RESEARCH ETHICS

Frank C. Jahn and Carol L. Mitch
1992 Woodrow Wilson Biology Institute

Rationale:

We hope to teach science as a process; to accomplish this we use experimental labs and independent research projects. When we discuss science as a process, we usually present an idealized view which holds that honesty is a necessary basis for scientific endeavor. Recently, the media have reported misconduct and fraud by scientists. A possible consequence is the inference by students that cheating is common among scientists and that there is little connection between what is taught in the classroom and what happens in the "real world." By extension, the body of knowledge developed by honest scientists is also suspect. By evaluating historical instances of misconduct and exploring the ethics involved in honest data analysis, it is hoped that students will begin to understand the consequences of scientific misconduct and the importance of self-regulation within the scientific community.

Intended Audience:

Objectives:

1. To develop students' ability to distinguish between simple error, misconduct, and fraud.

2. To lead students to consider a widening circle of the consequences of misconduct and fraud.

3. To understand the self-regulating nature of scientific research.

Procedures:

1. Provide students with a historical review of scientific fraud.

2. Involve students in discussion of the implications of scientific fraud.

3. Provide students with the opportunity to analyze experimental design and data in order to detect examples of error or misconduct.

4. Involve students in a role playing situation which presents an ethical dilemma in which honesty conflicts with personal gain.

Historical Review

Background Material:

Dishonesty in science may take several forms. It is important for the student to recognize the distinction between error and knowing dishonesty or misconduct. Strictly speaking, the latter is called fraud when there is intent to gain. Misconduct may take several forms including outright lies (inventing observations or experiments), plagiarism, "fudging" or "massaging" the data (manipulating the data to make them appear more convincing), or selectively choosing only data which fit the experimenter's hypothesis. Some of the most illustrious names in scientific history have been suspected of fraud. Galileo, Newton, Dalton, Mendel, and Millikan are sometimes included on lists of suspected cases of scientific misconduct (Broad and Wade, p225). In recent years a number of workers in the biomedical sciences have had their integrity publicly challenged. Examples include Elias Alsabti who apparently plagiarized as many as 60 scientific papers during the late 1970s (Miller and Henson, pp80-96); John Darsee whose scientific career in cardiovascular research appears to have included a series of fabricated experiments (Ibid., pp 55-79); and William Summerlin whose work in immunology in the early 1970s was challenged when skin grafts on mice were found to have been drawn with a black marking pen (Broad and Wade, p155). Two recent instances of suspected scientific fraud include Robert Gallo's claims to priority in the isolation and identification of the AIDS virus and David Baltimore's involvement in an immunological experiment that still cannot be replicated by other researchers.

The Baltimore case began in 1985 when Margot O'Toole, a researcher working in the laboratory of Thereza Imanishi-Kari at MIT, found that certain results of a key experiment described in a paper in Cell could not be replicated. Her complaints led to investigations which essentially absolved Imanishi-Kari of misconduct. The case was reopened by Ned Feder and Walter Stewart, described as "science fraud hunters" (Bell, p114), whose efforts led to investigations by NIH and the Secret Service (Bell, p114). In 1991, the NIH Office of Scientific Integrity found that significant portions of the data were fabricated and Baltimore, as co-author, was criticized for his role in supporting Imanishi-Kari. The Cell paper was retracted in 1991. In 1992 a Maryland prosecutor decided against prosecuting Imanishi-Kari; Baltimore responded immediately by announcing that the paper should be reinstated and that restitution should be made to Imanishi-Kari (Hilts, pC3).

Suggested Activities:

1. Assign research of an historical instance of scientific fraud. (See Background Material for some examples.)

2. Prepare a case study of a hypothetical case of fraud.

3. Ask students to respond either individually or in small groups to questions such as the following:

   Cite examples where the self-regulating nature of scientific research led to the exposure of error, misconduct, or fraud.

   What evidence suggests that the scientist is guilty of misconduct rather than simple error?

   What ethical principles were violated by the scientist's behavior?

   What are the possible consequences to the scientist of his/her behavior?

   What are the possible consequences of undetected misconduct to the following:

   • the scientist's colleagues and co-authors?

