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MUR ournalJVolume 8 Spr ing 2003


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Table of ContentsVolume 8 Spring 2003

MIT Science News in Review3 Architecture 3 Biological Sciences 7 Brain/Cognitive Sciences 7 Electrical Engineering and Computer Sciences 8 Materials Sciences 8 Mechanical Engineering 9 Physics

World Science News in Review11 Archaeology 11 Biological Sciences 13 Chemistry 14 Earth, Atmosphere, and Planetary Sciences 14 Materials Sciences 15 Mathematics 16 Physics

FeaturesAadel Chaudhuri

17 Factory Health Services: An Innovative Method ofProviding Health Care in Bangladesh

Kelley Rivoire 21 The Growing Threat to Research: Scientific Misconduct

W. Victoria Lee 27 Cloning: Where We Have Been and Where We

Will Go from Here

Farhan Merali 35 Stem Cell Research: The Debate that

Has Divided America

ReportsCameron Sadegh and Ronald P. Schreck

39 The Spectroscopic Determination of Aqueous SulfiteUsing Ellman’s Reagent

Oneil Bhalala 45 Detection of Cotinine in Blood Plasma by HPLC MS/MS

Alexander C. De Feo 51 Probing Solar Panel Design Systems

Angela S. Bassa 55 Environmental Accounting: Project Financing and

Strategic Interactions

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UNDERGRADUATE RESEARCH JOURNAL

Volume 8, Spring 2003

Editors in ChiefIssel Anne LimMartin Kurtev

Editorial BoardSid PuramIzzat JarudiShaudi HosseiniMin Zhang

Associate EditorsSamantha BrennerAadel ChaudhuriLouise GiamTian HeSherry KanMatthew KwanW. Victoria LeeFarhan MeraliBrett MitchellLakshmi NambiarLili PengLynn PunnooseKelley RivoireCameron SadeghMonica SircarVivek Venkatachalam

Research ContributorsAngela BassaOneil BhalalaAlexander de FeoCameron Sadegh

Advertising ManagerPatrick Stansbury

AdvisorLes Perelman

Design/ProductionLynn HorskyJocelyn HumelsineHeidi Wormser

For our online edition, visit:http://web.mit.edu/murj/www

Welcome to the Spring 2003 issue of the MIT Undergraduate

Research Journal. In this eighth issue of MURJ, we present

diverse research reports in fields ranging from biological

engineering to environmental accounting, in hopes that our readers will

delve into the world of the scientific unknown. Within these pages we

also include features examining topics such as stem cell research,

cloning, factory health services, and scientific misconduct.

Our reports and features, as always, are written in such a manner as

to make them accessible to all members of the MIT community. We hope

that this interdisciplinary journal will provide an opportunity for our

readers to learn of research in disciplines other than their own. We also

hope that this journal will serve as a forum for debate and discovery

through which our readers may explore arguments relating to the inter-

section of scientific research and public policy.

We extend our profound gratitude to those who have made this goal

possible. We acknowledge the assistance of our advisor, Dean Les

Perelman, the professors who reviewed our work, and the continued

support of The Tech. We would also like to thank Dean Larry Benedict,

the many academic departments at MIT, and the UROP and RLSLP

offices for their financial support of MURJ this semester.

If what you read piques your interest, we invite your input for the

Fall 2003 issue. Consider either joining the MURJ team or to submitting

reports on your own research. Submissions will be due by the middle of

September 2003. Please e-mail [email protected] or visit our Web

site, http://web.mit.edu/murj/www, if you have any questions or com-

ments.

Sincerely,

The Editors of MURJ

Massachusetts Institute of Technology77 Massachusetts AvenueCambridge, MA 02139

Cover photo ©University of Wisconson Board of Regents

PSB 03-02-0124 TP

[ A r c h i t e c t u r e ]

Building Energy-Efficient Homes BringsTogether Experts from MIT and China

Researchers from MIT’s Building Technology Programand Chinese universities are working with developing

companies to construct more comfortable yet energy-effi-cient living spaces for the Chinese population. The boom-ing economy and the energy consumption in China haveincreased people’s demand for more Western luxuries.Energy-efficient buildings have thus become the center ofattention. The researchers have demonstrated that a tradi-tional Chinese approach with additional building tech-niques can significantly improve the existing modernbuildings. Although the recently erected buildings in Chinaare mostly Western-style high-rises, the researchers haveshown that low-rise buildings allowing more access tocommunal green space can also be energy-efficient.However, insufficient financial resources and a lack ofworkers who are familiar with the new technology havepresented major setbacks. Lack of incentive to conserveenergy also exacerbates the energy crisis. But theresearchers on both sides are continuing their work todevelop energy-efficient designs and to make the technol-ogy accessible to the Chinese builders.

In the meantime, demonstrations of the designs and thetechnology studies are being prepared in three major Chi-nese cities. Among the researchers are Leon R. Glicksman,Director of the Building Technology Program and a profes-sor of building technology and mechanical engineering.

—W. Lee

[ B i o l o g i c a l S c i e n c e s ]

Of Mice and Men

The International Mouse Genome Sequencing Con-sortium recently produced a high-quality, publicly

available draft of the mouse genome. An effort manned byscientists at twenty-seven institutions in six countries,including the Whitehead Institute/MIT Center for GenomeResearch, produced the genome in less than two years. Thisdraft can now becompared with therecently sequencedhuman genome togain insight about thefunctioning of genes,likely improving theunderstanding andtreatment of diseases.

Already, interesting comparisons have been drawnbetween the mouse and human genome. Ninety percent ofthe mouse genome was found to have a correspondingregion in the human genome. Only 5 percent, however, hadexact groups of DNA bases that matched with the humangenome. This 5 percent is thought to represent importantfunctions such as protein-coding genes.

The mouse genome was determined to have fewerrepeated sequences of DNA than the human genome,making it 14 percent shorter in length. These repeatedsequences are not considered significant; they indicate thatthe mice have less extraneous DNA than humans do.Scientists also found a similar number of protein-codinggenes to that of humans: approximately 30,000.

The sequencing of the mouse opens many doors for fur-ther genetic research. The mouse can now be utilized as amodel for the study of a wide range of diseases. The newsequencing will also allow scientists to study the relation-ships between genes rather than only looking at each geneindependently.

The consortium now plans to produce a final, nearly100-percent-accurate sequence of the mouse genome.Other organisms such as the chimpanzee, cow, and honeybee will then be sequenced for further comparison betweenthe genomes of different species. —K. Rivoire

MIT Science News in Review Volume 8, 2003 3MURJ

MIT Science News in Review

Airflow analysis is an effective technique for guiding the design ofnew communities in China. Scheme (a) is a design prepared by aChinese architectural firm. Airflow analysis performed by MITresearchers showed that scheme (b) could provide the same livingspace while better blocking winter winds from the north, increasingpassive solar heating in winter and enhancing cross ventilation fromsoutherly winds in summer.

MIT Science News in Review4 Volume 8, 2003MURJ


High-Carbohydrate Dietary SupplementHelps with Weight Loss

MIT researchers recently reported that a high-carbohy-drate dietary supplement can help people who expe-

rience weight gain while taking antidepressants. Thesupplement includes a high-carbohydrate drink developedat MIT.

Even though the supplement altered serotonin levels inthe body, there was no change in effectiveness of the anti-depressant. The supplement also had a positive effect onnonmedicated obese individuals: During the twelve-weekstudy, every participant lost twelve to twenty-six pounds.

People who take antidepressant medication, whichincreases serotonin activity level in the brain, usually expe-rience weight gain by overeating sweet and starchy foods.The antidepressants block a serotonin receptor that alsoregulates protein and carbohydrate intake, which results infeelings of satiation. Because the receptors are involved inthe antidepressants’ therapeutic effect, the antidepressantsusually cause weight gain.

By giving participants a carbohydrate-rich beveragetwice a day, the researchers wanted to see whether increas-ing serotonin in the brain could reverse the obesity causedby antidepressants without affecting its therapeutic effects.

They found that obese individuals who had taken psy-chotropic drugs, such as antidepressants, were able to losejust as much weight as nonmedicated obese individuals.The treatment successfully increased serotonin levels with-out diminishing the therapeutic effects of the drug.

Co-author Judith J. Wurtman, visiting scientist at MIT’sClinical Research Center (CRC), examined the role of car-bohydrates in the brain and their role in weight loss. Sheshowed that carbohydrate craving is associated with sero-tonin-linked changes in mood and that woman with pre-menstrual syndrome (PMS) sometimes overeat carbohy-drates and gain weight. She also thought that overeatingincreases brain serotonin, which diminishes feelings ofdepression and anger.

The study, supported by a grant from the Center forBrain Sciences and Metabolism Charitable Trust, wasconducted at the TRIAD Weight Management Center atMcLean Hospital in Belmont, Mass. —M. Kwan

New Breast Cancer Treatment Enters FinalStage of Clinical Trials

The U.S. Food and Drug Administration (FDA) has givenMIT researchers approval to begin the final stage of

clinical trials for testing an innovative breast cancer treat-ment using microwave radiation. The randomized clinicaltrials include the participation of nearly 220 women with

early-stage breast cancer and began in October 2002, thefirst day of Breast Cancer Awareness Month.

The technology is based on radar research invented byDr. Alan J. Fenn, a senior staff member at the SensorSystems Division of the MIT Lincoln Laboratory. Fenndetermined that the focused microwave technology previ-ously studied for missile detection could possibly treat can-cer cells. The clinical trials, based on Fenn’s research, focusmicrowave radiation externally on the breast, heating andkilling internal tumor cells, prior to lumpectomy and radi-ation therapy.

The randomized clinical testing is expected to finish byFebruary 2004 and will be conducted at various hospitalsincluding Columbia Hospital at the University of Oklahoma(OU), Harbor-UCLA Medical Center at the Martin LutherUniversity in Halle, Germany, and the ComprehensiveBreast Center in Coral Springs, Fla. Additional sites haveapplied for Institutional Review Board approval. Past stud-ies of microwave heat therapy have been promising. Earlyresults from a previous phase II clinical trial showed sig-nificant tumor cell death in a majority of the patients priorto lumpectomy, which led to the FDA’s approval to beginthe final phase of clinical testing.

Dr. Robert A. Gardner, a breast surgeon at ColumbiaHospital’s Center for Breast Care in West Palm Beach, Fla.,and Dr. Hernan I. Vargas of Harbor-UCLA Medical Centerpresented the results of the phase II clinical trials at the2002 American Society of Breast Surgeons meeting inApril and in the May 2002 issue of the Annals of SurgicalOncology. The study is funded and led by CelsionCorporation, which has developed the clinical thermother-apy system and exclusively licenses the focused microwavethermotherapy technology from MIT. —C. Sadegh

Links: http://web.mit.edu/newsoffice/nr/2002/cancer.htmlhttp://web.mit.edu/newsoffice/tt/2002/may08/breastcancer.html

Engineering Adult Stem Cells

Dr. James L. Sherley, principal investigator at the MITBiotechnology Process Engineering Center, has engi-

neered cells that behave like adult stem cells. Because adultstem cells are capable of generating new tissue, scientistshope to use them for organ replacement for their ability todevelop into skin, red blood cells, and neurons.

Adult stem cells are unique in that they have immortalDNA; they pass on duplicates of their original DNA andthus avoid replication errors. However, these cells areextremely difficult to find because they appear in the bodyas normal cells.

The cells that Dr. Sherley created have the capability to divide like adult stem cells given a certain culture


What’s missing from MIT’sUndergraduate Research Journal?

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NEWS • FEATURES • REPORTS Submissions due September 2003 • [email protected]


condition or to divide like cancer cells when treated in adifferent environment. The cells that he made can revealbiological mechanisms that answer questions about cancerand aging. The next step is to understand the proteinsinvolved in stem cell division. —L. Giam

[ B r a i n / C o g n i t i v e S c i e n c e s ]

Slow-Wave Sleep Dreams in Rats Found toReplay Waking Tasks

MIT researchers reported in the December 19 issue ofNeuron that rats dream about their waking activities

during slow-wave sleep, not just REM sleep. The sleepcycle both in rats and humans is composed mostly of slow-wave sleep followed by sleep characterized by rapid eyemovements (REM).

This study was a follow-up to a landmark 2001 study,also conducted by Associate Professor of Brain andCognitive Sciences Matthew A. Wilson in MIT’s PicowerCenter for Learning and Memory, showing that animalshave complex dreams and are able to retain and recall longsequences of events, particularly during REM sleep.

In this study, Wilson and his co-author, graduate stu-dent Albert K. Lee, monitored the firing activity of collec-tions of neurons in the rat’s hippocampus as the rat ran ona simple track for a food reward. For each lap, differentindividual cells fired at different times, with each succes-sive lap producing the same sequence of firings.Subsequently, in the slow-wave sleep directly followingthe activity, the rats were found to replay in short, high-speed bursts brief memory sequences corresponding to sin-gle laps. A 4-second lap on the track would only last100–200 milliseconds in slow-wave sleep. This contrastsREM replay, which is played back in real-time.

Another difference was that the dreams found duringslow-wave sleep only seemed to occur in the period ofsleep immediately following the behavior. REM dreamsthat reactivate tasks, on the other hand, are detectable aslong as 24 hours after the activity was performed, sug-gesting a more gradual reevaluation of older memories.

Contrastingly, researchers have postulated that slow-wavesleep replay is part of the initial storage of memory pro-cessing during sleep.

Additionally, researchers found that the rats replayedonly the stretches of running or attentive behavior, not theinactive periods of waiting in between. Thus, memory maynot be as continuous as we tend to think it is. The authorshave suggested that this study will be useful to create amodel of how long-term memories are formed. “This mayrelate to work in humans that suggests that amount ofslow-wave sleep early in the night, as well as the amountof REM sleep later in the night, is correlated with subse-quent enhancement of performance on learned tasks,” saidWilson. —S. Brenner

[ E l e c t r i c a l E n g i n e e r i n g a n dC o m p u t e r S c i e n c e s ]

Virtual Touch of Loved Ones Now a Reality

On May 23, 2002, MIT and University College Londonresearchers collaborated on the first touch exchange

across the Atlantic. The MIT team led by Mandayam A.Srinivasan worked with software specialists at UCL totransform a commercially available, MIT-developedrobotic arm, the PHANToM, into a haptic (touch) machine.

UCL researchers worked on the software that runs therobotic arm and the networking communication involvedin the transatlantic experiment, while MIT engineersworked on adapting the PHANToM into a machine that cantranslate touch by exerting exact pressure onto the opera-tor’s fingers.

In the May experiment, the two participants on bothcontinents were simultaneously placed in a virtual roomon a computer. In the room was a box and pointers thatrepresented the participant’s location in the room. The par-ticipants then tried to work together to raise the box; eachparticipant’s individual motion affected the box’s positionand could be felt by the other.

But the experiment didn’t run perfectly. Due to networkdelay and Internet traffic there was a delay in the trans-mission of the real-time touch. The delay was only around150–200 milliseconds, but compared to a signal sent fromthe hand to the brain, which takes only 30 milliseconds,this delay is a major obstacle. As a result, the arm opera-tors had a hard time keeping in sync with each other.


The next step for the UCL and MIT collaboration is tominimize the traffic delay to less than 30 milliseconds andto develop a better touch algorithm for the promisingmachine. Just think, one day surgeons could operate onpatients thousands of miles away, and 8.02 students couldfeel the intranuclear forces inside atoms as part of their lab.

—T. He

Ultra-Thin Quantum-Dot LED Raises

Prospects of Better Flat-Panel Displays

MIT researchers have developed a quantum-dot-organic, light-emitting device (QD-OLED) using a

novel combinationof thin organicmaterials and high-performing inor-ganic nanocrystals.The research raisesthe possibility ofmanufacturing thesematerials to createthinner and brighterflat-panel electronicscreens to replacetoday’s popular liq-uid crystal displays(LCDs).

Quantum dots, orartificial atoms, arenanome t e r - s c a l eboxes that selectivelyhold or release elec-

trons. While LCDs must be lit from behind, quantum dotsemit their own light. Depending on their size, the dots canbe “tuned” to emit any color in the rainbow as well as inthe infrared and ultraviolet ranges. And the colors of lightthey produce are much more saturated than that of othersources. The QD-OLEDs created in the MIT study also havea twenty-fivefold improvement in luminescent power effi-ciency over previous QD-OLEDs.

This latest MIT QD-OLED contains only a single layer ofquantum dots sandwiched between two thin, organic films,whereas previous QD-OLEDs used ten to twenty layers. Theresearchers have demonstrated organized assemblies largerthan 1-square centimeter, and the same principle could beused to make bigger components.

Moungi G. Bawendi, professor of chemistry, andVladimir Bulovic, assistant professor of electrical engineer-ing and computer science, led the interdisciplinaryresearch on the hybrid optoelectronic structure for the QD-LED. The collaborative effort is supported by MIT’s Center

for Materials Science and Engineering (CMSE). Bulovic isalso affiliated with the Research Laboratory of Electronics.

In addition to being used for extraordinarily thin andbright flat-panel displays, the QD-OLEDs may also be usedin a variety of other applications: to calibrate wavelengthsfor scientific purposes, to generate wavelengths visibleonly to robot eyes, or for the miniaturization of scientificequipment. The MIT team’s method of combining organicand inorganic materials may pave the way for new tech-nologies and enhance understanding of the physics ofthese materials. Further understanding of the materialproperties of QD-OLEDs will open doors for the productionof flat-panel displays that are stable, simple to produce,high-resolution, and highly efficient.

The work, reported in the December 19 issue of Nature,is funded by the National Science Foundation’s MaterialsResearch Science and Engineering Center Program andUniversal Display Corp. —C. Sadegh

[ M a t e r i a l s S c i e n c e s ]

Lithium Ion Phosphate Batteries Take Charge

In a major breakthrough published late last year in NatureMaterials magazine, MIT Department of Materials

Science and Engineering (DMSE) researchers claimed to beone step closer to making inexpensive, safe, and recharge-able batteries a reality. The researchers proposed using cellsmade of a new lithium ion phosphate–based material,which is seen as the possible next generation for energycells.

For years, industries have been trying to find a new,safer, and cheaper rechargeable material for batteries thatis less chemically reactive than the current lithium-cobalt-oxygen cells. Such cells can overheat easily, causing pro-ducers to limit the size of such cells.

Researchers at the University of Texas-Austin first sug-gested looking into lithium ion phosphate as a potentialsubstitute. Although cheap and environmentally safe,however, the chemical has a very low electrical conductiv-ity so at the time it was impractical as a rechargeable cell.

Professor Yet-ming Chiang led the MIT research teamthat added some metals and developed a special process fortreating the compound, which increased its conductivityten-millionfold! As a result, the lithium ion phosphate bat-tery is quickly becoming a reality. Impending tests willallow scientists to scrutinize and verify the results, soexpect to see lithium ion phosphate batteries take chargeof the rechargeable battery market. —T. He


[ M e c h a n i c a l E n g i n e e r i n g ]

Helicopter Maneuvers Autonomously

MIT researchers developed a pilot model X-Cell 60 hel-icopter, which executed a complicated maneuver

autonomously: The split-S maneuver allowed it to reverseits direction in a limited amount of space.

In the past, complicated maneuvers required a skilledpilot, but now these advances will be applied to unmannedaircraft for use in national defense. The autonomous heli-copters can be used for collecting military intelligence andimagery when it is unsafe for other aircraft to enter the air-zone. These autonomous aircrafts are small, inexpensive,and easy to control, which make them suitable for militaryoperations. — L. Giam

[ P h y s i c s ]

Nuclear Fusion Envisaged as Major Sourceof Energy in Future

Nuclear fusion involves the joining of lighter elementsunder high pressure to produce heavier elements as

well as tremendous amounts of energy. Magnets dictate thebehavior of the plasma in which the fusion takes

place. Scientists at MIT’sPlasma Science and FusionCenter and the Departmentof Nuclear Engineeringhave created a cylindrical,150-ton magnet for theInternational Thermonu-clear Experimental Reactor(ITER). This magnet willform part of a bigger mag-net weighing 925 tons,which in turn will be partof a total magnet systemweighing some 10,000

tons. In recent years, a number of tests have been con-ducted on the 150-ton magnet, located in Japan. This mag-net can produce a magnetic field of 13 Tesla and can storeenergy of 640 megajoules at a current of 46,000 amperes;for reference, the current handled by typical householdwiring is around 20 amperes. It was also found that themagnet can be operated in pulses, and it can be brought upto 13 Tesla and back down in only a few seconds. Suitableranges for the magnetic field, temperature, and currentdensity continue to be defined as support for the initia-tive. This past September, a Department of Energy panelrecommended that the United States rejoin the ITER col-laboration along with Japan, Germany, and Russia. TheU.S. team involved in developing ITER is composed ofabout 20 researchers from the MIT Plasma Science andFusion Center, and the project is funded primarily througha multiyear grant from the Department of Energy to MIT.

— M. Sircar

First Rapid Detection Sheds New Light on“Dark” Gamma-Ray Bursts

The first X-ray image of a rare fast-fading “dark”gamma-ray burst was taken by the MIT-built High

Energy Transient Explorer (HETE) satellite, the first satel-lite dedicated to spotting gamma-ray bursts.

A signal for the birth of a new black hole, a gamma-rayburst is the most powerful type of explosion known, sec-ond only to the Big Bang in total energy release. “Dark”gamma-ray bursts are aptly named because prior to therecent X-ray image, they have had no detectable opticalafterglow. Other bursts have bright afterglows that lingerfor days or weeks, likely caused by the explosion’s shockwaves ramming into and heating gas in the interstellarmedium.

