscles was found to be much lower volume when compared to formalin fixed specimens. This may indicate that Thiel would be an ideal anatomical dissection model to be used in training of surgical dentistry procedures. All research was carried out within full HTA guidelines and with full ethical approval from the University of Liverpool.

PS2. 34 A link between worlds: Loss of cytoplasmic keratins downregulates nuclear lamins and compromises the nuclear lamina integrity. A murine study

Stenvall C. ,¹* Nyström J. ,¹* Butler‐Hallissey C. ,¹ Goldman R.² and Toivola D.^1,3

¹Cell Biology, Biosciences, Faculty of Science and Engineering, Åbo Akademi University, Turku, Finland; ²Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Illinois, USA; ³Turku Center for Disease Modeling, Turku, Finland and ⁴Indicates equal contribution

Keratins (K), cytoplasmic intermediate filament (IF) proteins found in epithelial cells, play a significant role in the maintenance of epithelial cell polarity and protection from mechanical stresses. Keratin filaments are formed of obligate heteropolymers of a type I (acidic) and type II (basic/neutral) keratin subunits. Keratin mutations have been associated with over 60 human diseases in the skin, liver and likely the intestine (IBD). Lamins, nuclear IFs, are responsible for nuclear structure and play a role in gene transcription, DNA replication and proliferation. Linker of Nucleoskeleton and Cytoskeleton (LINC) complexes composed of KASH and SUN domain proteins link cytoplasmic and nuclear IFs together. Keratin 8 (K8) is the main cytosolic IF expressed throughout epithelial cells in the colonic crypts. Using mouse models, it has been shown that absence of K8 leads to colitis, and hyperproliferation, decreased Na/Cl uptake, shifted differentiation and protein mistargeting in colonocytes. The aim of this study was to determine if cytoplasmic keratins affect nuclear lamins in colonocytes. The loss of K8 causes a significant downregulation of all major lamin proteins (A, B, C) in colonocytes, both in mouse colon and in K8 siRNA‐treated Caco‐2 cells. The lamin phenotype is likely not caused by the K8–/– colitis, since lamin levels were unaltered in the dextran sodium sulphate–colitis‐model. Broad‐spectrum antibiotic treatment, which ameliorates the colitis phenotype and removes most of the microflora, did not elicit a normalization of lamin levels in K8–/– colon. Furthermore, the lamin loss is colon‐specific, since lamin levels are unaltered in K8–/– small intestine, lung, liver and pancreas. In the absence of K8, colonocyte nucleoplasmic lamin levels are increased as determined by immunofluorescence microscopy and cellular fractionation, indicating a compromised nuclear lamina. In support of this, the lamin‐associated proteins emerin, LAP2α and pRb are downregulated in K8–/– colon. In conclusions, we show here a colon‐specific regulation of nuclear lamins and lamin‐associated proteins by cytoplasmic keratins, which may have consequences for colonocyte transcription and proliferation. All work was carried out with ethical approval of animal experiments to and under the supervision of Dr. Toivola .

PS2. 35 The use of chondroitinase ABC in an ex vivo rat model of spinal cord injury

Patar A. ,¹ Geraghty E. ,¹ Dockery P. ,¹ Howard L.² and McMahon S.¹

¹Dicipline of Anatomy Nui Galway and ²Regenerative Medicine Institute Nui Galway, Galway, Ireland

Chondroitin sulphate proteoglycans (CSPGs) are classed as the chemical barrier surrounding the glial scar and provide an environment that is repulsive to axonal regrowth. Chondroitinase ABC (ChABC) is an enzyme that is reported to neutralise the negative environment by degrading the glycosaminoglycan (GAG) side chains of the CSPGs. In this study, a transection ex vivo model of spinal cord injury (SCI) was used to study the cellular microenvironment of the glial scar in organotypic slice cultures of rat spinal cord. We examined if ChABC had any significant effect on the expression of the CSPGs in the microenvironment following SCI injury.

All animal experiments were granted ethical approval by the Animal Care Research Ethics Committee (ACREC) in the NUI Galway. Pregnant E14–E15 time‐mated females were maintained until their offspring pups reached postnatal day 4 (P4) age. P4 rat pup spinal cords were sliced in longitudinal sections at 350 μm thickness. A transection injury was performed using a double blade technique to separate two spinal cord pieces. The tissue was treated with ChABC (0.1 U/ml) in culture media and were fed every two days. Tissue was examined at day 1, 3, 7 and 10 post‐injury. Tissue was processed for immunohistochemistry markers of astrocytes (GFAP), CSPGs (NG2), microglia (IBA‐1) and neurons (βIII tubulin). The zones of interest examined were scar zone, near scar zone and far scar zone. Stereological analysis was carried out to calculate the volume fraction (Vv) of immunohistochemical staining in these regions of interest and the results were expressed as mean and SEM.

The ChABC treated group showed a significant reduction in Vv of all antigenic markers compared to injured and control groups, in particular at day 7. The stereological analysis also revealed significant differences in the Vv of all antigenic markers in all experimental groups.

The findings from this study suggest that ChABC may be beneficial in providing a positive microenvironment for axonal regeneration following SCI. This study also concludes that ChABC is effective at causing a significant change in expression of CSPGs and other cell types that are present in the injured microenvironment of the spinal cord.

PS2. 36 Anatomy and molecular composition of the spinal lesion site in zebrafish

Kassapis E. , Becker C. , Becker T. , Wehner D. and Tsarouchas T.

Univeristy of Edinburgh, Edinburgh, UK

Unlike mammals, zebrafish can successfully regenerate their spinal cord after sustaining a lesion. This interspecies divergence in regenerative capacity is partly attributed to differences in the anatomy and molecular composition of the lesion site, which render a more permissive environment in zebrafish, compared to that of mammals. A key determinant of the environment's conductivity to axonal regeneration is the composition of the extracellular matrix (ECM) in the lesion site. This is highly dynamic, undergoing dramatic changes in response to tissue damage to facilitate repair, and is modulated by fibroblasts, astrocytes and cells of the immune system. Growing evidence supports that the immune system is indeed a critical modulator of the quality of regenerative outcome in many regeneration systems. Previous studies have shown that in the absence of macrophages, axonal regeneration is strongly impaired in zebrafish, however, the exact mechanisms involved are not all clear. This has driven us to hypothesise that macrophages invading the lesion site promote axon regeneration by modulating ECM deposition in the spinal cord lesion site of zebrafish. We used in situ hybridization to investigate mRNA expression of 13 genes coding for ECM components, previously shown to be specifically upregulated in the spinal lesion site of zebrafish in response to lesioning, in wild‐type (wik) and irf8 ‐/‐ zebrafish larvae (do not develop mature and functional macrophages or microglia until after 31 dpf), before and after lesioning (under MS‐222 anaesthesia). We found no difference in expression levels of the selected genes between wik and irf8 ‐/‐ zebrafish larvae, thus we conclude that the pro‐regenerative effects of macrophages are not mediated through regulation of these genes, at least at the transcriptional level.

For the lesions, larvae were anaesthetized in 0.02% MS222 (Sigma‐Aldrich), and stabbing lesions were conducted using a 30 G needle loaded on a syringe, at the height of the anal pore of the dorsal trunk area, completely transecting the spinal cord. The larvae were killed by being initially anaesthetized in 0.02% MS222 (Sigma‐Aldrich), and subsequently fixed for 1 hour with 4% PFA‐PBS.

All experimental procedures were ethically performed and approved by the British Home Office.

PS2. 37 Highly regenerative panniculus carnosus skeletal muscle in the mdx mouse model of DMD

Bahri O. ,^1,2 Naldaiz‐Gastesi N. ,³ Wheatley A. ,¹ Izeta A.³ and McCullagh K.^1,2

¹Department of Physiology, School of Medicine, National University of Ireland‐ Galway, Ireland; ²Regenerative Medicine Institute, School of Medicine, National University of Ireland‐ Galway and ³Tissue Engineering Laboratory, Bioengineering Area, Biodonostia Institute, University Hospital Donostia, Donostia, Spain.

The sub‐dermal panniculus carnosus (PC) skeletal muscle layer found in most mammals with remnants in humans is unique in its highly regenerative capacity and, yet, its physiological function remains unclear. Here, we morphologically and functionally characterise the PC muscle in muscular dystrophic mdx mice compared to wild‐type mice. PC muscle was examined in 6 weeks, 12 weeks and 1‐year‐old mice. PC had 8% central nucleated muscle fibres (CNF), as compared to ˜1% CNFs in limb muscles in 6 week old normal mice. CNFs were elevated to 30% in 6‐week‐old mdx PC. The accumulation of CNFs further increased in PC with age in both wild‐type (12%) and mdx (53%) quantified at 12 weeks. Muscle fibre size distribution (minimal Feret's diameter) was heterogeneous in wild‐type PC and further accentuated in aged matched mdx mice, with a greater mean fibre size. PC vascular bed was examined in different aged mice by deriving capillary density and capillary to fibre ratio.

In order to examine the regenerative activity of dermal PC, we derived muscle precursor cells from the dorsal skin of wild‐type and mdx mice. Dermis‐derived PC cells were capable of forming striated contractile myotubes from both wild‐type and mdx mice. Myogenic activity was assayed by qPCR detection of myogenic factors Pax7, Myogenin and embryonic / adult myosin. Male mdx‐satellite cells‐derived myotubes exhibited the highest myogenic activity compared to wild‐type males and mdx females. This was also consistent with in vivo results where PC muscle's regenerative capacity, indicated by the percentage CNFs, was shown to be highly elevated in male mdx mice compared to male wild‐type and female mdx mice. We conclude that PC muscle is uniquely highly regenerative which is further perturbed in diseased muscle and makes for an ideal accessible tissue model to study aspects of the vascular‐myogenic niche and muscle satellite stem cell activity relevant to understanding DMD.

