Reporting ethical matters in The Journal of Physiology: standards and advice

Abstract

Reporting of ethical matters in The Journal is very important. To advise and assist authors, particularly those who may be less familiar with the legislation in the UK, this article sets out the basic principles and methods that should be used and provides many key web sources of information. It addresses the structure of regulations, and introduces the concept of research governance. The UK law is summarized. Advice is given on the format and description of experiments, and common problems addressed. Aspects of human studies are addressed. Ethical considerations of publication such as authorship and originality, and problems such as plagiarism and fabrication are described. Updates will be published regularly.

The Journal of Physiology has two specific features that require substantial details of the ethical aspects of studies. First, The Journal aims to meet the highest ethical standards. Since it is read by those with a general as well as a specific interest, reports need to be intelligible to the general reader. Second there are no strict restrictions to contribution size. For these reasons in particular, The Journal expects full and careful reporting of the ethical details of studies. The standards applied by The Journal are based on UK legislation. Nevertheless, the composition of the Editorial Board and the readership of The Journal are world-wide, and the policies that deal with ethical matters have to encompass this. This article explains and expands the guidance for authors, and indicates where problems frequently occur, but is not intended to replace the guidance. If necessary, reference to this article may assist authors to prepare a manuscript for submission.

Animal studies

Setting ethical standards in scientific research is not simple, and the use of animals in research is a contentious issue. The sources of our ethical beliefs are varied. Different views allow different degrees of ‘moral importance’ to be drawn by different individuals; indeed some views in society may be diametrically opposed. Chapters 11 and 12 of the independently commissioned publication The Use of Non-human Primates in Research give a useful summary and overview of these problems, although in a single specific context.

http://www.acmedsci.ac.uk/p99puid83.html

Although opinions differ, there are central principles of ethical conduct that are clear and should always be followed. Simply and practically stated, each study should be reported in a way that allows the general reader to appreciate that it has been carried out properly, without causing unnecessary pain and suffering to animals. (In the case of human studies, the reader should be told that proper consent was obtained.) In addition the reader must appreciate exactly which regulations were applied to the study, and that those regulations were properly supervised.

Similar ethical standards are regulated by law in many countries. Making sure that suitable ethical standards are not only regulated but are also met and followed is important in the publication of research findings. The Journal of Physiology has standards derived from UK legislation on animal work which are among the most exacting in the world. Clearly, scientific work done in other countries has to comply with the laws of that particular country. The Journal will need to know that the standards and regulations used are equivalent to those of the UK if the work is to be published in The Journal of Physiology. In fact, although the UK laws regulate much of the fine detail of conduct, the regulations are careful not to define the exact structure of regulation to be used. The law establishes the principles, and requires each scientific establishment to make suitable local provisions. The present article lists and explains the standards and processes that must be followed by work described in Journal of Physiology articles. The principal points covered are those summarized by our guidance to authors provided by The Journal. Aspects of human studies will be covered in a later section.

The use of animals for experiments can be considered in separate steps, although in many cases the same principles and the same law may apply to several of these. In fact the UK government emphasizes this concept: animal experiments are a process from study design, through the ethical aspects of the proposed study and the conduct of the experiment, to reflection on the results after the study is complete. The law does not state exactly how the process should be implemented, but it does define exactly the principles it expects to be followed. There are additional suggestions which can be helpful in specific circumstances such as protocols of minimal severity, which can be applied to procedures such as antibody production.

The UK law requires licences to conduct experiments:

• A personal licence for the researcher.

• A project licence, detailing the types and numbers of animals involved, the severity of the procedures, and the objectives of the research. Thus each separate experiment requires a separate application for a licence.

• A certificate for the establishment where the research is to be done.

The UK regulations apply to vertebrates and Octopus vulgaris, and provide a specifically greater level of protection to cats, dogs, primates and the horse family. For many species, the regulations only allow experiments on animals that have been bred at and obtained from a designated breeding establishment. They also require more regulation for experiments in which the animal is allowed to recover from anaesthesia, and experiments where neuromuscular blocking agents are used. Breaking the UK law can lead to a number of penalties, which can include imprisonment for up to 2 years for the most serious offences.

