Animal and Human Research Chapters 5 & 6 PDF

Use of Animals and Humans in Biomedical Research Keykavous Parang Chapman University Drug Discovery and Development Animal Research Human Research Cancer Drug Resistance, 2019, 2, 43 – 5. Chapte...

Use of Animals and Humans in Biomedical Research Keykavous Parang Chapman University Drug Discovery and Development Animal Research Human Research Cancer Drug Resistance, 2019, 2, 43 – 5. Chapter 5. Use of Humans in Biomedical Experimentation https://webcourses.ucf.edu/courses/1391134/pages/the-protection-of-human-subjects-in-research  An Example of Failure NIH conducted a study in Guatemala (1946-1948). Prisoners and soldiers were intentionally infected with syphilis and other venereal diseases for the purpose of studying the effects of penicillin. None of the research subjects provided informed consent. Documentation does not show evidence of treatment for the majority of those involved. The U.S. government officially apologized to the Guatemalan government in 2010.  Unethical experiments in Humans https://www.youtube.com/watch?v=0EtEFf6Bm_I&t=89s  Discussion Question What are different phases of clinical trials? Human Clinical Trials https://www.mdanderson.org/patients-family/diagnosis-treatment/clinical-trials/phases-of-clinical-trials.html  Discussion Question Would you volunteer to enroll in a clinical trial as a healthy volunteer? Why or why not?  Case 6 You have been attending a meeting on eukaryotic growth factors and have just finished listening to Dr. Sidney Wolanski give his keynote address. His overview involved some clinical studies, and he showed slides of patients undergoing procedures as part of an institutionally approved clinical trial. In all instances the faces of the patients were clearly visible. On two other slides there were clinical materials depicted and these were labeled with a tag that read “ Mrs. MacDonald.” After the lecture you leave to make a phone call. As you return to the lecture, you are intercepted by Professor Susan Jeris, a colleague you know casually from another institution. Susan confides in you that one of the slides shown by Dr. Wolanski was a picture of her stepmother, Shirley MacDonald. She is agitated and claims that Dr. Wolanski’s use of the picture and disclosure of her stepmother’s name are a violation of her stepmother’s privacy and in violation of accepted standards of clinical research. She claims that Dr. Wolanski’s presentation is an egregious violation of human subjects research practices and thinks he should be punished. She asks you what she should do about this situation. What advice and guidance will you provide to Dr. Jeris.  Some General Principles Informed Consent May not be coerced, free to withdraw at anytime. Minimize physical, mental discomfort, harm, and danger. Confidentiality Must honor any commitments made Researcher must detect and remove any undesirable consequence.  Some General Principles Every precaution must be taken to protect the privacy of research subjects and the confidentiality of their personal information. Participation by individuals capable of giving informed consent as subjects in medical research must be voluntarily. Every research study involving human subjects must be registered in a publically accessible database before recruitment of the first subject. https://www.clinicaltrials.gov/  Case 7 Professor Angela Durate is the course director of a physiology lab taught to medical students. One of the laboratory exercises involved students drawing blood from one another (under supervision) and using the serum to perform a variety of chemical and cellular analyses. The lab exercise is carried our successfully. At its conclusion, Professor Duarte announces to the class of 100 students that she would like to retain leftover blood sera. She informs them that some of the sera will be used individually while some will be pooled. In all cases, these sera will be used to gather baseline control data for a number of research projects. She asks if anyone wants to refuse to have his or her serum used for research but receives no objections. Are Professor Duarte’s actions appropriate? Is an IRB-approved protocol needed? Do the students need to give informed consent?  Protection of Human Subjects of Biomedical and Behavioral Research Respect for Person Must be treated as autonomous agents capable of making independent decisions 1. Informed consent process 2. Do not harm (minimizing risks and maximizing benefits) 3. Justice: The benefits and burdens of research should be equally distributed. No particular population is excluded from the potential benefits that biomedical research might bring to bear.  