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WUURD
wa s h i n gt o n
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u n i v e r s i t y
Undergraduate
Research Digest
Volume 2, No. 2
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S p r i n g 2007
[ Feature Articles ]
Homosexuality in Cameroon’s Public Sphere:
Rejecting Homosexuality as Protest against the Other Micah J. King
The Real Dramatis Personae: Researching and Creating
Characters for the New Play Chosen Noga P. Landau
You Are What You Eat: A Technique for Isotopic Analysis
of Archaeological Amino Acids Andrew B. Newman
Overexpression of Cx43 and NF200 in Hibernating Species
Citellus undulates During the Hibernation State Sangita Sudharshan
Analysis of DIAP2 Rescue of diap1 Mutant Cells in Drosophila
Lisa J. Sudmeier
The Georgian Feast and Azeri Carpet: A Cultural Exploration
through Metaphors Andrea S. Young and Margaret S. Fee
[
Summaries of Student Work
]
WUURD, the Washington University
Undergraduate Research Digest, is published
by the Office of Undergraduate Research once
a semester each academic year.
How to contact us:
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Campus Box 1026
Phone: (314) 935-7342
Fax: (314) 935-4384
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Web: http://ur.wustl.edu
Henry Biggs
Director of Undergraduate Research and
Associate Dean in the College of Arts & Sciences
[email protected]
Joy Zalis Kiefer
Undergraduate Research Coordinator, Editor
Assistant Dean in the College of Arts & Sciences
[email protected]
Kristin G. Sobotka
Co-Editor
[email protected]
Aline Boos Holz, Ph.D.
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Elizabeth Berkes, Ph.D.
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Emily Beck
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Issues of the print version may be obtained at the
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The primary requirement for inclusion in the Digest
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~
WUURD
wa s h i n gt o n
~
u n i v e r s i t y
Undergraduate
Research Digest
Volume 2, No. 2
~
S p r i n g 2007
Table of Contents
F O R EWO R D
3
Optimization of Design Parameters
for Future Gamma-Ray Telescopes,
Christopher Weaver
65
4
Regulation of the Urea Transporter
UT-A in Rats with Adriamycin Induced
Nephrotic Syndrome, Benjamin Nanes
66
The Limitations of a Mixed Phase
in Strange Star Crust, David Eby
68
Guanxi: A Reemerging or
Disappearing Key to China’s
Labor Markets?, Linda Zhou
69
Individualism, Economics &
Revolution: Socioeconomic Ideology
in Revolutionary France and America,
Andrew Schupanitz
70
A RT I C L E S
Homosexuality in Cameroon’s Public
Sphere: Rejecting Homosexuality as
Protest against the Other, Micah J. King
The Real Dramatis Personae:
15
Researching and Creating Characters
for the New Play Chosen, Noga P. Landau
You Are What You Eat: A Technique
for Isotopic Analysis of Archaeological
Amino Acids, Andrew B. Newman
26
Overexpression of Cx43 and
NF200 in Hibernating Species Citellus
undulates During the Hibernation State,
Sangita Sudharshan
36
Analysis of DIAP2 Rescue of
diap1 Mutant Cells in Drosophila,
Lisa J. Sudmeier
46
Efficacy of the Ketogenic Diet in
Children with Surgically Approachable
Epilepsy, Rebecca Stainman
71
The Georgian Feast and Azeri Carpet:
A Cultural Exploration through
Metaphors, Andrea S. Young and
Margaret S. Fee
53
Effects of Management on Garlic
Mustard (Allaria Petolata) Fitness,
Brittany Teller
72
Functional Domains of RNA Binding
Protein CUGBP2, Pavel Rodriguez
73
What Would Sex Education from
the Left Look Like?, Kristi Nigh
74
SUMMARIES OF
S T U D E N T WO R K
External Connections of the Dorsolateral
Prefrontal Cortex, Abhisek C. Khandai
62
The Role of JIP-1 in Axon Transport,
Pavan Bhat
75
Ecological Trap for Ovipositing
Mosquitoes Mediated Through
Preferences for Contaminated Habitat,
Julia Buck
64
Hopping Rates in Scandium Hydride
and Magnesium Hydride, Erik Carl
76
Noise Reduction by Spectral Contrast
Shaping, Nathan J. Killian
77
2
Pictures of the Mind: the Pictorial
Foundation of Contemporary
Neuroscience, Cara Dolan
78
Role of the Transcription Factor
HIF-1a in Epithelial Inflammation,
Rebecca Martin
91
Martin and Monradi: An Ethics of
the Ordinary, Maria Efimova
79
92
A New World Grain in an Old World:
A Study of Maize’s Impact on
Sustainability in Sub-Saharan Africa,
Margot Danker
80
Biodiversity and Energy Availability:
Testing the Evolutionary Rates
Hypothesis, Anne K. Greenberg
VERITAS Observation of GRB060501,
Rashied Amini
93
81
Lead Adsorption to Titanium Dioxide
Nanoparticles, Carolyn Maus
94
Elucidating Preconditioning-Induced,
AKT-Mediated Signaling Mechanisms
Operative in Cerebral Endothelium
that Protect the Microvasculature
from Stroke, Somalee Banerjee
Study of Decays from Excited States
of 10C, Kevin Mercurio
95
96
Neural Evidence of Syntactic and
Referential Competition During
Sentence Processing, Amir Francois
83
Rate of Hydrolysis of
4-(p-Nitrophenoxy)-2-Butanone
by Amine Base, Dafang Zhang
97
The Silent Victims of the AIDS
Epidemic: Rethinking the Illusion of
Monogamy and the Social Constructions
of Marriage, Emily R. Hawkins
84
Guatemalan Border Control and
Implications for the Trafficking of
Persons, Matthew T. Miller
98
Repetitious Element 1360 as a Target
for Heterochromatin Formation in
Drosophila Melanogaster, Alejandra
Figueroa-Clarevega
85
Mapping the MIA1 Gene in
Chlamydomonas Reinhardtii,
Michelle L. Miller
Public Policy and Poverty in
East St. Louis, Illinois,
Elizabeth Williams
99
Continuing the Technology Debate:
The Effects of Interactivity and Visual
Stimulation on Learning, Andrea Young
86
The Body Strikes Back: Media
Theory and AIDS Activism,
Ian Schatzberg
100
Design and Construction of a FormulaStyle Race Car, Melissa Holtmeyer
87
101
Difference-Blind Politics in a Multicultural State: The Political Implications
of Identity in Democratic Societies,
Marquita James
88
Binding Characteristics of the PAT
Protein Family - Perilipin, ADRP
TIP47, S3-12, and OXPAT, Michelle Mo
102
Bronze Age Steppe Dynamics and Stable
Strontium Isotopes, Taylor Hermes
89
Reading Islam in the Italian
Renaissance: Representations of
Islam and the East in 16th Century
Travel Narrative as seen in the
Itinerary of Ludovico di Varthema,
Jessica Pryde
Constructing Various Mutants
of Pathogenic Bioluminescent
Salmonella for a Murine Model
of Infection, Aasish Manglik
90
Exploring the Meanings of the
Huipil, Erendira Quintana Morales
103
U N D E RG RA D UAT E
R E S E A RC H PE E R
R EV I EW B OA R D
104
Foreword
The Washington University Office of Undergraduate Research is firmly committed to providing opportunities for undergraduates to work closely with a
faculty mentor in a research setting, not only to expand their learning experience, but also to allow them to be a part of the exciting process of contributing to the creation of new knowledge in their various disciplines. This edition
of the WUURD represents exciting growth for our publication in terms of
refining the quality of the feature articles presented and representing the
diverse areas of research in the Arts and Sciences and Engineering
We began in the fall semester accepting applications for our first WUURD
Peer Review Board. With over seventy applicants, the choice was a difficult
one. In the end, thirteen Washington University undergraduates became a
part of the first feature article review and acceptance process. Their names
appear at the end of this publication. We are very grateful for their diligence
and incredible efforts which contributed greatly to the quality of this publication, efforts which also aided our student authors in becoming better communicators of their research. We would especially like to recognize Morgan
Grossman-McKee, Sarah Frazier, and Olga Lozovskaya for their willingness to
take on the extra duties of final proof-reading and editing.
The feature articles represent a broad range of research interests. Micah King
from Anthropology explores the changing meanings of homosexuality in
Cameroon, West Africa. Andrea Young from Psychology and Margaret Fee
from French and International and Area Studies conduct an investigation of
metaphors as way of understanding the cultures of the Republic of Georgia
and Azerbaijan. From Drama, Noga Landau illuminates the process of historical research and its role in the creation of characters for a play based on real
events and people. An improved technique for isotopic analyses of proteins in
archaeological remains is introduced in the work of Andrew Newman from
Physics. Biomedical Engineering’s Sangita Sudharshan presents her investigation of underlying mechanisms that allow hibernating mammals to maintain
proper cardiac conduction and rhythm at extremely low body temperatures.
Finally, Lisa Sudmeier from Biology investigates the role of Inhibitor of
Apoptosis Proteins in cell death, which is essential to the development and
maintenance of healthy tissue in organisms.
3
This edition of the WUURD is a testimony to the active pursuit of
new knowledge by our undergraduate researchers, as well as by the faculty
who mentor them. We hope that you enjoy spring 2007 edition of the
WUURD and look forward to expanding our reach still further into all of the
undergraduate schools.
Respectfully,
JOY ZALIS KIEFER
Assistant Dean, Editor
Office of Undergraduate Research
KRISTIN SOB OTKA
Co-editor
HENRY BIGGS
Associate Dean
Director, Undergraduate Research
4
Homosexuality in Cameroon’s
Public Sphere: Rejecting Homosexuality
as Protest against the Other
Author:
Micah J. King
Micah is a senior and will graduate with a double major in
Anthropology and International
& Area Studies. He became
involved in undergraduate
research through a semester
in Cameroon with the School
for International Training. As
a result of that initial trip and
a second one funded by a
grant from the Office of Undergraduate Research, he became
more interested in global LGBT
issues and the intersection
of global and local sexual identity. Micah anticipates working
in the nonprofit sector before
resuming his education, continuing his research, and becoming a professor.
KEY TERMS
•
•
•
•
Cameroon
Homosexuality
LGBT
Public sphere
A B S T R AC T
This research examines the emergence of homosexuality into
Cameroon’s public sphere. It explores the “Top 50” list that named elite
Cameroonians as homosexuals, as well as other events, and their role
in the emergence of homosexuality into Cameroon’s public sphere. To
understand the List and its effects, this study examines the events leading up to the List and emergence. This research posits that the List is
the main catalyst for the emergence of homosexuality into the public
sphere, and that the resulting discourse has opened a space in which
Cameroonians critique homosexuality as a means of denouncing what
homosexuality has come to represent, namely Westernization, corruption and the elite class. This study builds upon Charles Gueboguo’s
work on homosexuality in Cameroon in order to create a basis for
understanding homosexuality in post-List Cameroon.
FA C U LT Y M E N T O R : R O B E R T L . C A N F I E L D,
P H . D. , P R O F E S S O R O F A N T H R O P O L O G Y
Professor Canfield’s early research was on the spatial conditions that
shaped the alignment of local factions and coalitions in the Hazarajat
of Afghanistan. But because Islamic categories provided the terms of
political identification in this area, he later began working on Islam as
the idiom of personal life and political affairs. Since the early 1980’s,
he has been working on the Islamic culture of the Central Asian region,
notably to understand how Islamic idioms have taken root in the
region to become the cultural forms through which individual experience has been understood and collective action justified and organized. He edited a book on the culture history of the eastern “TurkoPersian” Islamic world, and is co-editing a book on social affairs in
Greater Central Asia.
AC K N OW L E D G E M E N T S
A sincere thank-you to the Bea-Hop family for generously opening
their home to me, and to André-Guy Soh for being a teacher in Paulo
Freire’s sense of the term. I could not write this without thanking
Charles Gueboguo, Joel Nana, Steave Nemande and everybody at
Alternatives-Cameroun. This endeavor would not have been possible
without Dr. Victor Le Vine, Dr. Robert Canfield, Dr. Shanti Parikh and
Dr. John Bowen. All mistakes are my own. This research was supported through a generous Hoopes grant from the Office for
Undergraduate Research. A special thanks to Dean Henry Biggs and
Dean Joy Kiefer.
5
INTRODUCING THE EVENT
n January 2006, four fringe Cameroonian newspapers, most prominently La Météo
and L’Anecdote, published high profile lists of over 50 allegedly gay and lesbian
Cameroonians, among them governmental ministers, athletes, musicians, community
leaders, and members of the clergy. Initial copies of La Météo and L’Anecdote quickly
sold out, and newspaper vendors photocopied the originals to satisfy consumer
demand. Since the List, homosexuality has been the topic of numerous television and
radio shows, newspaper headlines, protests, and court cases. Nearly every participant
interviewed held the view that homosexuality is in Cameroon’s public sphere, and has
only existed in that capacity since the publication of the List, also called the le top
cinquante [the top fifty] or la liste rose [the pink list]. This shift is significant in that it
marks the first time homosexuality has been so publicly discussed in Cameroon.
Several questions guide this study. To begin, what is the cultural significance of the
emergence of homosexuality into the public sphere? Further, what are the catalysts for
the emergence of homosexuality into Cameroon’s public sphere, and what does
denouncement of homosexuality in this newly public discourse reveal about
Cameroonian culture?1
This study asserts that the publication of the List was a watershed moment, and
that the history of the factors leading up to its emergence must be analyzed. The publication of the List was not a singular event, but rather one resulting from decades of
history that represents many facets of Cameroonian culture. In order to understand
the emergence of homosexuality into the public sphere in Cameroon, the historical
context is presented, first with homosexuality in pre-colonial Cameroon and then in
contemporary Cameroon. To understand how homosexuality and the public sphere
converge, the events leading up to the publication of the List are examined. In chronological order, they are the liberalization of media laws, information from the Global
North, the arrest of eleven people on charges of homosexuality, the murder of a high
school student, the Yaoundé Archbishop’s Christmas 2005 homily and finally, the List.
Homosexuality as defined in Cameroon is different from Western definitions. In
Cameroon, allegations of homosexuality can be a means of attacking the elite while
working within acceptable modes of discourse. This is the case with the List and
rumors surrounding the death of a young man at the Hilton Hotel in Yaoundé. As
Sahlins notes, “Formal classification is an intrinsic condition of symbolic action,”2 and
attacking somebody as homosexual is not just an attack on real or perceived homosexuality. Indeed, to denote someone as homosexual in Cameroon carries with it the
active connotation of being corrupt, too elitist, or too Western.
I
RESEARCH SITE AND METHODS
Qualitative studies entail formal and informal interviews, with the former composed
of both open- and close-ended questions. Interviews were conducted with sociologists, journalists, editors, imams, priests, pastors, as well as with gay, lesbian and bisexual Cameroonians and with Cameroonians working for Lesbian, Gay, Bisexual, and
Transgendered (LGBT) rights. A quantitative component included the distribution
and collection of 270 questionnaires aimed at determining the links between informa-
6
tion flows and young Cameroonians’ perceptions of homosexuality. Questionnaires
were distributed and collected at the Université de Yaoundé I in two ways.3 In the first,
random responses were collected around the campus. In the second, more predominant mode, questionnaires were explained, distributed and collected during class
times. A less traditional mode of inquiry was utilized in the analysis of newspapers and
other media, a strategy that seems to have worked well for Kama.4 Sampling was primarily limited to Yaoundé, though information was also collected in Douala, Dschang,
and N’gaoundéré. The predominance of male participants is a consequence of contemporary Cameroonian society and an issue noted by Gueboguo.5 Temporal constraints were a substantial impediment to the depth of this research.
