. 2019 Nov 5;28(6):586–590. doi: 10.1159/000503547
Med Princ Pract. 2019 Nov 5;28(6):586.
Introduction
Fusidic acid (FD) resistance is a major problem in MRSA isolated in Kuwait hospitals. The resistance is mediated by 3 types of genetic determinants (fusA, fusB, and fusC) which vary in their genetic locations. The aim of this study was to investigate the genetic mechanisms of FD resistance in MRSA strains collected from Kuwait hospitals.
Methods
A total of 97 FD-resistant MRSA were obtained from clinical samples between January and June 2018. Isolates were tested for resistance to different antibiotics by the disc diffusion method. E-test was used to measure the minimum inhibitory concentration (MIC) of FD. DNA microarray was used to determine their genetic background and resistance mechanisms.
Results
The isolates were obtained from different samples, including blood (n = 17), nasal (n = 12), and wound (n = 11). The isolates were resistant to penicillin (97.9%), trimethoprim (48.4%), tetracycline (45.4%), kanamycin (31.9%), gentamicin (28.9%), erythromycin (27.8%), and clindamycin (27.8%). The MIC for FD ranged from 3 to >256 mg/L. fusA was associated with strains with FD MIC of >256 mg/L, whereas the MIC for fusB and fusC were variable. Of the 97 strains, 80 (82.5%) harbored fusA, 79 (81.4%) harbored fusC, and fusB was detected in 4 isolates. Sixty-four isolates carried fusA and fusC, and 2 isolates harbored fusA and fusB. Fifteen and 14 isolates carried only fusC and fusA, respectively, while two isolates contained only fusB. Clonal complexes (CCs) were determined for 83 of the 97 isolates. Eight CCs consisting of CC1, CC5, CC8, CC15, CC22, CC80, CC88, and CC97 were identified, with the majority belonging to CC5 (50.6%) and CC97 (13.2%).
Conclusions
The study revealed that fusA and fusC were the dominant genetic determinants of FD resistance. Whereas fusA and fusC were widely distributed among the CCs, fusB was limited to CC80. This study has increased our knowledge of the genetic mechanisms of FD resistance in MRSA in Kuwait hospitals.
Med Princ Pract. 2019 Nov 5;28(6):586–587.
Introduction
Physician burnout is important not only because of its personal implications, but also because it can interfere with patient care by impairing clinical judgement. This study assessed the prevalence of burnout and associated factors among surgeons in Kuwait.
Methods
A self-administered questionnaire was completed by 445 surgeons (181 general surgeons and 165 subspecialists of all ranks from 11 subspecialties) from all secondary and tertiary Ministry of Health hospitals in Kuwait. Burnout was assessed using the Maslach Burnout Inventory, which has 3 subscales: emotional exhaustion (EE, score range 0–54), depersonalization of others (DP, score range 0–30), and low personal accomplishment (PA, score range 0–48). Burnout is defined as an EE score ≥27, a DP score ≥10, or a PA score ≤33. A defining score in all 3 subscales was considered “severe burnout.” Associations of burnout or severe burnout with participant characteristics were assessed using multivariate logistic regression.
Results
Of the participants, 87.2% were male and 47.6% were Kuwaiti. The prevalence of burnout and severe burnout were 76.9 and 13.9%, respectively. The prevalence of high EE, high DP, and low PA scores was 44.7, 43.1, and 47.2%, respectively. The prevalence of burnout was highest among neurosurgeons (100%) and lowest among ENT surgeons (66%). After adjustment, burnout was associated with younger age (<33 vs. >40 years; OR 2.23; p = 0.007), lower income (<2,000 vs. >3,000 KWD; OR 1.88; p = 0.042), and more on-calls/month (>8 vs. <5 calls; OR 2.6; p = 0.01). Analysis of potential sources of stress also found a significant effect of work-life imbalance (p = 0.002) and case overload (p = 0.005) on burnout. The most common source of stress among surgeons was less time to spend with family (68.7%).
Conclusions
The prevalence of burnout among surgeons in Kuwait MOH hospitals is high. Occupational health programs should use these findings to design interventions to reduce burnout in this population.
Med Princ Pract. 2019 Nov 5;28(6):587.
