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The Scientific Method What is it? The scientific method is the experimental testing of a hypothesis formulated after the systematic, objective collection of data. A scientist who studies our immune system phrased this idea very well: I now appreciate how much I learn by being wrong. I can change my mind when confronted with a rational argument, without the need to have the change appear to be purely semantic or to hope it will pass unnoticed. What must it be like to be a priest, general, bureaucrat, lawyer, medicine person, or politician who is never permitted to be wrong? No wonder they learn so slowly. I am grateful to be in a profession where the realization of being wrong is equivalent to an increase in knowledge. -Melvin Cohn. Annual Review of Immunology 12, 2 (1994) The scientific method is often divided into steps. This is helpful for putting the method into context, but keep in mind that the key element of the scientific method is testing the hypothesis. In other words, can you prove that you are wrong? 1. 2. 3. 4. 5. 6. 7. Observe the situation Ask a question Turn that question into a testable hypothesis Predict the outcome of your experiment Perform your experiment Analyze the results Evaluate your hypothesis Evaluating science Mistakes made in applying the scientific method to real-world problems can result in unsupported, or even incorrect, conclusions. An example of a scientific conclusions based on insufficient scientific method recently occurred in the field of breast cancer research. Breast cancer is caused when normal cells change and produce a tumor. In a study published in the New England Journal of Medicine (336, 1269 (1997)), scientists from Norway studied the incidence of breast cancer in 25,624 women. Participants gave details about their height, weight, diets, and exercise habits. The results showed an over all reduction of breast cancer by 37% for women who exercise regularly. Women who are lean and exercise at least 4 hrs per week showed the lowest incidence. Image courtesy of WebPath Many newspapers picked up on the data with headlines stating that exercise prevents cancer, but an editorial in the same journal by Dr. Anne McTiernan put the results in proper perspective (ibid. p. 1311). She points out that women who exercise regularly have higher levels of education and income, smoke less, drink less alcohol, and consume fewer calories and less fat. She states that establishing a casual pathway between reduced breast cancer and physical activity will require exploration for biologic mechanisms and confirmation with clinical experiments. Her final conclusion states the situation very well. Should a woman exercise and will it prevent breast cancer? I recommend a resounding YES to the first question. Regular physical activity in women reduces overall mortality and the incidence of coronary heart disease, diabetes mellitus, stroke, osteoporosis, obesity, and disability, and it also lessens the impact of arthritis and cognitive decline. With respect to whether exercise reduces the risk of breast cancer, too many questions 1 remain for women and their doctors to make informed decisions on whether, how and how much to exercise. -Dr. Anne McTiernan, New England Journal of Medicine (336, 1311 (1997)) This example illustrates many of the problems associated with the science of biology and medicine. ACTIVITIES 1.- Study a serie of world maps and compare the differences between countries related with some parameters relate with the health. Write a short composition about it. 2.- We need a healthy food as the documentary “supersize Me” shows. State or research what do the students in our High School eat during the brake. Wonder around the playground during the braketime and write your observations, later, state a report with your conclusions. 3.- Analysing the composition of foods. 4.- Calculating the daily energy requirements. Page 91, 3 of Eso, Santillana. 5.- Evaluating your diet. “” 6.-Scandals about transplants, (Taken from the wipipedia). (To debate about them) In 2008, California transplant surgeon Hootan Roozrokh was charged with dependent adult abuse for prescribing what prosecutors said were excessive doses of morphine and sedatives to hasten the death of a man with adrenal leukodystrophy and irreversible brain damage, in order to procure his organs for transplant.[45] The case brought against Roozrokh was the first criminal case against a transplant surgeon in the US, and resulted in his acquittal. At California's Emanuel Medical Center, neurologist Narges Pazouki, MD, said an organprocurement organization representative pressed her to declare a patient brain-dead before the appropriate tests had been done.