Approve and oversee research on human subjects Five members and must be filed by Health and Human Services One nonscientific member (lawyer, ethicist, clergy) One nonscientific member not affiliated with the institution  Institutional Review board (IRB) The research to be ethically valid, it must first be technically and statistically valid. Clear endpoints so the IRB can determine their risk-to-benefit ratio Monitoring data to ensure the safety of subjects. Adequate plans to protect privacy and confidentiality Records containing identifying information should be maintained in locked locations with access restricted to those have a need to use the information and who are trained in medical confidentiality or privacy. Selection of subject must be equitable (not only for people who have health insurance) The Issue of Informed Consent Competent to consent: To understand the consequences and to make decisions Consent must be voluntary and free from coercion Pressure from a family member, researcher, physician, health care system Researchers are often reimbursed in clinical studies on a per-patient basis. The per-patient fee covers the experimental costs and often a portion of the researcher’s salary. Insurance companies may exclude experimental expenses. The result may be that even those willing to enroll in large peer-reviewed clinical trials may not be allowed to participate. Consent must be informed to make a valid decision. All the risks must be disclosed. Comprehension of the procedures is required. A relative or friend witnesses the signature on the consent form.  Discussion Questions Is it ethical to oversimplify an informed consent document so that in reality it is no longer scientifically accurate? Is it legal to do this?  Discussion Questions What are some examples of coercion that might come up in recruiting human subjects into human trials? Under what conditions should a human subject research study be immediately stopped?  Human Experimentation Involving Special Populations Incompetent Persons with mental illness: Consent must be provided by the legally responsible persons, and the study must be designed in such a way that adequate safeguards exist for the participant. Prisoners: Only certain types of federally sponsored research can be performed. Advantages gained by the prisoner must weigh the risks. Selection must be fair to all prisoners. Follow-up is required For example: Vaccine studies on hepatitis due to increased incidence of hepatitis in prisons. Children: Parents or guardians must give consent. If the child is capable, his or her consent is required. Fetal Tissue and Embryonic Stem Cell Research The use of human embryonic stem cells (hESCs) in research is governed by NIH guidelines. Online registry of hESC lines that are eligible for use in research supported by NIH funding. Payments to participate in research on fetal tissue for pregnant women are prohibited. Directed donations are prohibited. Use of fetal tissue or stem cells for transplantation, particularly for the treatment of Parkinson’s diseases and diabetes is controversial. There must be sufficient evidence from animal experimentation to justify the human risk. If a new therapeutic agent is considered necessary to improve a pregnant woman’s condition, her consent alone is sufficient even if the treatment poses greater than minimal risk to the fetus. Use of Animals in Biomedical Experimentation Chapter 6  Discussion Question How would you respond to an animal rights activist who says the use of animals in research is bad science? Can you describe an experiment in which a computer could effectively substitute for the use of an animal?  Beneficial Use of Animals The use of animals in biomedical and behavioral research played a critical role in understanding biological systems in translating basic research discoveries into products that benefit humans and animals alike. https://www.youtube.com/watch?v=iA_FfVuTfoM  https://fbresearch.org/ More than 95% of animals are used for food and clothing Research = small percentage https://speakingofresearch.com/facts/medical-benefits/ Why are animal used in research? Living organisms with complex anatomic systems may respond differently to chemical or biological entities than simple, single celled organisms Current regulatory guidelines require the use of animals for testing unless there is an approved alternative Some basic research can’t be done on a benchtop! Some advanced skills can’t be taught using non-animal models/dummies  History Early animal experiments were brutal. Dogs were nailed to boards by their four paws before being cut open. Animal Rights Movement How the animals used in biomedical research are being treated? Do the benefits derived from the biomedical research that is being considered offset the associated moral costs? Legislation In the early days of animal research: There was little restriction on scientists’ use of animals other than their own individual consciences. 1963: NIH published Guide for the Care and Use of Laboratory Animals. 1966: Congress passed the Laboratory Animal Welfare Act to minimize the number of instances of people’s cats and dogs being stolen and sold to research institutions. 