Introduction
The development of biofilms in the oral cavity is inevitable. In health, these biofilms are composed mostly of commensal oral flora. A shift in the microbial homeostasis towards the dominance of Gram-negative bacteria causes oral infections. Bacteria in biofilms are antibiotic resistant, and hence difficult to treat. Of these, elevated levels of Prevotella spp. in oral infections is well documented. Thus, the aim of our study is to characterize biofilms formed by Prevotella species and assess biofilm formation inhibition and detachment of preformed biofilms.
Methods
Biofilm formation potentials of Prevotella spp. were tested. Bacterial suspensions in brucella broth were inoculated into 24-well cell culture plates and incubated in anaerobic conditions for 3 days. The formed biofilms were washed with sterile H2O and quantified using crystal violet staining. The biochemical characterization included quantification of proteins and DNA in both biofilms and their extracellular matrix (ECM). Images of Syto® 9 green fluorescent-stained biofilms were captured using CLSM. Also, biofilm inhibition and detachment by proteinase and DNase was tested. The enzymes were added to the bacterial suspension for biofilm inhibition. Preformed biofilms were treated by the enzymes for 1 h at room temperature. Eventually, the biofilms were quantified as above.
Results
P. loescheii, P. oralis, and P. nigrescens showed the highest potentials for biofilm formation. Protein and DNA content were higher in biofilm than the ECM, with the highest protein value found in P. oralis biofilm and ECM. However, DNA content was higher in the biofilm of P. nigrescens and the ECM of P. loescheii and P. oralis. Proteinase showed a better effect on biofilm detachment than DNase. Biofilm formation inhibition was observed only in P. oralis with both proteinase and DNase.
Conclusions
Prevotella spp. demonstrate a biofilm formation potential that in some cases can be prevented by treatment with DNase and proteinase.
Funding Agency
SRUL 01/14.
Med Princ Pract. 2019 Nov 5;28(6):587–588.
Introduction
Inhaled bradykinin (BK) has been reported to both sensitize and induce cough, but whether BK can centrally sensitize the cough reflex is not fully established. In this study, using a conscious guinea-pig model of cough, we investigated the role of BK in the central sensitization of the cough reflex and in airway obstruction.
Methods
Drugs were administered to guinea pigs by the intracerebroventricular (i.c.v.) route. Aerosolized citric acid (0.2 m) was used to induce cough in a whole-body plethysmograph box, following the i.c.v. infusion of drugs. An automated analyzer recorded both cough and airway obstruction simultaneously.
Results
BK administered by the i.c.v. route dose dependently enhanced citric acid-induced cough and the enhancement in Penh (mean cough ± SEM: 8.8 ± 1.7 and 15.9 ± 4.5 for 0.03 and 0.06 nmol mL–1 BK, respectively, compared to vehicle-treated animals: 4.1 ± 1.3; p < 0.05; n = 5–9). This was reversed by administration of B2 receptor antagonist (mean cough ± SEM: 3.2 ± 1.3 and 2.0 ± 0.6 for 10 and 100 nmol mL–1 HOE-140, respectively, compared to vehicle-pretreated animals: 10.0 ± 3.6; p < 0.05; n = 6–8), TRPV1 antagonist (mean cough ± SEM: 13.0 ± 3.5 and 4.4 ± 2.6 vs. 17.7 ± 3.2 for JNJ-17203212, 1 and 3 µmol mL–1, compared to vehicle-pretreated animals, respectively; p < 0.05; n = 7–9), TRPA1 antagonist (mean cough ± SEM: 14.8 ± 4.7 and 4.5 ± 1.7 vs. 19.5 ± 4.5 for 60 and 150 nmol mL–1 HC-030031 compared to vehicle-pretreated animals, respectively; p < 0.05; n = 8–13), COX inhibitor (mean cough ± SEM: 11.2 ± 4.6 and 5 ± 1.7 vs. 17.1 ± 3.5 for 30 and 80 nmol mL–1 compared to vehicle-pretreated animals, respectively; p < 0.05; n = 5–10), and 5/12-LOX inhibitor (mean cough ± SEM: 14.2 ± 4.2 and 5.2 ± 4.0 vs. 19.8 ± 2.3 for 30 and 100 pmol mL–1 baicalein compared to vehicle-pretreated animals, respectively; p < 0.05; n = 5–6). However, administration of 15-LOX-1 inhibitor did not alter BK-induced sensitization of the cough reflex and airway obstruction.