In addition, the UK has established a separate body called the National Centre for the Replacement, Refinement and Reduction of Animals in Research: http://www.nc3rs.org.uk/

The major research funding bodies in the UK such as the Medical Research Council and the Wellcome Trust refer all proposals involving non-human primates, dogs and cats to this body for advice and comment.

Ethical review

Each application for a project licence to conduct a specific study must be reviewed by a local committee, usually called the welfare committee. This committee has to include animal care and welfare personnel and may include lay members. An important feature is that this committee, although established locally, must be independent of the researchers. More than half the members should not be currently involved in animal experiments, one should be legally qualified, and two-thirds should be qualified in medicine, veterinary medicine, or science. This committee should be named specifically when reporting matters such as approval.

The central concern of the welfare committee, when considering proposed research, is that pain and suffering should be as little as possible. If suffering is possible or inevitable, then the committee must assess the likely harm and suffering caused to the animals, and balance these against the expected benefits of the work. Some research, for example into pain mechanisms, may involve pain to the animal: this has to be balanced against the value of the observations.

The committees not only judge the ethics of proposed investigation but are also required by law to minimize the numbers of animals used, to suggest ways to reduce suffering, and to replace the use of animals when possible. These are the principles of ‘reduce, refine, and replace’ often known as the Three Rs. Consequently, to provide the best statistical power, aspects of study design may be as ethically relevant as the practical aspects of conduct.

Animal care and accommodation

The review committee also advises on the care and accommodation of laboratory animals. The care and use of animals is regulated by law, established by the UK government, in the Animals (Scientific procedures) Act 1986 (ASPA). Guidance on the act can be found at http://www.archive.official-documents.co.uk/document/hoc/321/321-xa.htm

Governance

The welfare committee has a responsibility to review the regulations set by the establishment, and to ensure that these rules are being followed and that all personnel involved are trained properly and are competent in animal care. In addition all bodies that carry out animal research are inspected to ensure that the law is followed, by the Animals (Scientific Procedures) Inspectorate. Such inspections can take place without prior warning. Satisfactory records should be kept to ensure that the process of animal care is followed, and the system should be regularly reviewed to assess the potential for improvement. This dual obligation, of suitable regulation and of ensuring that these rules are applied properly, is often expressed by the term governance.

Thus, in the UK the welfare committee establishes both the quality of animal use and care and also ensures that the institution's standards are followed. This leads to the requirement that all Journal of Physiology articles should state where the experiments are carried out and the exact names of the bodies

• that approved the experimental procedure,

• that set the regulations for the animal care,

• that governed the proper implementation of the rules concerning the conduct of the study and the acquisition and care of the animals concerned.

In some cases, these bodies may be the same. In the UK, the Home Office requires the local institution to set up the welfare committee, but appoints its own inspectors to ensure that the rules are applied. In other countries, there may be separate institutions, for example in the USA there may be Federal, state and local laws that govern animal care. Government institutions in the USA must follow the Public Health Service Policy on Humane Care and Use of Laboratory Animals, which includes the regulation of research establishments. This policy uses a Guide for the Care and Use of Laboratory Animals. This is a very comprehensive authority particularly on animal care and is available in several languages: http://www.nap.edu/openbook.php?record_id=5140

This policy requires institutions to have local Animal Care and Use Committees. Animal use in the USA is also regulated by the National Academy of Sciences, the Department of Agriculture, and by the Food and Drug Administration. In addition, the non-Federal organization, the American Association for the Accreditation of Laboratory Animal Care (AAALAC), inspects facilities of accredited institutions on a regular basis every 3 years.

In some countries the rules may be set by one body and governed by another, which is a good principle to ensure impartial governance. However in others, the legislation and governance may be by central government authorities, in which case this should be stated in the manuscript. If these regulations have been followed The Journal can be sure that the housing, care and use of animals has been carried out in properly set and regulated conditions, and the reader is clearly informed that the work has been carried out to a satisfactory standard.

