Research@UIC > OACIB > ACC > FAQs: Completing Protocols

Frequently Asked Questions: Completing Protocols

General Protocol Submission | Completing Protocols | Protocol-Funding Matches

1. What details do I need to include in Form A, item 10e?
2. When answering Form A, item 10f- Statistical justification -- What constitutes a pilot study? When do I need a power analysis and what is it?
3. What constitutes alternative procedures (Form A, item 15)?
4. Breeding Protocols Appendix 1- Which Appendix 1 do I need? Which animals do I need to account for and where should I account for these animals?
5. Using Hazardous Agents - What information should be provided in Appendix 2?

 

1. What details do I need to include in Form A, item 10e?

• All procedures involving the use of animals need to be addressed in this section including surgery, injections of all kinds, and any other manipulations (e.g., blood sampling, behavior tests, etc.).
• Be sure to include doses, routes and frequency of administration.
• Endpoints of the study should be well defined and if appropriate objective humane endpoints, which would indicate an early termination of experiment should be included.
• Clear experimental design (how many groups, doses, time points, etc.) with a timeline and/or table for experiments is helpful to the reviewer in evaluating the protocol.
• Do Not Include -- grant applications and extensive detailed descriptions of in vitro methodologies.
  

2. When answering Form A, item 10f - Statistical justification, What constitutes a pilot study? When do I need a power analysis and what is it?

A pilot study is a feasibility study. This is a study in which the investigator is asking whether the technique, idea, or manipulation will work. A pilot study does not require statistical justification; however, it does require that the PI indicate the main outcome measure being evaluated (e.g., reduction in tumor size) and state the percent of success that would indicate that the study is biologically or clinically worth pursuing and should graduate to a full study (e.g., 30% of those treated will show at least a 50% reduction in tumor size).

If the PI has proposed a specific number of animals for an experiment in order to conduct statistical analysis following data collection, statistical justification should be used to justify the number of experimental animals per group. The most appropriate tool for this is usually a power analysis. The goal of conducting a power analysis is to determine the appropriate number of animals per group to obtain statistically valid results. For statistical justification using a power analysis, the following information should be provided:

1. The statistical test that will be used to analyze the data and that was used to calculate the power analysis, as well as whether the test is one-sided or two-sided. If a one-sided test is used, adequate justification should be given for such a test. One of the most important factors in a power analysis is choosing the correct statistical test to use.
2. A reference to the method, book, or software used to calculate the power.
3. Significance level: The probability of a Type I error (a), rejecting the null hypothesis (usually finding a difference) when the null hypothesis is correct (there really is no difference).
4. Effect size: The magnitude of the effect (usually the difference) that the researcher is interested in detecting. It can sometimes be calculated from the difference between the means that are of interest, divided by the average of the two standard deviations.
5. Sample size: The number of subjects per treatment/group, plus the total sample size (the sum of the sample sizes for all treatments or groups).
6. Power: The probability of rejecting the null hypothesis (usually finding a difference) when the null hypothesis is incorrect (there really is a difference). The power is 1 minus the probability of a Type II error (b): Not rejecting the null hypothesis when it is incorrect. Depending on the study, a power of 0.8 (80%) is usually considered acceptable.

There are a number of commercially available statistical software programs that will run a power analysis.

3. What constitutes alternative procedures (Form A, item 15)?

Federally mandated policies and procedures, as well as an evolving interpretation of these documents charge the ACC and all principle investigators using animals in their research to conduct that research in accordance with the 3 R's.

1. Reduction in the number of animals used or needed in research.
2. Refinement of the techniques used in animal research to less painful and distressful methods.
3. Replacement of animals with other models for research.

The purpose of this question is to ascertain that the PI has looked for and considered alternatives to the painful and/or distressful procedures that are proposed in the protocol and if there are less painful alternatives or other models, justification of why these are not being used. According to USDA policy 12 on this issue, alternatives or alternative methods would involve some aspect of the 3 R's. Alternatives that do not allow for goals of the research to be obtained are by definition not alternatives.

4. Breeding Protocols Appendix 1 - Which Appendix 1 do I need? Which animals do I need to account for and where should I account for these animals?

There are three appendices used for breeding and depending on the research, more than one appendix may be necessary. Appendix 1a is specifically designed for use with studies involving establishment of new transgenic of knockout animals from the stage of embryo injection to identification of transgenic /chimeric offspring (founders). If this portion of the work is being conducted by the RRC or another core facility, appendix 1a does not need to be completed. Only investigators who are establishing their own transgenic/knockout strains should complete Appendix 1a. Appendix 1b is specifically designed for use with studies involving verification of germline transmission in heterozygous/homozygous transgenic/knockout animals starting with founder animals. If the transgenic or knockout animal is already established in the strain of interest, then Appendix 1b does not need to be completed. Appendix 1c is specifically designed for use with studies involving maintenance or established transgenic/knockout animals or for breeding other species.

All animals used for breeding or offspring used in experiments or sacrificed for inappropriate genotype (or as excess) must to be accounted for in Appendix 1, as well as, in Form A, item 7 with the use of each group of animals indicated. Collection of offspring as embryos should be indicated under the disposition of offspring in the appendices, but only the pregnant breeders need to be indicated in Form A, item 7, not the embryos. In addition, known breeding problems (e.g. embryonic lethal) should be indicated in the appendices.

In addition to breeding for experiments investigators must account for animals bred for maintenance of breeding colonies. The Animal Care Committee recognizes that difficulties exist in determining the precise number of animals necessary to maintain breeding colonies and produce the necessary number of experimental animals. Difficulties can include unpredictability in the number of pups born, the genotype of those pups, births occurring at times other than are optimal or usable for experiments, difficulties with breeders and the need to continuously breed to ensure line maintenance, etc. For each line being maintained, investigators are asked to account for an additional percentage of animals required to maintain the line. Whenever possible production of large numbers of excess animals should be avoided.

5. Using Hazardous Agents - What information should be provided in Appendix 2?

All hazardous material (chemicals, carcinogens, biohazards (tumor cells, human cell lines), radioisotopes, biological vectors, etc) that will administered to animals should be included in item 1 of Appendix 2. Be specific in answering item 2. Describe the precautions, containment facilities, protective devices, carcass, waste and bedding disposal, clean-up procedures, and other necessary safety procedures in place to protect personnel and prevent accidental animal exposure. If BRL staff will be responsible for handling any hazardous material, be sure to discuss this with the BRL veterinarians and establish a standard operating procedure (SOP) with the facility prior to initiation of the study. Indicate in Appendix 2 that a SOP has been established with BRL.

In addition, the Environmental Health and Safety Office or Radiation Safety Office (RSO) should be consulted for handling of chemical or radiation hazardous with animals. Use of radioactive compounds in animals requires prior approval from RSO. In addition, if radioactive animals will be housed at BRL or an approved satellite, this requires approval from the Director of the BRL.

Last updated: Monday, March 09, 2009.