Conclusions
Our findings show that central BK administration sensitizes cough and enhances airway obstruction via a B2 receptor/TRPV1 and/or TRPA1 channels which are coupled via metabolic products of COX and/or 12-LOX enzymes.
Funding Agency
This study was supported by the College of Graduate studies and by grant No. YP05/16 from Kuwait University Research Sector.
Med Princ Pract. 2019 Nov 5;28(6):588.
Introduction
Crush injuries occur from a traumatic compression of the nerve resulting in different degrees of neural damage. Significant cell death and axon degeneration occur as a result of this damage, leading to permanent functional deficits. Previous clinical trials showed that catfish skin preparations (CSP) have potent effects on chronic back pain and other neurological disorders. This study was designed to investigate the anti-apoptotic and neuroprotective effects of soluble protein fraction (SPF)-fraction C (FC) derived from CSP on the sciatic nerve crush injury. Methods: Adult male Wistar rats were randomly assigned into five groups (n = 8/group): SHAM, CRUSH, CRUSH+1.5 mg/kg SPF-FC, CRUSH+3 mg/kg SPF-FC, and CRUSH+4.5 mg/kg SPF-FC. Rats underwent sciatic nerve crush surgery, followed by treatment with SPF-FC administered intraperitoneally (IP) for 2 weeks, and sacrificed at the end of the fourth week. All animals were assessed for sensory and motor neurobehavioral tests throughout the 4 weeks. Peripheral axonal regeneration was assessed through whole mount staining of sciatic nerve using axonal markers. The neuroprotective properties of the treatment on the spinal cord neurons were assessed using Cresyl violet staining, while the apoptotic pathway was assessed using Western blot, immunohistochemistry, and TUNEL techniques.
Results
The results of this study showed that IP administration of different SPF doses significantly (p < 0.05–0.001) improved the neurobehavioral functional recovery of the nerve-injured groups. Visualization of sciatic nerve through whole mount staining revealed an increase in the axonal regeneration recovery with SPF-FC treatments. Moreover, SPF-FC treatments have neuroprotective effects on spinal cord neurons. Conclusions: Our results for the apoptotic pathway revealed that SPF-FC treatments reduce neuronal cell death resulting from sciatic nerve crush injury. The data suggest that SPF-FC reduces sensory and motor neurobehavioral deficits and enhances axonal regeneration.
Funding Agency
College of Graduate Studies.
Med Princ Pract. 2019 Nov 5;28(6):588.
Introduction
The 2016 WHO classification of brain tumors has incorporated molecular parameters in addition to histology to define diffuse gliomas. This integrated approach has important diagnostic and therapeutic implications. The aim of this study is to reclassify a cohort of diffuse gliomas, previously diagnosed on histological grounds alone, using the updated WHO system to evaluate its impact on the final diagnosis.
Methods
Gliomas, diagnosed between January 2015 and September 2018, were retrieved from the pathology database at Kuwait Cancer Control Center. The tumors were reclassified incorporating key genetic markers, e.g., IDH1/2 mutations and 1p19q codeletion. A glioma was labelled “not otherwise specified (NOS)” when molecular results were inconclusive or incomplete.
Results
Out of 87 gliomas, 56 (64%) had available molecular results (47 males, 9 females, mean age 41 years, range 12–69). The original diagnoses were as follows: diffuse glioma of “ambiguous” histology (n = 22), diffuse astrocytoma (DA = 5), anaplastic astrocytoma (AA = 3), oligodendroglioma (ODG = 7), anaplastic oligodendroglioma (AODG = 2), and glioblastoma multiforme (GBM = 17). IDH1/2 mutations were detected in 23/46 cases tested (50%), with R132H mutation in IDH1 being the most frequent, seen in 18/23 cases (78%). Following revision, all ambiguous cases were classified into specific categories mostly as astrocytoma (increasing the number from 8 to 27 cases in total) and into subcategories, e.g., IDH-mutant and IDH wild-type. Also, 3/7 original ODGs were reclassified into DA, being negative for 1p19q codeletion, highlighting the lower diagnostic threshold for this tumor type on histology alone. All GBMs remained as such but the diagnosis was refined into subcategories. Five cases were labelled as NOS due to limited molecular testing.