HOMOSEXUALITY IN THE CAMERO ONIAN CONTEXT
Despite colonial ideas that Africans could not be homosexual, and the common
Cameroonian assertion “l’homosexualité n’est pas Africain” [homosexuality isn’t
African], homosexuality has been documented in over 55 groups in sub-Saharan
Africa, and was documented in Cameroon starting with Günther Tessmann in the
early 20th century.6 Tessmann describes same-sex relationships among the Bafia,
including a ji’gele ketön [male-to-male intercourse] as common and acceptable for
youth up to the age of puberty. He also reported that, while much less common, there
were adult men known to be gay, those called a bele nuem e bongo [he has the heart of
a boy].7 Part of his work includes stories alluding to homosexuality, including “The
Four Suitors: A Pangwe Tail,” a story in which the male would-be suitors for a girl,
Akúkedanga, are thwarted in their plans and end up sleeping with one another.8
Charles Gueboguo describes homosexuality in pre-colonial Cameroon using analyses
of traditional initiation rites to build his assertion that homosexuality has existed in
Cameroon since well before the arrival of Europeans.9 In particular, Gueboguo studies the similiar mevungu rite of the Beti and the koo of the Bassa, rites which are, as
Laburthe-Tolra describes, “un célébration du clitoris et de la puissance féminine” [a celebration of the clitoris and of feminine power] in which menopausal women play the
role of men.10
Today, it is commonly asserted by Cameroonians that homosexuality is not
African, but rather a European idea brought to French West Africa by missionaries in
the 19th century as part of the mission civilisatrice [civilizing mission]. Another layer
to the Cameroonian concept of homosexuality is that of corruption, where homosexuality is seen as a means of social ascension. An additional layer is that of modernity
and influence from the Global North. There is a sort of social angst in this rapidly
changing country. Colonization, the breakdown of civil society, personal insecurity,
the influx of foreign media and cultural influences, and foreign economic intervention, along with other factors, have contributed to a sense of rapid change and loss of
the traditional. Namely, there is a perceived loss of the times when the chiefs and
lamibé were wealthy and their people sovereign within their respective group.11
Homosexuality is almost uniformly perceived as part of this foreign influx of ideas
and customs, where homosexuality is not African but rather synonymous with
Westernization, modernity, corruption, and the other. The musicians and popular fig-
7
ures named in the List are perceived as being strongly influenced by, or are even a part
of, the modern, Western world. Similarly, the governmental ministers and other members of the elite class are generally seen as corrupt. Rejecting the elite with allegations
of homosexuality is a means of rejecting those things that homosexuality means in the
Cameroonian context.
Homosexuality and LGBT people are commonly derided in Cameroon, both publicly and privately. Homosexuality is never just same-sex relations or sexual orientation or identity; rather, it carries with it a slough of connotations. Common conceptions of homosexuality by Cameroonians include: homosexuality as a means of social
and economic ascension; homosexuality as a form of sorcery, witchcraft and demonic activity; homosexuality as a form of corruption; and homosexuality as distinctly not
a part of African culture. Many participants suggest that gay men have sex with powerful men in order to gain employment and promotions. Many feel that homosexuality is found in several forms, including corruption, prostitution, witchcraft, and secret
cults (sic) such as Freemasonry and Rosicrucianism. Rumors relayed through informants, and previously documented by Gueboguo, include: soccer stars are homosexual,
(especially the biggest stars, such as Barcelona striker Samuel Eto’o Fils), numerous
powerful government officials are gay, certain priests are gay, and a choir member of
one of Cameroon’s largest churches was excommunicated for sex with another male
choir member. Further, the expression “promotion canapé” [couch promotion] refers
to the need to have homosexual intercourse in order to receive a job promotion. In
general, it is perceived that nkouandengué [homosexuality] brings with it economic
gains.12 Additionally, the Hilton hotel in Yaoundé – a towering symbol of foreign
wealth, influence, and the elite – is widely rumored to be a place where young
Cameroonians have sex with wealthy white men for money.
Cameroonian media outlets, including newspapers, magazines, television and
radio shows, overwhelmingly condemned homosexuality. The seven religious leaders
interviewed, including priests, pastors, and imams, were uniformly against homosexuality. Christian leaders were much more likely to decry homosexuality as a sin not
only against God and nature, but against African morals and the country as well.
COMING OUT INTO THE PUBLIC SPHERE
Because homosexuality is in the public sphere and used in public discourse,
Cameroonians can signal to each other their ideas and create common ground based
upon rejection of it. The public nature of this discourse means researchers can more
freely understand Cameroonian ideas of homosexuality; further, it brings with it the
opportunity to ask questions that would have been out of place, or even unacceptable,
prior to the List. The dialogue, conversations, and reader and media obsession with
homosexuality represents the emergence of homosexuality into Cameroon’s public
sphere, with the List being the catalyst of this emergence. Before analyzing the meanings of the List in detail, the events leading to emergence of homosexuality into the
public sphere will be described, beginning with the liberalization of the press.
8
The 1990 Liberalization of the Cameroonian Press
The List could not have been published and little of the Western media influence
would have arrived without the 1990 liberalization of media laws in Cameroon, a
result of internal and international pressure tied to foreign aid.13 Liberalization
brought with it foreign and privately owned media outlets, such as CNN, the French
TV5 Monde, Euro News Planet, the Senegalese channel RTS, a Brazilian soap opera,
and various other media. Domestically, it meant the creation of independent radio
stations, additional newspapers, and the inclusion of articles that the government
might find embarrassing, such as ones admitting the existence of homosexuality in
Cameroon. It is doubtful the List would have passed censor review before 1990. This
step primed Cameroon’s public sphere and legal environment for its arrival.
The Global North
Related to the liberalization of media laws is the role of information from the Global
North in affecting Cameroonian perceptions of homosexuality. These information
flows are relevant because, even before the List, they imported some level of information about LGBT movements and realities outside Cameroon. It must be noted that
the Internet has also been a source of outside information; a brief perusal of any cyber
café will reveal people browsing foreign news websites, dating service websites, and
reading and sending emails to friends, family, and colleagues outside of Cameroon.14
Most (81 percent) of questionnaire respondents report hearing about gay and lesbian
people or homosexuality through a Cameroonian television show, followed by 68 percent of respondents reporting exposure through European TV shows. When asked
how they were informed of the legality of homosexuality in the West, 72 percent of
respondents report that it was through a Western television show. Other reported
sources of information were Western films (71 percent), Western newspapers (47 percent), and Western radio stations (46 percent). The most common Cameroonian
source of this information was Cameroonian television shows with 36 percent. The
majority of university-attending Cameroonians from a variety of backgrounds heard
about homosexuality through the media, and the most common sources of information about homosexuality’s legality in the West came from Western media sources. The
information coming from America, France, and other areas in the Global North have
brought information about homosexuality to Cameroonians and helped bring the
issue into the public sphere, though in ways more subtle than the List.
The Arrest of Gay Cameroonians
The arrest of eleven men on charges of homosexuality and their subsequent court battles were widely covered by the Cameroonian press. The arrest of the Yaoundé 11 – as
they are called – while never creating the sort of controversy and discourse prompted
by the List, nonetheless marks another step on homosexuality’s path to emergence
into the public sphere. As was the case with many post-colonial governments,
Cameroon, under President Ahmadou Ahidjo’s regime, retained a version of the laws
against homosexuality that were imposed by its French and British colonial administrators. In 1972, that law was reaffirmed in article 347, ordinance 72-16 of Cameroon’s
penal code, which constitutes same sex contact as a crime punishable by six months to
9
five years in prison and a fine of 20,000 to 200,000 CFA francs (approximately $39 to
$394).15 This law was not strictly enforced until May 22, 2005, when 11 Cameroonians,
including three minors, were arrested at a nightclub in Yaoundé by gendarmes from
the Nlongka Brigade and detained at Kondengui Central Prison under harsh conditions. The plight of the Yaoundé 11 prompted discussion through media coverage and
was part of the increasing media attention and public discourse on homosexuality in
the months leading to the List.
The Murder of Jean Ndikumana
On December 5, 2005, Franck Abega fatally slashed the throat of Jean Alex
Ndikumana, his 17-year-old Burundian classmate, at the prestigious American School
in Yaoundé. The murder was a hate crime motivated by what Abega perceived as flirting.16 Ndikumana was murdered in a classroom as his peers looked on. The incident
garnered a large amount of publicity, both domestically and internationally, and over
19,000 people read the web version of the article on Cameroon-info.net. From the day
of the incident until February 2006, readers of the story posted nearly 345 comments
and reactions to the incident, with responses coming from people in Cameroon and
throughout sub-Saharan Africa, Europe, the Americas, and Asia.17
The Catholic Church Speaks Out
On Christmas day 2005, the title of Yaoundé Archbishop Victor Tonye Bakot’s homily
was “L’Archevêque s’insurge contre l’homosexualité” [The Archbishop takes up arms
against homosexuality],18 a sermon widely spoken about, mentioned and referenced to
in participant interviews, and noted in the reporting of the List by both the BBC19 and
Radio France Internationale.20 The homily became an instant best-seller in Yaoundé,
and according to a priest at the archdiocese, Christmas 2005 marked the first time that
the Catholic Church in Cameroon systematically denounced homosexuality.21 In his
homily, Archbishop Bakot denounced the movement in Europe and the United States
toward more equal rights for gay and lesbian people. In particular, he denounced the
1997 Amsterdam Treaty for its recommendation that countries within the European
Union recognize sexual orientations without discrimination.22 He pointedly rejected
gay and lesbian marriage and adoption rights, and stated that Africa was under attack
by modernity and by liberalism.23 His homily was especially remarkable as it was timed
to coincide with the attendance of elite Cameroonians, including politicians, for the
holiday. While the homily brought homosexuality into the public for some, it was not
the primary catalyst for several reasons. First, Cameroon is a nation of not only
Catholics, but also Protestants, Muslims, and Animists. The sermon greatly affected
Catholics, and to a lesser degree, Protestant Christians.24 However, Muslims that participated in this research never mentioned the sermon. Secondly, the sermon was not
as public as the newspaper headlines, and was not as gripping an event as the newspaper articles naming allegedly gay and lesbian persons in the highest echelons of
Cameroonian society. Nevertheless, the homily contributed to public dialogue of the
issue for a large number of people and was a step on the path to the emergence of
homosexuality via the List.
10
The List
The first paper to print the List was La Météo, followed quickly by L’Anecdote. For
L’Anecdote, the List was divided between two sequential printings, January 24 and 31,
2006.25 Both days the list commanded the front page title. An article with a pink background and pink title “Réseaux – modes d’emploi” [Networking – means of employment] accompanied the first printing of the List in L’Anecdote. The article asserted that
homosexuality was used by those at the summit of the state to gain more power, and
that wealthy gay men seduce young men who are attracted by the wealth and power
that homosexuality can bring.26 Placed immediately following the List was an article
“Quand la dénonciation vient de l’Église” [When the denouncement comes from the
Church], in which Charles Chime credits Archbishop Bakot’s homily, just a month
prior at this point, for denouncing homosexuality.27 The first printing of the List by
L’Anecdote was book-ended on both sides by prominently placed articles linking homosexuality to the elite, social ascension, and corruption. The List named musicians, bishops, governmental ministers and executives as homosexual. The people named included dead and living Cameroonians, colonial administrators, and French expatriates. The
claims of homosexual are true for some, and highly dubious for others. The only commonality the member of the List share is that they are elite. The List is significant
because only after the List was published was there a sustained national discourse on
homosexuality. The List brought homosexuality into the public sphere, allowing for a
better understanding of what homosexuality meant in the ensuing public discourse.
AT TACKING HOMOSEXUALITY AS A FORM OF PROTEST
Homosexuality is linked in the minds of many Cameroonians with corruption, the
elite and Western influence. The List and rumors surrounding the Hilton share the
trait of serving as a means to attack the elite. If the List brought homosexuality fully
into the public sphere, revealing Cameroonian attitudes and beliefs about homosexuality, an analysis of events surrounding the Hilton Hotel in Yaoundé reveals a compacted series of meanings that are representative of homosexuality in Cameroon. On
August 21, 2006, a thirty-year-old Cameroonian university student was found dead at
the base of the Hilton hotel, having fallen from a balcony at about the ninth floor. The
director of the hotel told the important daily newspaper, Mutations, that there was no
record of the young man being an occupant, nor was he an employee.28 Immediately,
rumors began. The most commonly accepted of these was that he was gay, and that he
was either murdered or committed suicide after having had intercourse with someone
of the same sex. In a special edition reflecting back on the most important events of
2006, Dorine Ekwè of Mutations relayed:
In the crowd, there were no doubts. The affair was linked to a network of homosexuals. Marks found on the body of the young man
support this hypothesis: The buttons on his pants had popped off,
these same pants had two large tears on the higher part of the thigh
and another between the legs. When he was undressed, you would
have noted traces of violence on his body. A gash between the anus
and the genitals, and marks from a hot iron on his back. 29
11
Other rumors included details about with whom the young man was sleeping
(most focusing on the president of the Chamber of Commerce) and a baggage handler who was supposedly stopped the night of the death by police at Nsimalen airport
in Yaoundé with a Cypriot visa and thousands of American dollars. The truth is not
certain, but the rumors, like shared beliefs, reveal that labeling someone as homosexual carries with it a complex set meanings.
The location of the Hilton is important as a symbol of wealth, Western influence,
and the elite class. There is perhaps no place that better represents foreigners and the
wealth that most Cameroonians know they will never have. There were a plethora of
rumors from many different participants that the Hilton is the place where wealthy
men, often foreign and white, corrupt the youth of Cameroon through enticing them
to have sex for money.30 From a Western lens, the outing of powerful or famous
Cameroonians through the List, like vicious rumors at the Hilton, may be interpreted
simply as an attack on gay and lesbian sexuality. Nevertheless, to understand why these
rumors are so pervasive, they must placed in the context of domestic concerns.
Cameroon is a notoriously corrupt country, where more than 50 percent of people
report that they or members of their household have paid a bribe in the past 12
months.31 Articles on homosexuality often create a link between members of the
regime, corruption, and homosexuality. For example, The Guardian Post alleged that
the 50 presumed homosexuals are gay in order to gain money through ministerial promotions, and for musicians, in order to gain a wider audience.32 The idea that homosexuality is a form of corruption and is used for social and economic ascension, is pervasive and deeply entrenched in Cameroonian culture.
Westerners, most notably the French, are viewed as bringing homosexuality to
Cameroon. France, and to a lesser degree other Northern powers, are seen as propping
up the corrupt government because of their economic interests in the country.
Cameroonian elite are seen as marionettes of former colonial powers—especially
France and increasingly the United States.33 Rejecting homosexuality is a means of
denouncing what it means in the Cameroonian context, something foreign, corrupt,
white, Western, and a related to the legacy of colonization and the rapid change of
Cameroonian cultures. The motivation for printing the List had not only to do with
financial incentives of the publishers, but naming people in the List was also a way of
attacking corrupt leaders and the elite.34
This research is not simply the story of homosexuality moving from the realm of
hushed conversations to national discourse in Cameroon, nor is it the story of persecuted LGBT and their fight for a safe space. When Cameroonians attack elite members
of society through the List or through rumors surrounding the Hilton, Cameroonians
are rejecting the layers of meanings that homosexuality has acquired. The List brought
homosexuality into the public sphere, opening a space for Cameroonians to join in
rejection of homosexuality, and along with it, the deeper issues it represents within
Cameroon, namely western influences and the corruption of elites. Further research
will illuminate the continuing process of the changing meanings of homosexuality
within the Cameroonian cultural context.
12
Notes
1
I will be using homosexuality to refer to any person who identifies or is labeled by their community as non-heterosexual. While this term is considered medical and antiquated in the West,
it is in line with the vocabulary used by Cameroonians themselves, including straight and
LGBT participants.
2
Sahlins, Marshall. 1985. Islands of History. Chicago: U of Chicago Press.
3
The universities of Yaoundé I and II are the state-run universities in the capital city.
4
Kama, Amit. 2000. From Terra incognita to Terra firma: the logbook of the voyage of
gay men's community into the Israeli public sphere. Journal of Homosexuality 38.4: 133-62.