[40] She refused. In September 1999, eBay blocked an auction for “one functional human kidney” which had reached a highest bid of $5.7 million. Under United States federal laws, eBay was obligated to dismiss the auction for the selling of human organs which is punishable by up to five years in prison and a $50,000 fine.[46] On June 27, 2008, Indonesian, Sulaiman Damanik, 26, pleaded guilty in Singapore court for sale of his kidney to CK Tang's executive chair, Mr Tang Wee Sung, 55, for 150 million rupiah (S$ 22,200). The 2 Transplant Ethics Committee must approve living donor kidney transplants. Organ trading is banned in Singapore and in many other countries to prevent the exploitation of "poor and socially disadvantaged donors who are unable to make informed choices and suffer potential medical risks." Toni, 27, the other accused, donated a kidney to an Indonesian patient in March, alleging he was the patient's adopted son, and was paid 186 million rupiah (20,200 US). Upon sentence, both would suffer each, 12 months in jail or 10,000 Singapore dollars (7,300 US) fine.[ 7.- Activitie about Asthma: Asthma is an inflammatory disease of the lungs in which the patient’s airways become temporarily constricted, making breathing difficult. Although the condition was virtually unknown in the early 1900s, today it affects 20.3 million people in the United States and kills 5,000 of them per year. From 1980 to 1994, the prevalence of asthma increased 75%. Asthma rates in children under the age of 5 have increased more than 160% in the same time period. The exact reason for the increased prevalence of asthma is not known, but the following observations offer clues: Asthma "attacks" can be triggered by the common cold, exercise, cold air, emotional stress, viral infections, aspirin, industrial air pollutants, smoking, obesity, ozone, and nitrogen dioxide. Having two parents with asthma increases a child's chances of having the disease. Approximately 40% of children who have asthmatic parents also develop asthma. The disease tends to cluster in England and English-speaking countries. Exposure of children to dust mites, cats, cockroaches, molds, and mildews increases their chances of getting the disease. Children who are exposed to common parasites seem to have a better-developed immune system than children not exposed to parasites. These exposed children do not suffer asthma as frequently. Exposure to occupational agents such as dyes may trigger the onset of adult asthma. Poverty seems to be a substantial risk factor for the development of asthma. Questions 1. 2. How have the factors that trigger an asthma attack changed since the 1900s? As a researcher, how would you put together the clues given here to explain the emerging epidemic? Why do you think Europeans—and especially the English—are most affected by asthma? Assigments 1. 2. 3. 4. 5. 6. Research the relationship between asthma and allergies. Describe how various agents trigger asthma or an asthma attack. Find out where in the world asthma is least prevalent, and hypothesize why this is so. Define the word "epidemiology." How do researchers use epidemiology to study a disease such as asthma? Research the CDC (Centers for Disease Control and Prevention), which is the epidemiological institution in the United States. Write a two-page paper on the different health problems the organization studies. When might common citizens need to use information from the CDC? Visit the CDC website on asthma at http://www.cdc.gov/asthma/default.htm and research the National Asthma Control Program. Find the name and address of agencies in your state that the CDC has funded to fight asthma. Contact one of those agencies to see what is being done in your area. References Asher, M.I., et al. 1998. Worldwide variation in the prevalence of asthma symptoms: The international study of asthma and allergies in childhood. European Respiratory Journal 12:315–35. Doyle, Roger. 2000. Asthma worldwide. Scientific American, June, 30. http://www.mayoclinic.com http://www.aaaai.org 3 8.- Activitie about Medical Benefits of marijuana. In 2004, Montana became the eleventh state to approve the limited use of marijuana for medical purposes. Previously, voters in Alaska, Arizona, California, Colorado, Hawaii, Maine, Maryland, Nevada, Oregon, Vermont, and Washington had also approved the medical use of this drug . However, the use of marijuana as both a psychoactive substance and a medicine is not new. Marijuana comes from the dried leaves and flowering tops of the hemp plant, Cannabis. As far back as the mid-1800s, both French and English medical literature reported the use of marijuana for such ailments as "spastic conditions, headache, and labor pains." Hashish, the concentrated resin from the flower of the hemp plant, was readily available in U.S. pharmacies until the