The orbiting HETE, which alerts scientists to gamma-ray bursts, spotted one on December 11, 2002, originatingsix billion light years away, and relayed its location to


observatories worldwide in 22 seconds. The ground-basedRaptor optical telescope, operated by the Los AlamosNational Laboratory, was the first on the scene, observingthe afterglow at 65 seconds. The afterglow was extremelyfaint after two hours and would have been missed andlabeled “dark” if not for HETE’s rapid detection.

“Perhaps none of these bursts is truly dark, providedthat we catch them fast enough,” said George Ricker, a sen-ior research scientist at MIT’s Center for Space Research,who leads the international team that built and operatesNASA’s HETE satellite. Some theorists have suggested thatdark bursts have no detectable afterglow because they areburied in thick dust and gas, which blocks the afterglow’slight from reaching Earth.

According to Ricker, the December 11 observation ofburst implies the opposite: “The burst may have occurredin a region with hardly any surrounding gas and dust, thusthe shock waves had little material to smash into to create

a prolonged bright afterglow.” The rapidly fading after-glow, in this case, may support the binary merger theory ofshort bursts. In the billions of years that old binary sys-tems, with a combination of neutron stars or black holes,took to form, they drifted outward to less dense regions of a galaxy. Thus, when they merge to form a black hole, there is little surrounding material to make a longafterglow.

HETE was built by MIT as a mission of opportunityunder the NASA Explorer Program. It is on an extendedmission until 2004. The HETE program is a collaborationbetween MIT; NASA; Los Alamos National Laboratory,New Mexico; France’s Centre National d’Etudes Spatiales(CNES), Centre d’Etude Spatiale des Rayonnements (CESR),and Ecole Nationale Superieure del’Aeronautique et del’Espace (Sup’Aero); and Japan’s Institute of Physical andChemical Research (RIKEN). The science team includesresearchers from the University of California (Berkeley andSanta Cruz) and the University of Chicago, as well as fromBrazil, India, and Italy. —C. Sadegh

For more information: HETE: http://space.mit.edu/HETE

Additional images and GRB021004 information:http://space.mit.edu/HETE/Bursts/GRB021211http://web.mit.edu/newsoffice/nr/2002/darkburst.html

World Science News in Review Volume 8, 2003 11MURJ

[ A r c h a e o l o g y ]

Fossils Found in China Could Be Earliest inEast Asia

In 1958, in thecounty of Liu-

jiang in southernChina, farm work-ers discovered sev-eral human bones,including a skull,while digging in acave. Because itresembles to-datefossils in Japan,scientists estimatethis Homo sapiensskull to be from20,000 to 30,000years old. However, according to an article in theDecember 2002 Journal of Human Evolution, the fossilsmight be much older than that.

The fossils probably came from sediment dating to110,000 to 139,000 years ago, estimates a team led bygeologist Guanjun Shen of Nanjing (China) NormalUniversity. He also said the fossil discoveries probablycame from either the cave deposit dating from 68,000years ago or one dating to more than 153,000 years ago.

The team concluded that if any of the estimates are true,“the Liujiang [specimen] is revealed as one of the earliestmodern humans in East Asia.” This would roughly coincidewith the earliest known fossil dates in Africa and theMiddle East.

Yet evidence of the skull may complicate the human-origins debate, creating problems for the out-of-Africatheory of human evolution—that modern humanity startedin Africa between 100,000 and 200,000 years ago and then spread elsewhere. If the fossils found in Liujiang aredated accurately, supporters of the out-of-Africa theorywill have to find even older African Homo sapiens fossils.Another possibility for the adherents would be showingthat modern humans migrated very quickly from Africa toEastern Asia.

The scientific accounts from 1959 and 1965 of the fos-sil discoveries led to the new determination of the likelyburial site of the fossils. Shen and his coworkers mappedvarious soil deposits in the cave and used uranium decaymethods to work out the age of the crystallized limestone.According to Shen, other sites’ uranium samples have beenanalyzed and support the idea of ancient human origin insouthern China. Teeth that were discovered in two othercaves have been estimated to be at least 94,000 years old,said Shen.

On the other hand, some anthropologists doubt the esti-mated age of the Liujiang skull. It’s still uncertain how theskull got in the cave and where it was originally buried,remarked Christopher B. Stringer of the Natural HistoryMuseum in London.

Milford H. Wolpoff of the University of Michigan in AnnArbor agreed. “I’d love for the Liujiang skull to be as oldas Shen proposes, but we’ll never know for sure withoutdirectly dating the specimen.”

Shen said he hopes he can get an agreement with theChinese officials to date the specimen directly. —M. Kwan

[ B i o l o g i c a l S c i e n c e s ]

Nervous System Drug Shows Promise forMale “Pill”

Researchers have discovered a new oral drug, known asN-butyldeoxynojirimycin or NB-DNJ, that prevents

sperm cells from developing normally in mice. Until now,the only FDA-approved male birth control methods havebeen condom use and vasectomy, though a practical birthcontrol pill for men has long been sought after.

Researchers at the University of Oxford in Englandstumbled upon NB-DNJ’s potential birth control abilitieswhile exploring its use in treating diseases of the nervoussystem, during which male mice exposed to the drug

World Science News in Review

World Science News in Review12 Volume 8, 2003MURJ

became temporarily infertile. “So, we started a study tolook at what this drug was doing to male-mouse fertility,”said Frances M. Platt.

She and her colleagues at Oxford spiked the mouse feedwith varying doses of NB-DNJ and found that the femalescoupled with the control males produced normal litters, butthe treated males, even those who were given low doses ofthe drug, could not impregnate the females.

After further investigation, the researchers found thatNB-DNJ damages the sperm mitochondria and nuclei, thusimpairing the sperm’s swimming ability. The drug also pre-vents the formation of a cap, called the acrosome, on thesperm’s head, which normally enables the cell to penetratean egg.

The effects of the drug are reversible: Sperm of the micetreated with any dose of the drug for up to six months(which is a significant portion of the average two-yearlifespan of a mouse) were able to regain fertility by threeweeks after researchers stopped administering the drug.

Though a version of NB-DNJ has been approved by theEuropean Union for the treatment of Gaucher’s disease, thedrug’s use as a male birth control pill will require furtherscrutiny of its known toxic effects, such as those affectingsperm internal parts.

NB-DNJ appears to be less risky than some other drugsbeing studied for their male birth control potential, statedRonald Swerdloff of the Harbor-UCLA Research andEducation Institute in Torrance, Cal. He added that NB-DNJ“is a long way from a final, tested product, but it seems tobe an exciting lead.” —F. Merali

Noroviruses Cause Many ShipboardOutbreaks

According to recent reports from the Centers for DiseaseControl and Prevention (CDC) in Atlanta, cruise ships’

sanitation practices do not reliably wipe out viruses thatcause diarrhea outbreaks. Although cruises are cleanertoday than in past years, stopping the viral scourge onships will require even more rigorous measures, includingcompletely scrubbing down affected vessels.

The viruses, calledNoroviruses, pervadeeasily through casualcontact and can last formany days in the openair. The viruses often setoff epidemics of diar-rhea and extreme vomit-ing in cramped spaceson ships, militarycamps, and overcrowded

areas. They also cause approximately 23 million illnessesin the United States every year.

In 2002, at least 23 outbreaks occurred on cruise ships,in which more than 2,000 people were sick on five shipsoperated by four different companies.

On three of the five ships, these outbreaks affected sev-eral consecutive voyages because the virus probably sur-vived onboard between scheduled cruises. The preventativemeasures used on the voyages ranged from scrubbing pub-lic areas to sanitizing poker chips. In one particular case,the virus struck Holland America’s Amsterdam on four dif-ferent voyages. The outbreak started to subside only whenthe ship was docked for ten days and underwent intensecleaning.

In 2002, the wave of viral outbreaks contrasted from anoverall downward trend in stomach infections on the ship.In the 1990s, inspectors reported improper handling ofwater on 55 percent of inspections and food on 62 percent.They also reported a 95 percent equipment maintenanceand dishwashing violation rate. Despite these scores, halfof all cruise lines received passing scores. —M. Kwan

Life under Thick Ice in Antarctica LakeIn 1996, the collab-

oration of Russian andBritish glaciologistshelped discover LakeVostok, one of thelargest and deepestlakes, below 4,000meters of ice inAntarctica. Three yearslater, the scientistswere able to map outthe dimensions of theterritory with advancedtechnology. The big-gest mystery that hasbewildered all scien-tists is whether lifeexists in the lake. In 1999, curiosity drove the NationalScience Foundation to initiate an airborne survey of thelake and later an actual expedition to the lake at the VostokStation. It is highly suspected that ancient microbes or lifeforms do dwell in the lake. Cylindrical ice samples havebeen pulled from the lake for examination, but the researchwas stopped in 1998 for fear of contamination. Yet fromwhat has already been extracted, scientists have found avariety of living organisms—from bacteria to algae—three-fourths of the way down the lake, and current researchersare working to find viable microbes even deeper into the


lake. Although the physical conditions at this level makethe chances of finding organisms slim, scientists expect tofind microbe existence. The study of Lake Vostok and itspossible inhabitants has also attracted NASA, whichplanned to use the lake as a practice site for an expeditionto Europa. —W. Lee

Medical Procedure Eases Pain in BoneCancer Patients

For most cancer patients, experiencing physical pain isperhaps one of the most difficult consequences of the

disease. However, scientists at a recent meeting of theRadiological Society of North America (RSNA) announceda new pain-reducing treatment for bone cancer patients:radiofrequency ablation (RFA).

Utilizing computer tomography (CT) and ultrasoundguidance, RFA transmits high-frequency electrical currentsthrough a needle to the targeted tissue, thus heating anddestroying a large part of the tumor. Since the process suc-cessfully eliminates a large portion of the lesion, RFA sub-stantially reduces the pain in patients. According to Dr.Matthew Callstrom of the Mayo Clinic in Rochester,Minnesota, RFA has been previously used to treat othercancers such as those in the lung and kidney tissue. Theablation procedure has recently been used to treat patientswith metastatic bone cancer. Metastatic bone cancer resultsfrom the spreading of a malignant tumor via the blood-stream to one or more bones near the body’s center, suchas the hip or spine.

Callstrom, who works for one of the nine research cen-ters in the United States and Europe participating in thestudy, confirmed that RFA has successfully eliminated painin patients with metastatic bone disease. Approximately 81percent of the patients attained “a high degree of painrelief,” and nearly half experienced “complete eliminationof pain” six months following treatment. Furthermore, thepatients reported a decrease in physical pain from 7 to 2on the standardized Brief Pain Inventory (BPI), a 10-pointscale with 10 being the highest level of pain. Taking these

results into account, Callstrom optimistically expressedthat RFA offers a safe and effective treatment option toreduce severe pain so as to “improve the quality of thepatients’ lives.”

However, what RFA can’t do is successfully eliminatepain in patients whose cancer has spread throughout theirentire skeletons—it can only help lesions. Despite thisdrawback, the outlook for this new treatment is optimistic:The U.S. Food and Drug Administration (FDA) officiallyapproved the use of RFA for treatment in December 2002.

—L. Peng

[ C h e m i s t r y ]

Findings of Acrylamide in Food May Leadto Health Risks

Those who love to eat French fries may be shocked tohear that what they are eating does not only contain

high fat content, but also contains acrylamide, an agentthat causes cancer in rats. According to the U.S. Food andDrug Administration (FDA) and Germany’s BavarianMinistry of Health, scientists have reported findings ofacrylamide in foods like French fries and gingerbread.

Acrylamide, a chemical substance used in the produc-tion of plastics, is created by high-temperature cookingmethods, especially in starches. The FDA has already initi-ated testing of acrylamide levels in starchy foods such asFrench fries and potato chips. The researchers first testedfor acrylamide concentration in French fries by bakingnumerous batches of different types of fries, from variouschain restaurants to various home-cooked (store-bought)types (see table). For restaurant-cooked fries, the acry-lamide concentration ranged from 117 ppb (parts per bil-lion) at KFC to 1030 ppb at Popeye’s. For home-cookedfries, Ore Ida brand registered at 616 ppb and Lamb WestonIsland Valley peaked at 1325 ppb.

RF ablation electrode with full deployment.


Acrylamide Concentrations (ppb) in French fries at different locations of chain restaurants

(FDA preliminary data)*

Batch Batch Batch Batch Batch Batch Batch1 2 3 4 5 6 7

Burger King 197 220 369

Checkers 257 407

Fuddruckers 346 452

KFC 117 162 270 313

McDonald’s 155 193 245 270 328 356 497

Popeye’s 301 484 610 1030

Wendy’s 157 169 254 260 302

Other foods that tested positive for acylamide concentra-tions were fish fillets (30 ppb), Pepperidge Farm CheddarGoldfish crackers (57 ppb), unbrewed Starbucks CoffeeColumbia Ground (175 ppb), Hershey’s Cocoa (909 ppb),and Lipton Soup & Dip Mix (1184 ppb).

What should one make of all these numbers? Eventhough these foods contain acrylamide, these results areonly preliminary and do not currently pose a major healthrisk to Americans, according to the FDA. More research isneeded to form a better conclusion about the potentialhealth risks that acrylamide might pose. —L. Peng

Source: Science News, “Acrylamide—from Spuds toGingerbread.”

[ E a r t h , A t m o s p h e r e , a n dP l a n e t a r y S c i e n c e s ]

Collisions Heat UpMars Climate andErosion Debate

Agroup of physics-drivenplanetary scientists pro-

pose that the mysterious ero-sion markings on Mars can beexplained by giant asteroidcollisions. The source of waterfor the massive amounts oferosion has remained a mys-tery for about thirty years.Climate models cannotexplain why the frozen planetwas warmer in its early years.However, a new physics-based approach is generatingvery convincing ideas andnumbers. The group calcu-lates that a relatively small

asteroid measuring 100 kilometers delivered about 4 x1,026 joules of energy to the planet.

The energy of the impact, combined with the deep layerof hot rock debris that covered the planet afterward,brought global surface temperatures to about 800 K. As aresult, the frozen planet thawed on its surface, at its polarice caps, and in its subsurface. A small 100-kilometerobject impact would result in 5 meters of water coveringthe planet for at least one year. A single 250-kilometerasteroid might have melted 50 meters of water for at leasta century. At least ten such objects collided with Mars,punctuating the planet’s cold, dry climate with episodes ofrain and erosion. According to the new research, the cumu-lative effect of the impacts can account for the massiveerosion.

Geologists point out that this new theory may explainthe earlier stages of erosion, but it cannot account for thelater valley formations and flooding of craters. The amountof erosion seems to have required kilometers of water,much more than the 50 meters produced by an impact. Toattract more supporters, the physics-driven planetary sci-entists need to develop a better model of the cumulativeeffects of impacts over time. —S. Kan

[ M a t e r i a l s S c i e n c e ]

Size Doesn’t Matter: Nanotubes Play BigRole in Strengthening Ceramics

University of California-Davis researchers, led byJoshua D. Kuntz and Amiya K. Mukherjee, have

managed to triple the fracture resistance of ceramics. Thescientists accomplished the feat by introducing smallamounts of carbon nanotubes into the mix of aluminacrystals to make a superstrong material.

Many researchers have tried to introduce carbon nan-otubes as the agent to eliminate the infamous brittleness ofceramics. The UC-Davis researchers strayed from tradi-tional approachesin two ways: thetype of nanotubesused and the tech-nique in harden-ing the ceramics.Previously, otherresearchers hadrelied on multi-walled rather thans i n g l e - w a l l e dnanotubes. Inaddition to using


the different type carbon nanotubes, the UC-Davis scien-tists also employed a low-temperature heating technique toharden the ceramic. They suspected that the previous sin-tering technique that operated at a higher temperaturedamaged many of the nanotubes.

The two changes caused a two-and-a-half-fold increasein fracture resistance compared to the old method. (Theprevious best had only decreased the brittleness of the alu-mina crystal ceramics by 24 percent.)

With this significant increase in shatter resistance, thefuture for the new ceramic material is boundless; it couldbe used for gears, industrial parts, and the like. But untilresearchers can find a way to lower the price of the expen-sive carbon nanotubes, the material will probably be usedonly in heavy industries, where production cost is not themain concern. —T. He

[ M a t h e m a t i c s ]

Mathematical Forays into MusicalCompositions

In Swedish composer Daniel Cummerow’s musical works,each musical fragment is determined by a mathematical

recipe through the use of a formula that links numericaldigits with musical notes. For example, the mathematicalconstant p, has an intricate, vaguely medieval correlate,whereas the decimal digits of the constant e, progress at arelentless, suspenseful pace.

A variety of different strategies are used by algorithmicmusic composers to get such interesting results. Onemethod involves converting prime numbers directly intotheir corresponding MIDI notes, at least up to 127, to get acuriously rising scale. The MIDI specification assigns anumber to each note on a keyboard (e.g., middle C is 60,C-sharp is 62, and so on, for a total of 128 tones) and isused in computer programs to represent musical notes.However, since there are an infinite number of primes, onecould continue by dividing each prime by a certain num-ber, then use just the remainder to assign the musicalvalue—an elegant use of modular arithmetic.

In some of his pi compositions, instead of mapping dig-its directly to their respective MIDI numbers, Cummerowconstructs the piece by assigning each digit from 1 to 8 toa note in a specific scale; 0 signals a pause; and 9 meanseither a pause or the repetition of the previous tone.

In yet another experiment, Cummerow uses a uniquemusical alphabet invented by French composer OlivierMessiaen, which extended the German names of the notesA to H by giving each letter of the alphabet its own pitch,octave, and note value. Cummerow paired the first 255

digits of pi and applied the above formula to those that fellbelow twenty-six. For the rest, he followed a differentrecipe.

Other techniques have yielded fascinating musicalpieces featuring the Fibonacci sequence, Pascal’s triangle,and intriguing structures that have been associated withchaotic dynamics, such as the Sierpinski triangle andLorenz’s butterfly.

Many other musicians have also delved into mathemat-ical music, exploring ideas such as the fractal notion ofself-similarity in which each component is a miniaturereplica of the overall structure.

Only rarely do composers enter the seemingly forbid-ding world of mathematicians and their abstruse concerns,though the recent frenzy of forays into the field may bejust the start of a beautiful marriage between mathematicsand music. —F. Merali

Intriguing mathematical music compositions of the fol-lowing artists can be found on the World Wide Web:Cummerow (http://www.geocities.com/Vienna/9349/);Chris K. Caldwell (http://www.utm.edu/research/primes/programs/music/listen/); and José Oscar Marques(http://www.midiworld.com/c/jmarques.htm).

A Step Closer to Finishing Pi

At the University of TokyoInformation Technology,

Yasumasa Kanada and hiscolleagues have been able tocalculate the value of p up to1,241,100,000,000 digits, thelast of which is 5. Thanks tothe improvements in thealgorithm to calculate π and602 hours of a Hitachi SR8000 computer’s time, the groupwas able to set this new record, surpassing their previousworld record in 1999 of 206,158,430,000 digits. Asexpected, initial statistical analyses indicate that each ofthe digits from 0 to 9 appears with the same fre-quency. Questions about the distribution and apparent ran-domness of π’s digits still remain: Do all digits appear infi-nitely often? At some point beyond the range of currentcomputations of π’s value, do its decimal digits revert to astring constrained of only digits of say 1 and 0 (or anyother number(s))? —M. Sircar


Survival of the Fittest Shoelaces

Computer scientist John Halton and Australian mathe-matician Burkard Polster recently investigated a prob-

lem to which even a small child can relate: how best to laceone’s shoes.

Though there exist more than 50,000 possible lacingmethods for a shoe with as few as five eyelets per side, overtime, humans have gravitated toward several common lac-ing patterns. The three most common lacings are criss-cross, also termed “American,” European straight, and shoestore straight.

Through mathematical modeling of the shoelace as twoparallel lines, each with n points, Halton and Polster wereable to evaluate the strongest and shortest lacing patterns.Halton discovered that the zigzag American patternrequired the least amount of lace for a given number ofeyelets, with the European straight next, followed by shoe

store straight, pro-vided that at leastfour eyelets arepresent on eachside of the shoe.

Polster investi-gated less tradi-tional lacings aswell, finding thata bow tie lacing infact uses the least

lace of all. When considering which lacing is strongest,however, Polster concluded that a “dense” lacing such asthe American style that crosses back and forth betweensides of the shoe is best. —K. Rivoire

[ P h y s i c s ]

Antineutrino Transformations Detected on Earth

Recent evidence gath-ered by an interna-

tional group using anunderground detectorknown as the KamiokaLiquid Scintillator Anti-neutrino Detector, orKamLAND, in Toyama,Japan, indicates thatelectron antineutrinos morph into muon or tau anti-neutrinos on Earth, as they do on the sun. The researchgroup determined this by monitoring light flashes causedby electron antineutrinos generated by a few dozen nuclearpower reactors in Japan and Korea. KamLAND, set up in amine one kilometer below ground level, contained thou-sands of tons of organic liquids that emitted light whenstruck by these electron antineutrinos. Because only 63percent of the expected number of electron antineutrinos,and thus light flashes, were detected, physicists concludedthat the electron antineutrinos had oscillated into the twoother possible forms—muon and tau antineutrinos. Theseearthly findings closely follow observations of transforma-tions of neutrinos and antineutrinos emitted by thesun. Giorgio Gratta of Stanford, the lead scientist of thestudy and a KamLAND spokesman, concluded that oscilla-tions are “a property of neutrinos and not that of the sun.”