5
Gueboguo, Charles. 2006. La Question homosexuelle en Afrique : le cas du Cameroun.
Paris: Harmattan, 13.
6
This number is taken from Stephen O. Murray and Will Roscoe, eds. 1998. Boy-Wives
Female Husbands: Studies of African Homosexualities. New York: St. Martin's Press.
7
Tessmann, Günther. 1998. Homosexuality among the Negroes of Cameroon and a Pangwe Tale.
Boy-Wives Female Husbands: Studies of African Homosexualities. Stephen O. Murray and Will
Roscoe, eds. Trans. Bradley Rose. New York: St. Martin's Press, 156.
8
Tessmann 1998:159-61.
9
Gueboguo, Charles. 2006. La Question homosexuelle en Afrique : le cas du Cameroun.
Paris: Harmattan.
10 Laburthe-Tolra, Philippe. 1985. Initiations et Sociétés Secrètes au Cameroun : Les mystères de
la nuit. Paris: Éditions Karthala, 327.
11 Lamibé is the plural form of lamido, the titular religious and political leaders of the Ful?e
(Peule or Fula) Muslims in Northern Cameroon and other Sahelian areas.
12 Gueboguo, Charles. 2005. Ce que parler veut dire : essai d’analyse de contenu de 10 rumeurs
circulant dan les milieux gais du Cameroun. Electronic document, http://semgai.free.fr/contenu/textes/gueboguo_ce_que_ parler.html, accessed Sept. 30, 2006. For more on
nkouandengué, see Gueboguo 2006:85.
13 The National Assembly passed the law granting freer speech, communication and media
as ordinance 90/052 on December 19, 1990 (Gueboguo 2006:152).
14 Information on homosexuality gained through the internet is not limited to Western sites.
The South African group Behind the Mask has an informative website, and for a time,
Joel Nana’s mygaypal.com provided information for and by Cameroonians.
15 While the law does not ban sexual orientation, but rather sexual contact between members
of the same sex, in recent years it has been enforced in terms of the former definition.
16 Tagne, Jean-Bruno. 2005. Yaoundé: Un élève égorge son camarade. Mutations, December 6.
Electronic source, http://www.cameroon-info.net/cin_reactions.php?s_id=17087, accessed
April 30, 2006.
17 Tagne:2005.
18 Unless otherwise noted, all translations are my own.
19 Row over Cameroon ‘Gay’ Witchhunt. 2006. BBC News, February 6. Electronic document,
http://news.bbc.co.uk/2/hi/africa/4685298.stm, accessed April 29, 2006.
20 Zinga, Valentin. 2006. Controverse sur une liste d'homosexuels. Radio France Internationale,
February 1. Electronic document, http://www.rfi.fr/actufr/articles/074/article_ 41571.asp,
April 29, 2006.
13
21 Interview with the author, Yaoundé, April 26, 2006.
22 Bakot, Victor Tonye. 2005. L’Archevêque s’insurge contre l’homosexualité, December 25.
Pamphlet. Yaounde: Cathédrale Notre Dame des Victoires, 8.
23 Bakot 2005:12.
24 The Presbyterian general assembly’s January 2006 denunciation of homosexuality was,
in part, a reaction to the Archbishop’s Christmas homily.
25 I was only able to secure L’Anecdote’s publications of the List.
26 Réseaux-modes d’emploi. 2006. L’Anecdote. Jan. 24:6.
27 Chime, Charles Chacot. 2006. Quand la dénonciation vient de l’Église. L’Anecdote. Jan. 24:9.
28 Ekwè, Dorine. 2007. Le mort qui a fait trembler Yaoundé. Mutations. January 2:7.
29 Ekwè 2007:7. Original text: “Dans la foule, il n’y a pas de doutes. L’affaire est liée à un réseau
d’homosexuels. Les marques trouvées sur le corps du jeune homme viennent étayer cette
hypothèse : boutons de son pantalon avaient sauté, ce même pantalon avait deux larges
déchirures sur le côté à hauteur de la cuisse et une autre entre les jambes. Lorsqu’il a été
déshabillé, on aurait constaté des traces de violence sur son corps. Une entaille entre l’anus
et le sexe, des traces de fer à repasser tout chaud sur son dos.”
30 The economic crisis, which started in the 1980s, and the resultant starvation for money is
often cited as a reason young Cameroonian males have sex with wealthy older men. This is
not the first time the Hilton Hotel as been perceived as a nucleus of gay life. As pointed out
in John Scagliotti’s Dangerous Living: Coming Out in the Developing World (After Stonewall
Productions, 2003), the Hilton in Cairo is seen in a similar manner to the one in Yaoundé.
31 Political parties are most corrupt. 2004. Transparency International. Dec. 9. Electronic source,
transparency.org, accessed Feb. 13, 2006.
32 Charles, Nji Blasius. 2006. Bishops, Ministers, GMs shortlisted amongst top 50! The Guardian
Post. Jan. 30 –Feb. 4, 2006:1-2.
33 A small but interesting facet of the debate is that The Guardian Post framed The List of alleged
gay and lesbian people within Anglophone problem rhetoric, stating, “It is also worthy of note
that no Anglophone featured in the List, which the newspaper (l’Anecdote) says is inexhaustive”
(Charles 2006:1). While Francophone papers were attacking the Cameroonian and Western
elite, this claim on the front-page of an Anglophone paper was a means for the Anglophone
minority to attack the francophone majority in general.
34 Interview with the author, Yaoundé, April 2006.
14
The Real Dramatis Personae:
Researching and Creating Characters
for the New Play Chosen
Author:
Noga P. Landau
Noga Landau is a junior and is
majoring in Drama. Noga
received research funding in the
summer of 2006 that gave her
the opportunity to immerse herself in the playwriting process,
from the initial research to the
completed draft. Noga plans to
pursue playwriting after college,
along with other roles in the
theater world.
KEY TERMS
• Dramatis Personae
A B S T R AC T
In 1972, eleven members of the Israeli Olympic team were taken
hostage and killed by terrorists during the games in Munich, Germany.
Numerous books, documentaries, and movies have been made detailing this event, and now, in an effort to present a more intimate, personal side to the story, a play. After conducting extensive interviews with a
man named David Pollak, a former wrestler on the Israeli Olympic
team, and carrying out background research to accurately portray his
story onstage, the author wrote a protagonist and cast of characters for
a play based on Pollak’s experiences, called Chosen. This study
describes the link between historical research and the process of creating characters for the stage. A person is not merely the facts that history books give, but these facts can be built upon to create a detailed,
fully-formed character who is brought to life solely through the words
he or she speaks. This is exemplified in the discussion of three specific characters from Chosen—the protagonist and two other major figures in the play. A theatrical character based on a real person will, in
final drafts, become the artistic product of the playwright based on the
initial research the playwright conducts. The final incarnations of these
characters are a mixture of known facts and artistic license.
FA C U LT Y M E N T O R : H E N R Y S C H V E Y, P H . D. ,
P R O F E S S O R O F D R A M A ; C H A I R , D E PA R T M E N T
O F P E R F O R M I N G A RTS ; P RO F E S S O R O F
C O M PA R AT I V E L I T E R AT U R E
Professor Schvey has lectured and published extensively in the areas
of modern European, British and American drama. He has directed
numerous plays in both Europe and the United States and more
recently, he has turned his attention to writing. His first play, Hannah’s
Shawl, first commissioned by the St. Louis Holocaust Museum and
Learning Center in 1999, was performed on Holocaust Remembrance
Day before more than one thousand people. A revised, two-act version of Hannah’s Shawl, under the direction of Annamaria Pileggi, was
staged at Washington University in 2000.
AC K N OW L E D G E M E N T S
To my department chair, Henry Schvey, who helped me turn one little
idea into a play. Thank you for everything—academic guidance, teaching me how to give a speech, and the cigars in Stratford. We’re going
to miss you so much next year. To everyone in the Office of
Undergraduate Research for giving me this unbelievable opportunity.
To my parents and sister who read rewrite after rewrite and helped me
to remember a time before I was born. Thanks DP.
Getting funding as a playwright is hard when you’re just starting
out, and I knew from the beginning that this play would require a period of full-time research. My department chair, Henry Schvey, encouraged me to apply for a research grant to write it.
15
t the very beginning of most written plays, before lines and stage directions
appear in the text, there is a page listing the dramatis personae, Latin for “the persons of the drama.” A page of dramatis personae is essentially a list of characters, each
provided with a few descriptive details such as age, ethnicity, and important personality traits. Such a list is nothing more than the skeletal properties of each character—
basic facts open to an infinite number of interpretations by directors, actors, and
designers. The human beings we see onstage are not brought to life by a mere list of
facts. As the contemporary playwright Charles Mee said, “To understand what a
human being is, you have to see a human being in the world.”1 To modify Mee’s statement, you have to see a human being interact with the world, and the artist who guides
this interaction is the playwright—an intermediate creator who works somewhere
between the dramatis personae and the actor, responsible for the words that bring each
character a little bit closer to life.
The playwright’s job is to compose dialogue that will bring each character far
enough in writing for an actor to take over and finish the job in action. Whereas a novelist may detail inner sentiments, minute actions, and physical sensations experienced
by his or her fictional characters, a playwright must transmit all of these details solely
through the words that each character speaks. A specific challenge for playwrights
comes when writing characters based on real people, whether they are famous historical figures or modern people still alive at the time the play is written. To turn a real
person into a character requires a perfect balance between accuracy and artistic
license. No written character can truly represent a real person, and no real person can
believably exist in the world of a play.
This was one of the challenges faced during the process of researching and writing
the original play Chosen. The piece is based on the real-life experiences of a man
named David Pollak who was a member of the Israeli Olympic team that lost its
coaches and teammates in the Munich Olympic Massacre of 1972. Pollak’s story began
in the summer of 1969 when he was a twenty-year-old college student from
Pennsylvania, studying economics at Wesleyan University. That summer he left the
United States and immigrated to Israel, a country that at the time was only five
months older than him. A prize wrestler in both high school and college, he quickly
joined a club team in Jerusalem and went on to win Israel’s southern regional championship that winter. The following spring he found himself at Wingate Institute
wrestling on the Israeli national team. Later that year he joined the army—as all Israeli
citizens do in their late-teens and early-twenties—and began training as a paratrooper. He continued to wrestle with the national team, and by 1972 he was preparing for
the Olympic games, knowing that his weight slot was open on the Israeli team.
These were the same Olympics that will forever be associated with images of terrorists leaning over balconies, crowds gathering outside of concrete dormitories, and
a burned-out helicopter smoking on a tarmac at daybreak. These Olympics became
the subject of books, movies, and political debate. It is because of the events that transpired at the 1972 Olympics that the name Munich has come to represent a moment
in history when modern terrorism went global and became much more than just news
reports about letter-bombs and jet hijackings.
A
16
However, this American-born paratrooper, then twenty-four, did not go to Munich
that year. As a paratrooper he was on active duty, and very close to the start of the
Olympic games, he abdicated his spot on the Israeli team. An eighteen-year-old
Greco-Roman wrestler named Mark Slavin took his place. When members of Pollak’s
wrestling team were taken hostage by Black September terrorists in the Olympic compound, he was staying with his platoon on an army base in Israel, and Slavin was
among the eleven athletes killed in Munich.
After conducting hours of interviews with Pollak about the members of his team,
his experiences as a wrestler, and the combat he was involved in as a paratrooper, it
became clear that the best way to tell his story was in the format of a play, where the
people he conjured through his memories could be given life onstage. The writing
process for the play actually began with the title, Chosen, which as a word captured
both the experience of success in sports (for example, being chosen for a select sports
competition) and the cynical way an individual can regard his existence after he is
granted life and his comrades are not. What followed was the creation of a protagonist
based on David Pollak. This character’s name became Daniel Politzer.
All the other characters in Chosen are based on real people—some still living and
some not. The cast is made up of eleven men and one woman. Each of the other characters was built using historical research and descriptive epithets told by Pollak. Most
of these epithets were nothing more than adjectives such as “soft-spoken,” “big,” or
“funny.” Historical research provided much of the same basic information—such as
“he was a mechanic,” “he was a shop owner,” “he lived in Jerusalem”—for history
books rarely contain more than simple facts about minor historical figures. The
quirks, habits, fears and pleasures of each character had to be obtained from a formula that integrated the empty shell of a textbook history figure and the wide array of
human traits that could fit into this shell.
To maintain the historical accuracy of the piece it was crucial that each character
resemble his or her real-life counterpart as closely as possible. People go to the theater
knowing that plays are fictional, but they sometimes leave with the notion that the
actors onstage are accurate representations of the real historical figures. This is often
a false notion, but not necessarily a negative one. For example, the novelist Rose
Tremain discusses Shakespeare’s creation of the title character for his play Richard III:
“Shakespeare simplified and played about with the life of King Richard III, but the
dramatic intensity of that play and the integrity—as a character—Shakespeare gives to
Richard make it feel like the real life of that king and it is upon this play and not upon
the more accurate historical accounts that many people base their knowledge of these
events.”2 Yet, Richard III lived six-hundred years ago; the characters in Chosen either
lived three decades ago or are still alive today. Therefore, any derivation from fact or
exaggeration in behavior of a given character had to be written out of the script early
on for these characters to play as true to life as possible onstage.
The play takes place in October of 1972, almost two months after the Munich
Olympic Massacre. Throughout the play the character of Daniel Politzer experiences
guilt and confusion over the loss of his teammates. At the age of twenty-four he has lived
through combat in Egypt during the War of Attrition, having lost many fellow soldiers
in the fighting. He experiences panic, flashes of anger, and frustration, which is contrasted by a numbed demeanor during most of his interactions with other characters.
17
The story opens on Moshav Bilu, a farming village in the center of Israel. Daniel
Politzer has arrived there to spend his weekend on leave with two friends, a man
named Jackie Marder and his wife Cheryl. The character of Jackie Marder is based on
a man of the same name who lived on Moshav Bilu and built chicken coops for a living. He was a good friend of David Pollak, whom Pollak talked about briefly during
his interviews. When asked to say more about his stay with Jackie and Cheryl Marder,
Pollak simply replied:
I remember coming out [of the army base], and then going
on leave, and I went and visited a young couple who were friends
of mine on a moshav in the center of the country. They had been
on Kibbutz Hulda originally, and she had been born in America
and he had been born on the kibbutz. And they had left the kibbutz and gone to this farming village. And I would stay there from
time to time, and I remember being there after all that [the
Olympic massacre].”3
Early in the script, Daniel and Jackie have a pivotal conversation about people they
have lost in recent wars. It is important to Daniel’s character development that the
audience knows from the beginning that he has recently lost his best friend and many
men from his platoon in combat, yet he is not the kind of character who would easily
share this information. It is only after Jackie tells a similar story that Daniel feels comfortable enough to tell his. However, having no knowledge as to whether the real Jackie
Marder ever lost a close friend in combat before, Jackie’s monologue is based on an
historical event that took place during the Six Day War. Being a quiet, uneducated
man, Jackie’s speech pattern is slow and careful without a lot of adjectives. His monologue is as follows:
Jackie
I always thought Cheryl would marry my brother....the one who
was killed in ‘67. Eitan. (pause) He was a paratrooper too, you
know? But we weren’t fighting together—I was in the infantry. We
grew up on Kibbutz Hulda—I’m sure you saw his picture there at
some point, in the dining hall? He fought at Ammunition
Hill....the fortified tunnels, trenches, outside the Old City that the
Jordanian troops built up. And Eitan’s unit attacked the fortified
position. Blew up the barbed wire, you know, getting to the top of
the hill, and they started clearing out the trenches. (pause) He got
hit a couple times, but I don’t think he noticed—or at least he kept
going. I mean, I guess he had to notice at some point, but he was
tough...We buried him back on the kibbutz, and I took Cheryl to
get some ice cream afterwards because she was really sad...she
liked Eitan. Had a crush on him. Don’t know why. He wasn’t very
handsome. Probably the, you know, whatever it is about you paratroopers that the women like. But, yeah. He was my brother.