Harrison Act in 1914, which attempted to regulate the traffic of opium. Somehow, by implication, the use of marijuana also came under regulation, although it is neither an opiate nor a narcotic. Virtually all use of the drug declined until the 1960s, when it became a popular recreational drug for the generation known as the baby boomers. The medical use of marijuana revived during the 1970s. In 1975, a small study at the Sidney Farber Cancer Institute published an article revealing that tetrahydrocannabinol (THC), a substance found in marijuana, could effectively lessen the nausea and vomiting associated with chemotherapy. Marinol, a pill form of synthetic THC, has been available in the United States since 1986. THC is also used to treat glaucoma, multiple sclerosis, and weight loss associated with AIDS. However, few wide-range, controlled studies on humans have been reported. In 1990, the first Cannabis receptor gene was cloned at the National Institute of Mental Health. The gene was found in rat brain tissue. The amino acid sequence of the protein receptor was named CB-1. When THC binds to a CB-1 receptor, the drug's effects are produced. Scientific studies show that this system of cannabinoid (marijuana) receptors is involved in the coordination of movement, short-term memory, and regulation of mood and emotion. In 1993, yet another receptor for THC was found at the MRC Laboratory of Molecular Biology. This receptor was named CB-2 because it was not found in rat brains but in macrophages from the spleen. CB-2 receptors have since been found on immune cells as well. Questions 1. 2. 3. 4. How would a "controlled" study be conducted in terms of human medical treatment? Why do you think few controlled studies of the effects of THC on humans have been conducted? Does marijuana work on the spleen as well as the brain? Does marijuana have an effect on the immune system? Why might pharmaceutical companies want to make an artificial, oral form of THC available? Do you think the U.S. government should relax current regulations on the growth of the marijuana plant? Why or why not? Assignments 1. 2. 3. 4. Define “cannabinoid,” and identify what type of organic molecule THC is. Can THC easily travel through membranes to the interior of cells? Why or why not? Why do cannabinoids and THC require receptors to produce their effects? Review the meaning of "cloning" as applied to the Cannabis receptor gene. How do scientists clone genes? Investigate the advantages and disadvantages of inhaling marijuana smoke compared to taking THC in capsule form. Write up an appeal for a Senate hearing defending the growth, sale, and use of marijuana based on its medical value. Use the scientific evidence presented above to support your appeal. You are suspicious that the pharmaceutical companies have used their vast lobbying machines to gain governmental support to keep marijuana use and growth illegal. In other words, the drug companies would prefer to manufacture an artificial substance to sell for medical purposes. What would pharmaceutical companies have to gain if the government did keep the growth, sale, and use of marijuana illegal? References Friedman, Hermand, and Thomas W. Klein. 1999. Marijuana and immunity. Science & Medicine. March/April, pp. 12-21. 4 Klein, Thomas W.; Cathy Newton; and Herman Friedman. Cannabinoid receptors and immunity. Immunology Today. Vol. 19:373-381. Watson, Stanley J. Jr., and John A. Benson, Jr. 1999. Institute of Medicine Report on the Medical Use of Marijuana. Washington, National Academy Press. http://www.rxmarihuana.com http://www.nap.edu/books/0309071550/html http://www.ecureme.com/emyhealth/natural/n_marijuana.asp http://www.drugpolicy.org/marijuana/medical/ http://www.time.com/time/covers/1101021104/tgood3.html 9.- Caffeine and the brain Ashley is about to take the final exam for her general biology class. She’s spent many long hours studying, and feels a little tired. On her way to class, she stops by the student center to grab a large cup of coffee. She hopes that the caffeine in the coffee will help her do better on the exam. Chemically, caffeine (C8H10N4O2) is a purine with two oxygens and three methyl groups attached. When ingested, it is rapidly absorbed and soon reaches all cells of the body, including those of the brain, where it acts as a stimulant. Caffeine is found not only in coffee, but also in tea, chocolate, and many soft drinks, including popular “energy drinks.” In addition, caffeine is present in numerous over-the-counter medications, such as headache remedies, diet pills, diuretics, and alertness aids. How much can Ashley count on coffee to help her on the exam? Without a doubt, caffeine does fight fatigue, so it will likely help Ashley avoid dozing off during the exam (surely among a student’s worst