   • the head of the lab in which the scientist works?

   • the institution for which the scientist works?

   • other scientists?

   • the general public?

   What are the possible consequences if the misconduct is detected? Use the same examples from the question above.

4. Assign students to prepare a brief for either the defending attorney or the prosecuting attorney in a case of science fraud.

Laboratory Exercises

To The Teacher:

The goal of this section is to give the student the opportunity to apply some of the scientific and ethical theory presented in the classroom. Students must integrate knowledge about the scientific method, biotechnology and ethical decision making in order to successfully complete the exercises. Dealing with fraudulent or misrepresented data is not a problem restricted to the scientific community but occurs throughout the business community as well. This series of labs follows a spiral approach, starting with a simple, straightforward experiment and culminating in a complex exercise requiring the student to call on both biological knowledge and ethical skills.

These labs are designed to be presented sequentially but not necessarily contiguously. Each exercise is most effective when used to reinforce related biological content. The ethical component of these labs also follows a spiraling strategy. The first lab demonstrates problems resulting from sloppy lab technique; the second focuses on the problem of misrepresenting data to promote a conclusion consistent with the hypothesis; the last involves the detection of deliberate manipulation of data to support desired results.

Laboratory One: The Importance Of Controls In Experimental Design

The first exercise places the student in the position of possibly reporting an erroneous conclusion because of faulty experimental design. The lab requires background knowledge of the scientific method, carbohydrate chemistry and enzyme action. The exercise consists of two very short and simple labs that effectively reinforce enzyme action content. The first day students learn the mechanics of identifying enzyme action and on the second day design a lab to demonstrate the action of the same enzyme on a second substrate. (In reality, the enzyme does not operate on the second molecule and a false positive is the result of poor exper imental design.)

Laboratory Two: The Importance Of Data Analysis

In the second lab, the student is presented with the dilemma of basing a conclusion on either reliable background information or on empirical data. The literature on dendrochronology seems to support overwhelmingly the hypothesis that the width of annual tree rings is closely correlated to the amount of rainfall. Exceptions are cited in the literature available to the student (see bibliography); however, most students overlook both exceptions and empirical data and present a conclusion conforming to the "opinion" of experts. After the completion of the lab, the concept of bias should be discussed and historical examples cited to illustrate the importance of objective data analysis.

Laboratory Three: Fraudulent Or Manipulated Data

The final experiment requires the student to scrutinize experimental data and apply critical thinking skills to the detection of fraudulent data. The student must then make a series of ethical decisions about how the data should be used and about truthfulness versus personal gain. The lab culminates with the student assuming the role of a salesman recently hired by an agrobiotech com-pany. As the salesman the student has been provided with background infor-mation, some suspect, most completely reliable (based on actual data from the literature) by the research staff of the fictitious company. The student must identify the fraudulent data and decide how to present the information in the context of a sales meeting with a potential client. This exercise not only helps the student clarify the ethical decision making process but also reinforces content material in the area of genetic engineering, cloning and transgenic organisms.

Protocol Appendix

Enzyme Action: The Breakdown Of Lactose By Lactase

Teacher Background:

The purpose of this lab is to enable students to investigate the breakdown of lactose, milk sugar, by the enzyme lactase, into two monosaccharides, glucose and galactose. A glucose indicator is used to confirm the hydrolysis by detecting the presence of the resulting glucose. Students can also test the breakdown of lactose in milk to better understand the causes of lactose intolerance.

Student Material (Per Lab Team):

• 2 - Glass slides

• 1 - Dropping bottle of milk

• 1 - Dropping bottle of water

• 1 - Dropping bottle of glucose solution

• 1 - 1/2 tablet of DairyEase, powdered

• 5 - Strips of Glucose Enzymatic Test Strips, such as Tes-Tape, 2cm long

• 4 - Toothpicks for stirring

• 1 - Spatula or wooden splint to deliver powdered lactase

Teacher Preparation:

Lactose Solution - Dissolve 5 grams of lactose in 100ml of warm water.

Milk - All types of standard milk have lactose.