In the UK, the Medical Research Council provides useful guidance on animal studies, including governance, welfare, and responsibilities of the involved participants: http://www.mrc.ac.uk/Ourresearch/Ethicsresearchguidance/Useofanimals/index.htm

Experimental procedures

The fundamental criterion is to give a description that would allow the study to be replicated by a similarly trained research worker.

Source of animals

In most cases, state the source of the laboratory animals used. For genetically manipulated animals, the name of the person who bred and supplied them, with citations of relevant work, may be necessary. In a similar way, the provenance of unique tissues, cells, or antibodies should be described. One of the understandings of publication of a paper is that the authors are prepared to supply unique reagents or cell lines used in the published work.

Animal care

Animals should be housed and fed in suitably regulated conditions. The relevant legislation and governance should be stated.

Numbers studied

In some studies, animals are used as a source of tissue or organs, and in others the individual animal is studied. Many of the latter type of studies are complex and can involve several groups of animals. The number of animals used in such studies should be given, to assure that the numbers are not excessive. Knowledge of the numbers used is important and also allows statistical evaluation.

Anaesthesia

The generic name, the doses (in mass units), the route of administration, the indications for giving supplementary doses and the amount of these doses should be stated. Methods of assessing the adequacy of anaesthesia should be stated. For example, reflex responses such as limb withdrawal to paw pressure may be used to indicate the need for a further dose of anaesthetic. The state of the animal should be charted regularly. This will allow the need for supplements and the number of doses used to be recorded. This can be usefully stated in the Results section, as it indicates how this requirement was met adequately.

A particular concern is the use of neuromuscular blocking agents. These agents are generally used for two reasons, either to allow a more stable preparation, or to allow light anaesthesia that will cause less suppression of neural activity. To establish that light levels of anaesthesia combined with blocking agents are required, deeper anaesthesia must be shown to interfere with the proposed study. The level of anaesthesia proposed must be shown to be adequate, without using neuromuscular blocking agents, for the procedures proposed. Preparatory surgery should always be done using full surgical anaesthesia. Residual pain must be prevented with long-acting local anaesthesia or analgesics before the level of anaesthesia necessary for the study is approached. Some methods of holding the head firmly can be painful, and alternatives should be used when light anaesthesia is used.

During neuromuscular block, there are few reliable indications of adequacy of anaesthesia. Cardiovascular autonomic responses to stimuli, when present, may indicate inadequate anaesthesia but the absence of such responses is not foolproof, as amply shown by human evidence. Other agents may affect useful signs, such as the size and reactivity of the pupil. Consequently careful monitoring of the EEG by an experienced observer may be the only indication of suitable depth of anaesthesia, and anaesthetic agents that have effects which allow reliable EEG assessment should be used. Agents such as chloralose and ketamine may cause effects that are more difficult to assess. An alternative approach is to periodically withdraw neuromuscular block and show that reflex responses are absent (Drummond et al. 1996).

If decerebrate animals are used, then the account should state the level of brain section. The site of the section must be sufficient to ensure complete lack of sentience. Generally rostral destruction of brain tissue is preferred.

Chilling is not an acceptable method for immobilizing cold-blooded vertebrates. In particular, an anaesthetic such as tricaine should be used to allow removal of oocytes from Xenopus. Chilling can be used for fetal or neonatal rats and mice.

Killing

The Journal of Physiology requires that authors state the method of killing animals at the end of an experiment. Studies may require the removal of organs and tissues from animals that have been humanely killed. The Animals (Scientific Procedures) Act 1986 has an appendix (Schedule 1) which lists methods that can be used (by appropriately trained staff) to obtain animal tissues that do not require a personal project licence. This code of practice describes methods that ‘may be performed consistently in a humane manner by someone with appropriate training’. (http://scienceandresearch.homeoffice.gov.uk/animal-research/).

The code describes such methods as anaesthetic overdose, and sets the size of animals that can be killed by physical methods such as neck dislocation. Fetal, larval and embryonic animals are also considered.

Tissues may be obtained from a regulated abattoir. Unless relevant, the method of killing does not need to be stated.