Animals used in this study were housed in the Bio Resource Unit, Biomedical Science Building, NUIG, according to the institutional guidelines. Experiments involving animals were strictly performed in compliance with the guidelines of NUIG, ethics and HPRA, Ireland.

PS2. 38 Using cadaveric dissection to validate high angular resolution diffusion imaging (HARDI) modelling of limbic white matter

Barry D. ,¹ O'Hanlon E. ,² El‐Kininy W.¹ and Roddy D.²

¹Department of Anatomy, Trinity College Dublin and ²Trinity College Institute of Neuroscience, Trinity College Dublin, Dublin, Ireland

Diffusion weighted (DW) MRI uses the diffusion of water molecules to generate contrast in MR images, which is beneficial in delineating large white matter tracts. However, concerns remain about the robustness and anatomical relevance of this technique in probing smaller or more convoluted tracts, such as those in the limbic system. Complex pathway commissures and methodological constraints, including the signal to noise ratio, are significant issues in generating images of clear and accurate limbic white matter. High angular resolution diffusion imaging (HARDI) is capable of discriminating multiple fiber populations across the same voxel. This study assessed the feasibility of comparing the accuracy of rendered limbic tracts, acquired using advanced HARDI tractography in DW MRI, to cadaveric dissection limbic tract measurements.

A human cadaver was scanned using a standard HARDI clinical research protocol; 3T Philips MRI scanner; Echo‐Planar Imaging, pulse sequence (TE = 52 ms, TR = 11.260 ms), 60 axial slices, 2 mm isotropic voxel size, in 61 non‐collinear gradient directions. Multiple b‐value acquisitions were acquired at 500, 1500 and 3000s/mm2 to facilitate Diffusion Kurtosis Imaging and assess the optimal b‐value for tract delineation. The approximate scanning time was 3.5 hours.

Using ExploreDTI software, full brain tractography was constructed using a Constrained Spherical Deconvulusion algorithm. Limbic system tracts, including the fornix, stria terminalis, stria medularis and cingulum, were extracted using standardised radiological gates. The brain was removed from the cranium and the above limbic tracts were identified and isolated. Tract length, diameter and relative position/distance to well‐defined structures were measured. These parameters were compared to the diffusion rendered images and metrics. Research consent was approved prior to donor donation and ethical approval has been granted to carry out this research.

This pilot study demonstrated the ability of HARDI to define limbic white matter tracts. Rendered metrics showed improved agreement with anatomical measurements after adjusting b‐values and significantly increasing the scan time, which is not possible in live patients. These findings are relevant to future research into neurological and psychiatric illnesses.

PS2. 39 Rescuing joint shape in immobilised chick embryos

Bridglal D. and Nowlan N.

Imperial College London, Department of Bioengineering, London, UK

A number of neonatal musculoskeletal conditions, including developmental dysplasia of the hip (abnormal hip joint shape leading to instability) and arthrogryposis (multiple joint contractures), are associated with reduced or abnormal movement in the womb. Research on animal models of fetal immobility have demonstrated that mechanical forces due to fetal movements are essential for skeletal development. Joint cavitation does not occur in immobilised chick and mouse embryonic limbs, and has been shown to be essential for subsequent joint morphogenetic events. In this study, we investigate if by applying external mechanical stimulation to an immobilised chick embryo model, we can induce cavitation and thereby enable more normal joint development.

Embryonic chicks were pharmacologically immobilised with decamethonium bromide between embryonic day (e) 5–8. No ethical approval was necessary as chicks were harvested prior to two thirds through gestation. At e8, when the hip joint normally cavitates, the uppermost hindlimbs were abducted and adducted 45° away from the midline five times. The unmoved, immobilised contralateral hip acted as a control. All embryos were harvested at e9. Optical Projection Tomography and Paraview were used to obtain morphological measurements of the hip joint (n=5 per group), for which a one‐way ANOVA with Tukey's test was performed. Histology was used to assess cavitation.

The effects of immobilisation on hip joint shape have previously been described and include smaller size, absent cavitation, a flattened femoral head, a shallower acetabulum and an abnormal angle between the femur and pelvis. The results of the current study indicate that partial cavitation occurred in the manipulated, immobilised hip joints, while no joint cavities were detected in the contralateral limbs. Our results also indicate that shapes of the manipulated hip joints were less severely affected than the contralateral joint (immobilised but not manipulated). Trends indicate that the height, width and depth of the acetabulum increased with manipulation, and that the femoral head was more pronounced in the manipulated joints.

This pilot study indicates the promising potential for the application of external forces to partially rescue the effects of fetal immobility on skeletal development.

No ethical approval was required for this study.

PS2. 40 Effects of Agmatine on stress induced on hippocampal morphology in rats

Yeğin B. ,¹ Alpay M. ,¹ Kacar S. ,¹ Ortadeveci A. ,¹ Meral C.C. ,³ Kilic C.S. ,⁴ Baskın V. ,⁵ Altuntas H.⁴ and Kevser E.⁴

¹Faculty Of Medicine/Anatomy, Eskisehir, Turkey; ²Faculty Of Medicine/Histology and Embrylogy, Eskisehir, Turkey; ³Faculty Of Medicine/Phisology, Eskisehir, Turkey; ⁴Faculty Of Medicine/Pharmacology, Eskisehir, Turkey and ⁵Faculty Of Medicine/Pharmacology, Ankara, Turkey

Agmatine, an endogenous amine derived from decarboxylation of L‐arginine catalyzed by arginine decarboxylase. Agmatine is reported to exhibit anti‐hyperglycemic, antioxidant and memory enhancing effects. Agmatine is an endogenous amine derived from decarboxylation of arginine catalysed by arginine decarboxylase. Agmatine is considered a novel neuromodulator and possesses neuroprotective properties in the central nervous system. The present study examined whether agmatine has neuroprotective effects against repeated restraint stress‐induced morphological changes in rat medial prefrontal cortex and hippocampus.

In the present study, we examined whether agmatine has neuroprotective effects against repeated immobilization‐induced morphological changes in brain tissue. Sprague Dawley male rats were used in the study. The animals were given enough food and water by keeping them in the light / dark cycle for 12 hours. The rats were divided into four groups: stress, stress+agmatine (30 mg/kg; intraperitoneal (i.p.)), stress+agmatine (100 mg/kg; i.p.) and control groups (total 4 groups and 24 animals). The rats were hungry for 16 hours before work. However, rats were allowed to drink enough water. Each animal was kept immobilized in a single animal cage and kept at 4 ° C for 4 hours. All tested compounds were administered i.p. in a single dose of Agmatine or saline. Animals were sacrificed by cetylamin‐xylazine anesthesia after removal from the cold room. After the animals were discontinued, the brain parts were removed rapidly and the fixation containing 10% formaldehyde was taken. Paraffin blocks were prepared after routine tissue‐typing procedures and 5 μm serial sections were stained with Hematoxylin‐Eosin for examination under light microscope. The hippocampal neurogenesis in stressed rats was measured by immunohistochemistry. Immunohistochemical staining with hematoxylen‐eosin showed that morphological alterations in the hippocampus. Results were compared with one‐way Anova following Bonferroni multiple comparison tests. Morphological changes in the hippocampus were examined by design‐based stereology. Total volume estimation of granul cell layer of the dentate gyrus and pyramidal cell layer of the hippocampus were not significantly between groups. Our results showed that Agmatine has no protective effects on parameters morphology of hippocampus. Ethical number:551.

PS2. 41 Dose dependent effect of Astaxanthin against ischemia/reperfusion injury on morphology of kidney, in rats

Yeğin B. ,¹ Alpay M. ,¹ Donmez D.B. ,² Oz S. ,³ Yucel F.¹ and Ozden H.¹

¹Faculty Of Medicine/anatomy, Eskisehir, Turkey; ²Faculty Of Medicine/Histology and Embryology, Eskisehir, Turkey and ³Departmant of Vocational School of Health Services, Eskisehir, Turkey

Astaxanthin (ASX), a carotenoid with potent antioxidant properties, exists naturally in various plants, algae, and seafoods. It is a strong antioxidant and exhibits other biological effects that have been well studied in various traumatic injuries and diseases. Ischemia‐reperfusion (I/R) causes oxidative stress and tissue damage by increasing free radicals. Previous studies showed that antioxidant substances decrease oxidative damages in tissues. In this study, we were aimed to evaluate the effect of ASX.

Wistar male rats were used in the study. The rats were divided into four groups: control, sham, experiment A (ASX 25 mg/kg; intraperitoneal (i.p.)) and experiment B (ASX 75 mg/kg; i.p.) groups (total 4 groups and 32 animals). Control group was given a single dose of dimethyl sulfoxide (1 ml/kg) i.p. 30 minutes before. The sham group was given a single dose of dimethyl sulphoxide (1 ml/kg) i.p. 30 minutes before. ASX was dissolved in dimethyl sulphoxide (soluble1 mg/mL). Experiment group A and experiment group B were given a single dose of ASX (25 mg/kg; i.p. and 75 mg/kg; i.p) 30 minutes before. Then common carotid artery was disrupted and clamped. After 15 minutes of ischemia and clamping, the muscle and skin were closed and perfused after 24 hours of reperfusion.

After rats were perfusion, the kidneys were removed rapidly and the fixation containing 10% formaldehyde was taken. Paraffin blocks were prepared after routine tissue‐typing procedures and 4 μm serial sections were stained with H‐E (Hematoxylin‐Eosin) for examination under light microscope. Using morphometric methods, the number of glomeruli per unit area and volumetric ratio of some histological structures were calculated. Results were compared with one‐way Anova following Bonferroni multiple comparison tests. Mean diameter of renal corpuscle and ratio of the Bowman's space to renal corpuscle were not significantly between groups. But the number of glomeruli per unit area was significantly between sham and experiment 2 (75 mg/kg) groups.