—M. Sircar

Features Volume 8, 2003 17MURJ

Factory HealthServices:

An Innovative Method ofProviding Health Care in Bangladesh

Aadel Chaudhuri

Health care in Bangladesh is a complicated issue. Despite having a land area of

merely 55,598 square miles, Bangladesh has a population of 133 million people,

making it the eighth most populated country in the world and the twelfth most

densely populated.1,2 Additionally, Bangladesh is one of the world’s twenty least-developed

nations.3 It comes as no surprise that due to the high population

density, coupled with the country’s low level of development,

health care for the masses is a major concern that is rather diffi-

cult to address.4,5 Funding agencies such as the United Nations

Children’s Fund (UNICEF), the United States Agency for

International Development (USAID), and the International Center

for Diarrheal Disease and Research in Bangladesh (ICDDRB) have

recognized this dilemma and have thus helped Bangladesh’s gov-

ernment and nongovernmental organizations (NGOs) provide

health care to the country’s great many people.4,5

Bangladesh’s Government Health SystemDespite the daunting challenges of providing health care,

Bangladesh’s government health system is very well-organized.

There are hospitals and clinics for every societal class. Big hospi-

tals cater to cities. More limited but still extensive clinics provide services to upazilas,

which are composed of a few districts. Small clinics cater to individual districts. Finally,

small doctor rooms or a couple of doctors take care of individual unions or villages. The

government provides its health services at very low costs, with the ultimate goal of pro-

viding free health care to all. Thus, the government attempts to target all members of the

population, even the poorest residents of the remotest villages.3

Aadel Chaudhuri

The World Bank is one of many organizations dedicated tohelping Bangladesh’s health program.

Features18 Volume 8, 2003MURJ Aadel Chaudhuri

Unfortunately, although planned well, thegovernment system has a number of draw-backs, one of which is inefficiency. Thereare simply too few doctors available to seethe patients that come in. This often forcespatients to wait exorbitant amounts of timebefore being able to see a doctor.6 Addition-ally, funds are often insufficient or misap-propriated, which results in half-constructedoperation theaters or unmanned examina-tion rooms in hospitals and clinics.6 As aresult of the government’s apparent ineffi-ciency in the health sector, only 30 percentof Bangladesh’s population utilizes the gov-ernment’s health services.7

A further disability of the government’ssystem is that it caters too specifically torural areas. The structured hospital/clinic

organization previously detailed seemsmostly to be designed with rural areas inmind.7,8 The lack of “remote areas” in large,dense cities such as Dhaka makes a tier sys-tem seem almost unnecessary because thelocation of the large city hospital may bejust as strategic as the site of the upazilahealth clinic, with both areas being equallydensely populated and publicly accessible.In short, the government’s health system hasnot been designed to specifically serve urbanareas, where the patient need is greatest.

Involvement of Nongovern-mental Organizations

Although the Bangladesh governmenthealth system’s weaknesses are unfortunate,NGOs have heroically stepped in to helpsupplement the ailing system. NGOs haveset up clinics in both urban and rural areasand have helped provide low-cost healthcare to the poor and the very poor.9,10 Therole of NGOs is especially apparent in urbanareas, where the government’s system is notable to sufficiently serve the large popula-tion.6,8,9 Additionally, NGOs are committed toteaching health awareness and providinghealth counseling.9,11,12 They have success-fully shown that they can improve people’shealth by suggesting lifestyle changes.9,10,11

The government does not provide and pro-mote such health awareness-related servicesas readily as the NGOs do.6,7,9 Thus, the NGOshave effectively filled in many gaps of thegovernment’s health system, supplementingand supporting it so as to provide more peo-ple with necessary health care.

Mary Stopes Factory HealthServices

Mary Stopes Clinical Society (MSCS), aBangladeshi NGO that is affiliated with MaryStopes International (based in the UnitedKingdom), has established clinics and miniclinics in Bangladesh’s major cities, focusingon reproductive health but also coveringgeneral health services.9,11,12 Other NGOs, suchas the Bangladesh Rural Advancement Com-mittee (BRAC) and the Bangladesh Associ-ation for Maternal and Neonatal Health(BAMANEH), are acting similarly as well asreaching into rural areas.10 Mary Stopes,however, is unique because it provides a

A Bangladeshi Slum Area.

A cook at work.

Features Volume 8, 2003 19MURJAadel Chaudhuri

service termed “factory health services,” inwhich Mary Stopes sets up and staffs a smallclinic room within a factory for the nominalfee of 12 Taka per worker per month ($0.21U.S. per worker per month). Factories thatchoose to purchase this service pay thisamount on behalf of the workers.9,11,12 Thebeauty of the service is that it allows a pre-viously neglected group (factory workers) toget quality, convenient health care.

The vast majority of participating facto-ries produce garments.9,12 Because of theirlong hours, garment factory workers havevery little time to go to hospitals or clinics,especially to government-run facilities,where a patient might spend an entire daywaiting to see a doctor.6 However, the MaryStopes factory clinic doctor schedules regu-lar checkups for each worker in addition tocheckups available on a walk-in basis.9,11

Workers from participating factories are alsoeligible to receive free services at any one ofthe many Mary Stopes clinics availablethroughout Bangladesh’s major cities andcan thus get zero-cost referrals to one ofthese larger clinics.9 Interviews with factoryworkers have shown that they very muchappreciate the health benefits received fromsuch a setup.13,14

Economic Viability of FactoryHealth Services

Mary Stopes is able to provide these serv-ices to factories in a manner that is eco-nomically viable to its own organization. Bycharging 10 Taka/worker/month prior to2002, it has been able to recover 70 percentof its expenses. In addition, it receives fund-ing from the Department for InternationalDevelopment (DFID), a United Kingdomfunding organization, although this fundingis scheduled to expire in 2004. As a result,Mary Stopes increased its customer costs to12 Taka/worker/month in 2002, with thegoal of 100 percent cost recovery, and thuseventual self-sustainability of the program.12

Appeal of Health Services toFactory Administration

Most factory administrators wholeheart-edly support the Mary Stopes health servicesprovided in their factories. They reason thatthe service cost is very low yet reaps huge

economic benefits.13,14 Workers are more pro-ductive because they take fewer days ofleave due to debilitating sickness. Workersalso feel that their work environment is safeand homey, existing for their benefit.13,14 Asa result, their on-the-job happiness remainsgreater. These two factors lead to incredibleattendance and productivity, as exhibited bythe 1 percent absentee rate observed in agarments factory run by director FaruqHussein, who contracts Mary Stopes factoryhealth services.13 From a less economic per-spective, many factory directors feel ahumanitarian obligation to help their poorworkers by providing them with health serv-ices. Thus, another garment factory director,M. Rahman Razu, began by hiring a doctorand buying medicine with his own moneyand that of fellow administrators in 1995.

Hundreds of villagers come to see the village doctor.

Dr. Shaukat Jahan, an American volunteer, sees patients in a poor village.

Later, when he heard of Mary Stopes factoryhealth services, his factory switched to it,like many other factory directors have for itslow cost, quality, and reputation.14

Factory administrators also enjoy MaryStopes because it helps protect the factoriesfrom quotas that are often imposed due tochild labor or poor working conditions.Since Mary Stopes is an internationalorganization, it is highly trusted by interna-tional buyers of Bangladeshi-made gar-ments.12,13 Additionally, by keeping thoroughhealth records of all workers, Mary Stopescan detail the health status of each andevery worker.11 Thus, if need be, such recordscan dispel myths of poor health and unsan-itary factory conditions. Given the preva-lence of quotas against Bangladeshi gar-ment factories, especially after September11, 2001, many factory directors are consid-ering health services such as Mary Stopeswith renewed interest.12

Appeal of Factory HealthServices to Government

In addition to factory workers andadministrators, the government approves ofMary Stopes factory health services. TheBangladesh commerce minister, Amir Kusir,looks very favorably on these services,12

mainly for assistance with quotas. Asdescribed previously, quotas are oftenimposed on factories when working condi-tions in the factories are suboptimal. Thelarge-scale effect of quotas and the resultingfactory closures and production reductionsare a general shrinking of the Bangladeshieconomy. Like any other government,Bangladesh wants to prevent such negativeeffects on its already frail economy.12 This,along with its general efforts to make avail-able better health services that reach agreater portion of the population, makes thegovernment a strong proponent of MaryStopes factory health services.

ConclusionMary Stopes factory health services, a

revolutionary system that benefits workersof a leading Bangladesh industry (gar-ments), factory administrators, the govern-ment, and the economy, is successfully serv-ing 73,000 factory workers.9,15,16 Health and

efficiency in participating factories isimproving dramatically for the minisculecost of approximately $2.50/worker/year.9,12

The success of the Mary Stopes programwith garment factories has helped it expandto other types of industry, such as fish-pro-cessing factories in the city of Khulna.12 Likegarment workers, these workers do not haveready access to health care. Customers, how-ever, do not pressure factories and govern-ments with quotas in any industry quite asmuch as they do the heavily export-orientedgarments industry.12 Thus, the need foroffering health services in other factoriesseems less pressing to factory owners. Thisnotwithstanding, most factory owners arequickly learning that proper health care candrastically improve worker productivity andlead to greater profits. In addition, otherBangladeshi NGOs such as Momota, whichprovides health services to some factories inthe city of Chittagong, are collaboratingwith Mary Stopes.12 Such cooperationamong NGOs as well as increased awarenessamong factory workers, administrators, andgovernment officials will allow factoryhealth services to multiply.

Features20 Volume 8, 2003MURJ Aadel Chaudhuri

References

1. The U.S. Agency forInternational Development(http://www.usaid.gov).

2. Geographic.org (http://www.geographic.org).

3. Bangladesh Ministry of Health.Overview of Health andPopulation Sector Program. 1sted. Dhaka: 1998.

4. ICDDR,B: Center for Health andPopulation Research. AnnualReport 2001. Dhaka: 2001.

5. United Nations Children’s Fund.The State of the World’sChildren. New York: 2002.

6. Kajal, Hasan Mahbub. PersonalInterview. 9 Jan. 2003.

7. Shafinar, Shumana. PersonalInterview. 9 Jan. 2003.

8. Dr. Modabbir. PersonalInterview. 18 Jan. 2003.

9. Rahman, Ayusu. PersonalInterview. 7 Jan. 2003.

10. BRAC (http://www.brac.net).

11. Choudhury, Samina. PersonalInterview. 8 Jan. 2003.

12. Choudhury, Wahiduzzaman.Personal Interview. 8 Jan. 2003.

13. Hussein, Faruq. PersonalInterview. 8 Jan. 2003.

14. Razu, M. Rahman. PersonalInterview. 7 Jan. 2003.

15. Waddell, Steve. “Strategies forCivilizing the Market: Four CivilSociety Approaches.” The ODPractitioner (2002).

16. Waddell, Steve. “Six SocietalLearning Concepts for a NewEra of Engagement.” Reflections3 (2002).

A Mary Stopes slum volunteer describes how to carefor a pregnant woman.

Inside the Mary Stopes Dhaka-2 Base Clinic.


The Growing Threatto Research:

Scientific Misconduct

Kelley Rivoire

In the past year, the news that a Bell Labs scientist performing Nobel-quality experi-

ments engaged in scientific fraud sent shock waves through the scientific community.

The work of 32-year-old physicist Jan Henrik Schön came under suspicion after he

published two papers with an identical figure, including noise patterns. The papers indi-

cated the figures to be measurements of two different phenomena from two different pieces

of equipment. One of the papers was published in Science, the other in Nature. Schön had

previously been publishing at the astonishing rate of roughly one paper

per eight days, gathering extraordinary results. “He discovered every-

thing in condensed matter physics in the last sixty years” in organic

materials, said Lydia Sohn of Princeton University. Later, an independ-

ent committee led by Stanford physics professor Malcolm Beasley

investigated Schön’s work and found Schön guilty of data falsification

in sixteen separate instances. 1,2,3 His many co-authors were absolved of

all charges, and nearly all of the questioned papers have since been

retracted.3,4,5 With the reproduction of his results having become a

“minor industry,” others’ careers are also at stake.2 Perhaps most impor-

tantly, however, Schön’s high-profile case has led to further discussion

of the troubling but prevalent issue of scientific fraud and misconduct

and the measures necessary to prevent and punish them.

Schön Not AloneSchön’s case is hardly an isolated example. In the past five years, instances of question-

able scientific research ethics at universities and other research institutions have been

numerous. In 2002, physicist Victor Nirov was fired from Lawrence Berkeley National

Laboratories (LNBL) in California after his computer analysis of the discovery of elements

116 and 118 was found to be fabricated. Nirov’s co-authors, like Schön’s, were cleared of

any misconduct. The news of his data fabrication also cast a shadow over the discovery of

Kelley Rivoire

“Nobody arrives

at fraud as the

first thing they

ever do....They

got there by doing

little things and

getting away

with it.”

Features22 Volume 8, 2003MURJ Kelley Rivoire

elements 111 and 112, to whose discoverieshe had contributed. The discovery of thoseelements, however, has not been refuted.6

Following are still more examples ofAmerican scientists engaging in scientific

misconduct. BiochemistRobert Liburdy, also atLNBL, had won morethan $3.3 million infederal money for hisresearch on a possibleconnection betweenelectromagnetic fieldsand cancer. In 1999, hewas found to have fab-ricated data. Heretracted his findingsand accepted as pun-ishment the loss of fed-eral funding for threeyears.7 In 2001, Harvardmedical researcherEvan Dreyer washanded a ten-year banfrom receiving NationalInstitutes of Health(NIH) grant fundingbecause of false datareported in papers andgrant applications.8

William Simmons, formerly of theUniversity of Texas Southwestern MedicalCenter, left his job in 1998 to seek employ-ment elsewhere, only to be called back thefollowing year because the postdoctoralfellow who replaced him could not dupli-cate his results. When redoing his experi-ments to prove their validity, he was spot-ted inserting material into the vials thatwould alter the results in his favor. His for-mer collaborator, Derry Roopenian, notedthat people in his laboratory “wasted a lotof time and money trying to reproduceresults that weren’t real to begin with,” acommon complaint of scientists who havebased their research on others’ fraudulentresults.9

Yet recent scientific misconduct has notbeen confined to the United States. Over-seas, allegations of misconduct are alsocommon. In the field of medicine, NeilMarshall of Britain’s General MedicalCouncil (GMC) notes that “we have seenenough complaints to warrant actionamong the whole profession.” In Britain, anarticle in the Journal of Bone and JointSurgery contained photographs stolen fromanother author’s work. When the author ofthe article submitted a replacement picture,claiming it as a correction, the new picturewas then found to be a combination of twophotographs, only one of them from theauthor’s own work.10 In China, a translatorof a 1998 American anthropology textbookdecided to publish it in Chinese as well—under his own name.11 Researchers fromtwo German universities, Professor Fried-helm Hermann of the University of Ulmand Professor Marion Brach of the Univer-sity of Lübeck, were accused in 1997 ofmanipulating data in more than thirty bio-medical papers. Hermann was later sus-pended from the university, and Brach losther position entirely.12 In Poland, authorMarek Wroñski, while writing a book aboutscientific misconduct in 1997, discoveredscientist Andrzej Jendryczko was guilty ofmore than twenty acts of direct plagiarism.Wroñski claims that an “old guys’ network”in Poland protects scientists like Jen-dryczko from prosecution or defamation. Infact, Wroñski was told that questioningJendryczko’s work “was going to destroyPolish science.”13 Clearly, the problem ofscientific misconduct is widespread.

What Exactly Is Scientific Misconduct?

One difficulty in prosecuting scientificfraud often is the lack of a precise defini-tion of the term. The United States hasadopted a definition produced by the Officeof Science and Technology Policy (OSTP),which states: “Research misconduct is

[Derry Roopenian]

noted that people in his

laboratory “wasted a lot

of time and money

trying to reproduce

results that weren’t real

to begin with,” a

common complaint of

scientists who have

based their research on

others’ fraudulent

results.

Features Volume 8, 2003 23MURJKelley Rivoire

defined as fabrication, falsification, or pla-giarism in proposing, performing, orreviewing research, or in reporting researchresults.” This definition, though quite nar-row, goes on to pinpoint specific acts thatviolate good scientific practice, clearing upsome confusion in the previous definitionsprovided by the National ScienceFoundation (NSF) and Health and HumanServices (HHS).14

Abroad, some countries also struggle todefine misconduct, while others merelyfocus on how to prevent it. Graeme Catto,vice principal of the University ofAberdeen, claims “there is little value inlengthy discussions about a definition ofscientific misconduct as done in the U.S. Abetter approach seems to be an emphasison implementing good research practiceguidelines.”15 The Wellcome Trust, thelargest biomedical charity in the UnitedKingdom, created its own definition, whichrecipients of its grants must follow. Thedefinition largely mirrors that of the OSTPbut is slightly broader. It is criticized fornot providing adequate protection forwhistleblowers, but this is partially due tothe United Kingdom’s strict libel laws.Wellcome hopes that its actions “canbecome the template for guidelines in otherfields of science.”16

What Measures Have Been Taken to Combat Scientific Fraud?

In 1989, the United States created theOffice of Research Integrity (ORI), a branchof the Department of Health and HumanServices, to investigate claims of fraud.17

The ORI receives more than 1,500 com-plaints a year and often requires ten totwelve months to investigate an allegation,resulting in what Barbara Mishkin, aWashington attorney who specializes indealing with scientific misconduct, calls a“black-hole effect.”18 More often than not,only lower-level researchers are convicted.

The ORI claims a 92 percent success rate,but it has failed several times in high-pro-file cases. These failures have includedfalse accusations, as in the case of a publi-cation in Cell by former MIT professorThereza Imanishi-Kari, who was supportedby Nobel Prize-winner David Baltimore,now president of the California Institute ofTechnology. Imanishi-Kari was accused offalsifying important data, and her caserequired a full decade to resolve before shewas cleared of wrongdoing.18,19 Nearly alluniversities and research institutions alsoconduct internal reviews whenever a sci-entist comes under fire. In addition, in the United States, the NIH now requiresmedical schools to provide training inresearch ethics, and some universities haveexpanded such classes to all graduatestudents.20,21

In Europe and Asia, methods of dealingwith scientific misconduct have also beendiscussed at length. The ChineseAssociation of Science and Technology hasissued a number of rules for science jour-nalists, requesting journals not to publishmultiple submissions or papers of poorquality and forbiddingthe publication of papersdealing with confidentialinformation.22 In theearly 1990s, Denmarkcreated the Committee ofScientific Dishonesty(CSD), which has consid-erable power over scien-tists because the Danishgovernment funds morethan 90 percent of thecountry’s research.15

Rather than develop anational body to investi-gate misconduct, other countries have pre-ferred to simply tie funding by nationalorganizations to compliance with miscon-duct procedures. In Australia, the NationalHealth and Medical Research Council has

“Many researchers

think that a high IQ

goes hand in hand with

high moral values.”

…[T]his correlation “is,

of course, absolute

nonsense.”

Features24 Volume 8, 2003MURJ Kelley Rivoire

guidelines to which grant-receivingresearch institutions must adhere.23 TheGerman Deutsche Forschungsgemeinschaft,the nation’s largest source of grants forbasic research, has proposed ways to pro-mote research ethics and deny funding tothose who do not comply. Among their rec-ommendations are requiring the storage oforiginal research data for ten years, offer-ing protection to whistleblowers, and elim-inating the practice of honorary co-author-ship. In addition, prominent research insti-tutions all over the world, such as the MaxPlanck Society in Germany, have beenpreparing their own procedures for dealingwith misconduct. President Hubert Marklhopes “universities or other institutionswill look closely at our new rules and per-haps use them as an example.”12 The onlyquestion that remains is how successful

these agencies can be atfinding and punishingmisconduct, as well aspreventing it.

Why WouldAnyone CommitScientific Fraud?

In the words ofStephen Lock, a pasteditor of the BritishMedical Journal, “Manyresearchers think that ahigh IQ goes hand inhand with high moralvalues.” However, hecontinues, this correla-tion “is, of course,absolute nonsense.”15

The high-pressure envi-ronment that today’syoung researchers entercan often tempt them to

practice poor ethics. More than 1 percent ofscientists report direct knowledge of aninstance of misconduct.21 The “publish-or-perish” mindset of research institutions

that often base promotions on the volumeof research published can lead a youngresearcher astray, particularly in theextremely competitive and lucrative bio-medical industry.17,24 Because of the hugevolumes of research produced in a year,many also believe their questionable activ-ities will escape notice. Computer programscan make falsifying images such as X-rayseasy and indiscernible from genuine ones.10

A person who successfully passes off smalldata manipulations may attempt miscon-duct on a larger scale the next time.“Nobody arrives at fraud as the first thingthey ever do....They got there by doing lit-tle things and getting away with it. Callingthem in might stop people from going offthe deep end,” said Paul Friedman of theUniversity of California at San Diego.17 Inaddition, the system of tenure in universi-ties can lead to a “cloak of academic free-dom,” where a professor’s research may notundergo the necessary internal scrutinybefore publication, because professors“invariably resist intrusion from their fel-low departmental colleagues.”25 The envi-ronment of the research setting seems topush researchers to produce results at anycost.

Who Is to Blame?The issue of responsibility for fraudulent

papers has been much disputed recently.Those often mentioned as culpable includeco-authors, supervisors, and peer review-ers. With multiple authors from multiplecountries working in cross-disciplinaryfields, it is hard to hold each personaccountable for every piece of data oranalysis reported in a paper. In the Schöncase, when co-authors were questioned asto the particulars of data measurements,they professed to know little. Yet it can beargued that, since each receives part of thecredit for a successful paper, each shouldbear responsibility for the contents. AsDonald Kennedy of Science remarked, “If

The high-pressure

environment that

today’s young

researchers enter can

often tempt them to

practice poor ethics.

More than 1 percent

of scientists report

direct knowledge of

an instance

of misconduct.