Glucose Solution - Dissolve 5 grams of glucose or dextrose in 100ml of warm water. A small amount of honey works just as well as reagent glucose for this test.

Lactase Source - Drugstores offer many over the counter digestive aids containing lactase, but for this lab it is important to use DairyEase. Use a mortar and pestle to powder the tablet.

Glucose Enzymatic Test Strips - There are many glucose indicators available without prescription to test the sugar levels of urine and blood. One of the least costly, simple and reliable is Tes-Tape. Tes-Tape contains glucoseoxidase and an indicator which changes from yellow to green in the presence of glucose.

Protocol Outline

1. Place two glass slides on a piece of white paper.

2. On one slide put the controls samples, glucose and lactase. Because the Tes-Tape requires a liquid, use the glucose solution and prepare a lactase solution by adding a drop of water to a small sample of the powdered DairyEase placed on the slide. Label the samples.

3. On the second slide place the experimental factors, lactose and lactose with the enzyme. Put a small samples of lactase powder on the slide; then put a drop of lactose solution on the slide (before) and on the lactase powder (after). Identify each by writing on the white paper.

4. Test the controls and experimentals by dipping the Tes-Tape in each solution. The color change from yellow to green, indicating a positive test for glucose, will occur within fifteen seconds if glucose is present.

5. Record the results on a data table.

6. The experimental procedure can be repeated using milk in place of the lactose solution to demonstrate the presence of lactose in milk.

Enzyme Action: The Breakdown Of Sucrose By Lactose

Teacher Background:

The purpose of this lab is to demonstrate problems encountered when poor laboratory techniques or deliberate oversights result in misleading data. The student is required to design an experiment to determine if lactase can catalyze the hydrolysis of a disaccharide other than lactose. This lab uses the disaccharide sucrose and another lactose intolerance treatment, Lactaid. Most students will follow the procedure from the model but many will try to shorten the process by eliminating the controls.

Since sucrose is a disaccharide consisting of glucose and fructose, a positive test for glucose would be an indication of enzymatic activity. Lactaid contains dextrose (glucose) as a filler and sweetener and therefore introduces glucose into the experiment causing a "false positive". The conscientious student will report that the control contains glucose and therefore the test cannot conclude anything about the activity of lactase on sucrose. Those students who short-cut the procedure report incorrectly that the glucose in the lactose/enzyme solution is a clear indication of enzyme activity.

The post-laboratory discussion should focus on the idea that a review of the poorly designed lab would alert the investigator to the possibility of an erroneous conclusion.

Student Material (per lab team):

• 2 Glass slides

• 1 Dropping bottle of water

• 1 Dropping bottle of sucrose solution

• 1 Dropping bottle of glucose solution

• 1 1/2 Lactaid tablet, powdered

• 5 2cm strips of Tes-Tape

• 4 Toothpicks for stirring

• 1 Spatula or wooden splint to deliver powdered lactase

Teacher Preparation:

Sucrose Solution - Dissolve 5g of sucrose in 100ml of warm water.

Lactase Source - In this experiment use Lactaid, rather than DairyEase. Again a mortar and pestle can be used to powder the tablet.

Protocol Outline

For this lab it is important that the student be responsible for the experimental design with as little help from the teacher as is practical for the level of students. Because the lactose/lactase lab is the model, it is essential that the students fully understand the preceding lab, especially the role of controls.The procedure identified by the student should resemble that in the previous experiment.

Annual Tree Rings: Correlation Between Rainfall And Tree Ring Width

Teacher Background:

The purpose of this lab is to give the student some practical experience with the concept of tree ring formation and dendrochronology. The students are asked to solve the problem, "Can annual tree rings be used to determine rainfall in the area?" At the same time students are confronted with an ethical dilemma: should they base their conclusion on the extensive background information which supports an hypothesis of correlation between variables or upon the empirical data which does not support that position? At the conclusion of the exercise students should be involved in classroom discussions regarding the importance of basing scientific conclusions on empirical data and not on bias.

There is extensive literature available to the student on dendrochronology and the influence of annual rainfall on the width of the tree rings. A bibliography of the more recent publications is listed at the end of this section. Students should be given a definitive article as well as one period for library research.