Morphometric examination showed that the glomerular dimensions may have grown and the kidney cortex may have shrunk. Together they can increase the number of glomeruli that are not actually present.

Ethical number: 544.

PS2. 42 Evaluation of toxicological effects of bufotoxin on histomorphology of testis of Wistar rat

Ihentuge C. ,^1,2,3 and Ugochukwu C.³

¹Howard University, Washington DC, USA; ²Abia State University, Nigeria and ³Imo State University, Nigeria

Toads secrete bioactive substances from skin and parotid gland, these components possess a range of medical properties. Twenty adult male Wistar rats, weighing between 100–150 g, were used in the study to evaluate the toxicological effects of Toad Parotid gland extract on Testis of wistar rats. They were divided into five groups of four animals each designated as groups A, B, C D and E. Group A served as Control group and was fed with diet and tap water only throughout the experimental period. Groups B, C, D and E were fed with standard diet and given a solution of extract from Toad Parotid gland (Toad Poison) at a single dose of 5 mg/kgbw, 10 mg/kgbw, 15 mg/kgbw and 20 mg/kgbw respectively (LD5040 mg/kgbw), the animals were handled in accordance with NIH guidelines for use of animals for experiment. Forty‐eight hours after the administration of the extract, rats were sacrificed by cervical dislocation after anaesthetized with chloroform and the testes were dissected out from each group, observed and processed for histological study. Results showed a decrease in physical activity of rats of groups B, C, D and E after thirty minutes to two hours of administration of the extract. Progressive weakness and staggered movement were also observed. Bleeding was observed from all the openings of the body including the conjunctiva from groups C, D and E twenty‐four hours after administration. Macroscopic examination of the testis shows no obvious pathology while histologically there was moderate to severe effect on the testicular tissue with necrosis of testicular cell (NTC), severe testicular atrophy (STA) and severe arrest of spermatogenesis (SAS). This results showed that toad parotid gland extract is capable of inducing pathological changes and could be a cause of male infertility.

PS2. 43 Clinical anatomy of bovine salpingitis – a new approach using live cell imaging

Owhor L. ,¹ Reese S.² and Kölle S.¹

¹School of Medicine, University College Dublin, Ireland and ²Institute of Veterinary Anatomy, Histology and Embryology, University of Munich, Germany

Salpingitis is a major cause for subfertility and idiopathic infertility. Inflammation of the oviduct is often associated with endometritis, but may also occur independently. The aim of this study was to elucidate the effects of mild, moderate and severe inflammation on tubal microarchitecture and function in the bovine model, with a special focus on investigating sperm transport and sperm‐oviduct interaction under near in vivo conditions. For this purpose, uterine tubes revealing mild (n=44), moderate (n=55) and severe (n=41) inflammation were removed from cows immediately after slaughter. Tissue samples with and without sperm and cumulus‐oocyte complexes were analysed by stereomicroscopy, light microscopy, scanning electron microscopy, histochemistry and live cell imaging. Our studies showed that moderate and severe inflammation caused a significant increase in the thickness of tubal folds (Anova post hoc Dunnett`s test, P<0.05). Severe inflammation was characterized by increased synthesis of glycoproteins resulting in luminal accumulations. Furthermore, enhanced apoptosis, shedding of the epithelial cells and accumulation of mucus obstructing the lumen were observed. Particle transport speed (PTS) was 31.3% (±19.2 μm/s) decreased in uterine tubes with severe salpingitis as compared to controls. Regarding PTS, the coefficient of variation was 11.5 times higher in oviducts with severe salpingitis as compared to the controls (Mann‐Whitney‐U‐test, P<0.05). Over all, the higher the grade of inflammation was, the lower was the ciliary beat frequency (CBF) (Spearman test, P<0.001). Spermatozoa were stuck in mucus and were therefore unable to form a sperm reservoir which led to reduced survival time and loss of fertilizing capacity. Cumulus‐oocyte complexes were also observed to be stuck to mucus which prevented intercellular communication between the cumulus cells and tubal cells.

Our results imply that salpingitis is a major cause of decreased bovine reproductive success. This is due to compromised gamete transport, decreased survival time of spermatozoa and impaired communication between the tubal epithelium and the oocyte. Thus, novel concepts for improving fertility should implement early diagnosis of inflammation and specific medications for treatment of salpingitis.

No ethical approval was required for this study as tracts were collected from the abattoir.

PS2. 44 Investigation of dose‐dependent effects of agmatine on rat kidney

Alpay M. ,¹ Yegin B. ,¹ Meral C. ,² Kacar S. ,³ Ortadeveci A. ,¹ Kılıç C. ,⁴ Baskın V. ,⁵ Altuntaş H.⁴ and Erol K.⁴

¹Faculty of Medicine/Anatomy, Eskisehir Osmazgazi University, Eskisehir, Turkey; ²Faculty of Medicine/Physiology; ³Faculty of Medicine/Histology and Embryology; ⁴Faculty Of Medicine/Pharmacology and ⁵Faculty of Medicine/Pharmacology, Eskisehir Osmazgazi University, Eskisehir Turkey

Agmatine (4‐aminobutyl guanidine) is a primary amine formed by the decarboxylation of L‐arginine by the enzyme arginine decarboxylase (ADC). Agmatine has been quantified in nearly all organs of the rat including kidney. It is metabolized agminal aldehyde and then guanidine‐butanoic acid in the kidneys by the help of diamine hydroxyciral enzyme. This molecule is used by many bodybuilders to push the limits because of its strong pain killer effect. Moreover, Agmatine is a very common and uncontrolled use as food supplement.. The purpose of this study was to investigate the effects of Agmatine on the kidneys.

Totally 24 male Sprague Dawley rats were divided into four groups (per contains 6 animals): (1) Stress rats (S), (2) control rats with single dose IP saline(C), (3) rats with single dose IP 30 mg/kg Agmatine (A30 mg/kg) and (4) rats with single dose IP 100 mg/kg Agmatine (A100 mg/kg). Stress rats were fasted for 16 hours before to the study. But we let them have enough water. Following this process, rats were individually placed in the immobilization cage and left to +4 degrees cold stress for 4 hours. Chemical substances (agmatine or saline) were administered intraperitoneally 1 hour before the animals were placed in the cold room. After the animals were removed from the cold room, their lives were ended by cetylamine‐xylazine anesthesia. Then 5 μm‐thickness sections taken by a systematic random sampling method from each kidney stained with Hemotoxylin‐Eosine. Alterations occurring in the kidneys were estimated by using morphometric methods. Results were compared with one‐way analysis of variance test following Bonferroni multiple comparison tests.

In the A100 mg/kg group, ratio of the Bowman's space to renal corpuscule was found significantly higher (24.49±7.41) than stress and A30 mg/kg groups. However, in healthy groups, no significant difference was observed. On the other hand, number of renal corpuscule per unit renal cortical area (Na) and the mean diameter of the renal corpuscule were not significantly different among groups.

These results suggest that Bowman's space expands with regard to the Agmatine and Agmatine's high dose affects glomerular filtration.

Ethical no: 551.

PS2. 45 Effects of agmatine on the histologic alterations of rat cerebellum

Alpay M. ,¹ Yegin B. ,¹ Meral C. ,² Ortadeveci A. ,¹ Kacar S. ,³ Kılıç C. ,⁴ Baskın V. ,⁵ Altuntas H.⁴ and Erol K.⁴

¹Faculty Of Medicine/Anatomy, Eskisehir Osmazgazi University, Eskisehir, Turkey; ²Faculty Of Medicine/Physiology; ³Faculty Of Medicine/Histology and Embryology; ⁴Faculty Of Medicine/ Pharmacology and ⁵Faculty Of Medicine/ Pharmacology, Eskisehir Osmazgazi University, Eskisehir, Turkey

Agmatine is used by many bodybuilders to push the limits on the basis of strong pain relief effects. However, this molecule, which is not known much about the effects on human health, can have adverse effects on athletic health or different uses. Some studies have pointed to constitute of schizophrenia‐like effects on the nervous system. The aim of this study was to investigate the effects of Agmatine on the cerebellum.

A total of 24 adult male Sprague Dawley rats were equally and randomly divided into four groups (per contains 6 animals) as follows: group 1 rats (Stress, S), group 2 rats with single dose IP saline (control, C), group 3 rats with single dose IP 30 mg/kg Agmatine (A30 mg/kg) and group 4 rats with single dose IP 100 mg/kg Agmatine (A100 mg/kg). Stress rats were fasted for 16 hours before to the study. But we let them have enough water. Following this process, rats were individually placed in the immobilization cage and left to +4 degrees cold stress for 4 hours. Agmatine was intraperitoneally administered to groups 3 and 4 consider their body weight. Beside this, group 2 received only i.p. saline at the same volume and time. And group 1 was not applied. This practise done 1 hour before the animals were placed in the cold room. At the end of the study, animals were sacrificed and bilateral brain hemispheres were removed for histological examinations. Alterations occurring in the cerebellum were estimated by using morphometric methods. Results were compared with one‐way analysis of variance test following Bonferroni multiple comparison tests.

In the control group, the mean diameter of the Purkinje cell was found significantly lower (15.20±0.38) than other groups. And this data showed no statistical difference between stress, A30 mg/kg and A100Mg/kg groups. On the other hand, ratio of the granular layer to molecular layer and ratio of the white matter to grey matter were not significantly different among groups.

In the light of these data, the mean diameter of the Purkinje cell enlarges with regard to stress and Agmatine does not affect this consellation.

Ethical no: 551.