(pause) Anyway, Cheryl was wearing this little kibbutz dress that
18
she’d gotten out of the laundry for the funeral, and it was so hot
that the ice cream melted all over our clothes, you know? I
just...well, it didn’t bother me, but Cheryl couldn’t stand the sticky
feeling. And...I guess at this point she just didn’t really care, so she
took off the dress and threw it in a trash can, and she walked the
whole way back in a cotton slip. (pause) Even though my brother
had just died and there was a war I had to go back to, all I could
think about was how I had to be with this woman.
Responding to Jackie’s story, Daniel talks about the death of his best friend, a fellow paratrooper named Shuki Attias. The real Shuki Attias died in 1973 during the
Yom Kippur War, so the date of his death in Chosen had to be moved back to 1970 during the War of Attrition in order to be included in the story. In the transcript of the
interviews conducted with David Pollak, he recounted Shuki’s death:
Pollak: I fought in the ‘73 war, but my regular army unit was down
south, on the Egyptian front. The unit I served on in the reserves,
we were on the Syrian front. All the guys in my unit were still there.
They walked into an ambush, it was one of the big major ambushes of the ‘73 war, the Yom Kippur war. In Sinai, a sector called the
Chinese farm, Hadar Sini. That was the military code name for that
particular sector. And the regular paratrooper brigade, they were in
that area and they were walking through it. The Egyptians had set
up a very effective ambush there. And guys were cut down...You
know, and here I was, I mean, I went through the war in combat
and everything else, but it’s like, it hit me like a ton of bricks when
I heard all, everybody who had been killed there.
Landau: Was that when Shuki was killed?
Pollak: This is how I heard when Shuki was killed. I had gotten my
first leave since the war began, it was like a 48 hour thing, I was living in Rehovot at the time...um, and, I had just gotten back that
night. I can’t remember, it was probably sometime in November
or December. And it was late at night, and I was going through the
main street on my way, I just hitchhiked there, and I was walking
to my apartment. I was walking quickly, and all the sudden,
parked on the main street there and standing by an army Jeep, I
saw one of the lieutenants that had been in my regular army unit
with two other sergeants whom I knew from the regular paratrooper unit. I stopped, we just were stunned to see each other.
Landau: They were waiting there for you?
Pollak: No, no, they had just happened to be there. Period. Why
they were there, I still don’t even know. And I said, you know, they
asked how I was, and then they started telling me all these guys
who had been killed in the Chinese Farm sector in this ambush,
19
and it was just like one name after another. And they were telling
me, you know—it was a hopeless situation, and this guy, these two
guys threw hand grenades and just charged, just, machine gunned
us. You know it was one guy after another, it was just, it was awful.
And, Shuki Attias, my best friend...I said, “What about Attias?”
And all three of them looked at each other, and it was like this
silence. And, you know, that’s like...I knew. Okay. I said, “What
happened?” And they said he wasn’t killed in the Chinese farm
sector. It was after the brigade had crossed the Suez Canal and
gone into Egypt...And they had just cleared out this fortified sector of tunnels...Shuki was going through the tunnel with another
guy picking up weapons from the Egyptians soldiers who were
dead, you know, throwing them out of the tunnel. And one of the
Egyptian soldiers was not dead and shot him. Hit him with a submachine gun blast to the gut. He died quickly.4
In the scene from Chosen, Daniel Politzer tells the fictional Shuki’s death as an
event that has occurred recently. His speech pattern is more intense and halting than
Jackie’s, and he uses several of the same phrases that David Pollak used to describe the
real Shuki’s death. Daniel’s monologue is as follows:
DANIEL
It reminds me of my friend, Shuki....we were in the same platoon.
You know, in the first four months of paratrooper training ...every
little thing we did wrong, they would punish us. I mean, like if we
had a spot of dirt on our guns, we got punished. If the platoon
didn’t line up in three rows in twenty seconds, we got punished.
And Shuki and I—man, we were such shits. Sometimes we would
just saunter out there late, acting like we didn’t care. And that
would make the sergeants so angry. And then we’d start smirking
or something while they yelled at us, and they’d just go off the
wall. They’d yell, “Politzer! Attias! Politzer get on Attias’ back, and
get your asses to the top of that hill!” You know, so Shuki would
have me in a fireman’s carry, with one leg in one arm, right? And
we’re in full gear, of course. My rifle, his rifle, ammunition, our
knapsacks, and then the sergeant yelling at us, “Run up that hill!
Come on! You think it’s so funny, give us a little entertainment
here!” And so Shuki would get to the top of the damn hill, put me
down—both of us panting and in pain—and then we’d look at
each other and start laughing again. Man, the looks on the sergeants’ faces...Or we’d be listening to a lecture, you know, about
gun maintenance or something, and we’d start nodding off. I
think we just assumed we were too tired to learn anything anyway.
So we’d be falling asleep, and suddenly they’d shout: “Ah! Politzer,
pick up Attias and take a lap around camp. Time to wake up. Let’s
20
go!” And it would start all over again. (pause) So we had each
other’s back...I mean, literally and figuratively, you know? (pause)
Two years ago my unit was down manning forts along the Suez
when they told me that I had leave for a month to visit my parents,
remember? So I flew to the States for three weeks, but right before
I got back to Israel my unit walked into an ambush. The soldiers
were doing a patrol, and suddenly they were attacked by Egyptian
commandos. It was an effective ambush. The guys were cut down.
At least ten guys from my paratrooper unit were killed in that
action. I mean, I’ve been through war combat and everything else,
but it’s like, it hit me like a ton of bricks when I heard everybody
who had been killed there. How could they all be dead? (pause) I
found out the night I got back to my kibbutz. I had just flown in
and hitchhiked from the airport. There was an army Jeep waiting,
parked near the dining hall there, and one of my lieutenants and
two other sergeants were standing next to it. And, you know, they
asked me how I was, and then they started telling me about all the
guys who’d just been killed along the Suez. (pause) They were saying, you know...it was a hopeless situation. The commandos threw
hand grenades and we charged their position, but they opened fire
on us. Machine guns...(pause) The first thing I said was, “What
about Shuki?” And all three of them looked at each other, and
there was just this silence. And, you know, it was like...I knew.
Shuki’s gone. I asked, “What happened?” (pause) Shuki survived
the ambush. It was afterwards when the remaining unit was clearing out the trench that the Egyptian commandos had fired from.
Shuki was going through the trench, and another guy was above
covering him. Shuki was picking up weapons from the Egyptians
soldiers who were dead, you know, throwing the weapons out of
the trench. The other guy was supposed to be watching him. And
one of the Egyptian soldiers was not dead. He hit Shuki with a
submachine gun blast, pointblank. Shuki died very quickly.
(pause) The other guy was obviously not watching Shuki like he
should have been, and if I’d been there, you know, I would have
watched him. I would have protected him, just like he would have
done for me. And this other guy...shit...if I had been there, Shuki
might still be alive. I can’t shake that feeling.
Daniel’s response to Shuki’s death is laced with frustration, whereas the frustration
expressed in Pollak’s interview only appeared in glimmers of regret. The same phrase
used one way in Pollak’s interview means something completely different in the context of Daniel’s monologue. For example, Pollak finished talking about Shuki’s death
by saying, “He died quickly.” It was a simple, heavy phrase to complete a story from his
past. Yet, Daniel uses a similar phrase, “Shuki died very quickly,” not to conclude the
story, but to work up to the final moment in his monologue when he expresses his
anger over not having been there to protect his friend. On top of adding that final bit
21
to the monologue in which Daniel expresses guilt over not having been with Shuki to
protect him, a background story setting up the relationship between Shuki and Daniel
was added at the beginning of the monologue to balance the journey Daniel goes
through during the monologue. With the added sections of the monologue a range of
emotions are shown by the character, rather than just a simple retelling of Shuki’s
death. In the course of this monologue, Daniel goes from amusement to sadness to
anger—a more thorough trip than the one taken by Pollak in his interview transcript.
In Chosen, monologues provide the audience a glimpse into hidden parts of specific characters. Not all the characters deliver one, but when monologues do appear in
the script they are always imbedded in dialogue between characters. Each monologue
represents a moment when one of the characters decides to let another character know
something secret or protected about him or her. This aspect of the monologues is
most important in the character of Eliezer Halfin.
The real Eliezer Halfin was David Pollak’s closest friend on the Olympic team, and
they wrestled in the same weight class for most of their time together on the national
team. They met when Halfin, the northern champion, beat Pollak, the southern champion, at the national wrestling championship in 1970. Pollak described Eliezer Halfin
simply as, “Such a nice guy. Soft spoken.” With so little information about the actual
personality and background of Halfin, his character had to be completely built from
Pollak’s simple description and the little that is written about Halfin in history books.
At some point in Halfin’s early adult life he had immigrated to Israel from the Russian
city of Riga. He was a national wrestling champion in Israel as well as a very successful wrestler back in the Soviet Union. When he was killed in Munich he was twentyfour years old.
In the script of Chosen, Eliezer is not very talkative or aggressive, and he mostly listens while other characters speak. Knowing that he was from Riga allowed for the possibility of writing a past for the character that could resemble the background of the
real Eliezer Halfin. After researching the history of Riga and Jews in the Soviet Union,
a scene between Eliezer and Daniel was added during which Eliezer explains to Daniel
why he left the Soviet Union. This is an excerpt from the scene, taking place on a kibbutz called Misgav Am during the summer of 1970:
ELIEZER
There was an empty lot close to our house in Riga that is now an
office building, and my mother would never let us walk by it. Ever.
We would take the long way home. I always thought it was because
she was worried about, you know, crime...So for years we didn’t
walk by this place. But I got older, and I started leaving the house
by myself, and just to be adventurous, I would walk by this empty
lot. One day I looked closely and saw that some of bricks there that
were lying on the ground were blackened. Like by flames. And this
was around the time they were starting to clear everything away to
start building the offices. And I asked one of the guys standing
there, “Did a building burn down here?” And he said to me, “What
year were you born?” And I said, “1948.” He said, “Seven years
22
before you were born this place burned down.” That was all he
said. The next day I went back, found the same man—he was an
investor for the building project, I think—and asked him what the
place used to be before it burned down. At first he shooed me
away. But finally, he told me. This place, this empty lot, used to be
a synagogue. And he asked me if I knew what that was, and I said
I did. Of course, I didn’t tell him I was Jewish. He explained that
the Germans occupied Riga in 1941, when there were still a lot of
Jews in the city. It was the summer of 1941. All the synagogues in
Riga except for one were burned down at this time, including the
empty-lot synagogue. But before they set fire to it, a large number
of people were rounded up and locked inside. Men and women
and their children. And they all died in the fire. (pause) But, see,
the Germans weren’t the ones who did this. It was the townspeople, the people from Riga, who killed all those families in the synagogue. And it occurred to me, suddenly, that this man—the man
telling me the story—could have been one of the people who did
this. Who, fifteen years later, was turning this place into office
buildings. (pause) That’s when I started to see the ghosts. Like my
mother must have seen. I would walk by the site and hear screams
and crying. I saw the flash of someone’s hair in the construction
scaffolding one day. Of course they were all just childlike hallucinations, I think. But I started dreaming about the fire, having
nightmares about being trapped in a fire. And no matter what I
did in the dream—how I tried to escape or find a way out—the
fire always won. It always won. (pause) So I started wrestling, and
wrestling became a lot more satisfying than dreaming.
Through the monologue delivered by Eliezer in this scene, the audience can begin
to identify the world and background that Eliezer comes from. It also provides clues
as to why Eliezer wrestles and feels most comfortable keeping a low profile. And while
it is very possible that the real Eliezer Halfin never visited the empty-lot synagogue,
he was surely aware of the bloody history of Riga, the town he grew up in and eventually left.
In the final stages of drafting this play, it has become clear that the research and
creation of each character deserves many more pages of analysis and explanation.
Perhaps the most bizarre phenomenon that occurs in the course of capturing a human
life on paper is the way each character begins to change and evolve as their story is told
more and more in depth. For example, the character of Daniel Politzer proved to be
much more moody and uncooperative in his earliest incarnations, the character of
Jackie proved to be much more demure, and the character of Eliezer proved to be
much less humorous.
The reason for these tweaks in each character’s personality was primarily due to
the fact that in between each script revision, more background research was carried
out and more facts were uncovered about the real people upon whom the characters
were based. In one instance, no first name could be found for the character of Mark
23
Slavin’s father, a man who appears in two short but critical scenes. In the list of dramatis personae he was just referred to as Mr. Slavin, an anomaly amongst a group of characters who all possessed first and last names. Subsequently, his character was dry and
uninteresting. Yet, as the research went more and more in depth, a picture was found
of Mark Slavin’s grave, which identified him as the son of Yaacov Slavin. With the discovery of Mr. Slavin’s first name, he suddenly became a more complex, nuanced character in the later drafts.
Similarly, before the real Eliezer’s profession was found in one of the many books
written about the Munich Olympic Massacre, he was described in the list of dramatis
personae as “man from Tel Aviv.” Without any knowledge about his day job, it was challenging to figure out both his economic and social background, the commodities he
would have been able to afford (such as a car), and his expectations for the future. As
soon as it was uncovered that Eliezer had been a mechanic his description changed
from “man from Tel Aviv” to “mechanic from Tel Aviv,” and his personality became
more casual and warm in later drafts.
There is a science to creating characters whose real-life counterparts have already
left traces of themselves in this world. As more factual information is discovered,
specificity in the characters increases. A mechanic would not drive a car he couldn’t fix
(as Eliezer did in the first draft of the play for a particular plot point involving a broken down car). Therefore, another character drives this car, and when it breaks down,
Eliezer is the character to fix it.
However, there is a point in character development when factual information isn’t
enough, and different kinds of specificity are needed for the sake of creating a complex character. Specificity of demeanor, of energy, of tempo, and—unique to any
ensemble piece like Chosen—specificity of alliances and rivalries, interpersonal
dynamics and tension are all set up by the playwright. To assemble facts is one distinct,
challenging task; to make the facts walk and talk is quite another.
24
DRAMATIS PERSONAE FOR CHOSEN (in order of appearance)
CHERYL MARDER (27), woman living on Moshav Bilu, American-born, seven
months pregnant.
DANIEL POLITZER (24), paratrooper in Israeli army living on Kibbutz Misgav Am,
American-born, member of Ha’Poel Yerushalayim and the National Wrestling Team,
middleweight.
YITZHAK “JACKIE” MARDER* (30), Cheryl’s husband, farmer, builds chicken
coops for a living.
ELIEZER HALFIN (24), mechanic from Tel Aviv, recent immigrant from Russia,
member of Ha’Poel Tel Aviv and the National Wrestling Team, middleweight, killed
in the Munich Olympic massacre.
AVRAM “UMALEH” ASHURI (30), mechanic on Kibbutz Misgav Am, member of
the National Wrestling Team, heavyweight.
MOSHE “MUNI” WEINBERG (33), coach of Ha’Poel Tel Aviv, coach of the National
Wrestling Team, killed in the Munich Olympic massacre.
GADI TSABARI (24), man from Tel Aviv, member of the National Wrestling Team,
survivor of the Munich Olympic massacre, lightweight.
IBRAHIM AZHAR* (26), man from Jerusalem, member of Ha’Poel Yerushalayim,
heavyweight.
YOSEF GUTFREUND* (40), coach of Ha’Poel Yerushalayim, international Olympic
wrestling referee, owner of a shop in Jerusalem, killed in the Munich Olympic massacre.
REFEREE*
MARK SLAVIN* (18), recent immigrant from Russia, member of the National
Wrestling Team, middleweight, killed in the Munich Olympic massacre.
YAACOV SLAVIN* (45), father of Mark Slavin.
* The following parts can be played by one actor instead of two:
Yitzhak “Jackie” Marder/ Referee
Ibrahim Azhar/ Mark Slavin
Notes
1
Scott C. Cummings, “History Plays,” The Boston Phoenix, February 3-10, 2000.