Features Volume 8, 2003 25MURJKelley Rivoire

the benefits of authorship are enjoyedjointly and severally by all the authors,shouldn’t the liability be shared in the sameway?”20

Supervisors and management are alsooften blamed for failing to catch errorsbefore an article goes to press. While someaccountability for the results produced isexpected of a supervisor, it is unreasonableto expect 100 percent overseeing of a proj-ect. Where should the line be drawn?Finally, no one has ever claimed that thepeer review process is perfect. Unless anarticle is in their particular field ofresearch, reviewers are unlikely to questionthe validity of a paper. Furthermore, whilepeer review is an anonymous process, areviewer would be identified and subject tolegal action if he or she were to accuse anauthor of misconduct.25 As Donald Kennedyof Science wrote, “A clever laboratory cookcan invent data that are immune to vigilantreviewers and to any diagnostic test, saverepetition, the only proven scientific rem-edy.”20 However, increasingly complexresearch means that duplicating another’sexperiment to ascertain its validity can bephenomenally costly and time-consuming,if not downright impossible.

What To Do Next?The ORI has several plans in the making,

but all have been met with stiff resistance.Its funded proposal to investigate thosefound guilty of scientific misconduct tolearn about their mentalities and whatcaused them to behave in such a way washalted by the Office of Management andBudget (OMB) after the completion of thefirst stage.26 The ORI also proposed a Gallupsurvey to send to 3,000 researchers to fur-ther define standards of ethical researchconduct. The survey required the scientiststo answer questions dealing with impropercitation of articles and poor research super-vision, but this approach has been attackedas too broad. Critics also feel that the ORI

may be exceeding its limits to areas notincluded in the OSTP definition of scien-tific misconduct.26,27,28 The president of theFederation of American Societies forExperiment Biology (FASEB), StephenTeitelbaum, calls the survey “a terribleinstrument” and its questions “outra-geous.”26

In Britain, the General Medical Councilplans to specify both the standards forresearch practice and the punishments forthose who violate them.10 Individualresearchers have also developed their ownways to deal with issues of fraud, such aschecking the work of their associates morecarefully.21 Some supervisors even feelunable to reveal the identity of reagents toresearchers working in their laboratoriesfor fear of data falsification.15 The Instituteof Medicine has also suggested continua-tion and expansion of research ethicstraining, though Arthur Rubenstein of theUniversity of Pennsylvania, the panel’schair, anticipates “quite a lot of flak” onthe controversial idea. Rubenstein evengoes so far as to recommend ethical con-duct as a part of promotion considerationsand research ethics programs as a require-ment for university accreditation.27 Evenwith ethics courses, as Hubert Markl of theMax Planck Institute remarked, “Oneshouldn’t expect too much from formal

References

1. Service, Robert F. “Bell LabsFires Star Physicist Found Guiltyof Forging Data.” Science. 2002;298(5591): 30–31.

2. Hellemans, Alexander. “IdenticalGraphs Chart a DubiousPicture.” IEEE Spectrum. July2002: 20–21.

3. Hellemans, Alexander. “SchönShown the Door.” IEEESpectrum. Nov. 2002: 20–21.

4. Service, Robert F. “More of BellLabs Physicist’s PapersRetracted.” Science. 2003;299(5603): 31.

5. Kennedy, Donald. “Next Steps inSchön Affair.” Science. 2002;298(5593): 495.

6. Dalton, Rex. “California lab firesphysicist over retracted finding.”Nature. 2002; 418(6895): 261.

7. Vergano, Dan. “EMF ResearcherMade Up Data, ORI Says.”Science. 1999: 285(5424):23–25.

8. Ready, Tinker. “Stiff penalty forvision researcher guilty of scien-tific misconduct.” NatureMedicine. 2001; 7(1): 8.

9. Malakoff, David. “TexasScientist Admits FalsifyingResults.” Science. 2000;290(5490): 245–246.

10. Williams, Nigel. “Editors Call forMisconduct Watchdog.” Science1998; 280(5370): 1685–1686.

11. Yimin, Ding. “Beijing U. IssuesFirst-Ever Rules.” Science. 2002;296(5567): 448.

12. Koenig, Robert. “Panel ProposesWays to Combat Fraud.”Science. 1997; 278 (5346): 2049.

13. Marshall, Eliot. “MedlineSearches Turn Up Cases ofSuspected Plagiarism.” Science.1998; 279(5350): 473–474.

14. Kaiser, Jocelyn. “A MisconductDefinition That Finally Sticks?”Science. 1999; 286(5439): 391.

15. Hagmann, Michael. “EuropeStresses Prevention Rather ThanCure.” Science. 1999; 286(5448):2258–2259.

16. Koenig, Robert. “Wellcome RulesWiden the Net.” Science. 2001;293(5534): 1411.

“If the benefits

of authorship are

enjoyed jointly and

severally by all the

authors, shouldn’t the

liability be shared in

the same way?”

Kelley Rivoire Features26 Volume 8, 2003MURJ

[ethics] courses; there are tons of lawyerswho finish their formal legal training andstill go work for the mob.” Instead ofmerely endorsing ethical practices, “Youhave to teach it by example; the young sci-entists have to see it day in, day out,”stated Lord Kilpatrick of Kincraig, a pastpresident of the British General MedicalCouncil.15

As to the best way to prevent and pun-ish scientific misconduct, there seems to beno clear answer. Awareness of the problemalone is a huge step forward, but it is cer-tainly not enough. Perhaps the best sugges-tion comes from Paul Grant, of Nature—for

colleagues to educate each other abouttheir work, to the level that any co-authorcan defend another’s work. Not only doesthis reduce likelihood of fraud, it alsoallows scientists to interact with oneanother to expand “technical vitality.”30

Without any additional supervision foreach and every researcher, the problem ofscientific misconduct will not go away. Inthe words of Cai Decheng, former vicepresident of the Chinese Science andTechnology Guide, “There must be no com-promise over dishonesty and no cover-up.Taking pity will harm the cause of sci-ence.”16

17. Check, Erica. “Sitting in judg-ment.” Nature. 2002; 419(6905):332–333.

18. Friedley, Jock. “ScientificMisconduct: ORI’s Self-Assessment: A Batting Averageof .920?” Science. 1997;275(5304): 1255–60.

19. Blau, Stacey. “Panel Clears MITScientist of Fraud: Imanishi-Kari, Baltimore Vindicated.” TheTech. June 26, 1996: 1.

20. Kennedy, Donald. “MoreQuestions about ResearchMisconduct.” Science. 2002:297(5578): 13.

21. Marshall, Eliot. “How Prevalentis Fraud? That’s a Million-DollarQuestion.” Science. 2000;290(5497): 1662–1663.

22. Wang, Justin. “Chinese JournalsPledge Crackdown.” Science.1999; 283(5407): 1427.

23. Dayton, Leigh. “Australia ProbesKidney Researcher.” Science.2002; 296(5567): 449.

24. Bostanci, Adam. “Germany Getsin Step with ScientificMisconduct Rules.” Science.2002; 296(5574): 1778.

25. Grant, Paul M. “Scientific creditand crediblity.” NatureMaterials. 2002; 1(3): 139–141.

26. Holden, Constance. “PlannedMisconduct Survey Meets StiffResistance.” Science. 2002;298(5598): 1549.

27. Kaiser, Jocelyn. “U.S.Universities Urged to Do aBetter Job.” Science. 2002;297(5580): 321.

28. “Soft responses to misconduct.”Nature. 2002; 420(6913): 253.


Cloning: Where We Have Been and

Where We Will Go from Here

W. Victoria Lee

In The Devil’s Dictionary, American author and satirist Ambrose Bierce defines admira-

tion as “our polite recognition of another’s resemblance to ourselves.” Indeed, we all

love ourselves. There are a few others whom we can love to the same extent: our par-

ents, our significant others, and our children. But can we also love our clones, exact genetic

replicas of ourselves? That is, if we were ever to have clones. This seems like a very distant

possibility. However, we may not be too far away from a world where we coexist with our

clones.

At the dawn of 2003, Brigitte Boisselier, a French chemist, claimed the

birth of the first human clone nickednamed Baby Eve. Boisselier was also

busy running the first human cloning company, Clonaid, and serving as the

bishop of the Raelian religion, a new religious sect that believes aliens cre-

ated life on earth, that sexual freedom is important, and that life can be

eternalized through cloning technologies.1 She managed to shake the scien-

tific community, add fuel to the already burning cloning debate in the U.S.,

and stir a bigger wave of controversy among religious groups with her evi-

dence-deficient claim before the festive New Year.

Scientific technology has advanced rapidly over the past six years since

the birth of Dolly, the first cloned mammal, in 1996. The question used to

be: “Is it possible to clone?” The question today is: “To clone or not to

clone?” In just a few years, an array of different animals has been cloned as

the technology has continuously improved. We can now look back at what

we have accomplished and ask ourselves, “Where we will go from here?”

The Journey So FarDolly: The Cloned Sheep

The hype about cloning began with the birth of Dolly, the first cloned mammal born on July

5, 1996, at the Roslin Institute in Edinburgh, Scotland.2 The project was headed by Dr. Ian

W. Victoria Lee

Although not an

easy thing to do,

cloning has proved to

be contagious. Now

there are hundreds of

cloned animals,

including sheep,

goats, mice, pigs,

cows, cats, and, most

recently, rabbits.

Features28 Volume 8, 2003MURJ W. Victoria Lee

Wilmut and his team of embryologists. Theirsuccess not only opened a new frontier inbiological possibilities but also initiated thefirst wave of debate on the ethics of cloning.

Dolly was cloned froman udder cell of a 6-year-old Finn Dorset ewe.3 Out of250 attempts and twenty-nine cloned embryos createdby the team, only Dollydeveloped successfully, afirst sign indicating thatcloning is not an easy busi-ness.4 Nevertheless, Dollywas able to mate normallyand give birth to healthylambs, proving that clonesare reproductively capable.A clone’s life, however, isnot easy. Besides beingunder constant attention,Dolly developed arthritis,obesity, and lung compli-

cations that rendered her a rather uncom-fortable life. As a result, Dolly was eutha-nized in mid-February this year at the age of6, about half of an average sheep’s lifeexpectancy.3

Before a postmortem autopsy is per-formed, it is hard to tell if Dolly’s declinedhealth was a result of her being a clone. Afterall, Dolly was not an ordinary sheep. As acelebrity animal, she spent most of her timeindoors; a factor that scientists now suspectmight have contributed to her lung failure.Her other health-related disorders such asarthritis and obesity have also been consid-ered side effects of her constantly beggingfor treats and standing on her hind legs.5

Some researchers, however, have foundsome scientific evidence suggesting that herarthritis and her lung problem, a commonlung disease seen in indoor elder sheep,were due to her being a clone.5 A study con-ducted when Dolly was 3 revealed that Dollywas in fact older than thought. At the tips ofthe chromosomes there is a region called thetelomeres, which usually becomes shorter ascells continue to divide and develop.Scientists found Dolly’s telomeres muchshorter than other sheep of her age.Although only 3 years old when the test was

conducted, her telomeres were similar tothose in a 6-year-old sheep, an indicationthat Dolly could have had a shorter lifeexpectancy.6 The decision to put her to deaththis past February because of her failinghealth might have validated this omen.

Cloned MiceAlmost two years after Dolly’s birth,

scientists in the laboratory of Ryuzo Yana-gimachi at the University of Hawaii success-fully created cloned mice. Even moreastounding is the fact that the cloned micewere produced using a different techniquethan that which produced Dolly, indicatingthat more than one way exists to clone ani-mals. A drawback to the technique is that itrequires types of cells found only in femalebodies. As a result, the technique only workson females. Like Dolly, the first clonedmouse that successfully reached adulthoodwas also given a name, Cumulina. Born onOctober 3, 1998, it was among more thanfifty cloned mice produced after hundreds ofattempts. Similar to Wilmut’s experiment,the success rate for cloning was much lowerthan one might have expected; only about 1to 2 percent of the clones were born.However, the team was also able to producesecond-generation cloned mice, clones ofclones.7

A more important aspect of the team’sexperiment is that the animal cloned was amouse. Experts previously thought thatDolly was a success because she was asheep. Many farm animals have cellularDNA intervention fairly late in embryonicdevelopment. The delay buys time for theimplanted genetic material “to be repro-grammed by the egg.”7 However, DNA startsdoing its work very early in mice andhuman embryonic development. The earlyintervention would require functional DNAthat has already been “properly pro-grammed.”7 The fact that cloned mice areattainable indicates that DNA can be repro-grammed in a short amount of time, whichsuggests that a cloned human can be created.

Cloned Human EmbryoA Worcester, Mass.–based company, Ad-

vanced Cell Technology (ACT), announcedin late November 2001 that it had success-fully cloned human embryos. By using the

The fact that cloned

mice are attainable

indicates that DNA

can be reprogrammed

in a short amount of

time, which suggests

that a cloned human

can be created.

Features Volume 8, 2003 29MURJW. Victoria Lee

standard cloning technique (as explained inthe next section), the company was able tocreate a six-cell human embryo.8 The newsinvited excitement, outrage, and skepticism.Many scientists thought the announcementwas rather hasty and premature, because amere six-cell embryo is a far cry from aclone. According to experts, an ovum iscapable of developing through the eight-cellstage “without any signals from the DNA inthe nucleus.”9 Therefore, it is possible thatwhat ACT observed is not the genuine suc-cess of cloned embryos.

ACT is not new to the cloning business.The company was known to have created anembryo by inserting human DNA into acow’s ovum. The high degree of reaction tosuch a quasi-minotaur creation is to beexpected. But the company only allowed theembryo to grow through five cell divisions,as the researchers’ goal was not to create anactual human clone but to experiment withpossibilities in stem cell research.10

The company also experimented withparthenogenesis, a process in which ahuman ovum is allowed to “divide into earlyembryos without being fertilized by a spermor being enucleated and injected with adonor cell.” The goal of this was to obtainstem cells to grow transplantable organsthat are less likely to be rejected by patients.Although previous experiments done byother experts with eggs from other animalshave proven successful, ACT was not able toproduce successful embryos that containstem cells.11

Cloned PetAlthough not an easy thing to do, cloning

has proved to be contagious. Now there arehundreds of cloned animals, includingsheep, goats, mice, pigs, cows, cats, and,most recently, rabbits.5 The famous firstcloned pet, a cat named Cc, was born at theend of 2001 in a laboratory in Texas. Cc wasthe only one to survive out of eighty-sevencloned embryos. Not unlike other cloningsuccess stories, Cc’s birth also generatedimmense excitement and controversy.Experts are now looking into cloning tech-nology to save endangered species; a type ofendangered wild cattle has already beencloned successfully. Businessmen are also

hoping that one day cloning technology canbe used commercially to allow pet owners tore-create their deceased beloved compan-ions.12 Although many types of animalshave already been cloned so, animals suchas dogs, certain monkeys, and human beingshave yet to be replicated.5

Cloned Human?Baby Eve is allegedly a clone of her 31-

year-old American mother made from herskin cell and an ovum. She was the first ofthe five cloned babies claimed by Clonaid tobe born.1 All babies were due to be born byearly February 2003, including the baby ofa Dutch lesbian couple. Although Boisseliermaintains that the birth of the cloned babiesis a new scientific breakthrough, most ex-perts are highly skeptical of her claim. Notonly does Boisselier have no evidence toprove the baby’s existence or the fact thatshe’s a clone, but she has also backed offfrom having the baby tested for authenticitybecause of her concerns for the safety of thebaby. The fact that Clonaid has not clonedanything before and that their success rateof the claimed human clone—five in ten—gave the public legitimatereason to doubt. But aswhat Robert P. Lanza, acloning expert with ACT,points out, “It may berather easier than any of usthought to clone a human.”13

Maybe, just maybe, we areall looking in the wrongdirection.

The ProcessCloning, another term

for making an exactgenetic replica, is notunnatural in the biologicalfield. Many organisms relyon cloning to continuetheir existence. Cloning,however, is an artificialnovelty in animals, espe-cially mammals. Decades ago, it was fic-tional or even fanatical to talk aboutcloning, making a genetic copy of an ani-mal, let alone a human. Are we magicians,are we miracle workers, or are we Dr.

With the low success

rate of producing a

viable cloned embryo,

it is legitimate to

question how we will

be able to obtain

hundreds of eggs just

to produce one or two

cloned embryos.


Frankenstein reborn? How does cloningreally work?

A generic cloning technology, somatic-cell nuclear transfer, provides the basicframework for other different cloning tech-niques. Somatic cells are the so-called bodycells. In the process of somatic-cell transfer,a somatic cell’s nucleus, the part of a cellthat contains the DNA, is extracted andinserted into an enucleated ovum. The com-bined entity will develop into an embryothat will then carry the DNA of the somaticcell and therefore the same gene as thedonor of the somatic cell in every one of thenew embryo’s cells.6 The exact process, ofcourse, is not so simple. The ovum, or egg asit is commonly known, first needs to beplaced in a culture dish until it matures,while being soaked in a chemical bath, andthen its genetic material is removed.Everything is also done on a microscopiclevel, which makes the process extremelypainstaking. In order to enucleate the ovum,the researchers have to first remove a smallportion of a protective layer, called the

“zona pellucida,” on theoutside of the ovum. To dothis, a microscopic needleis used to “drill” out thesmall piece of the layer,called a plug, while apipette is holding onto theovum. After the plug isremoved, the polar bodyand the genetic material inthe ovum are sucked outwith a needle. The nucleusor sometimes the entiresomatic cell is theninserted into the ovumthrough the “hole” cre-ated. The resulting entityis then placed in a chemi-cal bath that will stimulateit to divide and develop.14

Dolly was producedusing the somatic cell from a ewe. The spe-cific technique used by Wilmut’s team wasthe fusion of the entire udder cell and theenucleated ovum by electrical jolt. Thecloned mice, on the other hand, were pro-duced using cumulus cells, which nourish

and cling to the ova on the outside. In addi-tion, a cumulus cell was inserted into theovum by injection rather than electricalfusion. Replacing just any somatic cell witha cumulus cell is also significant in thatcumulus cells are found on human ova aswell.7

From the findings of cloned mice, ACTwas able to use two techniques to make thehuman cloned embryos. The companyinjected nuclei of skin cells into some ovaand cumulus cells into others. Like othercloning experiments, it took many attemptsbefore they were able to produce a success-ful cloned embryo from the ovum that wasinjected with cumulus cells.11

The Moral Dilemma We have longed to do it, and now we

have done it. But what remains is the moralquestion of the line between therapeuticcloning and reproductive cloning. Thera-peutic cloning explains why we wanted toclone in the first place.

The original goal of Dr. Wilmut and histeam of scientists was purely therapeutic.They made Dolly in the hope that the exper-iment would lead to the mass production ofgenetically engineered farm animals whoseorgans could be used for transplantation orwhose milk would have medicinal purposesfor sick patients.2 Yanagimachi’s team andACT’s researchers have also had similargoals in mind when they conducted theirstudies.

Therapeutic Cloning“Therapeutic cloning” is a term coined to

describe the process of harvesting stem cellsfrom a cloned embryo, which “is made usingthe DNA of a patient who could benefit froma stem cell transplant.”15 The harvested stemcells can then be developed into the specifictissue, organs, or even nerves that thepatient needs. A wide range of disorders canbe cured with the compatible body partsgrown from stem cells, including diabetes,Parkinson’s disease, Alzheimer’s disease,stroke, and epilepsy. It is even possible tocreate blood cells and bone marrow fromstem cells with proper cell differentiation.11

The ultimate goal is laudable, but it is theprocess of harvesting the stem cells that has

Dr. Jeffrey Kahn…

points out that the

majority of indignation

against therapeutic

cloning is because

most people think “it

is unethical to create

human life that will

be destroyed.”

Features Volume 8, 2003 31MURJW. Victoria Lee

generated an immense wave of controversy—a hot topic excessively discussed in theabortion issue, namely the debate ofwhether an embryo is a human being or justa bunch of cells.

Dr. Jeffrey Kahn, the director of theCenter for Bioethics at the University ofMinnesota, points out that the majority ofindignation against therapeutic cloning isbecause most people think “it is unethical tocreate human life that will be destroyed.”16

Indeed, the people who are enraged by suchscientific attempts range from religious topolitical leaders. When ACT first announcedits human embryo in 2001, President Bushhad already expressed his strong disap-proval. He maintained that “we should notas a society grow life to destroy it.”17 Al-though Archbishop Tarcisio Bertone praisedthe therapeutic aims of cloning, he insistedthat “if the process involves production anddestruction of human beings to treat otherhuman beings, then the end doesn’t justifythe means.”8

Other scientists and religious leaders,such as Cynthia Cohen, a researcher at theKennedy Institute of Ethics at GeorgetownUniversity, have come to the conclusion thatany embryo “younger than 14 days cannotbe considered human because cells have notformed a single individualized entity.”18 Forsome, then, research dealing with clonedembryos no older than 14 days old is there-fore acceptable and ethical.

As it turns out, some scientists also holda very skeptical view on how much embry-onic cloning will modernize stem cell ther-apy. There are millions of people sufferingfrom diseases that require various trans-plantations, but as Harry Griffin, assistantdirector at the Roslin Institute, points out,“There are simply not enough human eggsavailable” to produce cloned embryos fromwhich to harvest stem cells.9 With the lowsuccess rate of producing a viable clonedembryo, it is legitimate to question how wewill be able to obtain hundreds of eggs justto produce one or two cloned embryos.6

Reproductive CloningAnother aspect of cloning research that is

belittled by politicians and scientists alike isreproductive cloning. The process is essen-

tially the same as that of therapeutic cloningin terms of creating the embryo. In repro-ductive cloning, however, the embryo isplanted into a woman’s womb and devel-oped into a fetus. But with in vitro fertiliza-tion and other infertilitytreatment readily avail-able, why would one turnto cloning for babies? Itturns out that people whodon’t have ova, sperm, oreither can use cloning toproduce a child that isgenetically related (moreaccurately, identical) tothem by taking one par-ent’s somatic cell nucleusand inserting it into adonor egg that does notcontribute to the geneticmake-up of the embryo.6

Other candidates whomight benefit from repro-ductive cloning are lesbiancouples who don’t wanttheir children to carry thegenes of unrelated spermdonors. Couples who aregenetic disease carriersmight also want to usecloning to prevent havingchildren with a full-blownversion of their diseases that might resultfrom the genetic mixing during sexualreproduction. Furthermore, these parentscan have gene therapy applied to their dis-eased somatic cell before its nucleus isinserted into the egg to form an embryo,thereby forever obliterating the diseasedgene from the family tree.6

Although these applications of cloningmake the technology seem like a panacea toinfertility and genetic disease-related prob-lems, from a bioethical point of view, repro-ductive cloning is really the fountainhead ofall moral predicaments.