In the USA, an equivalent document that contains a great deal of useful additional information is available at: http://www.avma.org/issues/animal_welfare/euthanasia.pdf. Although there are some small differences, the recommendations are substantially similar and can be considered an equivalent authority. In the European Union, the document Aspects of the biology and welfare of animals used for experimental and other scientific purposes (Annex to the EFSA Journal (2005) 292, 1–136) provides current equivalent legislation for member states: http://www.animalexperiments.info/studies/welfare_efsa_2005.htm

The balance between suffering and science

Some experiments cannot avoid distress, pain, or discomfort to animals. Indeed the object may be to study the mechanisms of pain, itch, inflammation, and so on. The process of ethical review is to ensure that in such cases, the knowledge gained will justify the means employed. This balance is difficult, but the principles that can be applied are introduced by Porter (1992), and of course supported by the continued application of the three Rs: reduce the numbers, refine the methods, and replace with a more acceptable alternative whenever possible.

Common problems in submitted papers

The following topics are the most frequent cause of concern, and having to deal with these matters at a late stage of the editorial process can seriously delay publication.

Inadequate details

Some researchers appear to be unaware of the exact legal framework under which they operate, and refer to a ‘local committee’ in the mistaken belief that this will identify the institutional regulatory authority who approved the project. Please provide the names of these bodies, and of the authorities that regulate them. Many researchers do not specify the numbers of individual animals used, and may forget to state how they were killed at the end of the experiment. (The Journal prefers this expression rather than ‘sacrifice’, or variations on the word ‘euthanasia’.)

Refer to previous work

It is never sufficient to refer the reader to previous reports for details. Often the cited article contains little or no information, and a ‘daisy chain’ of citations can mean a prolonged search for the full details.

Use of neuromuscular block

As discussed above, use of agents to cause neuromuscular blockade is of great concern, and papers that describe their use are scrutinized very carefully. Recording and presenting the use of additional anaesthesia, given during the period of neuromuscular block, may be required. Anaesthetic requirements can vary considerably, for example some strains of a species may show different susceptibilities to anaesthetic agents, or different rates of metabolism.

Recovery studies

Analgesic and antibiotic therapy should be described. The conditions in which animals are kept should be stated, such as housing, nutrition, and social environment. Animals should be regularly monitored, and suitable measures used to recognize pain and distress.

Studies involving stress, pain or inflammation

Although some behaviour patterns are typical, recognizing animal behaviour that indicates suffering may require knowledge and training.

Helpful guidance on alleviating animal pain and distress is available in the National Research Council publication “Recognition and Alleviation of Distress in Laboratory Animals”http://www.nap.edu/catalog.php?record_id=11931

Differences among the UK, USA and other countries

Although the process of regulation in the UK and the USA is different, generally the principles are similar, although the UK regulation is more focused. In cases of uncertainty, The Journal will require convincing that the studies it reports from other countries would have been approved under UK legislation. In some circumstances the journal may seek informal advice from the Home Office or its advisors.

Human studies

The fundamental principle of research with human subjects is freely given informed consent, which can be withdrawn by the volunteer at any time. This incorporates two central ethical principles, truthfulness and autonomy. These principles are incorporated in the Declaration of Helsinki, which is issued by the World Medical Association, and is a code specifically established to govern medical research. The principles are generally followed by human research committees, although the entire Declaration is not accepted by some Governments. In addition, some revisions are accepted by some Governments, some by others, so the relevant version may vary from country to country. In the UK, medical research is also regulated by Research Governance Frameworks which established the National Research Ethics Service (in England): http://www.nres.npsa.nhs.uk/

The National Research Ethics Service has set up local National Health Service (NHS) Research Ethics Committees which are responsible for the approval of human studies if NHS subjects, staff, or resources are involved in the research. Research into medicines is subject to regulation by the Medicines and Healthcare products Regulatory Authority (MHRA).