1970, 1976, and 1985: Amended as Animal Welfare Act (AWA): Mandated human care and treatment for dogs, cats, rabbits, hamsters, guinea pigs, and nonhuman primates. Provided no protection for rats and mice. Animal welfare: The humane treatment of animals  Legislation 1985: Congress passed the Health Research Extension of Assurance obtained from research institutions stating that they are committed to Public Health Policy on the Human Care and Use of Laboratory Animals and the Guide for the Care and Use of Laboratory Animals. Public Health Service (PHS) laboratories and any institutions wishing to receive PHS funding must agree to comply with PHS policy and the Guide. Protection of Animal Facility and Personnel The law makes acts of violence directed against animal enterprises and individuals who are associated with them an act of terrorism. Biomedical facilities and researchers  Case 8 You are a graduate student working on a project that involved administering nerve toxins directly into the cerebrospinal fluid of rats by using a special infuser connected to tubing that you have surgically implanted into the base of each rat’s skull. Administering different nerve toxins to block specific effects of different types of drugs will help how the drug works. After surgery, the nerve toxin is given, and a few days later the investigational drug is given to determine whether it will have an effect. This protocol has been approved by the IACUC and is being funded by a grant from the Department of Defense. When you go to the vivarium to take the rats to the lab to administer the investigational drugs, you find that a number of rats are paralyzed or dead. The lab director is out of town, so you go to the lab’s senior graduate student, Tom, for advice. Tom will be able to complete his dissertation writing when this experiment is done and has made it clear that he wants his experiment to run without delay. You ask him whether you should stop the experiment to determine why some of the rats are dead or paralyzed. He responds that stopping the experiment now would waste several weeks of work and delay the completion of his dissertation. He suggests that the paralysis and death of some of the rats may be due to your inadequate experience in performing rat surgery or infusions, so your gaining further practice by continuing this experiment may result in better outcomes for the rest of the rats. What do you do now? Do you continue performing surgery and infusions on the rats, knowing that more rats may be harmed? Do you stop the experiment and inform the IACUC, which risks earning the disfavor of Tom, with whom you have to work? How would you explain each course of action to the IACUC? The 3 Rs: Planning an Animal Research Project  Reduction to use the minimum number of animals: Can statistical significance be achieved with 8 animals for each treatment instead of n 25 animals? If the group size is too small to permit any reasonable chance of demonstrating a statistically significant difference, then the entire experiment is a wasted effort. Refinement: Attempt to reduce the incidence or severity of pain and distress experienced by laboratory animals. Use of anesthetics and analgesics that are appropriate for the species Appropriate doses and intervals of administration Use of trained personnel Guide How animal research should be carried out within an institution, including recommendations for animal program oversight, animal housing, and environment, facilities management, and veterinary care? IACUC basics IACUC = Institutional Animal Care and Use Committee veterinary care The IACUC is responsible for evaluating the care, treatment, housing, and use of animals, including approval of research protocols. Comprised of at least 5 members (below) and their alternates o Community/ Non-affiliated Member (non-animal user) o Veterinarian o Scientist o Non-scientist (e.g., clergy member, lawyer, or ethicist) o Another person (duplicate any of the above) IACUC responsibilities Inspect animal care and use areas- all areas housing animals (at least) every 6 months Semi-annual program review (program of humane care and use of animals) Submit reports of reviews and inspections to the Institutional Official every 6 months Suspend activities not in compliance Investigate deficiencies in animal care and use IACUC responsibilities Review and Approve animal use protocols prior to research being performed Review all protocols annually and provide continuing oversight (post-approval monitoring) What does the IACUC need in order to do a thorough protocol review? https://iacuc.northwestern.edu/getting-started/review-process.html Protocol Review Criteria: Outlined in the Guide Protocol Review Criteria Scientific rationale and purpose of use A clear and concise sequential description of the procedures involving the use of animals that is easily understood by all members of the committee. Alternatives assessed (less invasive procedures, lower species, cell cultures) Justification of Species and number of animals, and experimental group size Adequacy of training and experience of personnel in the procedures used, roles and responsibilities of personnel involved Use of hazardous materials and provision of a safe working environment. Review Criteria, continued Impact of procedures on animal well-being Appropriate sedation, analgesia, and anesthesia Conduct