One potential problem has to do with theclone’s identity. Because the clone has thesame DNA as its somatic cell nucleus donor,the two are essentially identical twins. As aresult, one “could give birth to [one’s] owntwin, or the twin of [one’s] mother or

Although these

applications of cloning

make the technology

seem like a panacea to

infertility and genetic

disease-related

problems, from a

bioethical point of

view, reproductive

cloning is really the

fountainhead of all

moral predicaments.


father.”16 Although people are made up ofmore than genes and environmental factorsplay a significant role in their personalityand appearance, it is expected that clones,however different they might be from theiradult “originals,” would encounter difficul-ties in psychological development. Thecloned child might be expected to live a lifesimilar to the “original” and might be lookeddown on if he or she fails to do so. Thecloned children might also have low self-esteem because they are copies of someoneelse. They might lack motivation to controltheir own life because they already knowcertain traits from their adult “originals”that they are destined to bear, such as short-ness or baldness.

Furthermore, will we move from repro-ductive cloning to selective cloning? Willwe choose to clone only the beautiful, thehealthy, the strong, and the smart adultsamong us? As Robert Wachbroit points outin “Genetic Encores: The Ethics of HumanCloning,” we know some newborns will leaddifficult lives such as the children who areborn into poverty, “but we don’t therebyconclude that such children shouldn’t beborn.”19 These children might not have

material luxury, but they can still enjoyloving families. How do we decide whoshould be cloned? And by performing artifi-cial selection, are we meddling with MotherNature? What would become of humanevolution?19

These questions may appear far-fetchedand even unrealistic right now, but they arelegitimate concerns once we start cloninghumans as frequently as we use in vitro fer-tilization today.

But before we worry ourselves sick withthese ethical questions, we should ask ifhuman cloning is safe. As it turns out,human cloning might be a dream too goodto be true. Professor Rudolf Jaenisch at theWhitehead Institute for Biomedical Researchat MIT and his team reported in lastSeptember’s issue of Proceedings of theNational Academy of Sciences that “thecloning process jeopardizes the integrity ofan animal’s entire genetic make-up.”20

Having studied around 10,000 genes,researchers found that in cloned mice’s pla-centas as much as 4 percent of the genes isabnormal.20 Two other studies also showedthat the path to human cloning might not beas bright as we thought. Researchers fromthe University of Connecticut in Storrs havebeen studying the X chromosomes in unsuc-cessfully cloned cows and found that somegenes on the chromosomes are not ex-pressed. Researchers at the University ofPennsylvania have also discovered that afate-determining gene called Oct4 goes awryin more than 90 percent of the cloned mouseembryos.21

These findings not only put a stop sign inthe middle of the human cloning rush butalso raise doubts about the safety of stemcells harvested from cloned embryos. Re-searchers like Jaenish, however, have confi-dence in the safety of therapeutic cloningbecause tissues or organs grown from suchstem cells “would not contribute to thedevelopment of [the] whole organism.”21

What Do the Politicians Say? They say “no” to cloning, period. Repro-

ductive cloning has never received muchsupport, but people have also been lookingaskance at therapeutic cloning, trying tonudge it out of the picture inch by inch.

References

1. DeNeen L. Brown. “The Leaderof UFO Land.” Washington PostForeign Service. January 17,2003, C01.

2. Rick Weiss. “Scottish ScientistsClone Adult Sheep.” WashingtonPost. February 24, 1997, A01.

3. Emma Ross. “Dolly the ClonedSheep Put to Death.” LosAngeles Times. February 15,2003.

4. Lybi Ma. “Get Ready for Human2.0.” Discover. January 20,1999.

5. Janet Stobart and Rosie Mestel.“Sheep Dolly, 6, Dies; PioneerClone’s Birth Sparked Debate.”Los Angeles Times. February 15,2003.

6. Ronald M. Green. “I, Clone.”Scientific American. September3, 1999.

7. Rick Weiss. “Scientists CloneAdult Lab Mice.” WashingtonPost. July 23, 1998, A01.

8. “U.S. cloning advance shocksworld.” CNN: Technology.November 26, 2001.

9. Gary Stix. “What Clones?”Scientific American. December24, 2001.

10. Corey S. Powell. “BeyondCloning.” Discover. November12, 1998.

11. Jose B. Cibelli, Robert P, Lanza,Michael D. West, Carol Ezzell.“Exclusive: The First HumanCloned Embryo,” ScientificAmerican. November 24, 2001.

12. David Braun. “ScientistsSuccessfully Clone Cat.”National Geographic News.February 14, 2002.

13. Manuel Roig-Franzia and RickWeiss. “Religious Sect Says ItCloned Human.” WashingtonPost. December 28, 2002, A03.

14. “Therapeutic Cloning: How It’sDone.” Scientific American.November 24, 2001.

15. Jeffrey P. Kahn. “On the path tocloning?” CNN: Health.November 26, 2001.

16. “Dr. Jeffrey Kahn: Ethics ofhuman cloning.” CNN:Community. November 27, 2001.

Because the clone

has the same DNA as

its somatic cell nucleus

donor, the two are

essentially identical

twins. As a result, one

“could give birth to

[one’s] own twin, or

the twin of [one’s]

mother or father.”

W. Victoria LeeFeatures

Long before Dolly was born, the talk ofhuman cloning had already begun in thescientific community. By the end of 1994,the National Institutes of Health (NIH)decided to prohibit human cloning researchbut expressed desire to permit federal fund-ing for some human embryo research thatmight lead to the discovery of treatments forterminal diseases. President Clinton, how-ever, thought otherwise and prohibited NIHto fund human embryo research.2 WhenDolly’s birth was announced, PresidentClinton took it one step further and estab-lished a moratorium on all human cloningexperiments with federal funds. At the sametime, the National Bioethics Advisory Com-mission wanted a ban on private research aswell.6 When President Bush asked Congressto ban cloning in 2002, the House of Repre-sentatives agreed with him and introducedthe bill again as soon as Boisselierannounced Baby Eve’s birth. As of mid-February, the cloning ban has been endorsed

by the House Judiciary Committee, pendingfloor debate and vote.22

While lawmakers unanimously agree toplace restrictions on cloning research, theRepublicans seem to be more uncompromis-ing than their Democrat colleagues. They areseeking to pass a bill in the Senate that notonly puts a stop to cloning attempts but alsoto the creation of embryos for any purpose.Democrats, on the other hand, hope toexempt therapeutic cloning research in theirown bill.18

Cloning is such a familiar yet strangeword. We have come a long way, yet there isstill so much unknown. Surely scientists willone day figure it all out. The question is:How will we deal with the moral issues ofthe findings? And in the meantime, willlegal issues stand in the way? As Dr. Kahnpoints out, “It is not cloning techniques thatare unethical, but some of their potentialapplications.”15 Ultimately, what the futureof cloning will be is a question that onlytime can answer.

17. Rick Weiss. “Mass. Firm’sDisclosure Renews CloningDebate.” Washington Post.November 27, 2001, A03.

18. Bill Broadway. “Reports ofCloned Baby Overshadow SubtleShift.” Washington Post.January 11, 2003, B09.

19. Robert Wachbroit. “GeneticEncores: The Ethics of HumanCloning.” University ofMaryland: Institute forPhilosophy and Public Policy.Fall 1997.

20. Kristen Philipkoski. “Study:Humans Not fit for Cloning.”Wired News. September 9, 2002.

21. Josie Glausiusz. “The Woes ofthe Clones.” Discover. August2002, Vol. 23, No. 8.

22. “House Committee OKs CloningBan.” Associated Press. WiredNews. February 12, 2003.

Volume 8, 2003 33MURJ

Features34 Volume 8, 2003MURJ


Stem Cell Research: The Debate that Has

Divided America

Farhan Merali

A scenario:You have severe congestive heart failure. You need a heart transplant, but your doctor

tells you that due to the scarcity of replacement hearts, you might have to wait a year ormore. After the transplant, your body’s immune system might still reject the new heart. Evenif the transplant is a success, you will have to endure the effects of immunosuppressivedrugs for the rest of your life.

Another scenario, this one for the future: You have severe congestive heart failure. Your doctor admits you

to the hospital. During an operation later that day, she injects someheart cells into your heart, and after an integration period of a fewdays, your heart is healed substantially. You go home and lead ahealthy life.*

Doesn’t the second scenario sound better? What if you

knew that the heart cells were generated from human stem

cells, taken from human embryos? Depending on your

belief about when life begins, this might bother you a great deal or

it might not bother you at all.

Stem cells are the ordinary undeveloped cells of very early–stage

(no more than 64-celled) embryos.1 To put this in perspective, a

newborn baby is composed of billions of cells. Many of these

embryos have been grown in a laboratory from fertilized eggs; they

were produced for in-vitro fertilization but were later discarded or donated specifically for

research purposes. Embryonic stem cells are pluripotent—unlike more mature cells, they

hold the possibility of developing into any organ of the body. Scientists experimenting with

mice have introduced pluripotent mouse embryo stem cells into diseased organs. These stem

cells then begin to take on certain characteristics and functions of the organ cells. The stem

cells don’t actually develop into organs, but they do begin to resemble the organ cells; stem

cells introduced into a diseased kidney, for example, mime ordinary kidney cells. The other

kidney cells integrate the new cells until the organ is effectively regenerated.2

Farhan Merali

* Scenarios have been adapted from Horvath, 2003.

Microscopic 10x view of a colony of undifferentiated humanembryonic stems cells being studied in developmental biolo-gist James Thomson's research lab. The embryonic stem cellcolonies are the rounded, dense masses of cells. The flat,elongated cells in between the embryonic stem cell coloniesare fibroblasts that are used as a "feeder layer" on whichthe embryonic stem cells are grown. (Source: University ofWisconsin-Madison.)

Features36 Volume 8, 2003MURJ Farhan Merali

The second of the given scenarios is whatresearchers hope will become an everydayreality early in the 21st century, not only totreat diseased hearts but also to treat dam-aged livers, kidneys, and lungs as well asneurological diseases. By the very nature ofthe research, however, scientists have foundthemselves entangled in a moral and ethicaldebate that has divided America: Shouldstem cells from embryos that could poten-tially develop into living beings be used totreat a wide array of diseases? The answer iscontingent two main parts: When does afertilized human egg become a living per-son? and Do the benefits of the therapy war-rant the controversial use of stem cells?

Stem cells currently being used forresearch purposes and experimental studiesare usually derived from aborted fetuses orembryos that have grown in a laboratoryfrom fertilized eggs. The eggs used are nor-mally produced for in-vitro fertilization, aprocess that helps many women to conceivechildren. When doctors match sperm andegg to create embryos outside the womb,they usually produce more embryos than areplanted in the mother. These embryos arelater discarded or donated specifically forresearch purposes. Some Pro-Life activistsargue that an egg becomes a living personthe instant it is fertilized; some scientistsargue the opposite, using the following

logic: The fertilized eggs used for researchpurposes sit in vats of liquid nitrogen atsubzero temperatures until they are other-wise thrown out. The eggs have the poten-tial to become living beings, but they willnever be implanted inside a woman’s uterusto undergo the process of developing into aliving, breathing human being. How, then,can these be called “human beings”? Thesame holds true for aborted fetuses: Thedecision to prevent the fertilized egg frombecoming a living being has already beenmade.

Others argue that the question boils downto one of intent: What was intended tobecome of the eggs? If the egg in questionwere in fact implanted inside a woman, thenit would have the opportunity to undergothe process of development into a humanbeing. However, if the egg in question is oneof the 30,000 leftover eggs*1 which couplesdon’t need after they’ve had their child andwhich are set aside to be discarded everyyear, then the intended path is not one oflife. For such an egg, the decision regardingthe potential to become a living being hasalready been made. Why, then, scientistsargue, should such an egg be preventedfrom aiding researchers to find potentiallylife-saving treatments using stem cell tech-nology? The same holds true for eggsdonated for research, as well as abortedfetuses from which stem cells can be har-vested.

The end results of utilizing embryonicstem cell research are numerous and highlypromising: They could lead to life-savingtherapies for Alzheimer’s disease and dia-betes as well as to ways to prevent birthdefects and rebuild damaged organs. Forexample, in the 30 July 1999 edition ofScience magazine, scientists reported howthey were able to manipulate stem cells intoneural cells and inject them into fetal ornewborn rats who have a disease in whichthe myelin coating around nerve fibers ismissing.3 The cells had been developed intokey cells of the nervous system that wereable to promote the growth of myelin cov-ering to help nerves function normally.These tests show promise for a viable treat-

Culture trays containing human embryonic stems cells being viewed under a microscopeand studied by developmental biologist James Thomson's research lab. Photo by: Jeff Miller

* Figure represents numbers in the United States alone.

Features Volume 8, 2003 37MURJFarhan Merali

ment for multiple sclerosis (MS), a disease inhumans that parallels this demyelination inrats, for which there is currently no cure.

Geron, a small biotech firm in California,is working on a project that could pro-foundly change medical treatment by offer-ing patients customized parts to repair dam-aged organs. The Geron Corporation hasdeveloped a technology called “telomeraseexpression,” which it says allows cells tokeep replicating. With this technology, inconjunction with stem cell technology,Geron researchers would be able to grownew human tissue capable of repairing heartmuscle, bones, nerves, skin, and eyes.4

Though mainstream media sometimesgives the impression that the public willhave to wait years before revolutionarytreatments involving stem cell technologyfor human diseases become a reality, this isnot the case. This past July marked thetwenty-first month of “complete clinicalremission of [lupus]” in an 18-year-oldwoman who was so near death from the dis-ease that she needed life support because herlungs and kidneys were failing. Now—abouttwo years later—she is in good health andshowing no signs of the disease. The youngwoman became one of a handful of lupuspatients to undergo a stem cell treatmentthat may cure the disease.5

What if the government had decided tohalt stem cell research before this revolu-tionary treatment for lupus had come tofruition? Can one justify depriving an indi-vidual who has a life-threatening illness ofa potentially health-restoring treatment byutilizing an embryo that was set aside to bediscarded anyway? Again the question is,When does a fertilized egg become a humanbeing? Even if this line is unclear, can onejustify preventing scientists from conduct-ing research that could lead to and hasalready shown promise in treatments thatcan save thousands, if not millions of liveswith cells derived from these same eggs? Noone knows exactly how many lives could besaved, but if one examines the number ofpeople afflicted with diseases for which stemcell therapy could potentially be a cure, thenumbers are staggering: 1.4 million people

with lupus,5 about 1 million people* withParkinson’s disease,6 and approximately220,000 people with spinal injuries, withabout 10,000 new injuries per year.7 Thesefigures represent the number of peopleafflicted in the United States alone, and donot even include the countless other peoplewith disorders that could be cured orimproved with a regenerated body part ororgan.

One of the most promising frontiers ofscience has rocked America with one of thegreatest moral dilemmas: Should these“master cells” that are present only in early-stage human embryos be used for eventuallife-saving therapies for Alzheimer’s disease,Parkinson’s disease, multiple sclerosis, andother neurological and physiological disor-ders, as well as to prevent birth defects andrebuild damaged organs? Some people mayhave difficulty with weighing life-savingmedical benefits against moral costs, butsome argue that there does not need to beany moral cost. Advocates of this argumentposit that the much-debated “line” betweenwhen a fertilized egg becomes a living beingdoes not need to be delimited. Again, theysay the question boils down to one of intent.The implications of stem cell technology arevery real and have already saved several

After transplantation into the brains of young mice, the neural precursor cells give riseto functioning neurons (A) and astrocytes (B), a star-shaped cell of the brain and spinalcord. Photo courtesy of Su-Chun Zhang

* The number is approximate because many people, perhaps half of those affected, are thought to be undiagnosed.

Features38 Volume 8, 2003MURJ Farhan Merali

human lives as well as shown promise inanimal models. Halting stem cell researchcould potentially destroy the light of hopefor the millions of people in the United

States whose lives could one day beimproved or even saved through the use ofstem cell technology. Many agree that it iswrong to create human embryos in testtubes solely to experiment on them, but ifthe remaining eggs from an in-vitro fertil-ization are to be discarded regardless, theycould be used instead to cure a child withdiabetes, a 65-year-old with Alzheimer’sdisease, or a young father with a paralyzinginjury. An embryo has the potential for lifeat one point, but if this route is no longerthe intended one, stem cell technology givesan embryo the potential to save a life.Perhaps, then, we as a society must decidethe point at which the potential for life canbe outweighed by our need to survive. Then,if we found ourselves facing the openingscenario, would we change our minds?

Photos ©University of WisconsinBoard of Regents

References

1. Goldstein, Andrew. “What AreStem Cells?” Time MagazineOnline. 15 Jan. 2003.<http://www.time.com/time/2001/stemcells/#>

2. Horvath, Hacsi. “EthicalConcerns Surround Stem-CellResearch.” 14 June 1999. CableNews Network. 9 Jan. 2003.<http://www.cnn.com/HEALTH/9906/14/stem.cell.research/index.html>

3. “Stem Cells Show Promise inTreating Neurological Diseases.”29 July 1999. WebMd.com. 19Jan. 2003. <http://www.cnn.com/HEALTH/9907/29/stem.cell.advance/>

4. Knapp, Don. “Cell ScientistsHope to Grow Human SpareParts.” 22 May 1999. CableNews Network. 10 Jan. 2003.<http://www.cnn.com/HEALTH/9905/22/organs.to.grow/>

5. DeNoon, Daniel. Reviewed byGary Vogin, MD. “Stem CellsSave Another Lupus Patient.”WebMD Medical News. 17 Jan.2003. <http://webmd.lycos.com/content/article/1823.50659>

6. “About Parkinson’s: Who GetsParkinson’s?” Michael J. FoxFoundation for Parkinson’sResearch. 21 Jan. 2003.<http://www.michaeljfox.org/parkinson/index.php>

7. Palmer, Sara, Jeffrey B. Palmer,and Kay H. Kriegsman. SpinalCord Injury: A Guide for Living.Johns Hopkins University Press,2000.

Culture trays containing human embryonic stemscells being stored in heat-controlled storage andstudied by developmental biologist James Thomson'sresearch lab. Photo by: Jeff Miller

Reports Volume 8, 2003 39MURJ

The SpectroscopicDetermination of Aqueous Sulfite UsingEllman’s Reagent

Cameron Sadegh and Ronald P. SchreckAcademy for the Advancement of Science and Technology, Hackensack, NJ

Abstract

At room temperature and in N2-purged pH 8 aqueous phosphate buffer, the sulfite

ion (SO3-2) cleaves the disulfide bond in Ellman’s reagent, 5-5¢-Dithiobis-(2-

nitrobenzoic acid), and displaces one of the chromophoric anions, producing a

yellow hue. Absorption spectroscopy of the resulting chromophore, 5-mercapto-2-nitroben-

zoate, permits a quantitative assay of the initial sulfite concentration using a linear rela-

tionship in accordance with the Beer-Lambert law. This colorimetric method was validated

with several test reactions and was determined to be accurate to within a 2% average rel-

ative error. Its sensitivity has been demonstrated down to 0.8ppm sulfite (10-5M). One draw-

back of Ellman’s reagent is that it will react with other compounds containing thiol groups;

yet, in some interesting cases, such as in the water boiler industry, other such compounds

are absent and this technique would provide a reliable, inexpensive, and field-usable

method for determining sulfite concentration.

IntroductionSulfur-oxy anions play an important role in our nutrition and environment. One such

anion, the trigonal pyramidal sulfite (SO3-2), has several fascinating and uncommon uses.

Sulfites are currently used to control microbial growth, bleach certain food starches, and

prevent spoilage of certain perishable foods, beverages, and pharmaceuticals. Several exam-

ples of where sulfites are used include shrimp, dried apricots, dried raisins, lettuce and other

vegetables, and potato chips. Furthermore, their antioxidant and antimicrobial properties

play an important role in wine-making. The sulfites either inhibit or kill bacteria and wild

Sadegh and Schreck

Reports40 Volume 8, 2003MURJ Sadegh and Schreck

yeast, thus encouraging rapid and clean fer-mentation of wine grapes. The paper andpulp industries use sulfites as additives toimprove strength, increase brightness, andlower pulping energy. The photographicindustry uses sulfite for the testing of fixingbaths, stop baths, and developers. Sulfite isused in the water boiler industry as an oxy-gen scavenger, which binds well with excessoxygen to form sulfate. Yet sulfite concen-trations in boiler and process waters must bemonitored routinely to avoid overtreatment,which can lower the pH and cause rust.Waste treatment plants remove residualchlorine in wastewater by injection of eithersulfur dioxide gas or a solution of sodiumsulfite or sodium bisulfite (strong reducingagents). In practice, most plants subject todechlorination requirements run relativelyhigh-sulfite residuals to ensure completechlorine removal at all times. However,excess sulfite in wastewater effluents maybe harmful to marine life. Because sulfitesare widely used in industries, a method formonitoring its levels is a necessity for envi-ronmental safety.