The most difficult part of the exercise is the construction of the graph (if practical, students should be encouraged to use the computer to prepare the graph). The year is graphed on the X-axis and the Y-axis has two value scales, width of the tree ring and annual rainfall. This is a rather sophisticated graphing technique and should be discussed with the class before they begin graphing. Student anxiety is greatly reduced if a blank graph is provided with the axes labeled and the scale identified.

Student Material (per lab team):

• 2 - Copies of an article on dendrochronology, suggested material. Palmer, Alfred C. "What Tree Rings Tell." The Science Teacher. September, 1986. pp 70 - 73.

• 1 - Tree trunk slab, preferably conifer, wide tree rings, year round water.

• 1 - History of the tree slab, including type, general location and year felled.

• 1 - Metric Ruler

• 1 - Local annual rainfall chart, close to the source of the tree slab

• 2 - Graph paper, axes and increments labeled.

Teacher Preparation:

Tree slab - The two most important criteria for choosing tree slabs are ring width and proximity of the tree to a constant water supply. Trees from areas such as the West, with long growing seasons, provide samples with the widest rings (easier and more accurate measure-ment). Conifers show less correlation than hardwoods do to seasonal fluctuations, therefore conifers are a better choice for this lab. Trees growing near a year round water supply (along streams or under sprinklers) show the greatest consistency in ring width.

Tree slabs can be purchased from most biological supply houses but it is much cheaper to make your own. Contact the local forest service and explain the exercise, ask for the location of a down tree close to a weather recording station and for necessary permits. Often the annual rainfall records are available from the weather service. Cut 5cm slabs from the trunk and allow them to dry for at least a month. After sanding and a layer of wood sealer (a layer or two of polyurethane improves visibility of the rings and protects the wood from pencil marks) the slabs are ready to use. Often the wood shop is only too happy to help with the sanding and painting.

Protocol Outline

1. Calculate the life span of the tree by starting with the outermost ring as the year the tree was felled and counting toward the center to determine the range.

2. Construct a table to organize the year, rainfall and tree ring width.

3. Enter the rainfall amounts from the data provided.

4. Measure the width of each tree ring and enter the amount into the table.

5. Construct the graph and analyze the data by inspection.

Transgenic Plants: Business Or Science?

Teacher Background:

This exercise provides students with the opportunity to review information from classroom discussions of genetic engineering and gene expression as well as to gain experience in detection of fraudulent or misinterpreted data. Students will be asked to make ethical decisions regarding the disclosure or use of the data in a role playing situation. The entire exercise requires two and one half days to complete.

Teacher Preparation (per student):

1 - Set of Background Information Sheets (4 sheets)

1 - Dialog Interview Worksheet

Protocol Outline

• Day One (15 minutes) - Explain the simulation to students and distribute the information sheets which should be studied in preparation for a sales meeting.

• Day Two (One period) - Break the class into groups to study reports and graphs and to identify any areas of potential misrepresentation. After about 20 minutes, discuss the findings of the groups as a class. Inform students that they will be asked to respond individually to the questions of a potential client the next day.

• Day Three (One period) - Allow about 20 minutes for students to complete the Sales Meeting Simulation. For the remainder of the period discuss the ethical issues involved in using misrepresented data to sell a product.

Class Discussion:

The data in the sales review is accurate but some of the graphs display & quot;misrepresented data." For example, the information on insect mortality (Fig. 2) is misleading because every surviving insect may give rise to a toxin-resistant generation. Data on Insect Toxicity Levels (Fig. 1) uses " corrected" mortality which includes live but small larvae. Thus correctedmortality represents little impact on this year's crop however, if the larva matureto produce a resistant generation, they may have a profound impact in subsequent years. The biomass information (Fig. 3) is also misleading in implying that the difference in biomass represents a difference in growth rate. By keeping all external factors equal (using no pesticides), the biomass of the non-transgenicplants is reduced by insect predation while the transgenic plants have the advantage of reduced insect predation due to production of internal pesticide toxins.