PS2. 46 Evaluation of toxicological properties of bufotoxin on hematological parameters of male Wistar rat

Ihentuge C. ,^1,2,3 and Ugochukwu C.³

¹Howard University, Washington DC, USA; ²Abia State University, Aba, Nigeria and ³Imo State University, Owerri, Nigeria

Toad skin‐secretions (Bufotoxin) are potent source of drugs for treatment of various diseases. Twenty adult Wistar rats, weighing between 100–150 g, were used in the study to evaluate the toxicological effects of Bufotoxins on selected organs of male Wistar rats. They were divided into five groups of four animals each designated as groups A, B, C, D and E. Group A served as Control group and was fed with diet and tap water only. Groups B, C, D and E were fed with standard diet and given a solution of extract of Bufotoxin (Toad Poison) orally at a single dose of 5 mg/kgbw, 10 mg/kgbw, 15 mg/kgbw and 20 mg/kgbw respectively (LD50 is 40 mg/kgbw). The rats were treated in accordance with NIH guidelines for use of animals for experimental purpose. Twenty four hours after the administration, rats were sacrificed by cervical dislocation after anaesthetized with chloroform and organs (liver, heart, kidneys, lungs, testes and spleen) dissected out and processed for histological study. Blood was also collected for hematology and chemical analysis. Results showed Hemorrhage from all the orifices of the body including the conjunctiva from groups C, D and E twenty four hours after the last administration. Hematological results show a significant (P<0.05) decrease in total WBC count for experimental groups B (4.08±0.438), C (4.05±0.847) and D (2.33±0.125) when compared with the control group (7.58±0.334) while group E (10.85±0.068) was significantly (P<0.05) increased. The HGB and RBC levels and related parameters (MCV, MCH, and MCHC, HCT) were significantly (P<0.05) decreased in all the experimental groups (B, C, D and E) when compared with the control group (A). There was a dose dependent decrease in levels of platelets (PLT) in all the experimental groups (B, C, D and E) (251.50±55.720, 77.00±2.081, 55.60±8.580 and 48.50±11.528) which was significantly (P<0.05) lower when compared with the values of control group (288.50±7.511)

PS2. 47 The proliferative effect on the astrocytes of the brainstem of albino mice following long term ingestion of fresh and thermoxidized palm oil diets

Mustapha Peter T.

Human Anatomy Department, Gombe State University, Gombe State, Nigeria

The Astrocytic proliferation following chronic consumption of thermoxidized and fresh palm oil diets was studied on the brainstem of growing mice. Thirty mice were divided into three groups A, B and C. 15 g of thermoxidized palm oil was mixed with 85 g of mice chow (15%w/w) and given to the group C animals. 15 g of fresh palm oil was mixed with 85 g of mice chow (15% w/w) and administered to the group B animals while group A animals were fed with normal mice chow. The astrocytes in the brainstem of mice in the thermoxidized palm oil group significantly increased in number (Hyperplasia) and size (Hypertrophy) when compared with animals in the control and fresh palm oil groups due to the presence of concomitant evolution of toxic bye‐products (free radicals, peroxides etc). If the results obtained in mice are applicable to man, there is reason for concern regarding adverse consequences of chronic consumption of thermoxidized palm oil diet. This may be dangerous to health since it may result in Astrocytic proliferation in brainstem, thereby making the animals susceptible to loss of motor function like control of cardiovascular system, respiration, gastrointestinal tract, eye movement, etc.

Dr. Gedion is a Veterinary consultant from the State Ministry of Agriculture who certified the conditions of these animals and also gave clearance for the experiment, the procedure, the anesthetic injection and sacrificing the animals.

PS2. 48 Nrf‐2 and PGC1‐alpha deletion evoke ultrastructural changes joined to the alterations of oxidative stress and autophagy markers in aged retinal pigmented epithelium of knockout mice

Felszeghy S. ,¹ Viiri J. ,² Koskela A. ,² Paterno J. ,^2,5 Kettunen M. ,³ Koistinen A.⁴ and Kaarniranta K.^2,5

¹Institute Of Dentistry, University of Eastern Finland, Kuopio, Finland; ²Department of Ophthalmology, Kuopio University Hospital, Finland; ³A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland; ⁴SIB labs, University of Eastern Finland, Kuopio, Finland and ⁵Institute of Clinical Medicine, University of Eastern Finland, Kuopio, Finland

There is increasing evidence that NF‐E2‐related factor 2 (Nrf‐2) and peroxisome proliferator‐activated receptor‐gamma coactivator 1‐alpha (PGC‐1α) participate in age‐related retinal degeneration process. Our study aimed to provide de novo, in and ex vivo high‐resolution imaging of eyes from different degenerative mouse models mimicking cellular changes observed in aged retinal pigmented epithelium (RPE). The gross morphological alterations of eyes were analyzed by in vivo micro MRI. Oxidative stress marker (4‐HNE) and the different autophagy regulators (p62/SQSTM1, Beclin‐1, LC3, Ubiquitin) were examined with immunohistochemistry. Moreover on transmission electron microscopy ultrastructural analysis was carried out to monitor the organelle changes in the RPE obtained from Nrf‐2/PGC‐1α knockout mice. Collectively, age‐related autophagy decline and mitochondrial dysfunction was recorded in the RPE. Taken together, our experimental data highlight that deletion of Nrf‐2 and PGC‐1α appears to have potential and broad applicability for retinal aging research.

All animal protocols were approved by the Finnish National Animal Experiment Board and carried out in compliance with the European Community Council Directives 2010/63/EU and ARVO statement for the Use of Animals in Ophthalmic and Vision Research.

PS2. 49 Is there a future for emerging adult age‐at‐death estimation methods?

Radford D. and Chinnah T.

University Of Exeter Medical School, Exeter, UK

The ability to accurately identify human remains is vital in providing accurate reliable evidence in murder trials or in mass death scenarios. Formulating an age‐at‐death estimate for a set of human skeletal remains helps with forming the osteobiography of an individual. Available literature suggests that Suchey‐Brooks and Lovejoy et al methods are currently two of the most commonly used. These methods are based on analysing periodic changes in the morphological features of the pubic symphysis and auricular surface of the ilium respectively. However, there have been debates regarding their level of accuracy because of the subjective and qualitative nature of the analyses employed. These approaches have been suggested to introduce bias and conflicts of opinions. The conflicting accuracies reported by studies using these methods have raised debates among forensic anthropologists regarding the reliability of the results evidence. This literature review study examines the advantages of emerging methods for reliable accurate estimation of age‐at‐death.

Emerging new methods have been reported with potential advantages over current methods. One of these is mutated mitochondrial DNA (mtDNA). This is based on the ‘free radical’ or ‘mitochondrial’ theory associated with mtDNA mutations which increase throughout a lifetime and synonymous with ageing. This is being explored for accurate age‐at‐death estimation. However, the process has limited age range within which mtDNA mutations can be detected. Dual X‐ray absorptiometry (DXA) analysis is another emerging technique commonly used clinically to diagnose osteoporosis and has been considered for age‐at‐death estimation. DXA measures the quantity of hydroxyapatite in bone for bone mineral content (BMC) to calculate bone mineral density (BMD). BMD decreases with increase in age, and has been used for age‐at‐death estimation. Although, DXA method reduces observer bias, it requires a region at the neck of the femur (‘Ward triangle’) to be intact. BMD also has no set periodicity and data collection from a large sample size would be needed to establish a set of standardised values for classifying skeletal remains. These emerging new methods are more objective but raise questions about their future for improvement on accuracy for age‐at–death estimation.

No ethical approval was required for this study.

PS2. 51 Role of NRXN1α in early neuronal differentiation; a human study

Rosati S. ,¹ Boomkamp S. ,² Mc Donagh K. ,² Shen S.² and Dockery P.¹

¹Anatomy NUI, Galway, Ireland and ²Regenerative Medicine NUI, Galway, Ireland

Nrxn1, a cell adhesion molecule expressed at the presynaptic terminal, has been recently associated with autism spectrum disorders (ASD). Several studies have already showed the implication of NRXN1α in formation and stabilization of synapses, however its role during neuroprogenitor stem cells differentiation and neuronal maturation has been poorly investigated. In this study IPSC‐ derived neurons from patients and healthy donors were phenotyped and characterized in order to investigate and quantify cells with, neural stem cell marker, immature/mature neuron markers at different stages of neuronal maturation.

Methodology: Skin‐biopsies fibroblasts, obtained from patients carrying NRXN1α exon 5 deletion (n=3) and healthy donors (n=4), were reprogrammed into iPSCs using the episomal EPI5 kit. Validation of pluripotency was performed by alkaline phosphatase staining, by gene expression (Real time PCR) and pluripotent markers (Nanog, Oct‐4, Sox2, Tra1‐60, Tra‐1‐81, SSEA‐4) using immunohistochemistry (fig. 1). The iPSCs were differentiated into neuroprogenitor stem cells (NPCs) using Neural Induction Medium, and cultured for 20 and 40 days. Immuno‐histochemical methods were used to stain the following markers at two time points: 20 Days: PAX6, Nestin; 40 Days: β‐ Tubulin III, DCX and MAP2. The immunofluorescence was visualized with the Andor Revolution spinning disk confocal microscope, using Andor IQ2 software. Estimation of cell density (day 20) and neurite length per unit area (day 40) were obtained using some simple stereological methods employing a systematic random sampling protocol. This study had full ethics committee approval (C.A.750).

Data analysis: Mean ± SEM are presented in the bar charts. A one‐way analysis of variance was used to determine statistics significant using Minitab statistical software.

Results: The numerical density of NPCs was greater in control compared to patient‐derived NPCs at day 20. Total β‐Tubulin III stained neural length per unit area was higher in the control group after 40 days.

Conclusions: This study suggests: NRXN1α deletion may affect NSCs proliferation, commitment, and differentiation plus neuronal maturation. Further there is high variability between cell lines.

This initial quantitative exploration of the phenotype suggest that NRXN1α deletion may affect neurogenesis. This may contribute to our understanding of symptoms associated ASD.