Sulfite may also be harmful to somehumans with allergies. A 1985 study by theFederation of American Societies for Experi-mental Biology found that in some patientswith asthma, ingesting sulfites might lead toan acute and sometimes life-threateningattack of asthma. This study prompted theU.S. Food and Drug Administration (FDA) toissue regulations prohibiting the use of sul-fites on fresh fruits and vegetables that areusually eaten raw. In addition, the FDA nowrequires other foods, such as wine, to indi-cate the presence of sulfites on their labels.The FDA estimates that one in 100 people issulfite-sensitive to some degree, but for the10 percent of the population who are asth-matic, up to 5 percent are at risk of havingan adverse reaction to the substance.

Current methods of analysis of sulfiteinclude basic test strips, an iodide-iodatetitration,1 and sulfite acidification to sulfurdioxide gas. These kits vary in price andaccuracy. However, a spectroscopic methodwould be cheaper because it would onlyrequire a spectrophotometer, which isalready available in most scientific labs.

Further, it would be convenient, reliable,and accurate at low concentrations. The aimof the research was to determine a simplespectroscopic method to measure sulfite insolution using 5-5¢-Dithiobis-(2-nitroben-zoic acid), or DTNB (also called Ellman’sreagent). The method established in theresearch is also compared to previous stud-ies on DTNB.

Since the first synthesis of 5-5¢-Dithiobis-(2-nitrobenzoic acid), DTNB, byGeorge L. Ellman in 1959 for the quantita-tive analysis of mercaptans, “Ellman’sreagent” has been commonly used for thedetermination of thiols in biochemical sam-ples.2 An organic disulfide, DTNB reacts withaliphatic thiol compounds to produceequimolar concentrations of a mixed disul-fide and a luminescent thiol, 5-mercapto-2-nitrobenzoate (MNB), according to Equation1.

R¢SSR¢ + RSH fi R¢SSR + R¢SH (1)

The use of DTNB has been extended tothe reaction with sulfite ions (SO3

2-), whichdisplaces the thiol anion forming an organicthiosulfate, called a “Bunte” salt (Equation2).3,4,5,6

RSSR + SO32- fi RSSO3

- + RS- (2)

The reaction of DTNB with sulfite ionshas led to studies for a spectrophotometricdetermination of sulfite.3,4 While useful inbiochemical samples, the method for sulfiteanalysis proposed by Johnston et. al. maynot be effective in aqueous solutions. Sucha method may interest the boiler industry.Humphrey et. al. demonstrated the useful-ness of a similar method in aqueous solu-tions stabilized with the disodium salt ofethylenediamine tetraacetate (EDTA). Thiscompound was introduced with the inten-tion of stabilizing the readily oxidizable sul-fite in aerated solutions. The method pre-sented here demonstrates an effective sulfiteassay in aqueous solutions without the useof EDTA and outlines some of the parame-ters governing the reaction of DTNB withsulfite and with L-cysteine in the presenceof oxygen and ultraviolet light. A detailedcomparison of the present method with acommercially available titration kit demon-strates the enhanced accuracy (<2% avg.

Reports Volume 8, 2003 41MURJSadegh and Schreck

relative error) and range of analysis(<1ppm).

ExperimentalReagents. Ellman’s reagent, DTNB (99%),

was obtained from Aldrich Chemical Co. inMilwaukee, Wis.; laboratory-grade sodiumsulfite was obtained from Flinn Scientific,Inc., in Batavia, Ill.; L-cysteine (98+%) andTris-[hydroxymethyl]-aminomethane(99.9+%) were obtained from Sigma Chemi-cal Co. in St. Louis, Mo. These reagents wereused without further purification.

Solutions. The DTNB solutions were pre-pared 1.0 x 10-3M by dissolving the DTNBwith about 1mL 95% ethanol and dilutingwith N2-purged pH 8.0 0.1M Tris buffer.Further dilutions yielded 1.0 x 10-4M and 5.0x 10-5M DTNB solutions. The sodium sulfitesolutions were prepared in N2-purged waterand diluted in volumetric flasks to obtain10-4M samples. Aqueous 2.5 x 10-4M L-cys-teine solutions were similarly prepared.

Equipment. Absorption measurementswere made with a Perkin Ellmer ModelLambda 3B UV-visible spectrophotometer,using quartz cuvettes with 1cm pathlengths. Comparison testing was done with aHach Inc. titration analysis 1-200mg/L sul-fite detection kit (Model SU-5, Cat. #1480-02).

Procedure. For the reaction of DTNB andcysteine, the cysteine was added in fivefoldexcess relative to DTNB to ensure completedissociation of MNB. The cysteine wasdirectly introduced into the solutions. Theconcentration of DTNB was made in fivefoldexcess relative to the concentration of thesulfite solutions. The DTNB and sulfite werereacted in equal volumes of 5mL, and allabsorbance measurements were taken fromabout 2mL aliquots in triplicate. Referencesolutions included a 50:50 mixture of DTNBin pH 8 buffer solution and deionized water.

Results and DiscussionDTNB-Cysteine reaction. In order to

determine the em of MNB, a standard reac-tion was followed with DTNB and L-cys-teine. It was observed that the cysteine com-pletely cleaved the DTNB and formed doublethe molarity of the chromophoric thiol(MNB). Note that this is contrary to Equation

1; the cysteine is oxidized and forms cys-tine, while the DTNB is reduced and cleavedat the disulfide bond. Using a spectropho-tometer, the peak absorbance at 410nm of10–4M MNB was determined to be 1.660 with amolar extinction coefficient of 16,600cm-1M-1.This value does not equal the em previouslyreported.1,4,7 Humphrey et. al. documented15,500cm-1M-1 as the em value of MNB.Discrepancies in the em values of MNB arealso noted by Riddles et. al.8

DTNB-Sulfite reaction. It was hypothe-sized that a solution of 1.0 x 10–4M sulfitewould react similarly with DTNB to form 1.0 x 10–4M MNB (see Figure 1). This is inaccordance with Equation 2; the sulfite dis-places the chromophoric thiol, MNB. Theabsorbance measurements of this solution at410nm were near 1.600. This value is within4% of the absorbance from the DTNB-cys-teine reaction, justifying the reaction ofDTNB with sulfite as seen in Equation 2. Theabsorbance of MNB was then measured as afunction of the initial sulfite concentrationfor various samples. The data demonstrateda linear relationship between the absorbanceof MNB and initial concentration of sulfite(see Tables 1 and 2). The sensitivity of thismethod has been demonstrated as low as 10-5M sulfite.

Figure 1. MNB (5-mercapto-2-nitrobenzoate) isdisplaced by sulfite.

mL mL mL [SO32-] Avg.

MNB SO32- Tris Buffer (mol/L) Abs.

5 5 0 1.0x10-4 1.610

5 4 1 8.0x10-5 1.300

5 3 2 6.0x10-5 1.008

5 2 3 4.0x10-5 0.718

5 1 4 2.0x10-5 0.434

Table 1. Comparison of the concentration of sul-fite with the absorbance of MNB produced inthe DTNB-sulfite reaction.

COO -

O2N S S NO2

-OOC

O2N S

-OOC

S

OO

O

S+

O

O

O

SO2N

-OOC

+ pH=8.0 +DTNB

MNB - yellowsulfite

"Bunte" salt

Reports42 Volume 8, 2003MURJ Sadegh and Schreck

mL mL mL [SO32-] Avg.

MNB SO32- Tris Buffer (mol/L) Abs.

5 5 0 1.0x10-4 1.520

5 4 1 8.0x10-5 1.230

5 3 2 6.0x10-5 0.940

5 2 3 4.0x10-5 0.637

5 1 4 2.0x10-5 0.335

10 1 9 1.0x10-5 0.186

Table 2. Comparison of the concentration of sul-fite with the absorbance of MNB produced inthe DTNB-sulfite reaction with adjusted spectro-scopic reference solution.

Figure 2. Adjusted sulfite determination graph.

Sulfite Actual Calculated sample Mass (g) Mass (g) Error

A .1261 .1298 2.90%

B .0946 .0966 0.06%

C .0631 .0656 4.00%

D .0315 .0318 0.83%

Average % error — — 1.95%

Table 3. Calculated percent error from the DTNB-sulfite method.

Standard sample, Hach Inc.

Sulfite (ppm) kit (ppm) Error

16 14.7 8.0%

12.8 11.5 10.0%

9.6 8.3 13.3%

6.4 5.8 10.0%

1.6 1.9 20.0%

Average % error — 12.3%

Table 4. Calculated percent error for the HachInc. detector at low sulfite concentrations.

Validation. This method was validatedwith several standard sulfite samples anddetermined to be accurate to within a 2%average relative error (see Table 3) for diluteconcentrations of sulfite (10-5 – 10-4 M). Themethod was then tested against the accuracy

of a commercial sulfite detection kit. TheHach Inc. titration analysis sulfite detectionkit had an average relative error of 12% (seeTable 4) for dilute sulfite solutions.

Stability of Sulfite Solutions. It was ini-tially observed that the absorbance of MNBwas less than expected and decreased overtime after the DTNB-sulfite reaction. Thepresence of oxygen was found responsiblefor the rapid degradation of sulfite solu-tions, and all dilutions were then done withnitrogen-purged water. This differs from theEDTA in pH 7 buffer shown in the results ofHumphrey et. al. Johnston et. al. similarlyconducted their experimentation in pH 7buffer, but neglected to indicate sulfitedegradation. Nevertheless, there is stilluncertainty over the time dependency of thenitrogen-purged DTNB-sulfite reaction. Theabsorbance of MNB still decreased signifi-cantly after a period of about 20 minutes.There is a possibility that UV light willdegrade MNB or promote a side reaction. Ina DTNB-sulfite reaction placed in separatecuvettes, one in the dark, and one in UVlight, MNB in the cuvette in the presence of UV light had a significantly lowerabsorbance. Nevertheless, all reactions wereconducted expeditiously.

Application. Other existing techniquesfor sulfite analysis are either very expensive(such as analyzing the SO2 gas producedunder acid treatment) or have limited accu-racy and utility (such as iodine-iodate titra-tion). The present technique, however, couldbe translated into a simple color chart com-parison for field use. One drawback to theuse of DTNB is that it will react with anyother compounds present that contain thiolgroups; yet, in some interesting cases suchas in the water boiler industry, other suchcompounds are absent, and this techniquewould provide a reliable, inexpensive, andfield-usable method for determining sulfiteconcentration.

Boiler water is normally basic (pH 8–11),and the added sulfite (20–80ppm) and phos-phate (30–80ppm) prevent scaling or corro-sion on the walls of the boiler tanks. Thereducing agent, sulfite, acts as an oxygenscavenger, forming the sulfate ion and pre-venting electrochemical side reactions.

Plot of MNB Absorbance vs Sulfite Concentration

y = 0.186x + 0.0386

R2

= 0.9999

0.000

0.2000.400

0.6000.800

1.0001.200

1.4001.600

1.800

0 2 4 6 8 10

Sulfite (ppm)

MN

B A

bs.

(41

0nm

)

Reports Volume 8, 2003 43MURJSadegh and Schreck

Boiler water also does not contain any othercompounds that may skew the results. Themethod was used to test the boiler waterwith sufficient accuracy. Methods employedwere the filtration of the boiler water sedi-ment and dilution (by a factor of 10) tobring the sulfite levels in the desired rangeof 0.8–8ppm. The results of the DTNB-sulfitemethod were corroborated with the HachInc. detector kit, which is accurate at highersulfite concentrations. This showed that theDTNB-sulfite method is applicable andadvantageous in the water boiler industry.The method is currently being researched foruse in other industries, such as food andwine.

1. EPA Methods for ChemicalAnalysis of Water and WastesMethod. 377.1 (1983).

2. G.L. Ellman Arch. Biochem.Biophys. 82, 70–77 (1959).

3. D.L. Klayman and R.J. Shine.Quart. Rep. Sulfur Chem., 3, 190(1968).

4. R. E. Humphrey, M. H. Ward, W.Hinze Anal. Chem. 42, 698–702(1970).

5. J. B. Johnston, K. Murray, andR. B. Cain Antonie vanLeeuwenhoek. 41, 493–511(1975).

6. N. P. Luthra, J. D. Odom, and R.B. Dunlap Anal. Biochem. 117,94–102 (1981).

7. R.W. Huffman and D.M. Brown.J. Org. Chem. 56, 6,477–6,479(1991).

8. P.W. Riddles, R. L. Blakely, andB. Zerner Meth. Enzym. 91,49–61 (1983).

Reports44 Volume 8, 2003MURJ


Detection of Cotininein Blood Plasma byHPLC MS/MS

Oneil Bhalala

Abstract

Tobacco smoking is a major killer in the United States and is attributed to approxi-

mately 434,000 deaths per year. Primary and secondary exposure to tobacco and

tobacco smoke can be monitored by measuring cotinine levels in blood, urine, as

well as other matrices. This article describes a HPLC MS/MS assay to detect low concentra-

tion levels of cotinine in blood plasma. The assay was developed at Children’s Hospital,

Boston, and thus it was specifically designed for use with young children. This assay allows

for high throughput and turnaround because it does not use a column-based purification

process; it is also fairly inexpensive, using common laboratory reagents. Upon completion

of the study, the concentration ranges were found to be accurate from 0.l to10.0 ng/mL. The

limit of quantitation was calculated to be 0.2 ng/mL (CV% < 20%, accuracy range ± 20%).

The HPLC MS/MS assay is now ready for comparison tests with the ELISA test using patient

plasma samples.

Introduction

About one in five U.S. deaths are a result of tobacco usage.1 It is one of the main causes

of premature death in the country. Tobacco use, mainly in the form of cigarette smoking,

has been shown to increase the chances of various types of mouth and lung cancers, car-

diovascular disease, and emphysema.

The harmful effects of tobacco can be seen in both active and passive smokers. Active

smokers are those who smoke tobacco and tobacco products, while passive smokers are

those exposed to tobacco smoke. Exposure of nonsmokers includes second-hand smoke

from the air, as well as fetus exposure due to maternal smoking. Passive tobacco exposure

is a major health problem in the United States, and is classified as a Group A carcinogen

by the Environmental Protection Agency. Due to the magnitude of this health problem,

analysis of passive tobacco smoke has increased over the years.

Oneil Bhalala

Reports46 Volume 8, 2003MURJ Oneil Bhalala

Studies have shown that nicotine andcotinine are present in both active and pas-sive smokers alike. Nicotine (Figure 1) is anatural product of tobacco, and it is presentin the body with a half-life of 1–2 hours.Cotinine (Figure 2), on the other hand, is ametabolite of nicotine with a half-life of18–20 hours.2 The longer half-life makescotinine a more stable and prominent com-pound in the human body than nicotine.This makes it a more desirable compound, asit is easier to analyze. The objective of thisstudy was to develop an assay to detect andquantify cotinine levels in blood plasma.

The primary patient population ofChildren’s Hospital, Boston, consists of ado-lescents and young teenagers. Given thispatient demographic, any samples that thehospital receives would be from passivesmokers. Therefore, the cotinine levels intheir blood (<10 ng/mL) would be muchlower than in active smokers (≥10 – 15ng/mL).3 Previous cotinine assays were notsufficient because the extraction procedureswere too long and complicated, the limits ofquantitation were too high, or the samplematrix was incompatible.4 An inexpensiveassay with a high throughput, a low limit ofquantitation, and plasma as the samplematrix was designed to meet these needs.

Doctors at the hospital could analyzepatients’ cotinine levels quickly and accu-rately using the designed assay. By havingthe results back within a few working days,doctors could then promptly tailor treat-ments to the patient’s needs and condition.

In addition, this assay greatly enhances theability to detect cotinine levels in youngchildren and teenagers.

MethodA high-throughput, inexpensive, and

accurate assay was developed to meet theneeds of Children’s Hospital, Boston. Theassay utilized the PE SCIEX API 3000 withthe TubroIon Spray HPLC MS/MS (high-per-formance liquid chromatography tandemmass spectrometer) machine owned by thehospital to detect the cotinine levels inblood plasma. Before the sample could beinjected into the HPLC MS/MS, the cotininewas first purified and separated from therest of the plasma. The assay was dividedinto the following five steps: sample acqui-sition, standards creation, sample proteinprecipitation, sample reconstitution, andHPLC MS/MS configuration and injection.

Sample AcquisitionA blood sample was acquired from one of

the workers in the Hematology Lab atChildren’s Hospital, Boston. The blood sam-ple was centrifuged at 14,000 rpms for 10minutes to separate out the plasma. Theplasma was then transferred to a clean tubeand stored at -20° C for preservation. Beforeeach use, the plasma sample was thawed atroom temperature and certain amounts weretransferred to other tubes used for theexperiment. The remainder of the samplewas placed back into a -20° C freezer forstorage.

Standards Creation Blood plasma samples were equilibrated

with a D3 – cotinine Internal Standard(ISTD). 25 µL of plasma of various cotinineconcentrations were mixed with 25 µL ofISTD in 1.5mL Eppendorf tubes. The follow-ing are the cotinine concentrations thatwere used: 0.0, 0.1, 0.2, 0.3, 0.4, 0.5, 1.0,2.0, 5.0, and 10.0 ng/mL. These standardswere used to measure assay and machineaccuracy and recovery during HPLC MS/MSanalysis.

Sample Protein Precipitation500 µL of Acetonitrile was mixed into

each tube to precipitate out heavy proteins.The mixtures were vortexed for 30 seconds

N

NCH3

Figure 1. Chemical structure of nicotine.

N

N

0

CH3

Figure 2. Chemical structure of cotinine.


and then centrifuged for 4 minutes at14,000 rpms in order to form a cell debrispellet. 500 µL of the supernatant was thenplaced in clean glass test tubes and placed ina N2 vaporizer for 15 minutes to evaporatethe liquid from the supernatant.

Sample ReconstitutionEach glass test tube was reconstituted

with 150 µL of MeOH to dissolve the coti-

nine and any remaining proteins. The tubeswere again vortexed for 30 seconds to fur-ther precipitate out any remaining heavyproteins. The solution in each tube wastransferred to 150 µL vials for HPLC MS/MSanalysis.

HPLC MS/MS Configuration and InjectionThe buffer solution used for the assay was

10 mM Ammonium Acetate in 30% MeOH.

Oneil Bhalala

Figure 3. Calibration curve from 7-29-02. R-Value = 0.9993.



Figures 3 to 5 show the calibration curves from thethree analysis days. As demonstrated, all three curveswere linear from 0.0 to 10.0 ng/mL of cotinine con-centration. This range covered both the high and

low ends of cotinine concentrations. More impor-tantly, the R-value from each of the days was greaterthan 0.9990, as desired.


20 µL of each sample was injected into theHPLC MS/MS at a flow rate of 200 µL/min.The analysis time was set for 2 minutes. Thetemperature was set to 450° C. The Q1/Q3ratio for cotinine is 177/80, and the ratio forD3 – cotinine is 180/80.

ResultsSamples were prepared on 7-29-02 and

analyzed on 7-29-02, 7-31-02, and 8-1-02for accuracy and inter- and intra-day preci-sion. A set of samples was created for intra-day analysis and another set was created forinter-day analysis. This enabled us to testfor HPLC MS/MS accuracy as well as inter-and intra-day precision. The informationobtained from these analyses helped deter-mine the detection and quantification abili-ties of the newly developed assay.

Calibration CurveStandard samples were prepared with

cotinine concentrations of 0.0, 0.1, 0.2, 0.3,0.4, 0.5, 1.0, 2.0, 5.0, and 10.0 ng/mL. Theseconcentrations allowed us to test the accu-racy and precision of both the high-endconcentrations (5.0 – 10.0 ng/mL) and thelow-end concentrations (0.1 – 0.5 ng/mL) aswell as to identify the limit of quantitationof the assay. Each sample also contained10.0 ng/mL of ISTD (D3 – cotinine). Thisinternal standard aided in determining theaccuracy of both the assay and the machine.

The data from the standard samples wasused to construct a calibration curve foreach testing day. The calibration curve waslinear, with a “1/x” weighting. A regressionvalue (R-value) greater than R = 0.9990 wasdesired. Figures 3 to 5 show the calibrationcurves from the three days of analysis.

Inter-Day PrecisionOne sample each of 0.5 and 2.0 ng/mL

cotinine containing 10.0 ng/mL of ISTD wasprepared and run three times each in theHPLC MS/MS. The data was used to calcu-late the standard deviation, mean, and cor-relation value (CV%). The CV% was calcu-lated by the quotient of the standard devia-tion and mean. A CV% of less than 20% wasdesired. Table 1 contains the inter-day pre-cision data from the two types of samples.

Intra-Day PrecisionSamples were prepared with 0.5 and 2.0

ng/mL of cotinine containing 10.0 ng/mLISTD. These samples were run on 7-29-02,7-31-02, and 8-01-02 to determine theintra-day precision. The standard deviation,mean, and CV% were calculated from thedata. Again, a CV% of less than 20% wasdesired. Table 2 contains the intra-day pre-cision data for the two types of samples.

AccuracyThe recovery rate for the 0.5 and 2.0

ng/mL was calculated by taking the quotientof the measured cotinine concentration andprepared cotinine concentration for each ofthe samples. The desired recovery range wasbetween 80% and 120%. Samples betweenthis recovery range were deemed accurate.Table 3 provides the recovery range for eachof the samples.

Limit of QuantitationThe limit of quantitation is determined as

the lowest concentration of cotinine that hasa CV% of less than 20% as well as a recov-ery rate between 80% and 120%. The analy-sis showed that 0.2 ng/mL of cotinine wasthe limit of quantitation for this assaybecause it met these criteria. The only lowerconcentration, 0.1 ng/mL, did not meetthese criteria.

DiscussionThe goal of this project was to develop an

assay that would detect and quantify thelevels of cotinine in blood plasma. Giventhat the assay would be used at Childern’sHospital, Boston, it also needed to be sensi-tive—have a low limit of quantitation as wellas high throughput capabilities. We believethat an adequate assay was developed.