2
Rose Tremain, “Writing Historical Fiction,” 1 March 2007. <http://www.bbc.co.uk /dna/get
writing /module9p>.
3
David Pollak, personal interviews, March 2006.
4
David Pollak, personal interviews, March 2006
25
You Are What You Eat: A Technique
for Isotopic Analysis of
Archaeological Amino Acids
Author:
Andrew B.
Newman
Andrew is a senior and will graduate with majors in physics and
math. His first undergraduate
research experience was in the
lab of Professor Ken Kelton during the summer following his
freshman year and continued
through the fall of 2005. Since
then Andrew has worked at an
astronomical observatory in
Chile, a particle physics lab at
Columbia University, and an
archaeological science lab at
Oxford. “Undergraduate research
has allowed me to sample several
interesting fields and helped me
decide what to pursue after graduation.” Andrew plans to attend
graduate school in astrophysics
next year.
KEY TERMS
•
•
•
•
•
26
Chromatography
Collagen
Paleodiet
Radiocarbon
Stable Isotope
A B S T R AC T
Stable isotope analysis of archaeological remains offers a wealth of
information about the diets of ancient peoples. Many studies have
shown that both stable isotope and radiocarbon measurements of
individual amino acids offer greater insight than does measurement of
bulk protein alone. Therefore a particular interest exists in a process
that can separate amino acids in proteins, particularly human bone
collagen, while maintaining the integrity of the isotopic signal.
Previous methods have either undesirably used organic reagents or
have failed to separate several interesting amino acids. This paper
presents the development of a chromatography method that achieves
separation of 15 amino acids in a mock collagen sample, without
introducing any outside carbon. The method has also been successfully tested on an archaeological sample. Further development of this
method will allow for more routine and accurate isotopic measurements of individual amino acids in archaeological collagen samples,
which could greatly expand the data available for reconstructing paleodiets. Results for hair protein as an isotopic indicator of migration
are also presented.
R E S E A R C H M E N T O R : J A M E S S . O. M C C U L L A G H ,
P H . D. , C H E M I S T R Y R E S E A R C H L A B O R AT O R Y,
OX F O R D U N I V E R S I T Y
Dr. McCullagh currently works in two areas of application within mass
spectrometry. The first utilizes a forensic approach to archaeology at
the molecular level wherein stable isotope mass spectrometry is used
to develop new methods for tracking diet and disease markers in
human hair. The second area is a more conventional one and involves
the application of mass spectrometry to understanding the modification of proteins within the body.
AC K N OW L E D G E M E N T S
The work presented here was conducted at the Research Laboratory
for Archaeology and the History of Art (RLAHA) at the University of
Oxford during a six-week project made possible by an Israelow
Research Award, for which the author is grateful. The author thanks
Professor Robert Hedges for arranging the project and James S. O.
McCullagh and Bradley Hull for their mentorship and guidance. The
data presented here are the property of the RLAHA and may not be
reused or reproduced without permission. We are grateful to the Mary
Rose Trust for supplying pig bone.
1 INTRODUCTION
1.1 Isotopes in Archaeology
Within the past sixty years, archaeology has been revolutionized by advances in chemistry, physics, and biology. Since William Libbey’s discovery in 1949 of the potential of
radiocarbon to serve as an atomic clock, archaeologists and scientists have recognized
the power of conducting archaeology at the molecular and atomic levels. Today, an
array of scientific techniques is available to assess archaeological materials, providing
not only the age of artefacts and remains, but clues to the subsistence strategies,
migration patterns, economic systems, and technological development of ancient peoples. Archaeological science is devoted to developing such methods and applying them
in collaboration with field archaeologists. Combining scientific techniques with traditional archaeological methods often provides new light by which to examine the past.
Archaeology at the atomic level is primarily concerned with the isotopic composition of materials. Chemical elements naturally occur in a variety of isotopes which are
nearly identical chemically but differ in mass. Unstable isotopes decay radioactively,
while stable isotopes persist indefinitely. Heavier isotopes tend to react more slowly1,
which can cause the products to have varying isotopic compositions. This process is
called fractionation and is responsible for the meaningful information present in the
isotopic signal. As an example, consider carbon, which occurs in three natural isotopes: 12C, 13C, and 14C, occurring in the ratio2 100 : 1.1 : 10-12. The lighter isotopes are
stable, while 14C (radiocarbon) decays radioactively. The stable isotope 13C is central to
isotopic studies of paleodiet, because of the way it is fractionated by plants at the base
of the food web.
Plants may be classified according to their photosynthetic pathway. The C3 pathway,
so called because the first compound formed contains three carbon atoms, is the most
common and is used by wheat, rice, root crops, and legumes. The C4 pathway is an
adaptation to arid environments and is found in plants such as maize, sorghum, and
millet, all of which, fortunately, are highly archaeologically significant.3 The C3 and C4
pathways fractionate carbon differently. The degree of fractionation is measured by
where for carbon the standard is based on a reference fossil. For nitrogen, δ15N is similarly defined using the 15N/14N ratio and an atmospheric standard. Atmospheric CO2
has a δ13C of –7‰, meaning that its 13C/12C ratio is 7 parts per mil lower than that of
the standard. For archaeological C3 and C4 plants, the averages are –26‰ and
–12.5‰, respectively.4 Not only are the averages significantly different, but the ranges
observed in C3 and C4 plants show no overlap. Thus, types of plants may be distinguished based on their isotopic signatures. Furthermore, the isotopic signals of consumers reflect those of their diets5, making it possible to infer the plant diets of ancient
people by measuring the isotopic content of their remains.
Several stable isotopes have proven useful in archaeology6, but the most widely
studied applications use 13C and 15N to study paleodiet. Bone collagen is frequently the
organic material selected for analysis, because it is found well-preserved in many
27
archaeological settings and is representative of long-term diet. Taken together, δ13C
and δ15N data can discriminate among a wide variety of diets. Nitrogen complements
carbon by exhibiting a trophic level effect, in which the isotopic signal depends on the
consumer’s position in the food chain. It may be used to estimate the quantity of meat
in an individual’s diet, as well as the degree of marine protein consumption.7 8
Early archaeological applications of stable isotopes focused on maize domestication in the New World. This situation is ideal for isotopic study, because maize is a C4
plant, whereas almost all other North American plants are C3. Van der Merwe and
Vogel9 studied skeletal remains in North American woodland sites and found dramatically changing δ13C values beginning around AD 1000, allowing them to trace maize
domestication through time. By combining 13C and 15N measurements, Ambrose and
DeNiro10 showed that it is possible to distinguish marine from terrestrial consumers,
pastoralists from farmers, grain versus non-grain farmers, and camel versus capribovine pastoralists in Africa.
1.2 Amino acids in archaeology
There has been much interest in analyzing the isotopic contents of the individual
amino acids (AAs) from which proteins are built, which frequently vary from that of
the bulk protein.11 Examining samples at the AA level provides several advantages. It
provides a way to reduce contamination by studying only AAs likely to come from
collagen and not the soil.6 The most exciting possibility offered by the isotopic
analysis of individual AAs is the greater specificity of dietary information they provide. Amino acids are classified as dispensable if they may be biosynthesized and
indispensable if they must be provided by the diet. Since most dietary protein comes
from meat, the indispensable AAs should reflect the isotopic signal of the meat consumed. Several dispensable AAs are thought to be assembled largely from carbon
from non-protein sources, mainly carbohydrates.12 The isotopic signatures of dispensable and indispensable AAs, therefore, should reflect the isotopic contents of
different dietary sources.
To meaningfully interpret isotopic data, it is clearly important to understand how
dietary carbon is routed within the body, about which much remains unknown. In
fact, stable isotopes are often used to address this question by feeding animals isotopically controlled diets and measuring the resulting isotope levels in body tissues.3
The archaeologist, of course, must go about this process in reverse. In fact, dietary
routing in humans may be investigated by this reverse approach through a novel
application of radiocarbon. This method takes advantage of the marine reservoir
effect, the phenomenon by which marine carbon systematically appears about 400
years older in radiocarbon years than coeval terrestrial carbon, due to the slow mixing of dissolved marine CO2 with atmospheric CO2.13 Given collagen from an individual with high marine protein intake, AAs, which are either routed from or biosynthesized from dietary protein should appear older in radiocarbon years, in proportion to the amount of routing that occurs. To test this approach, it is first necessary
to develop a method for separating AAs in collagen while maintaining the integrity
of the isotopic signal.
28
2 SEPARATING AMINO ACIDS IN COLLAGEN
To test our ability to separate amino acids in archaeological collagen samples, we used
collagen extracted from pig bone that was excavated from the wreckage of the Mary
Rose. This vessel sank off the coast of England on 19 July 1545, overburdened by crew
and cargo, killing nearly all 600 persons aboard. Excavations before the 1982 raising of
the ship recovered barrels of beef, pork, mutton, and fish.14 The pig collagen is ideal as
a radiocarbon standard, because we know its exact age. To extract individual AAs, the
collagen sample was first hydrolysed (split into its constituent AAs) by an acid bath.
The hydrochloric acid was then evaporated, leaving the AA residue behind (see
McCullagh et al.12 for more detail). In order to conserve the archaeological sample, we
frequently tested and refined our method using a reference solution made from individual AAs. The AAs were mixed in the same proportions found in collagen, as shown
in Table 1, diluted to about 10 mg/mL.
2.1 Difficulties in Chromatography
The special demands of isotopic analysis, especially radiocarbon dating, impose constraints that render most chromatographic methods for separating AAs infeasible. Our
studies employed high performance liquid chromatography (HPLC), a separation
method in which the sample and a mobile phase acidic solution are forced at high pressure through a column containing a stationary phase, which consists of micrometersized beads that chemically interact with the sample. Different compounds in the sample are retained by the stationary phase for varying durations and hence elute from the
column at different times. Eluting compounds are detected by their absorption of
ultraviolet (UV) light. A sample chromatogram is shown in Figure 1. On-line chromatography techniques directly send the separated fractions to the mass spectrometer.
The most common problem with typical chromatographic techniques from an isotopic perspective is the addition of carbon. Any carbon introduced must either be
removed or corrected for by back-calculations. Neither approach is desirable, because
both decrease accuracy and introduce potential contamination sources, which can significantly alter radiocarbon dates. It is therefore desirable to effect separation without
additional carbon. Additionally, such a separation is attractive because the first commercially available, on-line system which links HPLC and isotope ratio mass spectroscopy (IRMS) has recently appeared12, but the technique requires that the mobile
phase be inorganic. (IRMS measures stable isotope ratios.) No satisfactory separation
method yet exists that satisfies these stringent demands.
2.2 High performance liquid chromatography
Our experimental setup consists of an automated Varian ProStar HPLC apparatus interfaced with a Varian UV detector operating at 205nm and a fraction collector. The core
component of any LC configuration is the column. We use the mixed-mode Primesep A
column manufactured by SIELC Technologies. This is a reverse phase (RP) column
which contains additional negative charges in the stationary phase, enabling an ion
exchange (IE) retention mechanism as well. In general, the less polar AAs are retained by
hydrophobic interactions with the RP surface, while the more polar AAs are retained by
the IE component.12 This column therefore allows the separation of the dispensable and
indispensable (generally more polar and less polar, respectively) AAs in a single stage.
29
The elution of AAs is highly sensitive to a number of parameters in the chromatography process. Clearly the choice of mobile phase is of utmost importance. Since we
seek to avoid adding organic substances, mineral acids are the natural candidates.
While hydrochloric acid (HCl) would be desirable because it is volatile and thus easily removed from the eluted solution, it is not compatible with our column. Instead, we
use sulfuric acid (H2SO4) for the mobile phase.
Both the IE and AA charge states are strongly affected by the pH of the mobile phase,
determined by the H2SO4 concentration. The ProStar system is capable of mixing reservoirs
of water and dilute H2SO4, so a gradient program may be specified. We experimented with
single-gradient programs at concentrations of 0.1% and 0.3% (by volume) and a variety of
dual-gradient programs. In general, we found that lower concentrations yielded better separation of the more polar AAs, which elute initially, while higher concentrations were more
favourable for the less polar AAs. We thus settled on a gradient program that begins with
pure water, increases to 0.1% H2SO4 linearly from 20-40 min., and further increases to 0.3%
H2SO4 from 56-60 min., where it remains for the duration of the program.
Further parameters include flow rate, sample concentration and volume, and temperature. Samples with concentrations of 10 mg/mL were found to separate well, while
performance suffered slightly at twice this concentration and severely at four times. For
each separation, 1 mL of sample solution was injected. Temperature was kept at 25°C,
and the flow rate was 6 mL/min. A chromatogram constructed from this program performed on the reference collagen solution is shown in Figure 1. Fifteen peaks may be
identified, exhibiting varying degrees of separation. (Other separation programs that
we tried yielded more peaks, but with less separation between several peaks of interest.)
In order to identify which peaks represent which AAs, we ran individual AA solutions through the program in the same concentration at which they are found in the
reference collagen solution. Ideally, the individual peak elution times should match
those in Figure 1, but practically, the solution concentration and interactions with
other amino acids apparently play a large role. Thus, elution times are often significantly shifted. Nevertheless, by comparing relative elution times of the AAs and peak
shapes, which sometimes deviate from the ideal, it is possible to identify many of the
peaks in Figure 1. We combined this data with experience from previous analytical
chromatography programs to produce the identifications shown.
The chromatogram for a sample of Mary Rose pig collagen is shown in Figure 2.
Relative to the reference solution in Figure 1, some peaks are time-shifted or overlap
other peaks. Nevertheless, 13 peaks can be identified in the Mary Rose sample. The separated AA fractions from this sample were collected at one-minute intervals. The oneminute fractions were combined into eight different collected solutions. Note that most
collected solutions contain more than one AA peak, even when the resolution is sufficient to collect single AAs. This is done for two reasons. First, entire peaks must be collected to obtain accurate isotope measurements, because the isotopic composition may
vary during the elution. Those peaks which do not exhibit complete separation must
therefore be collected together. Second, at least 1 mg of carbon is necessary for radiocarbon dating. The AAs that are present in lower concentrations in collagen may therefore
be combined with other peaks. (If it is scientifically interesting to examine such an AA
individually, fractions collected from several samples can be combined to achieve the
necessary 1 mg.) Isotopic analysis of the collected fractions remains for future work.
30
2.3 Removing the mobile phase
Amino acid samples are analyzed by IRMS in powder form. It is therefore necessary to
remove water from the collected solutions by lyophilization (freeze-drying). The
H2SO4 in the collected fractions is non-volatile, so lyophilization yields a concentrated mixture of acid and paste that is unlikely to combust well in the IRMS device. It is
thus desirable to remove the H2SO4. This must be done in a way which recovers most
of the AAs and introduces minimal fractionation.
One approach is to precipitate the sulfate ions (SO42-) by addition of barium
hydroxide (Ba(OH)2), which reacts as follows:
H2SO4 (aq) + Ba(OH)2 —>2H2O + BaSO4 (s)
Since barium sulfate (BaSO4) is insoluble, the solution may be centrifuged to recover
the AA solution while discarding the precipitate. Additionally, the pH is neutralized by
this reaction.
This idea was tested as follows. Four solutions were created containing the amount
of glycine (Gly) present in 1 mL of collagen solution at the concentrations used in this
study. In two of these solutions, the Gly was dissolved in 12 mL of mobile phase, while
in the other two 30 mL was used. These volumes were chosen to correspond to typical peak widths of 2 and 5 min, respectively. From the stoichiometry of the precipitation reaction, the necessary amount of barium sulfate octahydrate was calculated and
added to the solutions. Two methods for separating the precipitate from the supernatant were tested. In the first, both solutions were centrifuged for 20 min at 3000
rpm. The supernatant was decanted and the precipitate was rinsed with purified water.