Neither the MHRA nor the research governance framework regulate research into basic science, although local Research Ethics Committees established by these authorities can provide ethical approval for non-NHS establishments. The Medical Research Council has published Good Research Practice, which is a useful guide for all human studies.

http://www.mrc.ac.uk/Utilities/Documentrecord/index.htm?d=MRC002415

Reporting standards

For all human studies, details of methods of recruitment, including criteria for selection and exclusion, and the rates of recruitment, refusal, allocation, and follow-up should be stated. If the study is randomised, the Consolidated Standards of Reporting Trials (CONSORT) statement on reporting randomised trials should be followed, and trials of therapeutic agents should be registered: http://www.consort-statement.org

All human research should take place in suitably equipped and staffed locations. Contingency plans should be constructed to deal with possible emergencies, and local facilities should be accessible.

Coercion

Subjects should consent freely. Payment to volunteers is ethically problematic because it may be interpreted by some as coercion. If payment is made, this must be commensurate and not excessive, and should be stated. A more subtle form of coercion is when junior staff, students and people in a similar position feel obliged to take part. This is best avoided, but if this does occur, the ethics committee should pay particular attention to the circumstances and may require special methods for recruitment.

Consent must be given freely after full explanation. Children may consent if they are judged competent to understand the implications of taking part, but generally parental involvement is advisable. Individuals who are not competent to give permission can take part in research but there are complex regulations that govern these circumstances. The Medical Research Council provides two helpful guides: http://www.mrc.ac.uk/Utilities/Documentrecord/index.htm?d=MRC002430 and http://www.mrc.ac.uk/Utilities/Documentrecord/index.htm?d=MRC002409

For research involving ionising radiation, regulation is governed also by the Ionising Radiation (Medical Exposure) Regulations 2000, as amended by the Ionising Radiation (Medical Exposure) (Amendment) Regulations 2006, as well as any local regulations. This includes X-rays, including screening X-rays, and radionuclide imaging and tracer studies.

Data protection is an important aspect of human research. Participants should be informed that relevant data may need to be stored, and suitable secure methods should be used for this purpose. Data may not be released without the consent of the subjects, and if possible steps should be taken to keep the data anonymous.

Indemnity

Volunteers should be aware of mechanisms for compensation if they are harmed.

Declaration of Helsinki

As indicated, this set of principles is for guiding medical, rather than scientific, research. Consequently, some of the considerations may fall outside those relevant to research into physiology. For example, the use of placebo treatment has been controversial in considering revisions of the Declaration, whereas in basic scientific studies with volunteer subjects, the concepts of active therapy become less relevant, and placebos may be essential in the design of particular physiological studies. Nevertheless, some studies of physiology will necessarily involve patients, in which case the Declaration is often considered to take precedence over national law. If the Declaration is cited, then the revision used should be specified.

Human tissue act

UK legislation is complex, although the principles are those of freely given consent, and confidentiality. The Human Tissue Act (HTA) requires that informed consent is required to take tissue samples (this includes blood) wholly or partly for research unless the samples can be satisfactorily anonymised. Licences are required to take and store tissue. Again, the MRC provide helpful information: http://www.mrc.ac.uk/Ourresearch/Ethicsresearchguidance/Useofhumantissue/index.htm

The Human Fertilisation and Embryology Act 1990

The Human Fertilisation and Embryology Act 1990 (HFEA) regulates the creation, storage and use of embryos in research throughout the UK. A licence is required to conduct research, and the process is highly regulated.

The act can be seen at http://www.opsi.gov.uk/acts/acts1990/Ukpga_19900037_en_1.htm

but a more useful introduction is available at http://www.hfea.gov.uk/en/1179.html

Both the HTA and the HFEA are complex, and advice should always be sought before research using such material is planned. If research is governed by these acts, this should be stated in the methods section.

Ethics of publication

The Journal supports the guidelines on good publication practice laid down by the Committee on Good Publication Practice (COPE). These guidelines are primarily for editors but provide a useful indication of authors’ obligations: they are available at http://www.publicationethics.org.uk. A more detailed source is presented by Blackwell publishing at http://www.blackwellpublishing.com/publicationethics/

Authorship

Authors should have decided the authorship in advance, preferably when the study was planned. All authors should have had a material input into the generation of the paper, and be able to defend the contents. ‘Guest’ authorship is not infrequent in scientific publication, but is wrong and unacceptable. The Editorial Board endorses the general principles set out in Guidelines on Good Publication Practice produced by the Committee on Publication Ethics (COPE; http://www.publicationethics.org.uk/guidelines).