Conclusions
The integrated 2016 WHO criteria lead to more objective classification of gliomas and elimination of the “ambiguous” group. However, limited access to molecular resources is a major obstacle.
Med Princ Pract. 2019 Nov 5;28(6):589.
Introduction
The incretin hormones, glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1), are important regulators of many aspects of metabolism, including insulin secretion. Their receptors (GIPR and GLP-1R) belong to the secretin class of G protein-coupled receptors, which recently have been shown to form dimers. Since both GIPR and GLP-1R are expressed on pancreatic β-cells there is a possibility that they could form hetero-dimers. Our objective is to investigate whether these receptors can form hetero-dimers in vitro and to investigate the impact of receptor activation on dimer formation.
Methods
Receptor dimerization was investigated using a bioluminescence resonance energy transfer (BRET) saturation assay. GLP-1R was labelled at the C-terminus with a nano-luciferase (Nluc) and stably expressed in Flip-In HEK-293 cells. These cells were then transiently transfected with increasing concentrations of GIPR labelled at the C-terminus with a variant of yellow fluorescent protein (SYFP2). BRET saturation curves were generated following incubation in the absence of an agonist to monitor constitutive dimerization or in the presence of either 1 µm GIP or GLP-1.
Results
BRET saturation curves were plotted as a ratio of SYFP2 fluorescence to Nluc luminescence. In the absence of an agonist an exponential curve was generated, increasing and then reaching an asymptote, consistent with a saturable BRET signal. Treatment with GLP-1 resulted in a significant (p > 0.05) increase in the maximum BRET signal, whereas treatment with GIP resulted in a nonsignificant decrease compared to non-treatment (n = 3).
Conclusions
The results support the hypothesis that GIPR and GLP-1R form hetero-dimers. The increase in BRET signal observed with GLP-1 treatment can be interpreted as either an increase in the number of dimers formed or a change in receptor conformation. Future experiments will investigate the impact of receptor dimerization on cell signals.
Funding Agency
College of Graduate Studies, Kuwait University.
Med Princ Pract. 2019 Nov 5;28(6):589.
Introduction
Cancer patients are at high risk of developing anaemia either due to the nature of the disease or as a consequence of administering myelo-suppressive chemotherapeutic agents. This precipitates fatigue and general weakness. Cancer patients are seeking evidence-based knowledge about improving their dietary intake to manage chemotherapy side effects. Beetroot is a potent antioxidant and powerful source of dietary nitrates. Beetroot is considered a potential therapeutic treatment of heath conditions that are associated with inflammations and oxidative stress reactions. Also, it improves the endothelial function and iron level in the blood.
Methods
In a comparative prospective study, 30 breast cancer patients with invasive ductal carcinoma who were candidates for a standard chemotherapy protocol [doxorubicin/cyclophosphamide+paclitaxel] were included. Patients were allocated to either consume beetroot juice (250 mL/3–5 times weekly) or no intervention (control group). The baseline haemoglobin level was documented and monitored throughout the study.
Results
The baseline haemoglobin level was comparative in the two groups (Hb > 10.5 g/dL) indicating that all patients started as either non-anaemic or with mild anaemia (p value 0.1). However, post-chemotherapy, the haemoglobin level was significantly higher in patients who had regular consumption of beetroot juice (p = 0.00). While 13% of patients in the intervention group had mild anaemia (Hb 10.1–11.9 g/dL), 87% had moderate anaemia (Hb 8–10 g/dL). None of these patients developed severe anaemia. In contrast, the control group had 40% mild anaemia, 53% moderate anaemia, and 7% developed severe anaemia (Hb:6.5–7.9 g/dL).
Conclusions
Cancer patients undergoing chemotherapy are at a high risk of developing anaemia, which worsens their quality of life. Beetroot juice consumption has been shown to prevent severe or life-threatening anaemia by maintaining an acceptable haemoglobin level, and reduces the risk of progressing to mild-moderate anaemia.
Med Princ Pract. 2019 Nov 5;28(6):590.