nightmares). But will it help her recall the information she has learned? Most likely it will, because Ashley will be more alert. On the other hand, taking in too much caffeine could make her feel too “jittery” to think clearly. The mental effects of caffeine can be unpredictable. The rate at which an individual metabolizes caffeine varies by age and sex, and may be affected by the concurrent use of other drugs, such as alcohol and nicotine. Also, the amount of caffeine an individual ingests in a cup of coffee can vary quite a bit depending on the size of the cup, the variety of coffee beans, and the brewing method. How does caffeine exert its effects on the brain? Much of caffeine’s stimulant action is thought to be due to its blockade of adenosine receptors located in the membranes of the brain’s neurons. Adenosine is not considered a true neurotransmitter, such as acetylcholine, dopamine, or serotonin, because it is not stored and released at chemical synapses. However, adenosine is an important neuromodulator, a chemical that influences synaptic function. One of adenosine’s prominent roles in the central nervous system (CNS) is to promote sleep. When caffeine reaches the CNS, it binds to and blocks adenosine receptors, preventing adenosine from causing drowsiness. In 1999, Menahem Segal and Eduard Korkotian of the Department of Neurobiology at the Weizmann Institute in Israel reported an interesting discovery. They had removed the part of the brain called the hippocampus from rat embryos and kept the cells alive, cultured on microscope slides. The researchers then applied caffeine to the hippocampal cells and observed them under a microscope to see if there were any changes. Dosing the cells with caffeine caused a rise in cellular calcium levels—a phenomenon known to be related to learning and memory. Even more interesting was the observation that repeated caffeine exposure caused some of the spines on the neuronal dendrites to grow longer! Dendritic spines are protrusions that may be important in influencing the number of synaptic connections a neuron can receive. Thus, caffeine may influence learning and memory by inducing changes in the brain’s “wiring” at the microscopic level. Questions 1. Ashley is standing in line at the coffee counter in the student center. What advice would you offer regarding her impending use of caffeine? Explain your answer. 5 2. Track your caffeine intake for one day. Be aware that caffeine may be present in unexpected dietary sources, including certain herbal supplements and teas. Check food, beverage, and medicine labels for caffeine content. Did you take in more or less caffeine than you would have predicted? Assignments 1. 2. 3. 4. 5. Review how cells form adenosine. (Hint: What does “ATP” stand for?) Research recent work on adenosine and its role in sleep. What regions of the brain are involved in sleep? Review the role membrane receptors play at a chemical synapse. How do they produce their effects? What role do dendrites play in transmission at chemical synapses? Research caffeine’s effects on the body. What does it mean when we say that caffeine is a CNS stimulant drug? What are some other CNS stimulant drugs besides caffeine? Are they equally accessible and socially acceptable? Why or why not? Does caffeine affect other organ systems besides the nervous system? Consider the cardiovascular, respiratory, and urinary systems in particular. Besides the “caffeine jitters,” are there any adverse effects associated with ingesting caffeine? If so, what are they? Is there any evidence to support health benefits from the use of caffeine-containing beverages such as coffee and tea? If so, what? Find out how much caffeine is present in a typical cup of coffee. How does this compare to the amount in a cup of tea or a can of cola? Research the role of the hippocampus in learning and memory. Explain why Segal and Korkotian chose this part of the brain for their study of the effects of caffeine. References http://web.sfn.org/content/Publications/BrainBriefings/adenosine.html http://web.sfn.org/content/Publications/BrainFacts/index.html http://web.sfn.org/content/Publications/BrainFacts/index.html http://health.howstuffworks.com/caffeine.htm http://health.howstuffworks.com/sleep.htm http://my.webmd.com/content/article/46/1826_50681.htm http://my.webmd.com/content/article/17/1671_52718.htm http://my.webmd.com/content/article/11/1671_50418.htm http://my.webmd.com/content/article/11/1671_50455.htm http://www.weizmann.ac.il/neurobiology/labs/segal/segal.html http://health.howstuffworks.com/framed.htm?parent=sleep.htm&url=http://www.sciencenews.org/sn_arc97/5_24_97/fob2.htm Weinberg, B. A., and B. K. Bealer. 2001. The world of caffeine: The science and culture of the world’s most popular drug. Routledge. 6 7