of surgical procedures including multiple operative procedures. Post-procedural care and observation (e.g., inclusion of post-treatment or postsurgical animal assessment forms). Criteria and process for timely intervention, removal of animals from a study, or euthanasia if painful or stressful outcomes are anticipated. Method of euthanasia or disposition of animals, including planning for care of long- lived species after study completion (consider dogs, primates, birds, others) IACUC review of Painful Procedures Assume Human pain = Animal pain IACUC has the responsibility to verify that the Principal investigator has: o considered alternatives to animal use o use analgesics, anesthetics, tranquilizers (or justify their non-use) o consulted veterinarian for surgical protocols Lit Search Info Search Services can be provided by National Library of Medicine (NLM) They are free, and can provide an unbiased search Do-it-yourself lit searches: o Must include the refinements, reductions, and replacements o Written narrative- a paragraph o 99% of the time, there is an alternative- Explain why an identified, bona fide alternative method can’t be used Attending Veterinarian Direct housing and feeding of animals Provide adequate veterinary care Provide guidance to investigators Voting member of IACUC Guidance and oversight of surgery program and peri-operative care Ensure we follow current standards of animal care…… Principal Investigator Responsibilities Appropriate species, strain and model Minimize number of animals Minimize pain and distress Consider alternatives Determine appropriate study and humane end-points Use acceptable methods of euthanasia  Guide Details qualifications and training of the professional animal care staff and the establishment of an occupational health program to protect personnel who come into contact with the animals. Space: A 20-gram mouse be allotted at least 12 square inches of floor space in a cage that is at least 5 inches high. Temperature and humidity, noise levels, requirements for bedding, water, sanitation, waste disposal, separation by species, disease control, animal door sizes, ceiling heights, placement of floor drains, and suggested locations of storage areas for food and bedding. The institution provides training for those staff members involved in the care and/or research use of animals.  Case 9 Myron Castillo is a new graduate student in Dr. Jessica Ripka’s lab. Dr. Ripka instructed Myron to complete the university animal training program within his first 2 weeks of being in the lab. Six weeks after Myron begins his lab work., Dr. Ripka personally begins training Myron in performing specialized injections in rats to begin used in her research. During an injection, Myron is bitten by one of the rats, resulting in a significant laceration of his thumb. Dr. Ripka instructs Myron to go to the student health clinic where he was screened for possible animal allergies at the conclusion of his animal-use training. Myron confesses that he has not had time to complete the animal-use training, nor has he been screened for allergies at the student health clinic. Dr. Ripka chastises Myron for failing to tell her he had not completed the training and for not disclosing this before engaging in the specialized injection training with her. Dr. Ripka is worried that if Myron goes to the student health clinic, the report that will ensue from the visit will be filed with the Institutional Animal Care and Use Committee. She fears this will lead to sanctions being placed on her animal use authorization, thus impeding her research progress. She instructs Myron to get the bite wound sutured at the emergency room of a nearby community hospital. Later in the day, Dr. Ripka shares the incident and her response to it with you over a cup of tea. She asks you if she did the right thing. What is your analysis of the situation, and what do you tell her? If she has not acted appropriately, what should she have done?  Graduate Students New graduate students should be sure that they attend these training sessions as early as possible. Ensure that the animal does not suffer any unnecessary pain or distress Students need to look at the protocols. Make suggestions to submit amended protocol if needed and use nonanimal alternative techniques  Case 10 You are a graduate student in behavioral pharmacology, and your lab is conducting a drug discrimination study, an operant procedure in which rats are trained to identify drugs with stimulus properties similar to those of a training dog. The primary goal of the present study is to test several experimental compounds for their similarity to clozapine, an important treatment for schizophrenia. The compounds to be tested have been sent to your advisors part of a contract awarded from a drug company. The generalization testing portion of the study is nearing completion, with only one dose-response curve left to obtain. During routine feeding, you notice that 8 of the 10 animals in the study have developed tumor- like growths at the site of injection on the stomach. Additionally, these