PS2. 52 Telocytes – novel cell type that may play a crucial role in regeneration and reparation of the myocardium after infarction: a study with human telocytes

Varga I. ,¹ Klein M. ,¹ Miko M.¹ and Danisovic L.²

¹Institute Of Histology And Embryology, Faculty of Medicine, Comenius University In Bratislava, Slovakia and ²Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, Slovakia

Telocytes represent a recently discovered cell population with a potential to be the game‐changer in our understanding of etiology and pathogenesis of various diseases. Not only that, telocytes are also in the spotlight because of their possible breakthrough contribution to the advances in the field of regenerative medicine. Telocytes are highly scrutinized, not only in terms of continuous influx of new studies regarding their different roles in various organs, but also in an ongoing disagreement among scientists, whether they really represent a new and unique cell population, or they are only a highly specialized cells that are well‐known, like fibroblasts. Nevertheless, telocytes are intensely studied in different organs, including the heart. Particular attention in the research of cardiac telocytes is given to their possible role in the regeneration and reparation of damaged myocardium after infarction. Our preliminary study focused on the immunohistochemical description of telocytes in human hearts after infarction. Myocardial tissue was obtained during necropsies of a group consisting of 14 subjects. We used antibodies against c‐kit (CD 117), as we consider this antigen as suitable for telocytes′ detection. We used diaminobenzidine as a chromogen. Our result shown presence of c‐kit positive cells (probably telocytes) in myocardial tissue except the site of infarction. That lead us to conclusion that telocytes probably perished as a consequence of necrosis. Interestingly, a few weeks after infarction in patients who survived, we recognized a significant rise in c‐kit positivity. This may suggest, that telocytes started to migrate at the infarction site, what may be an evidence of their role in regeneration and reparation processes. The exact role of telocytes and the consequences of their decreased number in diseased heart is still largely hypothetical, calling for further research. This study was supported by Grant of the Slovak Research and Development Agency No. APVV‐0434‐12.

This study was approved by the local ethical committee of the Health Care Surveillance Authority, Slovakia.

PS2. 53 Neuronal polarity in Drosophila: a role for ankyrin?

Theodorou P. , Wingate R. and Williams D.

MRC Centre for Neurodevelopmental Biology, King's College London, London, United Kingdom

For many neuronal cells, two very different types of processes protrude from the cell body: axons and dendrites. These have been described – both anatomically and physiologically – as the major output (axon) and input (dendrites) devices. This functional division of labour is reflected at the molecular level. By thinking about how proteins that are destined for axons or dendrites are sorted, a means of answering the question of how neuronal polarity is established may be provided. The majority of work carried out in vertebrates has focused on multipolar hippocampal pyramidal neurons, but can ʻatypicalʼ cell types reveal underlying molecular mechanisms involved in the establishment and maintenance of neuronal polarity? Firstly, we established a system for exploring this in Drosophila – an identifiable monopolar interneuron where the presynaptic active zone protein Bruchpilot (homologue of ELK/CAST), can be clearly visualized. Secondly, the function of Ankyrin proteins was examined by a series of loss‐of‐function experiments. These findings suggest a role for Ankyrins in the proper dendritic exclusion of Bruchpilot, raising questions about their role in the establishment and maintenance of neuronal polarity as a whole.

No ethical approval was required for this study.

PS3. 1 Distinctive copy number alteration of gene encoding histone H2AX in human breast cancer

Passos J. ,¹ Dodson H.¹ and Flaus A.²

¹Discipline of Anatomy, School of Medicine, NUI Galway and ²Centre for Chromosome Biology, School of Natural Sciences, NUI Galway. Ireland

Histones are basic proteins responsible for the packaging of DNA in eukaryotes. Chromatin compaction is undertaken by canonical histones, whereas a number of variant isoforms contribute to specific functions involved in chromatin transactions.

H2AX is a H2A variant that plays an important role in the DNA damage response. Unrepaired or poorly repaired breaks can lead to genomic instability, which is a hallmark of cancer and impacts on cancer progression and treatment.

The direct contribution of H2AX abundance is not well understood, therefore we have been exploring copy number, expression and abundance of H2AX in breast cancer. Interestingly, the H2AFX gene encoding H2AX has two transcripts which encode the same protein. Absolute quantitative PCR of a panel of breast cell lines shows that the H2AX short transcript is more abundant than the long transcript. The ratio between the amount of the short and long transcript is constant across the cell lines tested.

Analysis of the METABRIC primary tumour dataset shows that H2AFX copy number alterations involving loss of one allele are very common (41% of 1905 samples) and correlate with genome instability. 2% of the samples show H2AFX copy number amplifications, and no diploid deletions are observed. In this dataset, H2AX transcript levels separate into two groups based on PAM50 breast cancer classification: Low H2AX mRNA levels are seen in normal‐like, luminal A and claudin‐low sub‐types, whereas high H2AX mRNA levels are seen in luminal B, HER2‐enriched and basal sub‐types.

These results suggest that H2AX expression can be regulated in different ways, and that the role of H2AX in genome stability may be more complex than as a simple chromatin‐incorporated signal. Based on its high abundance and variability between cancer sub‐types, we are investigating the potential of H2AX as a breast cancer biomarker.

No ethical approval was required for this study.

PS3. 2 Angiogenesis in uterine tube and uterine horn in domestic cat during prenatal development; SEM study on vascular microcorrosion casts

Prozorowska E. and Jackowiak H.

Deapratment of Histology and Embryology; Poznan University of Life Sciences, Poznan, Poland

Angiogenesis is the process of blood vessels development and play a significant role in organogenesis. There are two different types of angiogenesis that can be defined during the prenatal vascular development, i.e., sprouting and intussusceptive angiogenesis. This study aims to describe the development of intramural vascular system in paramesonephric ducts and also maturation of this vascular system in the uterine tubes and uterus in domestic cat fetuses, with particular emphasis on the identification of sprouting and intussusceptive angiogenesis.

The research material was 25 domestic cat female fetuses aged 36th – 63rd day p.c. The observations were conducted on the microcorrosion vascular casts of paramesonephric ducts (Mullerian ducts) and uterine tubes and uterus, in accordance with the guidelines set out by the Local Ethics Commission.

The vascular corrosion casts observations on the 36th day of prenatal development (p.c.) revealed that vascular system of paramesonephric duct is simple and consists of the longitudinal supplying and collecting blood vessels and circularly arranged capillaries in the mesenchyme. Both, in tubal and uterine segments of paramesonephric ducts, the intussusceptive angiogenesis is the main process of growth of capillary bed, while sprouting angiogenesis occur less often and only locally participating in the formation of the new branches of vascular network.

The intramural vascular system both, in uterine tubes and uterine horns consists of longitudinal supplying arteries and collecting veins, circumferential arterioles and venules, subserosal capillary networks and mucosal subepithelial capillary network. Observations of the microcorrosion vascular casts show that in the uterine tubes the intussusceptive angiogenesis is observed between 46th – 55th day p.c. mainly in the subserosal capillary network. On the 52nd day p.c. the intussusceptive angiogenesis is observed in the circumferential arterioles and venules of the uterine tubes. The sprouting angiogenesis in uterine tube is rare. The intussusceptive angiogenesis in the uterine horns wall is detectable between 46th – 63rd day p.c. on the subserosal capillaries. Additionally, around 50th and 56th day p.c. this process is visible on longitudinal arterioles and venules. Sprouting angiogenesis in uterine horns is noticed on the 50th and 54th day p.c. on capillaries of the subserosal network.

PS3. 3 Anatomical variation in branching pattern of brachialplexus and its clinical significance

Khan G. , Yadav S. , Gautam A. , S Shakya S. and Chetri R.

Chitwan Medical College, Bharatpur, Nepal

Anatomical variations in the branching pattern of the brachial plexus have been described in humans by many authors; however these have not been extensively catalogued. The aim of the study was to describe and observed anomalies in distribution of the branch (branching pattern) derived from the cord of brachial plexus, both in its supraclavicular and infraclavicular parts.

This study included thorough dissection of 60 brachial plexuses which belonged to 30 cadavers (male: female ratio = 28:02) with age range of 20–60 years, obtained from the Department of Anatomy, College of Medical Sciences (CMS‐TH), following standard guidelines.

Out of 60 limbs dissected in present study, Normal branching pattern of the posterior cord was encountered in 52 (86.67%) limbs, the remaining 8 (13.33%) being variants in one form or the other. The upper subscapular nerve, the thoracodorsal nerve, the lower subscapular nerve and the axillary nerve were found to arise normally in 91.66%, 96.66%, 96.66% and 98.33% of the limbs respectively.

The present study carried out on adult human cadavers revealed some rare variations in the branching pattern of the brachial plexus. These variations are of clinical significance are very useful for the anatomists, radiologists, anesthesiologists, neurosurgeons and orthopedic surgeons and general surgeons.

No ethical approval required for this study.

KEYWORDS: Brachial plexus, branching pattern, Anatomical variation.

PS3. 4 The human thyroidea ima artery: an uncommon encounter

Nawab F. , Ahmed U. , Smith B. , Hunter A. and Ellis H.

Department of Anatomy, Kings College London, UK

The highly vascular thyroid gland is supplied by two paired arteries. The anterosuperior aspect of the thyroid gland is supplied by the first branch of the external carotid artery, the superior thyroid artery, and the posterointerior aspect supplied by the largest branch of the thyrocervical trunk, the inferior thyroid artery. Reportedly, 3–10% of the population have a further unpaired thyroidea ima artery. This most commonly arises from the brachiocephalic trunk and enters the thyroid gland at the inferior border of the isthmus. It may also branch off from the common carotid artery and/or aortic arch. It usually but not always arises from the right side, and when present, may function as additional arterial supply to the thyroid gland. Its variability and low percentage of occurrence have been emphasized by many reports since it was first described in the eighteenth century by Neubauer.