Table 1. Inter-Day precision of 0.5 and 2.0 ng/mLof cotinine. Table 1 provides the overall valuesobtained from the inter-day analysis of 0.5 and 2.0ng/mL of cotinine. As shown, the CV% is well belowthe 20% maximum for both samples. The 0.5 ng/mLsample has a CV% of 3.75, while the 2.0 ng/mLsample has a CV% of 11.35%.

Sample 0.5 ng/mL 2.0 ng/mL

S.D. 4.93E+05 1.25E+06

Mean 1.32E+07 1.10E+07

N 3 3

CV% 3.75 11.35

Reports Volume 8, 2003 49MURJOneil Bhalala

The developed assay uses a HPLC MS/MSwithout the need of a column to purify thesample. This significantly reduces the timeneeded per sample. If a column was to beused, the run time for each of the patientsamples would easily triple or quadruple,because the sample would enter and exit thecolumn and the column would need clean-ing between sample runs. Previous assaysperformed in the laboratory demonstratedthat a column did not improve the resolu-tion or sensitivity of the assay. Therefore, acolumn was not included as part of the pro-posed assay.

In addition, blood plasma is the mostcompatible matrix for the target patients(young children and teenagers) at the hospi-tal; it is very hard to obtain enough salivaor urine samples from infants and toddlers.Also, this assay requires the use of only 25µL of plasma, thus only a small amount ofblood needs to be drawn from the patient.

The sample matrix also needs to be puri-fied before it can enter the HPLC MS/MS.The extraction method that was developedfor the assay is sufficient in its purificationabilities. It precipitates out the heavy pro-

teins with the use of Acetonitrile, a commonand easily obtainable laboratory reagent.The other steps in the extraction processfurther purify the sample, such as the N2

evaporation. This assay does not damage theHPLC MS/MS machine by injecting unpuri-fied samples, thereby safely allowing forrepeated analyses.

The results showed the assay’s limits ofquantitative capabilities. An upper bound of10.0 ng/mL and a lower bound of 0.2 ng/mLof cotinine were established. These were theupper and lower limits in which the assaywould quantitate effectively; any concentra-tion outside of this range will produce unre-liable results using the developed assay.Because the hospital deals primarily withchildren, a modified upper bound of 2.0ng/mL was created—only significant passivesmokers contain over 2.0 ng/mL of cotininein their blood, as demonstrated by previousstudies. An example of a significant passivesmoker is one who is constantly exposed toenvironmental smoke, such as a waitress.The hospital is mainly concerned with rela-tively low passive smokers who have plasmacotinine levels much lower than 2.0 ng/mL.

Table 2. Intra-Day precision of 0.5 and 2.0 ng/mL of cotinine. Table 2 provides the overall valuesobtained from the intra-day analysis of 0.5 and 2.0 ng/mL of cotinine. As shown, the CV% values for each ofthe samples were under the maximum level of 20%. These matched the desired results for this analysis.

Sample 0.5 ng/mL 2.0 ng/mL

S.D 1.04E+05 1.62E+04 6.64E+05 1.05E+05 2.53E+05 5.07E+05

Mean 5.49E+05 1.92E+05 4.77E+06 2.59E+06 2.47E+06 4.81E+06

N 7 9 9 7 9 9

CV % 18.94 8.42 13.91 4.05 10.25 10.54

Table 3. Concentration of cotinine and recovery rate for 0.5 and 2.0 ng/mL of cotinine. Table 3 showsthe recovery rates of the 36 analyzed samples. As shown in the table, all 18 of the 0.5 ng/mL cotinine sam-ples had a recovery rate between 80% and 120%. Based on the criterion established before, these 18 com-pounds were determined to be accurate. For 2.0 ng/mL cotinine samples, 15 of the 18 compounds had arecovery rate between 80% and 120%. Therefore, 15 of 18 2.0 ng/mL cotinine samples were accurate.

0.5 ng/mL (N=18) 2.0 ng/mL (N=18)Conc. Recovery Conc. Recovery Conc. Recovery Conc. Recovery

0.464 92.80 0.425 85.00 1.410 70.50 1.870 93.50

0.456 91.20 0.433 86.60 1.470 73.50 1.920 96.00

0.468 93.60 0.438 87.60 1.430 71.50 1.900 95.00

0.505 101.00 0.493 98.60 1.870 93.50 2.030 101.50

0.492 98.40 0.489 97.80 1.850 92.50 2.050 102.50

0.509 101.80 0.487 97.40 1.810 90.50 2.140 107.00

0.493 98.60 0.477 95.40 1.680 84.00 2.030 101.50

0.470 94.00 0.483 96.60 1.700 85.00 2.040 102.00

0.491 98.20 0.492 98.40 1.750 87.50 2.110 105.50


It is the hospital’s goal to identify thosepatients who have a low, yet significant,exposure to environmental smoke. Thisassay allows the hospital to do exactly that.

The experimental data showed that thesamples were linear within the range of 0.2to 2.0 ng/mL of cotinine. This is substanti-ated by the R-value of the calibration curvebeing greater than 0.9990 and the CV%being less than 20%. From the data, we con-clude that our assay is also accurate from0.2 to 2.0 ng/mL.

The data gathered from the analyses indi-cates that we accomplished our goal ofdeveloping an assay that is inexpensive,sensitive (low limit of quantitation), and hashigh throughput capabilities.

However, before this can be used as astandard patient assay in the hospital, acomparison test needs to be carried out.First, samples need to be collected from hos-

pital patients. The cotinine concentrationsshould then be quantitated using the devel-oped assay and the ELISA test, the standardassay currently used. If this comparisonshows that the newly developed assay is justas accurate as the ELISA test, then thehospital will be able to quantify cotininelevels in patient samples using the newassay. This study supports the outcome thatthe developed assay will be as accurate asthe ELISA test.

AcknowledgmentsI would like to thank Dr. Nader Rifai,

director of Clinical Chemistry at Children’sHospital, Boston, who allowed me to spear-head the project. I would also like toacknowledge the help of Charlie Bu, TerryLaw, and Masa Sakamuto as well as Dr. NealLerner for his help in editing this paper.

References

1. Centers for Disease Control andPrevention. Cigarette Smoking –Attributable Mortality and Yearsof Potential Life Lost – UnitedStates, 1990. Morb Mortal WklyRep 1993; 42: 645–9.

2. Benowitz NL, Jacob P. Nicotineand Cotinine EliminationPharmacokinetics in Smokersand Nonsmokers. ClinPharmacol Ther 1993; 53:316–23.

3. Watts, Langone, Knight et al.Cotinine Analytical WorkshopReport: Consideration ofAnalytical Methods forDetermining Cotinine in HumanBody Fluids as a Measure ofPassive Exposure to TobaccoSmoke. Environ Health Perspect1990; 84: 173–82.

4. Bernert, Turner, Pirkle et al.Development and Validation ofSensitive Method of SerumCotinine in Smokers andNonsmokers by LiquidChromatography/AtmosphericPressure Ionization TandemMass Spectrometry. Clin Chem1997; 43: 2281–2291.


Probing Solar PanelDesign Systems

Alexander C. De Feo

Current drawbacks of utilizing solar energy are high initial costs and the inconven-

ience of high maintenance. Although the use of solar energy has been steadily

growing, it is only 0.08 percent of the energy-producing market within the United

States.1 One of the growing areas in solar research has been the mounting

brackets of solar arrays. The goal of this design project is to develop inno-

vative methods for producing an inexpensive mounting system to reduce

initial costs of a photovoltaic solar grid. Limited engineering research in

this area has kept these panels retailing at the average cost of $600 per

panel, while the mounting system costs per panel are over $100. Thus, this

article will discuss possible cost-effective solutions for mounting systems.

In addition, experimental methods for testing the panels will be suggested.

After reviewing current designs available on the market, an improved

design was established by following these specific functionality require-

ments: (1) a mounting system to install on commercial and industrial flat

roofs; (2) a simple and flexible design to facilitate mass production; (3) injected molded

recycled plastic rather than traditional steel structures, to reduce corrosion on the system;

(4) a simple and quick installation method such that individual panels can be easily

removed; (5) a freestanding system that eliminates the

need to penetrate roof seals; and (6) a wind rating to con-

form to building code specifications.

The following sections will discuss four possible

designs of solar mounts for different applications. The

Modular design meets all of the above functionality

requirements and is intended for heavy winds. The sec-

ond design, Corners, is more flexible; however, it will

not protect the panels under heavy loads. For greater

solar conversion efficiency, the Tilt design yields the best

results. The last design, Awnings, aims to be visually

exposed and uses novel materials.

Alexander C. De Feo

Image courtesy of SOHO/EITconsortium. SOHO is a project ofinternational cooperation betweenESA and NASA.

“Solar energy has

an untapped potential

…current efficiency

conversion from

sunrays to electricity

is only 12%"

Reports52 Volume 8, 2003MURJ Alexander C. De Feo

Design I — ModularThe initial design is simple yet robust,

and is composed of only two elements, afoot and slat. As shown in Figure 1, the pan-els are mounted to slats that are joined byfeet. The grooves in the foot componentshown in Figure 2 constrain the slats fromrotation. A single stainless steel bolt con-strains the slat in the vertical as well as lat-eral axes.

As with traditional mounting systems, thepanels are glued to the slats. The installationcosts are greatly reduced because there is noon-site customization for oddly shapedroofs; that is, the panels can be arranged inseveral configurations to adapt to roofobstructions. When mass-produced, the footand slat components cost less then $3 each,and the total cost per panel is about $18. Forsmaller-scale production runs, a wood/plasticcomposite lumber could be used as an alter-nate material to injected molded plastic. Thiscomposite material is corrosive-resistantand can be cut and drilled similarly to wood.

This design meets all of the functionalityrequirements named previously, except forthe 90 mph wind rating of Massachusettsbuilding codes, which has yet to be proven.Due to the complications in theoreticallymodeling the system, an experimentalmodel has to be made of air flowing overthe top of commercial buildings, and the liftand drag forces on the system must bemeasured. Because the weight of the panelsis not sufficient for keeping the system sta-tionary, the corners of the array have to beconstrained by cable stays that can bemounted to the side walls of the roof. The

aims of this experimental test are to deter-mine the necessary constraining forces aswell as to find any system complicationssuch as resonance frequencies that mayarise in storm conditions. Once the forcesare calculated, finite element analysis soft-ware can be used to specify the contactstresses in the glass panels.

The high wind-speed conditions on theroof of a commercial building will be simu-lated at the MIT water tunnel. The schematicin Figure 3 shows a delrin structure simulat-ing the flow of air over a commercial build-ing. The tunnel is 1.2 meters long and has across section of 0.5m x 0.5m. The waterflows over the bulkhead to simulate thewalls around the roof of a commercialbuilding. The one-third-scale panel in theexperiment is a stainless steel plate ofdimensions 0.64m x 0.32m x 0.008m. As thewater flows over the bulkhead, water canflow above and below the panel becausethere is a clearance of 0.01m to simulateventilation under a panel. To vary the posi-tion of the panel, the distance between the

Figure 1. Modular array of seven panels.Figure 2. Parts identification of foot (A), slats(B), bolts (C), and panels (D).

Figure 3. Experimental schematic cutaway oftunnel section. The water flows from left toright at a maximum velocity of 5m/s. The dis-tance between the panel and bulkhead can beadjusted up to 0.64 meters.

Reports Volume 8, 2003 53MURJAlexander C. De Feo

panel and bulkhead can be adjusted up to0.64 meters.

Using nondimensional analysis, themedium and scaling properties can be simu-lated for the actual conditions. The Reynoldsnumber in Equation 1 can be used to equatethe actual and simulated conditions.

Re = r × v × Lm

Equation 1. Reynolds number, where r is thedensity, v is the velocity, L is the width of thepanel, and m is the viscosity.

The forces on the panel will be measuredusing a six-degree-of-freedom dynometerthat is positioned below the water tunnel.The measured vertical and horizontal forcescan be scaled appropriately by using thecoefficient of lift and drag in Equations 2and 3, respectively:

CL = FL

0.5 × r × v2 × A

CD = FD

0.5 × r × v2 × A

Equations 2 and 3. Lift and drag coefficients,where FL is the vertical force, FD is the horizon-tal force, v is the velocity, and A is the area ofthe panel perpendicular to the force.

Design II — CornersFor every design there are revisions for

different applications. In this case, the cus-tomer has dictated the creation of the threesubsequent designs. MIT facilities recentlyreceived a grant to install solar panels oncampus.2 The supplier of solar panels usesvarious panel sizes that are not compatiblewith the Modular design. It would thereforebe necessary to have shorter or longer slatsdepending on the specific size of the panelas there is not one industry standard. Thisissue spurred the idea of having a configu-ration that is not constrained by the specificdimensions of the panel. Removing the slatsand modifying the foot component devel-oped a design whereby the panel is onlyconstrained in the corners. As shown inFigure 4, the corners of the panel are sand-wiched in between two symmetric parts.

To reduce possible concentrated stresseson the extremely inelastic glass panels, athin layer of neoprene can be insertedbetween the plastic and glass. The installa-tion efforts are also reduced with this designbecause the gluing phase is eliminated. Thedrawback is that a small portion of the pho-tovoltaic cells are blocked off from sunlight.In addition, the system is not as robust asthe first: The slat backing does not fullysupport the panels. Under high winds, vibra-tions may dangerously strain the panels,reducing the life of the photovoltaic cells.

Design III — TiltThese first two designs were economi-

cally driven, streamlined systems. This nextdesign aims to variably tilt the panels to“follow the sun,” which can increase photo-voltaic efficiency up to 40 percent. As thesun travels from east to west, tilting thepanels in one axis will increase efficiency by20 percent. In the northeast region of theUnited States, the sun also moves north andsouth, yielding another 20 percent increasein efficiency if the panels are tilted in a

Figure 4. Depiction of bottom corner componentwith panel. The corresponding symmetrical cor-ner component is not shown.

Figure 5. Array of panels with corner designdepicting partial installation.

Reports54 Volume 8, 2003MURJ Alexander C. De Feo

second axis. The modular design with slatscan be altered by adding two moving actu-ators in adjacent corners of a square array.These actuators could be comprised of eithera lead screw configuration or an air pistondriven by an air compressor, depicted inFigure 6.

The tracking control system could becomposed of either an optical tracker or ascheduled position based on the given dayof the year. Because these panels are ele-vated, there are additional bending stressesdue to the weight of the panels. This willdictate brackets in addition to the foot com-ponents or even slats that extend the lengthof the array. The limiting factor of this moreglamorous design is based on the addedfixed cost of the movable parts as well asthe marginal costs of the added energy con-sumption.

Design IV — AwningsBesides the economic barriers, solar

energy awareness has been extremely lim-ited. One of the aims of the MIT solar panelproject is to make the panels visible toeveryone on campus. Consequently, the firstthree designs are limited to isolated roofs. Afourth design would place the panels on thesouth sides of buildings to resemble awn-ings. The construction would be a wood/plastic composite lumber as supportingbrackets, with alternative foot junctions.

ConclusionThe creation of better solar panel mount-

ing systems has been a growing develop-ment in the solar energy industry becausethey provide electricity during peak middayusage and can be installed on small scales.This independence decreases the strain on

electricity grids and eliminates the energyloss of distributed energy.

The current efficiency conversion fromsunrays to electricity is only 12 percent, andonce more research is invested in develop-ment this efficiency can increase. If the ini-tial cost of installing a solar panel array isdecreased due to improved mounting sys-tems, more solar packages will be pur-chased, increasing the market size. As thesolar energy industry expands, more profitscan be cycled back into R&D to increase thephotovoltaic efficiencies. The MassachusettsRenewable Energy Trust has given MITfacilities a grant to install solar panels onand off campus to increase the efforts ofpromoting solar energy. Ultimately, produc-ing a solar panel package that is competitiveto fossil fuels is a long-term goal that canonly be achieved once this novel technologybecomes mass-produced.

AcknowledgmentsI would like to thank Island Energy

Solutions for the inspiration of improvedsolar panel mounting systems. The initialfunding for this project was graciously pro-vided by the Paul E. Gray (1954) EndowedFund for UROP. Advising Professor Alex-andra Techet and water tunnel engineerRichard Kimball gave me invaluable guid-ance. Laxmi Rao from MIT Facilities (Massa-chusetts Renewable Energy Trust) inspiredthis journal article.

References

1. Energy InformationAdministration. RenewableEnergy Annual 2000. March2001.

2. MIT News Release. Grant fundssolar power project on and offcampus. Oct. 24 2002.

Figure 6. Schematic of a four-panel tilted array.Pneumatic actuators are positioned in two of the cor-ners (right side).

Figure 7. Awning mounting system installed onthe south side of a concrete building for maxi-mum sun exposure and visibility.


EnvironmentalAccounting:

Project Financing and StrategicInteractions

Angela S. Bassa

Abstract

This paper discusses the Kyoto Protocol propositions for reducing global warming by

financing appropriate projects in developing countries through the Clean

Development Mechanism. The paper shows that care must be taken to avoid the

global loss of social well-being.

Introduction

The Kyoto Protocol, agreed upon in December 1997, sets legally binding limits on green-

house-gas emissions for developed countries.4 Each country must achieve at least a 5 per-

cent reduction from 1990 levels between 2008 and 2012, a significant departure from cur-

rent trends. The protocol indicates specific areas for action and sets up a number of mech-

anisms for international cooperation. One such mechanism is called the Clean Development

Mechanism (CDM). The CDM is intended to attenuate the foreseen economic impact of an

abrupt reduction in the levels of emissions and to encourage cost-efficiency by establish-

ing an international market for the negotiation of Greenhouse-Gas Emissions (GGEs)

between developed and developing countries.

The CDM, as proposed in the protocol, allows for transactions of Credits of Emissions

Reductions (CERs). These credits result from exchanges between countries and may be used

by developed countries to achieve their GGEs reduction quotas. A list of the developed

countries that were original signatories and were part of the Kyoto Protocol’s Annex-I can

be found in the Appendix, together with their relative levels of GGEs in 1990.

The Kyoto Protocol foresees that developing countries will participate solely through the

CDM. Because the greenhouse effect results largely from the emissions of the developed

countries, there is an effort to guarantee that the economic growth of these nations will not

be curtailed by restrictive environmental policies. Also, imposing environmental controls

Angela S. Bassa

Reports56 Volume 8, 2003MURJ Angela S. Bassa

that result in economic costs to the develop-ing countries would be impractical withoutsome type of compensation.

CERs may be bought by a developedcountry or by a firm with headquarters in adeveloped country. Given the large volumeof greenhouse gases emitted by these coun-tries (about 80 percent of the total), it isapparent that, economically, the greaterbeneficiaries of the CDM will be developingcountries with industrial parks that are notsubject to strong environmental legislation.These developing countries could generategreat volumes of CERs competing directly inthe International Trading Quotas market (ITQ),the future emissions quota internationalmarket. The great appeal of these CERs isthat the costs of emissions reduction indeveloping countries, it is assumed, are sub-stantially lower than in developed countries.

Intuitively, the primary way in whichthese exchanges take place may seem to bevia direct CER trading. However, direct CERtrading with developing countries is not theonly way that CERs could be passed ontodeveloped countries. Firms located in devel-oped countries may also finance their emis-sions-reduction projects directly, instead oftrading quotas in an international market.The economies of the developing countriesinvolved would have subsequent advantagesfrom directly participating in a partnershipproject with a foreign firm from a developedcountry. Firstly, capital and technologywould be attracted and would finance partof the development cost. Moreover, the proj-ects would lead to an improvement in theenvironment of the affected areas. By takinginto consideration the nature of developingeconomies and the large volume of CERspossibly demanded by developed countries,the question of who finances the projects foremissions reduction becomes fundamental.Another aspect to consider is that whenfirms from developing countries come tofinance their own projects of emissionsreduction to get CERs and offer them forsale, the developing economies are the onesfinancing the environmental policies indeveloped countries. This financing is amor-tized by the payment of the CERs.

There may be additional costs imposed onfirms in developed countries for implement-ing this alternative solution. Contrary to ini-tial expectations, as Tietenberg points out,it is possible that the firms headquartered indeveloping countries have no interest inallowing their emissions to be reduced (evenwithout cost), because these firms expecttheir market share and their profits to in-crease due to the change in cost structure ofthe other firms that reduce their emissions.7

This paper develops a model in whichsuch a result always emerges, and it does soas a consequence of definitive strategies oftwo firms: Firm 1 and Firm 2. Firm 1 islocated in a developed country, and may ormay not be interested in developing a proj-ect through the CDM. Firm 2 is located in adeveloping country which may or may notaccept this partnership. The paper alsodevelops a means of incentive in which thefirm that proposes a project may induce abetter situation for itself, affecting the deci-sion of the other firm. With such a mecha-nism, the proper agency of environmentalcontrol in a developed country can deter-mine a priori whether or not the project canimplemented.

In Section 2, a few possible hypothesesare presented and refuted; one hypothesis isaccepted and carried out. In section 3, theaccepted model is established and itsassumptions explained. In Section 4, theeffects of the model are analysed and pre-sented and possible consequences evaluated.In Section 5, an alternative conjecture forfuture work is proposed; it takes into con-sideration the possibility of externalresource financing.

HypothesisConsider an economy with two countries

and two firms. Say Firm 1 is located inCountry A, Firm 2 is located in Country B,and both firms are able to do business in thetwo markets. Country A is developed,whereas Country B is in development. Theimpact of the CDM on the strategies of thefirms is considered here, where the chosenstrategy defines the production level. Beforethe adoption of the CDM, both firms aresupposed to be maximizing profits given thecompetitor’s strategies. To simplify, assume

Reports Volume 8, 2003 57MURJAngela S. Bassa

an economy in which information is full,perfect, and symmetrical—a reasonable andconvenient assumption in the Internet age.