Evaluation:

Student responses in the simulation should be evaluated in different ways depending upon the nature of the question. Questions about content may be graded objectively; those which call for judgment and ethical decisions should either be ungraded or evaluated on the basis of the number and quality of considerations rather than on the final decision itself.

References:

Historical

Bell, Robert 1992 Impure Science: Fraud, Compromise and Political Influence in Scientific Research John Wiley & Sons, New York [includes discussion of flaws in current systems for uncovering scientific misconduct as well as an extensive bibliography]

Broad, William and Nicholas Wade 1982 Betrayers of the Truth Simon and Schuster, New York [describes numerous cases of scientific misconduct prior to 1991; style willprobably appeal to high school audience; an appendix lists cases of known or suspected science fraud from the time of the Greeks through 1991]

Hilts, Philip J. 1992 "Researcher Accused of Fraud in Her Data Will Not Be Indicted" The New York Times, July 14

Kohn, Alexander 1986 False Prophets Blackwell, New York [highly readable account of specific examples of science fraud]

Miller, David J. and Michel Hersen (Eds.) 1992 Research Fraud in the Behavioral and Biomedical Sciences John Wiley & Sons, Inc. [series of essays on various aspects of misconduct in the sciences; interesting to contrast the account here (by Arthur Jensen) of the Cyril Burt case with that in Broad and Wade; also includesan extensive bibliography with each essay]

National Academy of Sciences, National Academy of Engineering, and Institute of Medicine 1992 Responsible ScienceL: Ensuring the Integrity of the Research Process Vol. I National Academy Press, Washington, D.C. [a government study of the practice of science with recommendations for public policy; bibliography includes many government publications]

Tree Rings

Stahle, M.K., et al. "North Carolina climate changes reconstructed from treerings: A.D. 372 to 1985." Science: 1(240). pp 1517-1520.

Trefil, James. "Concentric clues from growth rings unlock the past." Smithsonian: July, 1885. pp 381 - 395

You have recently been hired for the sales staff of Greengene, a small agricultural consulting company. Greengene recently acquired an emerging biotechnology company that produces transgenic plants, most notably insect resistant clover. As the most junior member of the sales team you have a low salary and you must rely on commissions for the bulk of your income. You will shortly be meeting with an important client and in preparation you requested and received summary reports from the research and development scientists.

(The summary reports are essentially based on material from the current biotechnical literature. In order to conform to a simulation format, names, places and situations must be "contrived" for story continuity. Names, places and graphs are all fictitious; other "altered" information will be identified by italics.)

Recent advances in biotechnology have enabled scientists to insert genes from one organism into the genome of another organism. The resulting organism, with functioning foreign genes as well as its own, is known as a transgenic organism. Scientists have developed transgenic bacteria, animals and plants, including a number of transgenic plants, such as rice, tomatoes, apples and corn, resistant to insect attack. We at Greengenes have recently developed a transgenic clover, resistant to larval butterflies/moths (Lepidopterans), some beetles (Coleopterans), and roundworms (Nematodes). These insect resistant plants were developed by inserting toxin producing genes from the bacterium, Bacillus thuringiensis, into the plant cells.

Bacillus thuringiensis (B.t.) is a spore forming bacterium which produces a number of endotoxins proven effective in controlling many plant pests.Topical sprays of B.t. have been successfully used for over 30 years in pest control. Extensive testing has shown the B.t. toxin to be harmless to non-targeted organisms, including humans. In target pests, the endotoxin is ingested and dissolves in the mid gut where it is digested into toxic polypeptides. The polypeptides complex with receptor sites on the cell membrane of cells lining the midgut. This causes an osmotic imbalance in the cells resulting in their death. The organism stops feeding and eventually dies. There has been little widespread resistance to B.t.-based bioinsecticides apparently because there are over 42 varieties of toxins produced by 14 different genes.

The transgenic clover produced by Greengene was the result of standard genetic engineering techniques used in the production of other transgenic plants, including varieties of corn and potatoes. Below is basic outline of the procedure used at Greengene:

1. The genes that produce the B.t. endotoxins are isolated using recombinant DNA techniques.

   A) Plasmids are fragmented by using restriction enzymes.