During dissection of a thyroid gland and its associated neuro‐vasculature, in a formalin‐fixed cadaver, we found a symmetrically enlarged thyroid gland extending retrosternally. A pyramidal lobe was discernible but the isthmus absent. Both superior thyroid arteries were large and visible, as well as both normal sized inferior thyroid arteries. We incidentally found a further large thyroidea ima artery which in our case arose from the brachiocephalic trunk coursing superiorly towards the thyroid gland, and entered on the medial border of the right lobe.

Clinically, knowledge of the presence of the thyroidea ima artery is important, particularly for clinicians performing invasive procedures such as tracheostomy and thyroid or parathyroid surgery. Presence of a thyroidea ima artery would be a contra‐indication to performing a percutaneous tracheostomy, a procedure which is increasing in popularity. During thyroid, parathyroid and midline neck surgery, as well as certain surgical approaches to the trachea, early identification of a thyroidea ima artery would help to prevent significant intra‐operative haemorrhage.

No ethical approval was required.

PS3. 5 Relationship between cranial length, cranial width, cranial index and body mass index in nigerian children

Musa M.A. ,¹ Ahmed H.² and Zagga A D.¹

¹Department of Anatomy, Faculty of Basic Medical Sciences, College of Health Sciences, Usmanu Danfodiyo University Nigeria and ²Department of Pediatrics, Faculty of Clinical Sciences, College of Health Sciences, Usmanu Danfodiyo University, Nigeria

Measures of physical growth in children are very essential to pediatricians and others who provide healthcare to infants and young children to aid recognition of morphological variations and clinical abnormalities at an early stage. Evaluation of craniofacial parameters in growing children is a well‐documented approach to well‐being. The aim of this study was to investigate the relationship between Cranial Length, Cranial Width, Cranial Index and Body Mass Index in 153 children from Sokoto, Nigeria. Ethical approval for this study was obtained from the ethics committee of the Institute of Child Health, Usmanu Danfodiyo University Teaching Hospital, Sokoto and ethical consideration included taking informed consent from parents/guardians of study participants. Cranial Length is defined as the longest dimension of the skull vault along the anterior‐posterior axis while Cranial Width is the widest transverse dimensions of the skull vault between the two parietal bones. The children were aged 2 – 8 years with mean ages of 4.25 ± 2.12 years for boys and 4.80 ± 2.21 years for girls. Cranial Index and Body Mass Index values were calculated according to sex and Pearson's product‐moment correlation coefficients between variables were computed using Microsoft Excel Spreadsheet. The mean value for cranial length in boys was 15.99 ± 1.28 cm and in girls 16.17 ± 0.86 cm, while cranial width mean values in boys and girls were 12.50 ± 1.14 cm and 12.99 ± 0.85 cm respectively. The mean Cranial Index in boys was 78.16 ± 3.17% and 80.34 ± 3.17% in girls. The Body Mass Index in boys and girls was 13.98 ± 2.65 kg/m² and 14.44 ± 1.95 kg/m² respectively. From this present study, there was positive correlation between Cranial Index and Body Mass Index in both boys and girls.

PS3. 6 The incidence of accessory foramina for the vertebral artery in the posterior arch of the human atlas

Dhumale S. and Alashkham A.

University Of Edinburgh, Edinburgh, UK

The vertebral artery provides blood supply to the spinal cord and brainstem. Disruptions to its course and flow usually result in rapid hospitalisation, if not death; and therefore variations in its course are of great importance. The aim of the study is to examine the incidence of accessory foramina in the posterior arch of the atlas, which have been shown to accommodate the artery in previous literature. The passage of the artery through additional ossified tunnels may lead to clinical consequences.

The bones are from the University of Edinburgh's anatomy department, regulated by the Anatomy Act 1984.

A total of 60 atlases were analysed of unknown sex nor gender. The accessory foramina were classified into distinct types; first they were identified as either unilateral (left or right) or bilateral, and then either complete or partial. Measurements of the accessory foramina were taken using Fischer Scientific digital callipers.

Out of 60 atlases, 15% (n = 9) were found to have a complete accessory foramen (bilateral or unilateral), with two presenting bilateral complete accessory foramina. Three showed right‐sided complete foramen with either no foramen or partial foramen on the left, and only one showing a complete left‐sided foramen with no presence of a foramen on the right. The average width of the nine foramina was 5.46 mm, with the average width of the right‐sided (5.55 mm) being slightly larger than the width of the left‐sided foramina (5.28 mm).

Limitations: The age, sex, and previous medical history for each of the 60 individuals were not known.

This study has found a significant number of a population demonstrating accessory foramina in their C1 vertebrae. Some studies have linked the accessory foramen to vertebral artery tethering, leading to subsequent dissection following repetitive trauma from neck movement. Less traumatic symptoms have been recorded linking accessory foramina to neurovascular problems, including frequent headaches and neurosensory deficits. Its incidence in the posterior arch of the atlas is significant, particularly for medical specialists investigating cervical trauma or interpreting medical imaging.

PS3. 7 Investigating the role of the mastoid process in sexing British DryHuman skulls

Dhumale S. ,¹ Hubbard S.² and Alashkham A.¹

¹University Of Edinburgh, Edinburgh, UK and ²University of Dundee, Dundee, UK

Sexing human remains is a common initial step in forensic investigation that usually relies on intact skeletal material. The mastoid process is a feature of the dense temporal bone protected at the inferolateral aspect of the skull and is therefore more resistant to damage during traumatic events. Recovering this aspect of the skull may help identify the sex of an individual, particularly when the more typically used pelvis is damaged beyond use. Here, we investigate the significance of mastoid length in sexing British skulls.

All skulls utilized in this study are from the University of Edinburgh's anatomy department, regulated by the Anatomy Act 1984.

A total of 78 (53 male, 25 female) skulls were used. Each skull received a code number to minimize operator bias. Two raters measured 13 skulls to test for inter rater reliability. The osteometrics of the mastoid were measured bilaterally using Frankfurt's plane. Inclusion criteria: (1) skull of known sex; (2) suitable cranial structure to identify the orbital margin and external acoustic meatus. The length of the mastoid was then measured from this line to the tip of the mastoid process using a digital calliper.

All analyses were conducted in R (R core team 2017).

General linear models were used to check for differences between raters in the skull measurements, and then for differences between sexes. Analysis showed no difference between the two raters (F1,1 = 2.876; P = 0.0965), but a highly significant difference between the measurements for male and female skulls (F1,1 = 15.451; P = 0.000127). The final model was checked for constancy of variance and normality in the model residuals, and these were found to be satisfactory.

Limitations: The age of skulls was not known, as well as cause of death.

The results suggest the mastoid process is a cranial feature which can be useful for anthropological and forensic investigation. The sexual dimorphism presented by the mastoid is reliable enough to use confidently in sexing British human skulls.

PS3. 8 Anatomical variations of the suprascapular nerve and notch: a cadaveric study with implications for pain management of the shoulder joint

Love A. ,¹ Michla Y.² and Matthan J.³

¹Newcastle Medical School, Newcastle University; ²Trauma and Orthopaedics Department, Sunderland Royal Hospital, City Hospitals Sunderland Foundation Trust and ³Anatomy and Clinical Skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle, UK

The suprascapular nerve arises from the brachial plexus before passing posteriorly through the suprascapular notch to supply the supra‐ and infraspinatus muscles and a large proportion of the sensory fibres to the shoulder joint. Ultrasound‐guided suprascapular nerve blocks (SNB) into the notch are effective in analgesic control of painful shoulders but landmark‐guided blocks into the supraspinatus fossa are less effective however ultrasound machines not readily available in clinics. A relatively fixed, palpable, anatomical landmark for the suprascapular notch would allow SNBs to be administered more accurately into the notch without ultrasound guidance for better analgesic outcomes. This preliminary cadaveric study assessed the consistency of the supraclavicular notch in relation to palpable bony landmarks.

Seven cadavers, prepared using several embalming methods, were dissected using a posterior approach to the shoulder. The supra‐ and infraspinatus muscles were identified and reflected, the suprascapular nerve was identified deep to these and followed superiorly through the notch. Three pre‐existing prosections were also assessed. The notch was identified on all specimens and measurements were taken from the centre of the notch to four easily palpable bony landmarks: the superomedial border of the scapula (MBS), the acromioclavicular joint (ACJ), the medial border of the scapular spine (MSS) and the lateral edge of the acromion (LEA).

Ten shoulders were assessed in total, six left‐sided and four right‐sided specimens. Due to the variation in specimen type and time‐since‐death, sex was known only in six of the ten cases. The median distances measured were as follows: MBS 61 mm (Interquartile Range (IQR), 58–65 mm); ACJ 52 mm (IQR, 46–52 mm); MSS 78 mm (IQR, 73–83 mm); LEA 67 mm (IQR, 59–72 mm).

The variation in distance from all the landmarks was relatively large, with little anatomical consistency in the position of the suprascapular notch relative to these bony landmarks. However, when gender was taken into account in this small sample, there appeared to be more consistency, especially in the distance from the LEA. A large‐scale study is required to gauge the consistency of the landmarks before widespread use of landmark‐guided SNBs are performed routinely. No ethical approval beyond the remit of the Human Tissue Act was required.

PS3. 9 Mapping the location, dimensions and prevalence of the thyroid isthmus: a cadaveric study with surgical implications

Peng Yong W. , Lloyd T. , Walker A. , Pilarski A. , Lunn J. and Brassett C.

Human Anatomy Teaching Group, Dept. of Physiology Development and Neuroscience, University Of Cambridge, UK

Detailed understanding of thyroid morphology is critical to both emergency and elective surgical procedures of the neck and airway. In standard anatomy textbooks, the thyroid isthmus is stated to lie over the 2nd to 4th tracheal rings anteriorly. The location and morphology of the thyroid isthmus were investigated in this cadaveric study to aid planning in surgical approaches.