Changes of authorship

Investigators should obtain the full written agreement of all parties before changes are made to the authorship of a paper. If the original authorship agreement is revised it is advisable to retain records of this in case of dispute.

Corrigenda/errata

The convention of The Journal of Physiology is to designate corrections initiated by the author as corrigenda and those initiated by the publisher as errata. If the authors are aware of errors in published papers, they must inform the Editorial Office and provide material to allow a corrigendum to be published. If the publishers recognise any mistakes in the published material they will publish an erratum.

Retraction

If a serious error is found, retraction of the paper may be necessary. In addition, if misconduct is suspected, The Journal will issue a Notice of Concern. If the misconduct is confirmed, a full retraction will follow.

Author contributions

The author submitting a manuscript must ensure that all authors listed are eligible for authorship. Each author should take responsibility for a particular section of the study Authorship credit might include contributions to concept/design, acquisition of data, data analysis/interpretation, drafting of the manuscript, critical revision of the manuscript and approval of the article. Each author's contribution should be described briefly in a statement added before the Acknowledgements section. Acquisition of funding or the collection of data alone does not alone justify authorship. Such contributions to the study should be included in the Acknowledgements to appear immediately before the references. Contributors who do not qualify as authors should be listed and their particular contribution described. Further information on authorship and contributorship may be found at <http://www.icmje.org/#author Please also indicate where the experiments were done.

Conflict of interest

Authors should declare conflicts of interest. For a good definition of conflicts, which may not be only financial, please see http://journals.plos.org/plosmedicine/competing.php

Other links such as family or employment may also be relevant. Authors may wish to disclose conflicts of interest in an appendix to the manuscript, but they must always be declared to the Editor, and may be required to be disclosed in a published article.

Originality

Material submitted for publication should be original. Prior publication includes presenting material on a website, other than as an abstract. A manuscript which is merely an expanded version of work published elsewhere is not acceptable, unless this is an abstract that does not exceed about 400 words.

Copies of any material published in the preceding year that overlaps the content of the manuscript should be submitted with the manuscript. This includes notes, communications, abstracts, chapters or reviews. Articles submitted for consideration elsewhere or ‘in press’ should also be provided. This material should be submitted as ‘Supplemental files’ in PDF format, and specifically mentioned in the authors’ covering letter.

Plagiarism and redundancy

Unattributed use of previously published work is absolutely unacceptable. If you are in doubt, please indicate to the Editor that the material has been used before. Some authors are concerned that text that contains an exact repetition of previously used words, particularly in a methods section, will be detected by software designed to detect text that is common with previously published work and they will be accused of plagiarism. Plagiarism is usually more extreme, and a simple citation of the original text is sufficient to dispel the possibility.

Fabrication, falsification, manipulation

Altering images, manipulating photographs, cosmetic changes to blots, and so on are fraudulent and unacceptable. Editors may request to see original data. If there are concerns, such matters are always referred to the Publication Ethics Committee of the Physiological Society for further action, following rules suggested by the Committee on Publication Ethics. Scientific misconduct of any type is regarded very seriously.

The MRC has set rules for management of suspected misconduct which can be a useful general guide: http://www.mrc.ac.uk/Utilities/Documentrecord/index.htm?d=MRC002454

In the UK, an independent Panel for Research Integrity has now been established to address misconduct in research. It will provide advice and guidance. http://www.ukrio.org.uk/sites/ukrio2/uk_research_integrity_office__ukrio_/index.cfm

Summary

This article has been written to help researchers report the ethical aspects of their studies completely and correctly, and also as a guide to some of the more complex aspects of physiological research. Comments and criticisms are welcomed to allow further publications to be incorporated into revisions of this article.