The process was then repeated. In the second method, both solutions were placed in
Centriprep tubes which contain inner and outer tubes separated by a filter. Ideally, the
filter would block the crystalline BaSO4 from entering the inner tube. The Centriprep
tubes were centrifuged by the same process, with outer tube rinsed after the first spin
and the contents of the inner tube collected twice. All samples were then frozen,
lyophilized, and prepared for IRMS analysis.
IRMS measurements show that the filtered samples clearly suffered from a large
amount of fractionation, which renders this method unsuitable. The unfiltered samples
were much less fractionated, though still too much for radiocarbon dating, and too little glycine was recovered. It seems that the filter may have contaminated the solution
with its own carbon. Measurements of C:N ratios support this conjecture. The filtered
samples were vastly altered, suggesting a large addition of carbon. This example illustrates the great care which must be exercised in each stage of sample preparation.
A second approach for removing sulfate ions involves passing the collected solution through a column of ion exchange resin. We chose the DOWEX 66 resin, a weak
anion-exchange resin that is capable of removing strong acids from solution. A small
column of approximately 2 cm diameter and 6 cm height was used to test this method.
The plastic column and filter were thoroughly rinsed. The ion exchange resin was
added and thoroughly rinsed before being generated with 1 M NaOH solution, after
which it was again rinsed. A reference collagen solution was passed through the column and collected. The column was rinsed with approximately ten column volumes
and the eluent collected in order to recover the maximum amount of glycine. The col-
31
lected solution was concentrated to the same level as the reference collagen solution.
Addition of Ba(OH)2 showed that the sulfate ions were successfully removed, but
HPLC analysis revealed that the column also retained glycine.
3 HAIR ISOTOPES AND GEO GRAPHIC MOVEMENT
In addition to bone collagen samples, several hair samples were analyzed. Although
some preliminary work has been done on individual AA analysis for hair keratin, these
samples were analyzed in bulk. Because hair grows relatively quickly, it offers isotopic
information on timescales much shorter than that of bone collagen. It may therefore
be used to detect abrupt changes in diet, such as those made possible in modern times
by long-distance migration. An earlier study showed that the hair keratin of individuals in the US is depleted in 13C relative to individuals in the UK, and that migrants
from one country to the other equilibrate to the 13C of their new home in about 4
months.15 The authors of this study ascribe this difference mainly to livestock feeding
differences. In North America, livestock are fed high quantities of maize, a C4 plant,
while in Europe C3 grains are used.
To investigate correlations between hair isotope values and unusual diets or migration, hair samples from 10 donors were collected along with self-descriptions of diet.
For each individual, the hair was kept intact or cut into two or three pieces, depending on the mass of hair available. The samples were submitted for 13C and 15N IRMS
analysis. The hair samples exhibit a range of carbon and nitrogen isotope values, ranging from δ13C = -21.7 to -19.2 and δ15N = 8.7 to 9.7. The clearest trends are found in
two migrant donors, whose δ13C data are shown in Figure 3. Donor 1 moved from the
US to the UK, and Donor 7 moved from Estonia to the UK. These results indicate that
hair may be used as a tracer of migration between countries with isotopically distinct
diets. No correlations between isotopic measurements and self-reported anomalous
dietary practices (e.g., high meat intake) were found, although the sample size was
quite small.
4 CONCLUSIONS
Fractionation within food webs creates distinct isotopic signatures for different
dietary patterns, which are recorded in the bodies of consumers. Although analysis of
bulk proteins in archaeological remains provides much useful information, isotopic
studies of amino acids promise new insight into ancient diets. In order to perform
such studies, it is necessary to have a technique that can separate many amino acids
without fractionation. We have developed an HPLC method to separate 15 amino
acids with minimal fractionation. It has been successfully tested on both a mock collagen solution and archaeological pig collagen. A procedure for removing the acidic
mobile phase from the collected fractions without fractionation is still necessary and
will be the focus of future work. Additionally, hair keratin seems to serve as a migration indicator in some dietary contexts.
32
Amino Acid
Glycine (Gly)
Proline (Pro)
Alanine (Ala)
Hydroxyproline (Hyp)
Glutamic acid (Glu)
Aspartic acid (Asp)
Argenine (Arg)
Serine (Ser)
Lysine (Lys)
Leucine (Leu)
Valine (Val)
Threonine (Thr)
Phenylalanine (Phe)
Isoleucine (Ile)
Histidine (His)
Tyrosine (Tyr)
Methionine (Met)
% (by number)
31.87
12.29
11.39
10.36
7.19
4.70
4.70
3.60
2.80
2.60
2.40
1.80
1.40
1.30
0.60
0.50
0.50
Table 1. Amino acid composition of human bone collagen used to create the reference solution described in the text. Percentages are by number of amino acid
molecules. Hydroxylysine, a small component of real collagen, was not included.
Figure 1. Chromatogram showing the separation of amino acids in mock collagen solution using the separation program described in the text. The vertical
axis shows the signal from the UV detector, and the horizontal axis shows the
elution time. In this example, 15 peaks corresponding to amino acids are identified. (The spikes at the beginning and end are artifacts.) The amino acids are
identified by comparing the peak positions with those in single-AA solutions
subjected to the same HPLC program, as described in the text.
33
Figure 2. Chromatogram showing the separation of amino acids in pig collagen
recovered from the Mary Rose wreckage. The agreement with Figure 1 is fairly
good, indicating that the collagen extraction yielded a pure sample.
Figure 3. Variation through time of d13C values in the hair keratin of migrant
individuals. Donor 1 moved from the US to the UK, and Donor 7 moved from
Estonia to the UK. Hair was divided into three lengths to obtain measurements
of the three temporal periods shown. The total time span represented in the
hair growth is several months. The typical measurement error is 0.1‰.
34
Notes
1
Gannes, L. Z., C. M. del Rio, and P. Koch. 1998. Natural abundance variations in stable
isotopes and their potential uses in animal physiological ecology. Comparative Biochemistry
and Physiology 119A: 725-737.
2
van der Merwe, N. J. 1982. Carbon isotopes, photosynthesis, and archaeology. American
Scientist 70: 596-606.
3
Ambrose, S. H. and L. Norr. 1993. Experimental evidence for the relationship of the carbon
isotope ratios of whole diet and dietary protein to those of bone collagen and carbonate.
In Prehistoric Human Bone: Archaeology at the Molecular Level, eds. J. B. Lambert and
G. Grupe, 1-37. New York: Springer-Verlag
4
DeNiro, M. J. 1987. Stable isotopy and archaeology. American Scientist 75: 182-191.
5
DeNiro, M. J. and S. Epstein. 1978. Influence of diet on the distribution of carbon isotopes
in animals. Geochimica et Cosmochimica Acta 42: 495-506.
6
Tripp, J. A. and R. E. M. Hedges. 2004. Single-compound isotopic analysis of organic materials
in archaeology. LC-GC Europe (June 2004): 358-364.
7
Schoeninger, M. J., M. J. DeNiro, and H. Tauber. 1983. Stable isotope ratios of bone collagen
reflect marine and terrestrial components of prehistoric diet. Science 220: 1381-1383.
8
Schoeninger, M. J. and M. J. DeNiro. 1984. Nitrogen and carbon isotopic composition of bone
collagen from marine and terrestrial animals. Geochimica et Cosmochimica Acta 48: 625-639.
9
van der Merwe, N. J. and J. C. Vogel. 1978. 13C content of human collagen as a measure
of prehistoric diet in woodland North America. Nature 276: 815-816.
10 Ambrose, S. A. and M. J. DeNiro. 1986. The isotopic ecology of East African mammals.
Oecologia 69: 395-406.
11 Hare, P. E., M. L. Fogel, T. W. Stafford, Jr., A. D. Mitchell, and T. C. Hoering. 1991. The isotopic
composition of carbon and nitrogen in individual amino acids isolated from modern and fossil
proteins. Journal of Archaeological Science 18: 277-292.
12 McCullagh, J. S. O., D. Juchelka, and R. E. M. Hedges. 2006. Analysis of amino acid 13C abundance from human and faunal bone collagen using liquid chromatography/isotope ratio mass
spectrometry. Rapid Communications in Mass Spectrometry 20: 2761-2768.
13 Ascough, P., G. Cook, and A. Dugmore. Methodological approaches to determining the marine
radiocarbon reservoir effect. Progress in Physical Geography 29.4: 532-547.
14 Tripp, J. A., J. S. O. McCullagh, and R. E. M. Hedges. 2006. Preparative separation of underivatized amino acids for compound-specific stable isotope analysis and radiocarbon dating of
hydrolyzed bone collagen. Journal of Separation Science 29: 41-48.
15 McCullagh, J.S.O., Tripp, J.A. & Hedges, R.E.M. 2005. Carbon isotope analysis of bulk keratin
and single amino acids from British and North American hair. Rapid Communications in Mass
Spectrometry 19: 3227-3231.
35
Overexpression of Cx43 and NF200 in
Hibernating Species Citellus undulates
During the Hibernation State
Author:
Sangita Sudharshan
Sangita is a senior majoring
in biomedical engineering. She
decided to try undergraduate
research at Washington University because she wanted to gain
experience working at the cutting-edge of biomedical engineering. She was interested in
learning about the entire research
process and her mentor Professor Igor Efimov helped her
gain the experience she sought.
Her research in cardiac electrophysiology has affected her
future plans to study cardiology
after medical school, which she
will enter in the fall. Sangita also
now realizes that although she
will not pursue research full-time,
she plans to continue to do
research at some level throughout her medical career.
KEY TERMS
• Torpor
• Fluorescence
Immunohistochemistry
• Epicardium
• Mid-myocardium
• Endocardium
A B S T R AC T
Most mammalian species experience cardiac arrhythmias in hypothermic conditions. Hibernating mammals, on the other hand, are able to
maintain proper cardiac conduction and rhythm even at -2o C body temperature. The purpose of this study was to understand the underlying
molecular mechanisms utilized by the hibernating species Citellus undulatus (Siberian ground squirrel, GS) that allow the GS to maintain proper conduction under extreme hypothermic conditions by looking at protein expression in the left ventricular free wall. This study aims to quantify expression of connexin 43 (Cx43), the main ventricular gap junctional protein, and neurofilament 200 (NF200), a marker of intracardial nervous cells. Fluorescence immunohistochemistry was performed on transmural sections of the left ventricular free wall of summer active (SA,
n=2), winter hibernating (WH, n=3), interbout arousal (IBA, n=3), and
entrance into torpor (ET, n=2) ground squirrel (GS) heart. Quantitative
analysis indicates that there is significant overexpression of Cx43 in WH
epicardium and endocardium compared with ET and SA. NF200 expression in the endocardium was found to be higher in WH GS than in ET,
IBA, and SA. Citellus undulatus can maintain proper heart rhythm during
extreme hypothermic conditions through overexpression of Cx43 in ventricular myocytes. For the first time, the pattern of autonomic innervation
was characterized in endocardial ventricular tissue of a hibernating
species through NF200 immunostaining. Overexpression of NF200 in
WH indicates an important role of autonomic regulation in ventricular
function during hibernation. Long-term goals of this research include
contribution to treatments that allow for extended organ storage prior to
transplantation, as well as development of anti-arrhythmia therapies.
FA C U LT Y M E N T O R : I G O R R . E F I M O V,
P H . D. , A S S O C I AT E P R O F E S S O R O F
BIOMEDICAL ENGINEERING
Professor Efimov’s research is focused on advancing the understanding
of the fundamental mechanisms of arrhythmogenesis, disturbances of
cardiac rhythm that can result in sudden cardiac death, and on developing lifesaving antiarrhythmic therapies including pharmacological agents
and implantable devices. The goals of the Efimov lab include providing
the best possible research environment for the education of future biomedical engineers and to conduct research investigations in the field of
cardiac bioelectricity for the advancement of our understanding of heart
disease and the development of better therapies.
AC K N OW L E D G E M E N T S
Thank you to the Efimov Lab for providing the materials and guidance
throughout the summer. Also, thank you to the Hoopes Undergraduate
Research Grant for its financial support. Support for the research conducted was provided by the Stanley and Lucy Lopata Endowment and
the Russian Foundation for Basic Research.
36
INTRODUCTION
ibernating mammalian species have evolved to survive extreme conditions which
would result in cardiac arrest in humans and other non-hibernating species1-3.
During the hibernation period, hibernating mammals are adapted to tolerate severe
hypothermia, hypoxia and ischemia, without succumbing to cardiac arrest. Even at 2oC, hibernating species of ground squirrels (GS) maintain adequate sinus rhythm
and cardiac conduction2-4. However, for non-hibernating mammals, including
humans, the heart goes into cardiac arrest due to ventricular fibrillation (VF) at 10°C
to 27°C 1-3.
Mammalian hibernators do not have cardiac dysfunction and/or malignant
arrhythmias during either the onset of hibernation, which is a relatively slow gradual
process, or during arousal, despite the fact that the body temperature may increase or
decrease by 30oC and metabolism can change by more than 100 times3. Although the
application of this physiological phenomenon to clinical medicine is strongly desired,
it has been prevented by a poor understanding of the hibernation mechanism.
The purpose of this study was to understand the underlying mechanisms utilized
by the hibernating species Citellus undulatus (Siberian ground squirrel) that allow
them to maintain proper heart rhythm under extreme hypothermic conditions.
During hibernation (or torpor), ground squirrels go through periods of arousal that
occur about every two weeks in between torpor bouts and last for approximately 12
hours. This study aimed to look at the change in protein expression during the hibernation season, by examining ground squirrels during four states: winter hibernating
(WH), entrance into torpor (ET), interbout arousal (IBA), and summer active (SA).
Figure 1 below shows the body temperature changes that GS undergo during the physiologically distinct states.
H
Figure 1: Body temperature Tb as a function of time (months) in a golden-mantled ground squirrel (modifications on reproduction by Carey et al. 1)
37
Mechanisms of hibernator resistance to spontaneous hypothermia-induced conduction block and tachyarrhythmia, including VF, remain unclear. Onset of cardiac arrest
is frequently associated with the following factors, which must be considered as contributing to the difference in propensity to VF between hibernators and non-hibernators: dispersion of repolarization/refractoriness and of ventricular conduction, adrenergic innervation density (previous studies have reported that ventricular muscle
fibers in hibernators lack sympathetic innervation), metabolic factors (different temperature activity curves in hibernators compared to non-hibernating mammals) and
ultrastructure (different density of connexins in ventricular muscle at different hibernation states). This study concentrated on two of the possible factors.
1.Gap junction proteins, called connexins (Cx), form channels that facilitate cell-cell
communication. Connexins are used to pass along the signal for synchronous contraction through the cardiac cells. Cx43 is the most abundant connexin in the ventricle and was a major focus of this study. It is hypothesized that the level of Cx43
increases during hibernation in order to maintain proper cardiac rhythm. Duker
et al. suggested that hibernators are able to withstand hypothermic conditions
because of changes in conduction velocity 5. Increased Cx43 would provide an
increase in conductance, which could protect the heart from arrhythmias.
2.Autonomic innervation consists of both sympathetic and parasympathetic nerves,
which influence heart rate and cardiac muscle contraction. Neurofilaments are the
main structural components of nerve fibers, and are composed of three units with
different molecular weights. Neurofilament 200 (NF200) is the heaviest unit and is
a protein indicator of all autonomic nerve fibers. We used NF200 staining to study
how autonomic innervation changes during the stages of hibernation.
Immunohistochemistry and density quantification were performed in transmurallysectioned left ventricular free wall tissue to study expression of Cx43 and NF200. The
results of the immunohistochemistry and quantification give insights into the molecular mechanisms employed by hibernators to protect themselves against arrhythmias
during hibernation.
If successful, this research could lead to identification of new pharmacological targets for anti-arrhythmia therapy in settings of hypothermia, ischemia and hypoxia. It
could also contribute to treatments of trauma, low-temperature surgical procedures
and extended organ storage prior to transplantation.