animals have begun losing weights. Finally, you note that the animals do not exhibit any behaviors suggesting that they are experiencing any discomfort. Concerned, you mention the growths and weight loss to your advisor, who instructs you to continue with generalization testing. He is concerned that having to train a new set of animals in order to test one drug would waste large amounts of research time and resources and may cause problems in interpreting the results. He further states that the animals will be euthanized as soon as the testing phase of the study is completed in less than a month and that the animals will be fine until then. Is your advisor’s suggested course of action legally and ethically appropriate? If not, what should be done in this case? What are your obligations in this situation? Scientific Record Keeping Chapter 10  Discussion Questions If an NIH-funded principal investigator takes a new position, does he have the right to take all of his data and data books with him to his new institution? If not, what policies or ethical issues apply to such a situation? Could sloppy and incomplete record keeping ever qualify as scientific misconduct? Explain. Do you think that electronic record keeping will either increase or decrease our ability to detect scientific misconduct? Why or why not?  Case 11 Ming Shu, biochemistry graduate student from China, can speak and write in English, but her speed at each task is relatively slow. Every few weeks Dr. Andrekia Keys, Ming’s graduate mentor, meets with her to discuss research and to generally inquire as to how her transition into academic life in the United States is progressing. This week Dr. Keys asks Ming to leave her research notebooks out so that she can stop by the lab and inspect them. When Dr. Keys arrives at the lab, Ming has left for the day, but she finds the notebooks lying on her desk. Upon inspection, she finds that several of the notebooks are written in Chinese. Upon further investigation, she discovers that Ming is recording her initial observations in Chinese in one notebook and then translating them into English at a later time. Furthermore, when Dr. Keys examines both sets of notebooks, she observes that the current English notebook is approximately 3 weeks behind the Chinese counterpart. Dr. Keys comes to you for advice. Is Ming guilty of improper notebook-keeping practices? Can the notebooks written in English be considered valid records of her research thus far? What advice on how to handle this situation do you have for Dr. Keys?  Case 12 The research laboratory of a faculty investigator has begun using a new electrophoresis technique. The technique works well in the hands of the laboratory investigators. A field service representative from the company that manufactures the apparatus asks several of the workers in the laboratory if she may borrow some of the photographs of their results to show them to potential clients. In return, she offers to take the whole lab to dinner at an expensive restaurant. The lab members comply, and the whole group goes to dinner. You, as laboratory director, are told of these events after the fact. Comment on the implications of this scenario for data ownership and laboratory record keeping. What action, if any, will you take?  Case 13 Joshua Hanani is preparing his first major paper as an independent investigator. His faculty colleague Dr. Ellen Fang reviews the manuscript, and they meet over coffee to discuss her critique. She has some questions about the image of a DNA gel analysis in the paper. She says part of the gel image appears deliberately altered. Joshua admits this to be true and offers the following explanation. The gel contains the products of PCR-amplified whole-cell DNA. The amplified DNA fragments are arrayed linearly from largest to smallest on the gel. The experiment produced the expected results but also contained some unexpected, exciting findings. Specifically, two small DNA fragments suggest an exciting hypothesis that will likely take 6 to 8 months to test. Worried that readers would recognize this and “scoop” him, Joshua used an image editor to “underexpose” those fragments so they were no longer visible. He mentions to Ellen that he included an explanation of this in the figure legend, writing: “Minor signals of unexplained origin were present in the lower molecular size range in this experiment, but they are not visible in the photograph.” This, he claims, will protect him from the competition of major labs working in the field. Ellen cautions him that his reasoning is flawed and he is deliberately falsifying data. She offers an alternative solution, saying he should just electronically crop the image so that it includes just the expected DNA fragments and not the unexpected ones. She says this can be easily done since the unexpected fragments are at the edge of the image. Then, she argues, no explanation about the image will be needed. Comment on Joshua’s actions and Ellen’s alternative solution. If they came to you for your opinion and advice, what would you tell them?