Anterior neck dissection was undertaken in 38 formalin‐fixed cadavers to reveal the thyroid gland and associated structures. Pins were inserted into the trachea at the superior and inferior borders of the thyroid isthmus. The isthmus was subsequently divided in a longitudinal midline plane. Three measurements were taken for length (superior‐inferior distance, mean calculated) and one for depth (anterior‐posterior distance). Isthmus length was mapped in relation to the corresponding tracheal cartilages at the level of the pins (first tracheal cartilage = 1, second = 2 etc.; half and quarter measurements denote interval distances). Tracheal diameter was also measured. Where there was uncertainty regarding agenesis or the presence of a pyramidal lobe, the sample was sent for histology. All measurements were taken using calibrated digital vernier calipers and all donors had provided informed consent for anatomical research, in accordance with the Human Tissue Act 2004.

The 38 cadavers analysed comprised 17 males and 21 females. A pyramidal lobe was present in 8/38 (21.05%) subjects, with 2/38 (5.26%) exhibiting isthmus agenesis. Mean isthmus depth was 4.56 mm, (Range 0.92–20.61, SD 3.59). There was no significant difference between male and female cadavers (P = 0.1295).The median total number of tracheal cartilages covered was 2 (range 0.5–4.5, IQR 1.25). The thyroid isthmus was more superior than textbooks suggest, with 64.86% covering the first tracheal cartilage, 89.19% covering the second, 64.86% covering the third, and only 16.22% covering the fourth. Mean tracheal diameter was 20.58 mm, with male tracheas being significantly wider (22.90 vs 18.79 for females, P < 0.0001). There was no significant correlation between isthmus depth and tracheal diameter (P = 0.3134, rho = 0.1704).

The results of our study suggest the thyroid isthmus is located more superiorly than defined textbook values, which may have important implications in planning emergency and elective surgical procedures.

PS3. 10 Cystic ovary disease and its effect on the bovine oviduct: new concepts

Scully D. ,¹ Kölle S.¹ and Reese S.²

¹University College Dublin, Ireland and ²Ludwig‐Maximilians University, Munich, Germany

Cystic ovary disease (COD) is a major factor contributing to poor reproductive efficiency of lactating dairy cows. However, the precise effects of follicular and luteal cysts on oviductal form and function are still unknown. As the oviduct is pivotal for gamete transport, fertilisation and early embryonic development, the aim of our study was to analyse the effects of ovarian cysts on the functional integrity of the oviduct using stereomicroscopy, light microscopy, scanning electron microscopy and quantitative and qualitative live cell imaging. Oviducts were excised from female bovine genital tracts collected immediately after slaughter at a local abattoir. Our results showed that 90% of the genital tracts collected revealed COD on the right ovary. The oviductal mucosa from cystic cows revealed edematous enlargement of the primary longitudinal folds in the ampulla which are important in orchestrating gamete and embryo transportation. The ampullar epithelium from cows with luteal cysts revealed a constant predominance of ciliated cells lacking the ability to form secretory cells required for embryo nutrition after fertilization. Additionally, synthesis of glycoproteins and acidic mucopolysaccharides was increased within the ciliated cells of the ampulla from cystic cows as compared to controls. Using live cell videomicroscopy we found that the attachment of the cumulus‐oocyte‐complex to the ampullar epithelium and the formation of the sperm reservoir in the isthmus was not negatively affected in cows with ovarian cysts. However, particle transport speed was significantly (P < 0.05, T‐test) decreased in the tubal ampulla as compared to controls. Our results imply that ovarian cysts impair the functional integrity of the oviduct which is pivotal for successful gamete transport, fertilization and early embryogenesis. Tubal occlusion by edema and mucus, as well as reduced secretory and ciliary activity result in compromised gamete and embryo transport. Thus, innovative therapeutic strategies for treatment of ovarian cysts do not only require successful induction of ovulation and fertilization, but also optimization of nutrition and transport of the early embryo in the oviduct.

No ethical approval was required for this study.

PS3. 11 Variations in origin and course of the dorsalis pedis artery in humans

Keogh M. and Cheung C.

School of Biomedical Sciences, University of Edinburgh, Edinburgh, UK

Dissection for anatomical study of a female cadaver identified an aberrant right dorsalis pedis artery originating from the peroneal (fibular) artery as a continuance of the perforating branch as opposed to the usual anterior tibial artery. The remaining structures within the lateral and anterior compartments were observed to run standard courses as described in the literature. Comparison was made to 5 other adult cadaveric limbs undergoing anatomical dissection and study.

All specimens had specific consent to be reproduced as photographic images of human tissue in addition to consent for anatomical study under the body bequeathal scheme and Anatomy Act (2006) for Scotland.

The formaldehyde fixed specimens were further dissected along their distal and proximal courses, measured in diameter and length and photographed with markers. The right anterior tibial artery was found to be hypoblastic and terminated 65 mm proximal to the talocrural joint. The posterior tibial artery gave origin to the peroneal artery 50 mm distal to the inferior border of popliteus. The perineal artery (with a diameter of 3.80 mm at origin) descended inferolaterally between tibialis posterior and flexor hallucis longus parallel to posterior tibial artery to supply the lateral compartment of the leg; giving rise to muscular, nutrient, communicating and a final perforating 6.3 cm from the talocrural joint. This perforating branch (diameter 2.24 mm) pierced the interosseous membrane entering the anterior compartment of the leg passing deep to the flexor retinaculum continuing as dorsal pedis (diameter 2.28 mm) between the tendons of extensor hallucis longus medially and extensor digitorum laterally.

Peripheral pulse detection is crucial in cardiovascular, peripheral vascular disease, hypertension medication monitoring and diabetes.. Surgically the vascular anatomy is pivotal in burns cases and plastics where foot skin flaps are required. In addition monitoring of the dorsalis pedis and posterior tibial pulses are essential in orthopaedics where external fixation may compromise vascular integrity. Abnormal arterial morphology can efect pulse width and velocity making understanding of anatomical varitions clinicaly pertinent

No further ethical or other consent was required in addition to standard body donation requirements. Specific consent for the photographic reproduction and use of anatomical images was documented and referenced.

PS3. 12 Three‐dimensional arrangement of collagen fibres in the human zygomaticotemporal suture

Robertson K. , Muir S. , Wight L. , Gregory J. and Grӧning F.

School of Medicine, Medical Sciences and Nutrition, University Of Aberdeen, Aberdeen, UK

The zygomaticotemporal suture is located on the zygomatic arch, a thin bone which endures high forces during mastication from the attached masseter muscle and temporalis fascia. Analysing the collagen fibre arrangement of the suture can help determine the forces that act upon this suture. Most research in this area has been conducted using 2D analysis in animal models whose skulls are different in shape to humans and thus the results may not be transferable. This project aimed to determine the 3D orientation of collagen fibres in the human zygomaticotemporal suture.

Zygomatic arches were obtained from human cadaveric specimens, decalcified and cryosectioned. Brightfield microscopy at low magnification was used to identify sections in which the suture was visible. The sections of one specimen fulfilled this criterion and were stained for confocal microscopy at ×20 and ×40 magnification. Zeiss Zen software was used to visualise collagen fibre arrangement in 2D and 3D.

Consistent long parallel fibres, oblique in orientation relative to the bone surface were visualised in one individual. The angle of the fibres became less acute at the insertion sites of the suture to bone. This arrangement was consistent throughout the suture despite variations observed in the 3D topography. Further histological study showed variation in acuteness of the fibre angle from the lateral to medial surfaces and between the superior and inferior boarders of the suture. However, it was unclear in this study if this was consistent with the 3D topography of the suture. These results are suggestive of mainly compressive forces but high levels of variation may exist between different individuals. To our knowledge, this is the first research to have successfully visualised the 3D arrangement of the collagen fibres in the human zygomaticotemporal suture.

This work was funded by an Anatomical Society Undergraduate Student Summer Vacation Research Scholarship. We are grateful to the individuals who facilitated this cadaveric study by their generous body donations. No ethical approval was required for this study.

PS3. 13 Defining the connections of the left ascending lumbar vein and their significance in laparoscopic live‐donor nephrectomy

Walker A. ,¹ Slim N. ,¹ Nicholson M.² and Brassett C.¹

¹Human Anatomy Teaching Group, Anatomy Building, Department of Physiology, Development & Neuroscience, University of Cambridge, Cambridge, UK and ²University of Cambridge Department of Surgery, Addenbrooke's Hospital, Cambridge, UK

The advent of laparoscopic live‐donor nephrectomy for renal transplantation in recent years has prompted the need to define the precise anatomical relations of the left renal vein (LRV) and its tributaries. The left kidney is preferred as the greater length of the LRV facilitates implantation in the recipient. While previous studies have described variations in the LRV system, the connections between the left ascending lumbar vein (LALV) and LRV tributaries have been less well‐defined. This study aims to further characterise the LALV and proposes a novel classification for its relation to other veins. Dissection of the LRV system, including the left suprarenal vein (LSV), left gonadal vein (LGV) and LALV, was performed in 38 cadavers. Their drainage points into the LRV were recorded, and measurements taken of the distances from these points to the junction of the LRV and inferior vena cava (IVC). In 35 (95%) cases, the LRV was anterior to the aorta, entirely posterior in 1 case, and circumaortic in 2 cases. Duplication of the LSV and LGV occurred in 6 (16%) and 10 (27%) cases respectively. A direct posterior connection between the LALV and LRV was identified in 32 (86%) cases. The drainage point of the LALV into the LRV lay between the IVC and LGV in 8 (25%) cases. In 20 (63%) cases, the drainage points of the LALV and LGV were equidistant from the IVC; and in 5 (16%) cases, those of the LALV and posterior branch of the LRV were equidistant from the IVC. In these two groups, the vessels shared a confluent trunk in 10 and 4 cases respectively. In 3 cases, connections were observed between all three vessels (LALV, LGV and posterior branch of LRV). No confluence trunk was shared by the LALV and LSV. These results confirm the high incidence of communicating LALVs, which represent a troublesome source of operative bleeding if unrecognised. Confluent venous trunks may also present difficulties during vessel ligation prior to nephrectomy. It is suggested that a novel classification of the relation of the LALV based on these findings may assist in surgical planning and reduce complications.