Actinomycosis is an uncommon, chronic granulomatous disease caused by Gram-positive, anaerobic bacteria. These bacteria normally colonize the bronchial system and gastrointestinal tract in humans. The most common diseases associated with actinomycosis are orocervicofacial, thoracic, and abdominal infections. It can be easily mistaken for other clinical conditions, including malignancy, because of its rarity and its different clinical presentations. We report a case of a young 26-year-old Sri Lankan lady who presented with abdominal pain and was found to have multiple liver and mesenteric cystic lesions as well as a para-splenic cyst. She was suspected of having a parasitic or protozoal infection or an intra-abdominal malignancy with metastasis. However, after being thoroughly investigated, she was discovered to have actinomycosis secondary to an intrauterine contraceptive device (IUCD) with subsequent multiple abscess formation. The rare diagnosis of actinomycosis should be considered in patients with abdominal pain and fever, especially if a previous history of IUCD usage is known.
Med Princ Pract. 2019 Nov 5;28(6):590.
Introduction
According to the World Health Organization the prevalence of overweight and obesity in children increased globally from 4% in 1975 to 18% in 2016. Both genetic and environmental factors are involved in the development of obesity. The common fat-mass obesity (FTO) gene polymorphism (rs9939609) has been found to be associated with obesity in both children and adults worldwide. However only a limited number of studies on children have been conducted. Our aim is to investigate for the first time the FTO gene polymorphism in children from Kuwait. Methods: Categorical body mass index (BMI) values of 705 school children (n = 509 Kuwaiti and n = 196 non-Kuwaiti) aged 9–14 years were available. The FTO gene polymorphism rs9939609 was genotyped for all children and the relationship with categorical BMI was analyzed using logistic regression. Results: We observed an association between the common A-allele of the FTO gene polymorphism with obesity in Kuwaiti children (OR 1.53; 95% CI 1.19–1.98; p = 0.0009). No association was observed in non-Kuwaitis (OR 0.94; 95% CI 0.64–1.39; p = 0.77).
Conclusions
This is the first study in the region to observe an association between the common FTO gene polymorphism and obesity in Kuwaiti children. Our findings are consistent with other publications and support that genetics contribute to the development of obesity. Moreover, this finding supports the importance of public health awareness of the role of our genes in disease susceptibility.
Med Princ Pract. 2019 Nov 5;28(6):590.
Introduction
Type 2 diabetes mellitus (T2DM) affects 14.7% of the Kuwaiti population. Recent research shows that diabetes is associated with an increased risk for developing Alzheimer's disease (AD), a neurodegenerative disorder, characterized by hippocampal inflammation and synaptic loss. However, the mechanisms through which diabetes may mediate these pathological changes remain unclear. This study investigated putative long-term effects of T2DM on oxidative stress, inflammation, and synaptic integrity in the hippocampus.
Methods
T2DM db/db (leptin receptor knockout obese mice) and lean control mice were segregated into young (3–4 months old), and aged (9–11 months old) groups. The hippocampus was assayed for PSD95, a marker of synaptic integrity. A cytokine array was used to estimate the inflammatory cytokines in the hippocampus. The hippocampal microglial count was also performed by immunolabeling IBA1. The oxidative stress status was assessed by 8-oxo-dG immunohistochemistry.
Results
PSD95 expression levels were unaltered in young T2DM mice, but decreased in aged T2DM mice compared to those in control mice (P<0.05). The levels of proinflammatory cytokines IL-1α and IL-1β were increased and anti-inflammatory cytokines IL-10 and IL-13 were decreased in aged T2DM mice (p < 0.05). Aged T2DM mice also showed a significantly higher microglial count in the hippocampus than in controls. The 8-oxo-dG expression was also increased in aged T2DM mice.
Conclusions
T2DM in the hippocampus of aged mice causes post-synaptic degeneration as indicated by reduced PSD95, and these findings are consistent with those in AD patients. Only prolonged T2DM causes these changes as young diabetic mice do not show post-synaptic degeneration. These findings appear in the hippocampus in association with increased oxidative stress, inflammation, and microgliosis. These results provide evidence of age-dependent propagation of AD-like pathology in the hippocampus of T2DM mice.
Funding Agency
ZM03/16 SRUL02/13.