   B) B.t. chromosomes are cut with restriction enzymes and pieces spliced into plasmids with a ligase.

   C) Plasmids with the B.t. DNA fragments are transformed into the bacterium E. coli.

   D) Bacteria containing the desired toxin-producing genes are isolated and cloned.

2. Identified B.t. gene-containing plasmids are transformed into special bacteria, Agrobacterium. Agrobacterium naturally transfers DNA segments into chromosomes of the plant cells.

3. The plant cells infected with the desired B.t. genes are then isolated, cloned and grown in a culture solution. These initial plants are propagated and tested for expression of the B.t. genes.

All the strains of transgenic clover have been tested for lethality to Coleopterans (beetles), Lepidopterans and Nematodes. The results were compared with lethality of non-engineered parent plants of the transgenic clover. The results are striking; mortality rates for beetles and lepidopterans approach 100% and for nematodes near 85%. Those insect larva not killed within the first four days of testing were severely stunted compared to larva feeding on the control tissue. Surviving nematodes showed no morphological effects of the toxin (Fig. 1).

The mortality rates compare favorably with the results attained with conventional chemical spraying without the risks of releasing potentially dangerous chemicals into the environment. Reduction in the use of externally applied pesticides is sound from an economic standpoint and also reduces health risks to farm workers associated with the spraying of chemical pesticides.

We have also conducted resistance studies and after six generations we have noticed only a slight drop in mortality in the third generation (Fig. 2). The reduction then stabilized and remained the same for the next three generations.

BIOMASS PRODUCTION OF TRANSGENIC CLOVER

Prepared By

Dr. Hamilton Burr, Botanist

Growth rate of the Greengene transgenic clover clearly exceeds that of other commercially available strains. Study plots consisting of four one- hectare sections were located at the Greengene experimental station. Alternate sections were planted with the Greengene clover and 'Good Luck' (the most widely sold strain of feed grade clover). One meter square samples were taken on a weekly basis to determine dry weight biomass. All factors were closely controlled to insure similar biotic and abiotic influences (Fig. 3).

Setting:

A table at the town's most expensive restaurant. You arrived about 10 minutes ago and the president of the company, Bob Kesham, arrived only minutes before. Mr. John Spa, owner of Lakewood Farms, is coming through the door and is headed for your table. Lakewood Farms raises show horses and most of the feed for the horses.

Mr. Bob Kesham: Well, here he is now. John, I'd like you to meet __________ __________________ (your name), our newest addition to the sales department.... quite a biologist, too.

Mr. John Spa: Nice to meet you. You must like working for Bob here, he's a great fellow. I've done business with him for over ten years.

You: ___________________________________________________________________________

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Mr. John Spa: Well, while we're waiting tell me how you guys invented these insect eating plants.

You:

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Mr. John Spa: Not quite science fiction, but sure close. It sure would be nice to cut back on pesticides, but do those little mutant clovers really grow to be ten times normal size like Bob says? That would sure increase profits. You:

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Mr. John Spa: Is it true that these plants can really kill any insect that tries to eat it or is this another one of Bob's tall tales?

You:

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Mr. John Spa: You know, about six years ago we started using a new kind of pesticide spray and everything went fine that first year -- killed every one of those pesky little bugs. The next year, great! but the third year we noticed that some of the bugs were surviving the spraying and we had a little damage, but nothing major. Last year they were drinking that poison like it was "sody pop". That's when ol' Bob came out and tested the bugs. "Resistant to the toxin", that's what you said, right Bob?

Mr. Bob Kesham: Sure did. That's when I suggested our transgenic clover.

Mr. John Spa: Then why can't the same thing happen with these plants?

You:

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Mr. John Spa: OK! OK! OK! Just one more thing. How safe are these plants for the farm workers? I don't want to get a whole lot of static from the union.

You:

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Mr. John Spa: Well I've got to do some thinking. I'll let you know by Friday. See you, Bob. You'll hear from me.

Mr. Bob Kesham: Good talking to you again, it's always a pleasure. Just let us know what you decide and we'll do a good job for you.

You:

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