MATERIALS AND METHODS
Preparation of Tissue Sections
Left ventricular tissue from each heart was imbedded in OCT and cryosectioned
transmurally in the region near the apex. Sections of thickness 16 µm were taken from
the following ground squirrels (GS): WH (n=3) SA (n=2), ET (n=2), and IBA (n=3).
38
Immunohistochemistry
Immunohistochemistry was performed using the protocol shown below.
Fluorescence Immunohistochemistry Protocol
DAY 1
• Wash sections in 4% Formaldehyde 6 minutes at room temperature
• Wash slides 3 times for 5 minutes each in Phosphate Buffer Saline (PBS)
• Dry off slides with Kimwipe and circle sections with PAP pen
• Apply 0.10% Triton X-100 to each section for 15 minutes
• Remove excess Triton
• Wash slides 3 times for 5 minutes each in PBS
• Pour 10% Normal Horse Serum into container for 1 hour
(36 mL PBS+4mL NHS for each container)
• Dry off slides with Kimwipe
• Apply diluted (in 1% PBSA) primary antibodies overnight in 4ºC
DAY 2
• Remove excess Triton
• Wash slides 3 times for 10 minutes each in PBS
• Dry off slides with Kimwipe
• Apply diluted (in 1% PBSA) secondary antibodies (concentration 1:1000)
2:30 hours at room temperature
• Remove excess antibody
• Wash slides 3 times for 10 minutes each in PBS
• Apply DAPI mounting solution and coverslip
• Seal with clear nail polish
Expression of Cx43 in GS Left Ventricular Free wall
Immunohistochemistry was performed following the protocol described above. In
order to measure the density of Cx43, GS tissue was stained for both Cx43 and alphaactinin. The purpose of alpha-actinin staining was to produce a background staining of
the entire tissue. The primary antibodies used were Sigma Rb-Cx43 (1:1000) and Sigma
Ms-alpha-actinin (1:1600). The fluorescent secondary antibodies were Alexa555 Gt
anti-Rb and Alexa488 Gt anti-Ms. Following staining, fluorescence pictures were taken
using the Nikon confocal microscope. Four or five pictures from each location (epicardium, mid-myocardium, and endocardium) were taken for each GS. The epicardium was defined as the first 100µm from outer layer of connective tissue. Mid-myocardial pictures were taken from the region adjacent to the epicardial layer. The endocardium was defined as the first 100 µm from the inner layer of connective tissue.
39
Expression of NF200 in GS Left Ventricular Free wall
Immunohistochemistry was performed following the protocol described above. In
order to measure the density of NF200, GS tissue was stained for both NF200 and
alpha-actinin. The purpose of alpha-actinin staining was to produce a background
staining of the entire tissue. The primary antibodies used were Chemicon Ms-NF200
(1:200) and Sigma Rb-alpha-actinin (1:1000). The fluorescent secondary antibodies
were Alexa594 Gt anti-Ms and Alexa488 Gt anti-Rb. Following staining, fluorescence
pictures were taken using the Nikon confocal microscope. Four pictures from the
endocardium were taken for each GS.
Quantification of Cx43 and NF200 densities in GS Left Ventricular Free wall
Cx43 and NF200 tissue density was measured using a custom MATLAB program written
in the Efimov Lab. The density was found by calculating the areas of Cx43 and NF200, as
well as the area of alpha-actinin in each picture. Then, the area of Cx43 or NF200 was
divided by the corresponding alpha-actinin area to calculate the total density of the protein in the tissue. An example of the calculation procedure for Cx43 density is shown
below in Figure 2. The same method was used for NF200 density measurements.
(a): confocal composite picture
(b): Cx43 area
(c): alpha-actinin area
Figure 2: Calculation of Cx43 using a custom MATLAB program was performed
by dividing the white area in (b) by the white area in (c)
Statistical Analysis
T-tests were performed on the quantification data and P<0.05 was considered statistically significant.
RESULTS
Expression of Cx43 in GS Left Ventricle
In our study of the changes in Cx43 expression at different hibernation/active states,
we observed different levels of Cx43 in the epicardial, mid-myocardial, and endocardial layers of left ventricle of hibernating GS. Select pictures of Cx43 expression from
each hibernation state can be seen in Figure 5. Figure 3 shows a graph of Cx43 densities in each layer for all of the different hibernation/active states.
40
Figure 3: Histogram showing the results of quantification of Cx43 density in epicardium, mid-myocardium, and endocardium in GS LV free wall. Values obtained
from confocal images. Three animals of IBA and WH were sampled, and two
animals of ET and SA. In the epicardium, 5 sample pictures were taken from each
animal, except from IBA (4 pictures). In the mid-myocardium, 5 sample pictures
were taken from each animal. In the endocardium, each animal had 4 sample
pictures. The data from each group are expressed as mean±SEM. In the graph,
* indicates significant difference between groups marked by bar with P<0.05.
** indicates significant difference between group marked by bar with P<0.01
In the epicardium, the order of mean Cx43 density in different states from highest to lowest was WH, ET, SA, and IBA. T-tests were performed to compare the results from WH
and each of the other states. Results indicate statistical significance for all differences
In the endocardial layer of the left ventricular free wall, WH GS still had the highest density of Cx43, but there was a different order for the other three states. WH GS
was followed by IBA, SA, and ET. T-tests were performed to compare the results from
WH and each of the other states. They indicate that the differences between WH and
IBA and WH and ET were significant. The difference between WH and SA does not
appear to be statistically significant.
Quantification of Cx43 density in the mid-myocardium suggested that SA GS had
the greatest amount of Cx43. This was followed by WH, ET, and IBA. T-tests indicate
that the differences between WH and ET and WH and SA were statistically significant.
The difference between WH and IBA does not appear to be statistically significant.
T-tests were also performed to study the statistical significance of the change in
Cx43 in different locations within the same hibernation state. The results are shown
in Table 1 below. Statistical significance is shown between the epicardium and midmyocardium of WH and SA GS, between the epicardium and endocardium of IBA GS,
and between the mid-myocardium and endocardium of SA and IBA GS. The density
of Cx43 in ET GS did not appear to vary significantly between myocardial layers.
41
T-Test Probabilities
WH-WH
SA-SA
IBA-IBA
ET-ET
Epi-Mid
0.049
0.001
0.441
0.730
Epi-Endo
0.803
0.750
0.000
0.638
Mid-Endo
0.350
0.003
0.000
0.946
Table 1: Table showing t-test statistics for the change in Cx43 in different
ventricular locations within a hibernation state. Values with P<0.05 were
considered statistically significant.
Expression of NF200 in GS Left Ventricular Endocardium
Heterogeneity of NF200 expression was found in the endocardial layer of the left ventricular free wall. Select pictures of NF200 expression can be seen in Figure 6. NF200 was
seen in the epicardium and mid-myocardium of some GS, but the results have not been
quantified. In the endocardium, NF200 density was highest in WH GS, followed by ET,
SA, and IBA. Figure 4 shows a graph of mean NF200 density in different states of GS.
Figure 4: Histogram showing the results of quantification of NF200 density in
the endocardium of GS LV free wall. Values obtained from confocal images.
Three animals of IBA and WH were sampled, and two animals of ET and SA.
Each animal had 4 sample pictures. The data from each group are expressed
as mean±SEM.
42
WH
SA
IBA
ET
Figure 5: Cx43 Staining in Epicardium (left), Mid-Myocardium
(center), and Endocardium (right)
Figure 6: NF200 Staining in Endocardium of WH (left),ET (center-left),SA
(center-right),& IBA (right) GS
43
DISCUSSION
Expression of Cx43 in GS Left Ventricular Free wall
The results of our study suggest that there is heterogeneity of Cx43 expression in
regions of left ventricular free wall. In the epicardial and endocardial layers, WH GS
had the highest level of Cx43 density. Saitongdee et al. found similar results that suggested an increased size and number of Cx43 plaques in hibernating compared to
active hamsters, although their study did not discriminate between the tissue layers 6.
Up-regulation of Cx43 in those layers during hibernation might indicate that in conditions that would normally decrease conductance in gap junctions and lead to a
breakdown in proper heart functioning, hibernating species are able to compensate
and maintain sufficient cell-cell communication. This would allow the heart to continue beating with normal rhythm even at low temperatures.
There was a significantly higher Cx43 density in the mid-myocardium of SA GS
than in the epicardium and endocardium. This could reflect on the different functions
of the ventricular layers, but the trend seen in SA GS was not seen in the other GS
states. There was no significant difference between SA GS and WH GS in the midmyocardium, so it appears that hibernators do not require additional Cx43 in this
region to protect themselves from hypothermia-induced arrhythmias. Surprisingly,
however, our data suggests that Cx43 density is significantly higher in hibernating GS
than in ET GS. It is possible that the ET animals in our study were sacrificed at the
onset of their entrance into torpor, and not immediately before they entered steady
state hibernation. Protein production may not have been completed at the time the
animals were sacrificed.
Previous immunostaining and western blotting has shown that there is up-regulation of Cx45 in WH GS ventricle and that this could be a mechanism for protection7.
Future studies will include immunohistochemical staining and western blotting analysis in order to quantify and compare Cx45 density and Cx43 density.
Expression of NF200 in GS Left Ventricular Endocardium
It well known that sympathetic stimulation can promote VF induction. The cardiac
adrenergic innervation of certain hibernators shows significantly different distribution from that of non-hibernators. 3 Nearly total absence of ventricular adrenergic
nerves was shown in ground squirrels and bats 8. The lack of adrenergic innervation
of the myocardial muscle fibers in hibernators may offer a protective mechanism
against VF induced by low body temperature.
For the first time, we characterized the pattern of autonomic innervation in the
ventricle of a hibernating species. The results of our NF200 density quantification suggest that there are different levels of innervation during the different states of the GS
hibernation cycle. It appears that WH GS had the highest level of innervation in the
endocardium, suggesting that greater autonomic innervation might contribute to protecting the GS heart during the hibernation state. However, NF200 does not differentiate between sympathetic and parasympathetic nerve fibers, so more protein examination is necessary.
44
In order to quantify and compare sympathetic and parasympathetic innervation in
GS, specific protein markers for the two autonomic types of innervation must be identified. Possible proteins to use as markers are tyrosine hydroxylase (TH) as a sympathetic marker, and choline acetyl-transferase (ChAT) as a parasympathetic marker.
Immunohistochemical and western blotting analysis of the protein markers will be
performed in order to understand the mechanisms involved that result in the change
of NF200 expression seen during the hibernation state.
NOTES
1
Mattu A, Brady WJ, Perron AD. Electrocardiographic manifestations of hypothermia.
Am J Emerg Med 2002;20:314 –326.
2
Wang SQ, Lakatta EG, Cheng H, Zhou ZQ. Adaptive mechanisms of intracellular calcium
homeostasis in mammalian hibernators. J Exp Biol 2002;205:2957–2962.
3
Johansson BW. The hibernator heart–nature’s model of resistance to ventricular fibrillation.
Cardiovasc Res 1996;31:826–832.
4
Burlington RF, Milsom WK. The cardiovascular system in hibernating mammals: recent
advances. In Comguihem B, Malan A, editors. Living in the Cold. London. Jon Libbey
Eurotext Ltd., 1989:235–243.
5
Duker GD, Olsson SO, Hecht NH, Senturia JB, Johansson BW. Ventricular fibrillation
in hibernators and nonhibernators. Cryobiology 1983;20:407– 420.
6
Saitongdee P, Milner P, Becker DL, Knight GE, Burnstock G. Increased connexin43 gap
junction protein in hamster cardiomyocytes during cold acclimatization and hibernation.
Cardiovasc Res 2000;47: 108–115.
7
Fedorov VV, Li L, Glukhov A et al. Hibernator Citellus undulatus maintains safe cardiac
conduction and is protected against tachyarrhythmias during extreme hypothermia:
Possible role of Cx43 and Cx45 up-regulation. Heart Rhythm. 2005; 2:966-975.
8
Nielsen KC, Owman C., Acta Physiol Scand. 1968; 316 (Suppl):1-30
45
Analysis of DIAP2 Rescue of
diap1 Mutant Cells in Drosophila
Author:
Lisa J. Sudmeier
Lisa knew that she wanted to
do research before she came to
Washington University and was
fortunate to have had the
opportunity to get “started
right away in her freshman year
in a great lab” with Professor
Ross Cagan who fueled her
love for research and helped
her grow as a scientist. She is a
junior majoring in Biology and
Spanish and eventually “would
like to get an MD/Ph.D...the
best of both worlds!”
KEY TERMS
•
•
•
•
•
Apoptosis
Caspases
IAP’s
FRT/FLP system
UAS/GAL4 system
A B S T R AC T
Programmed cell death, known as “apoptosis,” is an essential part of
development and the maintenance of healthy tissues in organisms.
Mutated or harmful cells also self-destruct via apoptosis, thereby preventing their own proliferation and the development of tumors. In fact,
many tumors are characterized not only by the over proliferation of
damaged cells but also by the lack of sufficient apoptosis. Interference
with the activation and/or success of the cell death machinery can
occur at many different steps in the apoptotic pathway.
In this study, Drosophila melanogaster was used to analyze the
role of Inhibitor of Apoptosis Proteins (IAPs) in the cell death pathway.
IAPs are believed to be one of the last steps in the prevention of cell
death by the organism. These proteins bind caspases, which are
responsible for cleaving the proteins within a cell and causing apoptosis. Caspases may only be activated when not bound by an IAP.
Drosophila have two known IAPs, DIAP1 and DIAP2. Without a functional DIAP1, cells are generated but soon after, die by apoptosis.
Conversely, cells not expressing DIAP2 are viable and healthy but are sensitized to stress-induced cell death. In this study, mitotic recombination
clones of diap1 homozygous mutant cells overexpressing DIAP2 were
generated in the Drosophila eye to test whether DIAP2 overexpression can
rescue the diap1 mutant lethal genotype. Rescue was tested in both the
adult eye and larval imaginal disc. While significant rescue was observed
in the adult eye, no rescued cell clones were present in the larval imaginal
disc. This suggests that DIAP2 overexpression may play a role in cell
death rescue by allowing diap1 mutant cells to survive longer during early
development. In this way, the rescued cells have more time to send out
proliferative signals to neighboring wild type cells, encouraging the wild
type cells to divide and compensate for the death of the mutant cells.
R E S E A R C H M E N T O R : PA S C A L M E I E R , P H . D. ,
T E A M L E A D E R O F T H E A P O P TO S I S L A B O R ATO RY
AT T H E B R E A K T H R O U G H R E S E A R C H C E N T R E
The aim of Dr. Meier’s research is to uncover the precise mechanisms
that regulate apoptosis, in order to determine why cancer cells don’t die
when they should. The work is being carried out at Breakthrough, the
United Kingdom’s leading charity committed to fighting breast cancer
through research, campaigning and education.
AC K N OW L E D G E M E N T S
I am extremely grateful to my mentor Francois Leulier and the Apoptosis
Lab group and Dr. Pascal Meier, who worked tirelessly with me this past
summer. I would also like to extend much gratitude to Professor Ross
Cagan and the members of the Cagan lab. Finally, I would like to thank
the Office of Undergraduate Research and the Hoopes Undergraduate
Research award for funding this study.
46
INTRODUCTION
rogrammed cell death, or “apoptosis,” is an essential part of development in multicellular organisms. Young organisms produce many more cells than they will
actually need as an adult. As these new cells specialize, unnecessary cells die allowing
for continued growth and development of the organism. Apoptosis is also the process
by which mutated or harmful cells self-destruct, thereby preventing their own proliferation and the development of tumors. Caspases are the key proteins responsible for
carrying out apoptosis. Caspases, when activated, are the proteins within cells that
cleave polypeptides necessary for cell function, which ultimately results in cell death.
The activity of caspases begins with activation of initiator caspases, which then activate effector caspases that are responsible for substrate cleavage. Cell destruction rapidly follows caspase activation making it absolutely necessary that this activation be
carefully regulated. Inhibitor of apoptosis proteins (IAPs) are largely responsible for
caspase regulation and have been shown to be overactive in many types of cancers.