In order to study the impact of a jointproject of emissions reduction on the strate-gies of the firms, consider the following sit-uation. Given the necessity to control spe-cific GGEs associated with the production ofsome good in Country A, the agency ofenvironmental control in this countrydecides to transfer the reduction costs toFirm 1, which is responsible for the GGEs.Firm 1 is assumed to be using the best pos-sible technology for reducing emissions.Two possibilities exist: The agency charges atax on Firm 1 to finance the purchase ofquotas in the ITQ market; or Firm 1 devel-ops a project in partnership with Firm 2, inCountry B, that is not using the best tech-nology available for emissions reduction.For simplicity, it is assumed that any projectbeing considered for implementation willgenerate the exact amount of credits neces-sary to meet the expected reductions inCountry A.

One hypothesis is that the market struc-ture forces the production decisions to betaken in independent and simultaneousform, in a classic model of Cournot compe-tition.6 The existence of technological differ-ences and input barriers cannot explain amarket structure of this type when bothfirms directly compete in the markets ofboth countries. The only difference it allowsbetween the firms is the cost due to pollu-tion control. This counterexample can befound, for instance, in the paper and cellu-lose industry.1

Another hypothesis is that technologyand plant scale are distinct in such a waythat Firm 1 has a bigger plant and is moretechnologically intensive, with inferior mar-ginal costs. In this way, Firm 1 could exertmarket pressure to determine, directly orindirectly, the prices of the good in the mar-ket. This situation is interpreted through thetypical Stackelberg competition model6

instead of a Cournot model. This second hypothesis occurs even when

the technologies are similar, as long as theproduction capacities of the firms are dis-tinct. For instance, even though production

costs are basically identical, one of the firmsis able to raise offers in both markets inorder to keep equilibrium marginal pricesbelow production costs in a strategy ofdumping. An example of these practices canfrequently be found in the Chinese footwearindustry, which counts on the state tofinance its actions.2

This kind of situation is not an anomaly.Moreover, this is a limit-situation: One firmimposes leadership in the market due solelyto its ability to use predatory behavior.However, strategies of dumping can becometoo costly, and their success depends oncosts and profits determined in a monopolistmarket. Therefore, it is probable that firmsthat operate such strategies do not do so,although they do exert some sort of leader-ship in the market, either in price or quan-tity, as treated in this paper.

In general, leadership in the market isjustified by scale economies or technologi-cal innovations. If such factors were incor-porated into the hypotheses, then theywould reduce the relative profits that Firm 2would get once Firm 1, the leader, reducedits participation in the market due to theimposition of environmental policies inCountry A. The model presented in Section3 uses limit-situations where market leader-ship can be exerted. In these situations, anequilibrium that benefits both countries canemerge by considering the strategic interac-tion between the firms.

ModelConsider Firm 1 to be a leader company

in a developed country, and Firm 2 a fol-lower in a developing country. Their strate-gic interactions in the market are treatedthrough a Stackelberg duopoly model.6 Theleader firm directly decides its profit maxi-mization strategy by predicting the reactionfunction of the other firm. The latter, inturn, maximizes its profits, given theleader’s chosen strategy.

Some assumptions are necessary so thatthe payoffs for both firms are determined:

1. The fixed costs for both firms areirreversible.8

2. The cost for Firm 1 to carry on the project inpartnership with Firm 2 is also irreversibleand equal to k.


3. The tax charged by the environmentalagency is applied to the level of productionof Firm 1 and is equal to d, where 0 £ d£1.

4. There is some type of restriction hinderingthe firms from establishing plants of produc-tion in the other country.

5. Both firms are profit maximizers.

Given the imposition of the emissionscontrol agency, Firm 1 in Country A con-templates the following strategies: Eitherpay the agency tax or implement the projectin partnership with Firm 2 in Country B.Firm 2, in turn, also contemplates twostrategies: Accept or reject a partnershipwith Firm 1. Given Assumption 4 above, thepossible strategies for Firm 1 are signifi-cantly reduced. In fact, as Motta and Thisse5

indicate, the imposition of environmentalstandards in a country can lead to the relo-cation of the plants to countries where envi-ronmental policy practices are less restric-tive. However, this situation only occurs inspecial circumstances, such as when thecosts of implementing a new plant are rela-tively low in relation to the costs of theenvironmental policies. The capital-inten-sive nature of most firms makes this situa-tion improbable.

For each country i, let Qi be the total pro-duction, let Pi be the price established forthe good being produced, and let ai be thestand-alone consumption. According toPindyck and Rubinfeld, the typical demandfunction is then:

We can assume that aA is greater than aB

because consumption in developed CountryA is likely to be larger than that of develop-ing Country B. The costs of production areconstant and equal to c for both firms.

The nature of a Stackelberg game isdynamic (Firm 2 plays after Firm 1), but theassumptions that the information is full,perfect, and symmetrical allow the payoff ofeach type of approach, by each firm, to becalculated. The game can then be reduced toits standard form, a reduced 2 x 2 payoffmatrix, through a refinement known asretroactive induction. The payoff matrix of atwo-firm game has rows labeled by Firm 1’sstrategies and columns labeled by Firm 2’sstrategies. The xyth entry of the matrix is the

payoff that accrues to each firm in the eventthat the “row player” uses strategy x and the“column player” uses strategy y. When ana-lyzing any game, we make the followingassumptions about both firms:

1. Each firm makes the best possible move.2. Each firm knows that its opponent is also

making the best possible move.

The strategy for Firm j is an ordered pairof quantities (qjA, qjB) sold in Countries Aand B, respectively. The reduced payoffmatrix is given by the profit associated withthe chosen strategies. The possible strategypairs related to the environmental policiesin Country A and the choice of Firm 2 re-garding the implementation of an emissionsreduction project are shown in Table 3-1.

Table 3-1. Dynamic Strategy Choices

Case 1 Firm 1 chooses to pay the taxFirm 2 would accept the projectimplementation

Case 2 Firm 1 chooses to pay the taxFirm 2 would not accept projectimplementation

Case 3 Firm 1 chooses to propose partnershipFirm 2 does not accept projectimplementation

Case 4 Firm 1 chooses to propose partnershipFirm 2 accepts project implementation

For comparison purposes, the equilibriumsolutions are given here prior to the imple-mentation of environmental policies. Sincethe algebra of the standard Stackelbergmodel is simple, only the strategies and theprofits associated are presented. Addition-ally, the production cost c is such that theamounts of equilibrium are strictly positive.

Each firm presents the following profitfunctions pi in their initial configurations:

(3-1)

(3-2)

Profits associated with equilibrium strate-gies pI* for each firm are given by

(3-3)

(3-4)

π∗1 =

(αA − c)2

8+

(αB − c)2

8,

π2 = PAq2A + PBq2B − c(q2A + q2B).

π1 = PAq1A + PBq1B − c(q1A + q1B),


Quantities Qi sold in each country i aregiven by

(3-5)

Case 1 in Table 3-1 occurs when Firm 1prefers to pay the tax, even when Firm 2would accept the implementation of a jointproject of emissions reduction. In this case,the profits pi are given by Equations 3-1aand 3-2a.

(3-1a)

(3-2a)

Recall that d is the tax charged by theenvironmental agency. Thus the updatedprofits for each firm associated with theequilibrium strategies pI* are given byexpressions 3-3a and 3-4a.

(3-3a)

(3-4a)

Given that in Case 1 Firm 1 pays the taxd, the amounts negotiated in each country iare given by

(3-5a)

By comparing Equation 3-5 with Equa-tion 3-5a, when Firm 1 opts to pay an envi-ronmental tax, the equilibrium quantitiesare reduced in both countries, and the pricesmust be raised relative to the original equi-librium. More importantly, Firm 1’s profitmay or may not decrease, while Firm 2’sprofit certainly increases.

Case 2 occurs when Firm 1 pays the tar-iff and Firm 2 does not accept project imple-mentation. The strategies and payoffs asso-ciated with this case are the same as in Case1. Therefore, when Firm 1 chooses to paythe tax, Firm 2’s decisions do not affect thefinal result of the game.

Case 3 occurs when Firm 1 prefers toimplement the project of emissions reduc-tion, and Firm 2 accepts the partnership.Firm 1 then pays for all costs of the projectk if there is a reduction of emissions inCountry B. In this case, the profit functionsare given by Equations 3-1b and 3-2b.

(3-1b)

(3-2b)

Because Firm 1 pays all the costs of theproject, which are fixed and equal to k, thestrategies and equilibrium payoffs do notdiffer from the initial situation, except inrelation to the profit of Firm 1. Profits foreach firm associated with the equilibriumstrategies pi* , taking into account this cost,are given by Equations 3-3b and 3-4b.

(3-3b)

(3-4b)

It follows that Firm 1’s choice modifiesneither the prices nor the equilibrium quan-tities in either country.

Case 4 occurs when Firm 1 prefers toimplement the project, but Firm 2 does notaccept the partnership. In this case, Firm 2’s

π1 = PAq1A + PBq1B − c(q1A + q1B) − κ,

Qi =34(αi − c) − δ

2.

Table 3-2. Payoff Matrix

Case 1

Firm 1: Pays tax

Firm 2: Accepts project

Case 2

Firm 1: Pays tax

Firm 2: Refuses project

Case 3

Firm 1: Proposes project

Firm 2: Refuses project

Case 4

Firm 1: Proposes project

Firm 2: Accepts project

π∗1 = (αA−c)2

8 + (αB−c)2

8 − κ

π∗2 = (αA−c)2

16 + (αB−c)2

16

π∗1 = (αA−c)2

8 + (αB−c)2

8 − δ2 [αA + αB − 2(δ + c)]

π∗2 = (αA−c)2

16 + (αB−c)2

16 + δ4 [αA + αB + 2(δ − c)]

π∗1 = (αA−c)2

8 + (αB−c)2

8 − δ2 [αA + αB − 2(δ + c)]

π∗2 = (αA−c)2

16 + (αB−c)2

16 + δ4 [αA + αB + 2(δ − c)]


decision affects game payoffs because whenit refuses the project, the best (and only)option for Firm 1 is to pay the tax. Thestrategies and equilibrium payoff in thiscase are identical to those indicated in Case 1.

Once payoffs for each case have beenestablished, the payoff matrix can beexpressed as shown in Table 3-2. Note thatthis matrix is formed using the assumptionthat the information is perfect, symmetrical,and full, and in this case, that each firm cananticipate the results that will take place,given its choices.

If there is a set of strategies with theproperty that no player can benefit fromchanging strategies mid-game, then that setof strategies and the corresponding payoffsconstitute a Nash equilibrium.9 Thus, theresultant Nash equilibria from this analysisare perfect in subgames. Each cell presentsthe profits of Firms 1 and 2, respectively,associated with each of their strategychoices.

The payoff matrix in Table 3-2 revealsthat the dominant strategy for Firm 2 is torefuse the partnership exactly when Firm 1considers the project without any type ofcost for itself. The strategy for Firm 1depends on the cost of the project in relationto the total cost of the tax: The bigger thedifferences between the emissions reductiontechnologies for the firms, the smaller thetax. A reduction in d, in turn, reduces k.Conversely, the bigger the differencesbetween k, aA, and aB, the bigger thecharged tax d. Still, no matter how muchFirm 1 earns with the implementation of theproject, the Nash equilibrium is achievedexactly when Firm 1 pays the environmen-tal tax and Firm 2 benefits from the smallermarket share taken by Firm 1, raising itsprofits in relation to the original equilib-rium.

Therefore, a clear loss of well-beingoccurs in Country A as a result of the reduc-tion in the profits of Firm 1. In Country B,there is a loss due to the price increase. AsFirm 2’s profits increase, it is necessary toverify which of the effects predominates.Since d is less than 1, its dimension relativeto consumption ai has to be significantlyreduced, indicating that the ensuing social

costs of demand reduction and priceincrease also have to be reduced. Only Firm2’s shareholders benefit from this result.Moreover, the increase in profit by Firm 2 isinferior to the profit loss of Firm 1; hence,the imposition of environmental policies inone of the countries causes a global loss ofsocial welfare. These results indicate that arestrictive environmental policy in thedeveloped country, in addition to a mecha-nism such as the CDM, instead of inducinga cost-efficient solution, would lead to asolution where there is loss of well-being.

The possibility that Firm 1 generatesemissions reduction savings in a less tech-nologically advanced country adds value tothe excess emissions of Firm 2; Firm 2 is theone that is on the receiving end of the proj-ects. In the model used here, this value isgiven by the extra profit that Firm 2 gets byinducing Firm 1 to pay an environmentaltax. Firm 2 only accepts the project if Firm1 offers a greater value than this extra profitfor the right to implement a project of emis-sions reduction. In this case, it can be saidthat Firm 2 sells its excess of emissions toFirm 1.

Incentive Strategies’ EffectsWe now consider strategies that induce

Firm 2 to cooperate when requested. As seenin Section 3, Firm 2 does not have anyincentive to cooperate and allow the imple-mentation of a project of emissions reduc-tion. The imposition of an environmentaltax on Firm 1 reduces its participation in themarket, and part of this loss is reverted asprofit to Firm 2. Allowing the project to becarried through, however, does not increaseits profit.

For Firm 1, however, the choice betweenthe project and the tax depends on the costsof each one. Inequality 4-1 indicates thatFirm 1 will always prefer the project thathas the lesser impact on its profit:

(4-1)

Inequality 4-1 does not have any effecton the result of the game; therefore, it isFirm 2’s choice that determines the resultantequilibrium. Note that the application ofInequality 4-1 determines the resultant pair


of strategies, regardless of the fact that thepayoffs associated with both of the twostrategies are the same. In order to incorpo-rate an incentive restriction to the problem,assume that Firm 2 agrees to allow theimplementation of a project, as long as thetax loss of its extra profits is compensated.Thus, Firm 1 could add the extra profit ofFirm 2 to the costs of the project k when itpays the environmental tax. Firm 1 wouldconsider the viability of the project depend-ing on the tax payment according toInequality 4-2 or, alternatively, Inequality4-2a.

(4-2)

(4-2a)

The idea is that Firm 2 accepts the projectwhen Inequality 4-2a is true. The mecha-nism of incentive shown above completelymodifies the resultant equilibria. The domi-nant strategy for Firm 2 is now to allow theimplementation, and Inequality 4-2a deter-mines game payoff. Assume that, for Firm 2,the implementation of the project withoutthe loss of extra profits is preferable to theextra profits alone, either because of localemissions reduction or for a better image ofthe firm in the market. In the case that Firm1 does not request the project, payoff forFirm 2 is indifferent.

When the inequality given by 4-2 or 4-2ais true, Firm 1 proposes the project to Firm2. For such a proposal to carry through, thedifferences between project cost and thecomponents dimension of stand-alone con-sumption for both markets ai, in addition tothe value of the environmental tax to becharged d, are determining.

For simplicity, denote the left side ofInequality 4-2a as f (d, k, aA, aB, c), or f̂ inshort. Whenever f̂ is less than 0, Firm 1 con-siders the project. An important pointbecomes visible from the graph of f̂ whenonly the environmental tax is fixed, that is(d1 < d2 < d3 < dn), as seen in Figure 4-1. Thesmaller the tax charged for particular valuesof k, the larger the probability that Firm 1prefers this mechanism.

In Figure 4-1 the parameters that deter-mine the shift to the right are the smaller

size of the components of stand-alone con-sumption, aA and aB ,and the higher costs forthe implementation of the project. Thus,once the environmental tax is fixed, Firm 1observes the associated f(d). It then decideswhether to propose an environmental proj-ect to Firm 2 or whether it should pay thetax to Country A. The resultant equilibriumis then known. For example, if the cost ofthe project is very small relative to the com-ponents of stand-alone consumption, thenthe curve is similar to that of d1, where evenwith lower taxes, the environmental projectis the more viable alternative for Firm 1.

Figure 4-1. Effects of deferment taxes on projectimplementation likelihood.

An alternative use of Inequality 4-2 is totrigger the accomplishment of a project ofenvironmental control in Country B or totrigger the reduction of local GGEs inCountry A. Note that with payment of thetax, the amounts produced by Firm 1 arereduced and, consequently, its emissionsdecrease as well. Because all information iscommon knowledge, the agency has knowl-edge of Inequality 4-2 and would establish atax in compliance with its objectives. There-fore, let d be such that f̂ = 0, as variationsof this tax immediately affect the result ofthe game. In the specific case treated here,this tax is given by

(4-3)

Another result of using an incentivemechanism, such as that indicated byInequality 4-2, is that the resultant finalequilibrium can by and large be determinedin the beginning of the game. Equation 4-4is comparable to Equations 3-3, 3-3a, and3-3b. The predicted profits for each firmassociated with the equilibrium strategies

δ =αA + αB − 2c −

√(2c − αA − αB)2 − 96κ

12.

Reports62 Volume 8, 2003MURJ


can then be expressed by Equations 4-4 and4-5.

(4-4)

(4-5)

An Alternative Conjecture forFuture Work

The final report of the Third Conferenceof Parts (COP3), carried through in Kyoto in1997, considers four basic mechanisms forthe global control of GGEs.4 The approachintended in these mechanisms is guidedtoward the use of environmental markets,where the main contributory element of costefficiency of this global policy is the ITQmarket.

In order to finance the implementation ofenvironmental control programs in develop-ing countries, the CDM was included, bywhich the decurrent credits of reductionseffected from designs implemented in thesecountries can be used by developed coun-tries to their own advantage. Initially, thismechanism may be the one used because ofthe great cost savings it can generate,because environmental policies in develop-ing countries are, in general, less rigid. It isalso probable that these designs will bedirectly financed by developed countries orby firms located in these countries.

However, as Hahn and Stavins point out,the role of domestic politics is decisive forthe success of the program, as a good part ofthe GGEs come from industrial processes. Asdiscussed in the previous section, the impo-sition of one specific environmental policyin a developed country can generate,through the CDM, value for the excess ofemissions in developing countries. Theresponsibility for the fulfillment of quotasagreed upon in Kyoto is transferred to thefirms headquartered in developed countries;these can negotiate the right to carrythrough GGE reductions in developingcountries.

Under particular hypotheses, the marketis able to provide the necessary instrumentsto avoid compromising the efficiencyderived from the propositions put forth inthe Kyoto Protocol. This model, as presented

in this paper, is only possible given theassumption regarding the nature of theinformation between the countries involved.Such assumptions indicate the fundamentalrole of local environmental authorities inpropitiating conditions so that marketcrashes do not compromise the effectivenessof the program.

Hence, despite the fact that CDM presentsa series of advantages for developing coun-tries, some aspects of the partakers’ behav-ior must be considered in relation to thepracticed environmental policies in theircountries. Individual actions by firms can-not harm the well-being of the population.Alternatives are the immediate mapping ofthe sources of GGEs in these countries andthe imposition of referring external costsprovoked by this type of pollution, in orderto induce local firms to accept the imple-mentation of designs of emissions control,financed by external resources.

Angela S. Bassa Reports64 Volume 8, 2003MURJ

References

1. Confederation of EuropeanPaper Industries. “CEPI’s Viewson the Emission Trading-Responses to the Commission’sGreen Paper on EmissionTrading.” Online Document,http://www.cepi.org, 2000.

2. The European Commission,European Communities. “TradePolicies.” Online Document,http://europa.eu.int/comm/enter-prise/footwear, 2000.

3. Hahn, R. W., and R. N. Stavins.“What has Kyoto Wrought? TheReal Architecture of Inter-national Tradable PermitMarkets.” The AEI Press:Washington, D.C. (1999).

4. “Kyoto Protocol.” OnlineDocument, http://cop3.unfccc.int/resource/protintr.html: 1997.

5. Motta, M., and J-F Thisse. “DoesEnvironmental Dumping Lead toDelocation?” EuropeanEconomic Review:1994.

6. Pindyck, R. S., and D. L.Rubinfeld. “Microeconomics.”Prentice Hall: New Jersey(2000).

7. Tietenberg, T. “EconomicInstruments For EnvironmentalRegulation.” Oxford Review ofEconomic Policy. Vol. 6; No. 1:March 1990.

8. Tirole, J. “The Theory ofIndustrial Organization.” MITPress: Cambridge, Massachusetts(1989).

9. Varian, H. R. “IntermediateMicroeconomics: A ModernApproach.” W. W. Norton &Company: New York (1999).

Appendix

Table A-1. A list of Annex-I countries, together with their relative greenhouse-gasemissions levels in 1990, taken from endnote 4.

Country Emissions (Gg) Percentage of Total Emissions

Australia 288,965 2.1

Austria 59,200 0.4

Belgium 113,405 0.8

Bulgaria 82,990 0.6

Canada 457,441 3.3

CzechRepublic 169,514 1.2

Denmark 52,100 0.4

Estonia 37,797 0.3

Finland 53,900 0.4

France 366,536 2.7

Germany 1,012,443 7.4

Greece 82,100 0.6

Hungary 71,673 0.5

Iceland 2,172 0.0

Ireland 30,719 0.2

Italy 428,941 3.1

Japan 1,173,360 8.5

Latvia 22,976 0.2

Liechtenstein 208 0.0

Luxemburg 11,343 0.1

Monaco 71 0.0

Netherlands 167,600 1.2

Norway 35,533 0.3

New Zealand 25,530 0.2

Poland 414,930 3.0

Portugal 42,148 0.3

Romania 171,103 1.2

Russia 2,388,720 17.4

Slovakia 58,278 0.4

Spain 260,654 1.9

Sweden 61,256 0.4

Switzerland 43,600 0.3

United Kingdom 584,078 4.3

United States of America 4,957,022 36.1

Total 13,728,306 100.0


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