No ethical approval required for this study.

PS3. 14 Multilevel oblique corpectomy on a human cadaveric specimen: Precarotid and retrocarotid surgical approaches may have different patient outcomes

Haynes E. ,¹ Tykocki T. ,² Eltayeb M.² and Matthan J.³

¹Newcastle University Medical School; ²The Newcastle upon Tyne NHS Foundation Trust and ³Anatomy and Clinical Skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle Medical School, Newcastle University, UK

Various approaches and techniques are utilised for surgical treatment of cervical spondylotic myelopathy (CSM) and cervical spondylotic radiculopathy (CSR); to date, anterior or posterior approaches have been the mainstay. When less than three vertebral levels are affected, an oblique corpectomy, using a lateral approach, is a viable alternative. First performed in 1992, the technique has grown in popularity and is indicated in cases of predominant anterior cord compression with no associated spinal instability, a kyphotic or straight spine and dehydrated or collapsed disc/s. Many benefits are incurred for surgeons and patients, not least that the approach helps maintain vertebral stability, even when performing a multilevel oblique corpectomy (MOC). We aimed to establish whether precarotid and retrocarotid approaches are practical approaches, to pave the way for both techniques to be adopted routinely.

Bilateral oblique corpectomies, both precarotid and retrocarotid approaches, were performed on an 86‐year‐old male human cadaveric specimen. Vickers embalming fluid number 4 was administered by gravity feed via a cannula inserted into the common carotid artery for embalming purposes. This process rendered the specimen more challenging to dissect, as it increased rigidity and made smaller structures difficult to identify. A burr drill and retractors were used to perform the technique, rather than the commonly employed surgical equipment. No pre‐operative imaging was available.

The lateral aspect of the vertebrae (C6 and C7) were approached, the vertebral artery visualised and protected. The lateral segment of the pathological disc was removed and a vertical trench drilled in the lateral aspect of the vertebral body to a depth of the posterior cortex. Horizontal drilling extended to the contralateral pedicle. The posterior cortex and posterior longitudinal ligament were removed to release the spinal cord, with no sign of instability.

Previous research suggests MOC is a valuable alternative procedure. We were able to demonstrate safe and viable precarotid and retrocarotid approaches on a human cadaver. Further dissections on fresh‐frozen cadavers would ensure accurate measurements of distances between key landmarks to definitively establish the safety of both approaches. This project was covered by the HTA process and required no further ethical approval for its completion.

PS3. 15 Proposal for a diagnostic algorithm of pediatric patients with Hirschprung disease with other concomitant associated congenital anomalies

Varga I. ,¹ Zabojnikova L. ,² Slavikova T.^1,2 and Miko M.¹

¹Institute Of Histology And Embryology, Faculty of Medicine, Comenius University In Bratislava, Slovakia and ²Children's University Hospital and Faculty of Medicine, Comenius University In Bratislava, Slovakia

Abnormalities of neural crest development are responsible for as much as one third of developmental disorders. Neural plexuses of the gastrointestinal tract are also derivatives of neuroectoderm of the neural crest. Hirschsprung disease is the most common congenital anomaly of the intestinal motility, caused by abnormal organization of the enteric nervous system. Characteristic feature of the disease is the absence of neural ganglia (aganglionosis) most commonly in the distal parts of the gut in variable length. In addition, more than 20% of patients with Hirschprung disease have other associated morphological congenital anomalies due to disrupt development of other organs associated with neural crest‐derived cells. Based on our previous results, we set a diagnostic algorithm, applicable in daily practice in patients with Hirschprung disease to detect other associated anomalies. We tested the algorithm in a pilot study. Twenty patients from the study went through internal pediatric examination, anthropological examination, ultrasound examination of the thymus, thyroid gland and urinary system and genetic testing. We tested function of the thyroid gland by laboratory tests as well, and we also implemented immunological examination focused on lymphocytes population. Although the study group was too small to obtain any statistically significant outcomes, we noticed significant decrease of the CD8 +  cells in all tested patients, and significant insufficiency of the antibodies (agamaglobulinemia) on half of the patients. We suppose that these results are due to disrupt development of thymus (because the developing thymic primordia are under the influence of neural crest derived mesenchyme). During genetic testing we focused on diagnosis of RET proto oncogene mutations in pediatric patients with Hirschsprung disease, which may be associated also with multiple endocrine neoplasia type 2 (MEN2). DNA analysis was conducted in 5 patients – direct sequencing of exon 8, 10, 11, 13–16 of the RET gene. We found 2 variants in RET gene – c.1648 + 84G>A (IVS8 + 84G>A) and c.1648 + 88delC (IVS8 + 88delC), heterozygous form in 4 patients and homozygous in 1 patient. These variants are not associated with higher risk of MEN2 (according to the databases of The University of Utah). Acknowledgment: This study was supported by Grant VEGA No. 1/0086/17. This study was approved by the local ethical committee of Children University Hospital in Bratislava, Slovakia. Parents of pediatric patients provided written informed consent prior to participating in the study.

PS3. 16 Study of morphological variations in renal artery anatomy in human cadavers can aid the development of fenestrated grafts utilised in aneurysm repair: anatomical literature review

Ogbuagu C. ,¹ Altahir A.¹ and Matthan J.²

¹Newcastle Upon Tyne NHS Foundation Trust Hospital, Newcastle and ²Anatomy and Clinical Skills Department, School of Medical Education, Faculty of Medical Sciences, Newcastle Medical School, Newcastle University, Framlington Place, Newcastle upon Tyne, NE2 4HH, UK

Abdominal aortic aneurysm (AAA) accounts for ca 6,000 deaths per year in England and Wales; around 80,000 people are estimated to have a pre‐existing AAA. Risk of death from a ruptured AAA is high, at 80%, with most deaths being pre‐hospital. While open abdominal graft repair remains the gold standard procedure, endovascular aneurysm repair (EVAR) has gained popularity, particularly for patients at risk of surgery due to age, comorbidities or previous abdominal surgery.

EVAR has successfully been employed in the management of infra‐renal AAA; management of juxta‐renal AAA remains challenging owing to the need for renal artery (RA) stenting during the procedure. Current practice involves customisation of a stent based on CT measurements, a process taking 6–8 weeks and greatly increasing mortality risk for patients with a ruptured juxta‐renal AAA, particularly for those at risk of open repair. A ready‐made fenestrated graft for aneurysm repair (FEVAR) may mitigate the mortality risk associated with this condition.

Existing anatomical literature on RA variations and RA's relation to the superior mesenteric artery (SMA) was reviewed. Level of origin of RA, angle of origin in relation to midline and vertical distance between the SMA and RA ostia were examined. The majority (96%) of RAs arise between the upper border of the first lumbar vertebra (L1) and the lower border of the second lumbar vertebra (L2) (range T12 to L3 vertebra).

Different mean angles of origin for right RAs were reported, the mean of the reported mean angle origin RA was 25.82^® (−26 to 70^®). The mean value of the reported mean angle origin of left RA was −15.5 (−75 to 38^®). Other dimensions were right RA ostial diameter (mean diameter of 4.82 mm [2.30 to 9.40] and 4.89 mm [2.60 to 7.60] for left RA), and SMA to renal ostia distance (SMA to RA 15.9 mm [0 to 118], SMA to LA 17.55 mm [4 to 60]).

An off‐the‐shelf stent graft for FEVAR is possible; more research is required on morphological variations of the RA in a sample population, as precise measurements are required for cannulation of the RA. No ethical approval was required for this review.

PS3. 17 Variation between the brachial plexuses of a human subject due to the presence of a unilateral cervical rib

McMahon D. ,¹ Love A. ,¹ Haynes E. ,¹ Creissen A. ,¹ Matthan J.² and Michla Y.³

¹Newcastle Medical School; ²Anatomy and Clinical Skills Department, Newcastle Medical School and ³Trauma and Orthopaedic Department, Sunderland Royal Hospital, Newcastle, UK

Cervical ribs are found in approximately 0.5% of the population; in half of these cases their occurrence is unilateral. The presence of such an anatomical variant is associated with variation in the anatomy of the adjacent brachial plexus, knowledge of which is clinically relevant for surgeons operating in its environs; clinical effects may also arise from compression of the plexus trunks by the aberrant rib and/or associated fibrous banding. Dissection of a human cadaver revealed a left‐sided Type III cervical rib, the subject of this study.

Initial discovery of the cervical rib occurred following dissection of the left shoulder of a female cadaver, deceased within the previous 26 months, aged 88 years at death and preserved using Vickers Embalming Fluid Number 4, administered via gravity‐feed without drainage. The brachial plexus was identified and mapped and the presence of the cervical rib noted. The right plexus was similarly mapped and dissection performed to assess for presence of a further cervical rib. Dissection of each side was undertaken by two dissectors, both independently confirmed their findings; subsequent independent confirmation was sought from three other dissectors.

The right side of the subject, lacking a cervical rib, was found to have an anatomically normal brachial plexus. In contrast, the left brachial plexus showed abnormalities of the median and musculoskeletal branches, with both arising from a confluence between the medial cord and two divergent partitions of the lateral cord; a variation was also seen with the presence of an extra division reconnecting the posterior and anterior divisions of the middle trunk. The remaining plexus was otherwise normal.

Embryologically, the formation of cervical ribs follows from the development of the plexus in the limb bud, the arrangement of the plexus determining whether the potential rib develops or regresses. For well‐developed cervical ribs, as in this instance, it is expected that the T1 contribution will be absent or minimal. However, this was not apparent in the specimen, with the diameter of T1 comparable to that of C8, potentially suggesting a different embryological aetiology.

No further ethical approval beyond the remit of the Human Tissue Act was required.