Drosophila melanogaster contains two IAPs, DIAP1 and DIAP2. It has been shown
that DIAP1 is essential for cell viability in Drosophila melanogaster. Without a functional DIAP1, cells are generated but die shortly thereafter by apoptosis. On the other
hand, in wild type conditions, cells not expressing DIAP2 are viable and healthy but
are sensitized to stress-induced cell death. It has been shown that DIAP1 binds to the
Drosophila effector caspases drICE and DCP-1 via its BIR1 domain as well as the
Drosophila initiator caspase Dronc at its BIR2 domain. In contrast, DIAP2 only binds
active drICE, but not Dronc or DCP-1. Like DIAP1, DIAP2 can inhibit cell death
induced by many different triggers, and recently DIAP2 overexpression has been
shown to rescue the lethality of cells where expression of diap1 is knocked-down
through RNA interference. These results demonstrate that DIAP2 can function as a
cell death regulator in vivo and suggest a functional redundancy between DIAP1 and
DIAP2 in the regulation of cell survival.1 To further address this functional redundancy and confirm that DIAP2 can substitute the loss of DIAP1 in vivo, this study investigates whether overexpression of DIAP2 can promote the survival of cells homozygous for diap1 loss-of-function mutation.
P
MATERIALS/METHODS
A. Rescue of diap1 mutant cell survival by DIAP2 overexpression
during eye development: Adult eye read-outs
To obtain DIAP1 homozygous mutant cells overexpressing DIAP2, mitotic recombination clones were produced using the yeast FLP/FRT system. In this system, the flipase gene (FLP) is under the control of a tissue specific promoter, eyeless, which
drives its expression in the developing fly eye. FLP is a recombinase that activates
recombination at FRT sites during mitosis. Instead of generating two identical cells,
this artificially induced recombination produces two different types of daughter
cells.2 In this experiment, using mother cells heterozygous for diap1 mutations,
daughter cells were generated that were either homozygous for the diap1 mutation
(lethal) or wild-type (see Figure 1). The diap1 gene (also called thread, th) is located
47
on the left arm of the Drosophila third
chromosome. Therefore, the flies used
in this experiment were carrying a third
chromosome with the FRT80B site
recombined to different diap1 alleles
(FRT80B,th). To test if DIAP2 overexpression could rescue the lethality of
diap1 mutant cells, the UAS/GAL4 system was combined with the FRT/FLP
technique to simultaneously overexpress DIAP2 with the flp gene.
DIAP2 overexpression was achieved
by using a UAS-diap2 transgene in
which the diap2 cDNA is under the control of a minimal promoter fused to the Figure 1 Generation of mitotic clones
3
upstream activating sequence (UAS). with the Flp/FRT system
These UAS sequences are target sites for
the yeast transcription factor GAL4.
Another transgene, eyeless-GAL4, which
drives the expression of GAL4 very early
and throughout the eye development in
Drosophila (see Figure 2), was used in
this experiment. Both the flp gene and
diap2 are under the control of UAS
sequences. Therefore, both are regulated
by the same driver (ey-GAL4).
To increase the probability of observing the effect of DIAP2 overexpression
on the viability of diap1 mutant cells, it
was ensured that most of the cells generated in the eye by mitotic recombination
would be diap1 mutant. To achieve this,
Figure 2 The GAL4-UAS system for
mother cells were transheterozygous for
directed gene expression3
diap1 and Rps17 (diap1/Rps17). Rps17 is
a mutation on the left arm of the third
chromosome (the same arm as diap1) that affects a subunit of the ribosome. Cells heterozygous for this allele are viable but slow growing, while cells homozygous mutant
for Rps17 are lethal. When mitotic recombination of diap1/Rps17 mother cells occurs,
two types of daughter cells are obtained: one that is homozygous for the diap1 mutation and overexpressing DIAP2, and another that is Rps17 homozygous and therefore
dies. The cells that do not recombine are Rps17 heterozygous and therefore grow slower than the other cells. Consequently, if DIAP2 overexpression rescues the diap1 mutant
cells, these cells will grow faster than the cells that did not recombine.
If DIAP2 overexpression fails to promote the survival of some diap1 mutant cells,
the end result of these cellular events during development would be a small eye in flies
composed of only slow growing Rps17 heterozygous cells (cells that have not under-
48
gone mitotic recombination). diap1 mutant cells would have died without DIAP2
overexpression along with the Rps17 homozygous cells. However, if DIAP2 overexpression rescues diap1 mutant cells, a larger eye would be expected. This is because
these rescued cells would not contain the Rps17 mutation and would therefore grow
faster than the Rps17 heterozygous surrounding cells.
For this experiment, five different diap1 alleles were used. The th11-3e and th9 alleles
give rise to proteins with a mutated BIR1 domain. The th109.07 and th117 alleles result in
no or truncated proteins. The th21-4s allele produces a protein with a mutation in the
RING finger domain, which is required for autoubiquitilation of the IAP and ubiquitilation of the caspase to which the IAP is bound.
To generate flies of the appropriate genotypes, males w; (UAS-diap2/CyO);
(FRT80B,th/TM6b) were mated with virgin females w; (ey-GAL4,UAS-FLP/CyO);
(FRT80B, Rps17/TM6b,Tb) at 25∞C. The markers associated with the CyO and TM6b
balancer chromosomes (curved wing and hairy shoulder respectively) were used to
select the correct F1 adult progenies. The eyes of the progenies of the following genotypes w; ey-GAL4,UAS-FLP/CyO; FRT80B, th/FRT80B,Rps17 and w; ey-GAL4,UASFLP/UAS-diap2; FRT80B, th/FRT80B,Rps17 were compared.
Pictures of adult eyes were taken under a stereomicroscope using a Nikon D1 camera. Figures were made with Adobe Photoshop.
B. Rescue of diap1 mutant cell survival by DIAP2 overexpression
during eye development: larval eye discs read-out
After analyzing adult eyes, survival of larval diap1 mutant cells expressing DIAP2 was
assayed. To achieve this, larval eye imaginal discs expected to contain clones of “rescued” diap1 mutant cells were dissected and examined. To obtain larvae with these
clones, flies of the genotype w; ey-GAL4, UAS-FLP/ey-GAL4, UAS-FLP; FRT80B,
UbiGFP/FRT80B, UbiGFP were mated with w; UAS-diap2/ UAS-diap2; FRT80B,
th/TM6b,Tb and w; FRT80B, th/TM6b,Tb flies. The two types of cell clones produced
from recombination in the imaginal eye discs of the progeny of interest are wild type
green fluorescence protein (GFP) expressing cells and homozygous diap1 mutant cells
(not expressing GFP). If DIAP2 overexpression rescues diap1 mutant cells, clones of
cells without GFP (the “rescued” cells) should be present among the cells expressing
GFP. To illustrate this system, flies of the genotype w,eyFLP/w,eyFLP;
FRT42D,UbiGFP/FRT42D,UbiGFP were mated with w; (FRT42D/FRT42D) flies.
Although this test examines the recombination of the second, rather than the third
chromosome arm, it was useful for visualizing non-GFP expressing cells in an otherwise GFP positive field.
The progenies of the genotype w;ey-GAL4,UAS-FLP/UAS-diap2;FRT80B,th/
FRT80B,UbiGFP were selected using the larval marker Tb (short larvae) associated
with the TM6 balancer. Larvae were dissected in PBS at the 3rd instar stage. The larval eye imaginal discs (the larval tissues giving rise to the adult eye) were fixed in 4%
paraformaldehyde for 20 minutes. There were then washed three times (5 minutes
each time) with PBS and mounted with Moviol, an anti-bleaching/mounting solution,. Using a confocal microscope with a UV light, the discs were observed and analyzed for the presence/absence of GFP expressing cells.
49
C. Using MARCM clones to examine DIAP2 overexpression rescue of DIAP1 mutants
A third attempt was made at observing DIAP2 overexpression rescue of diap1 mutants
with the MARCM system (Mosaic Analysis with a ReCessive Marker). This technique
allows the experimenter to specifically overexpress a transgene of interest in a clone of
mutant cells in an otherwise heterozygous animal. This technique combines the
FRT/FLP and UAS/GAL4 /GAL80 systems, but allows the UAS transgene to be
expressed only inside the mutant clone generated by mitotic recombination. The
GAL80 molecule is a strong repressor of the GAL4 transactivator. In this experimental setting, GAL4 is under the control of a ubiquitous promoter (Tubulin). Only the
diap1 mutant cells will not express the GAL80 transgene, meaning that only these cells
will have the GAL4 transactivator functional to express the UAS-diap2 transgene. In
addition, a UAS-GFP transgene is present in order to visualize the cells where the
GAL4 driver is functional (i.e. the diap1 mutant cells). The GAL80 transgene is inserted on the left arm of the third chromosome containing a FRT80B site, so when this
arm of the chromosome recombines with the FRT,th chromosome under the action of
the FLP, the daughter cells generated are either diap1 mutant (with no GAL80 transgene so GAL4 is active) or homozygous for the GAL80 transgene (GAL4 is non-functional). The cells that did not recombine retain one copy of the diap1 mutant and one
copy of GAL80 transgene (GAL4 is non-functional). In this setting, the flp gene is
under the control of a heat-shock inducible promoter.
Mitotic recombination was achieved by incubating (‘heat-shocking’) F1 first instar
larvae 1 hour at 37∞C. Four days later, third instar larvae were selected and wing
imaginal discs were dissected, fixed and observed as in part B.
To perform this experiment hsFLP, UAS-GFP/hsFLP,UAS-GFP; TubGAL4/TubGAL4;
FRT80B,TubGAL80/TM6b,Tb flies were crossed with w; uasdiap2/ uasdiap2;
FRT80B,th/TM6b,Tb flies. The Tb marker was used to select larvae of the following genotype for dissection: hsFLP,UAS-GFP/+; TubGAL4/UASdiap2; FRT,th/FRT,TubGAL80. If
DIAP2 overexpression rescues survival of diap1mutant cells, clusters (clones) of cells
expressing GFP in a wild type, non-GFP background would be present in this wing tissue.
RESULTS
A. Rescue of diap1 mutant cell
survival by DIAP2 overexpression during eye development:
Adult eye read-outs
Significant rescue of the size of
the adult fly eye was observed
when DIAP2 was overexpressed
in th11-3e, th117 (see Figures 3 and
4), and th9 homozygous cell
clones. A weaker rescue was
observed with the th21-4s and th109.07
alleles. None of the rescues were
especially strong, but all eyes
50
Figure 3: DIAP2
rescue of diap1 mutant
Figure 4: diap1
mutant ey-GAL4,UASFLP/cyo;
ey-GAL4,UASFLP/UASdiap2;
FRT80B,Rps/FRT,Th117
FRT80B,Rps,/FRT,Th117
with DIAP2 overexpressed in diap1 mutant clones were larger than the diap1 mutant
condition alone. This suggests that indeed DIAP2 is able, at least partially, to rescue
survival of some diap1 mutant cells (data not shown for other th mutants).
B. Rescue of diap1 mutant cell survival by DIAP2 overexpression
during eye development: Larval eye discs read-out
The larval clone studies showed that mitotic recombination events can be detected
successfully, as clusters of cells expressing GFP and clusters of cells not expressing GFP
were visible in the eye disc from the control cross where the right arm of the second
chromosome recombined (see figure 5).
However, when the imaginal eye discs
from the progeny of the other crosses with
diap1 mutant alleles were dissected, all
cells appeared to be expressing GFP (see
figures 6 and 7). This suggested that, when
the diap1 mutant clones were produced,
the overexpression of DIAP2 either failed
to rescue the death of these cells or did not
promote their survival long enough to be
observed in this experimental setting. It
was also noticed that the eye discs containFigure 5: Larval eye disc control
w,eyFLP/+;FRT42D, UbiGFP/FRT42D Top arrow points
ing diap1 mutant clones with DIAP2 overto non GFP-expressing cells. Bottom arrow points to
expressed had patches of homozygous
GFP-expressing cells.
UAS-GFP cells (two doses of GFP
expressed, meaning the daughter cells
were made by mitotic recombination) and patches of heterozygous GFP cells (only
one dose of GFP expressed meaning the mother cell did not flip). In contrast, the
mutants without DIAP2 overexpressed had mostly homozygous GFP cells.
Figure 6: Larval eye disc with
only homozygous GFP-expressing
cells (diap1 mutant cells died)
w; ey-GAL4,UASFLP/+; FRT80B,th117/FRT80B,UbiGFP
All cells appear to be expressing 2 copies of GFP.
Figure 7: Larval eye disc with
homozygous and heterozygous GFPexpressing cells (diap1 mutant cells
died) w; ey-GAL4,UASFLP/UASdiap2;FRT80B,th117/
FRT80B,UbiGFP Top arrow: homozygous cells. Bottom
arrow: heterozygous cells.
51
C. Using MARCM clones to look for DIAP2 overexpression rescue of diap1 mutants
No GFP-expressing cells were observed in the wing discs of the dissected progeny from
the MARCM crosses. Importantly, the wing discs from the progeny of the control cross
also did not have any GFP-expressing cells, indicating that the system did not function
as expected. Therefore, no conclusions can be made from this particular experiment.
CONCLUSION/DISCUSSION
The results of this study are optimistic about the potential ability of DIAP2 overexpression to rescue diap1 mutant cells, but many questions remain unanswered. In the
larval experiment, no rescued cells are observed, suggesting that DIAP2 does not rescue diap1 mutants. However, the experimental adult progeny have a significant rescued eye phenotype, suggesting the opposite. Together, these results indicate that a
phenotypic rescue does occur with DIAP2 overexpression, but the mechanism by
which the DIAP2 achieves this rescue is not yet understood.
One possible mechanism is that the DIAP2 rescue may only help the diap1 mutant
cells live for a very short time after cell division rather than completely rescuing them
throughout development, as previously expected. However, this rescue may still be significant because it allows more time for cell-to-cell signaling, which would encourage
the surrounding cells to proliferate more as the mutant cells die. In this way, the surrounding cells would compensate for the death of the mutant cells, resulting in a phenotypic rescue. Without DIAP2 overexpression, mutant cells die shortly after cell division leaving little time for cell-to-cell signaling. Consequently, a smaller adult eye is
observed in the control.
Follow up experiments would begin with determining why the third experiment of
this study involving the MARCM system in the larval wing disc did not work. A functional experiment with a MARCM assay would be very valuable in this study for elucidating the rescue ability of DIAP2. This system, using a heat shock promoter, would
allow the experimenter to induce mitotic recombination at a finite time and then analyze the tissue for the presence of rescued cells at specific time intervals following the
recombination event. Had this experiment been successful, it would have provided
valuable insights into the mechanism by which DIAP2 overexpression rescues diap1
mutant cells.
Notes
1
Leulier, F et al. 2006. Systematic in vivo RNAi analysis of putative components
of the Drosophila cell death machinery. Cell Death and Differentiation: 1-12.
2
Greenspan, Ralph J. Fly Pushing: The Theory and Practice of Drosophila Genetics.
Cold Springs Harbor: Cold Springs Harbor Laboratory Press, 2004.
3
St. Johnston, D. 2002. The Art and Design of Genetic Screens: Drosophila melanogaster.
Nature Reviews (3): 176-188.
52
The Georgian Feast and Azeri Carpet:
A Cultural Exploration
through Metaphors
Authors:
Andrea S. Young
& Margaret S. Fee
Andrea is a senior majoring in
Psychology. Andrea first began
conducting undergraduate research as a Mellon fellow in her
sophomore year researching the
effects of video games on learning. It was then that she discovered and developed a passion for
research. Andrea became interested in this particular research
topic through a freshman FOCUS
class called “Global Culture and
the Individual.” Andrea will be
enrolling in a doctoral program
in Clinical Psychology in Fall 07.
Margaret is a junior majoring in
French and International and
Area Studies. As an International
and Areas Studies major,
Margaret has always been very
interested in the idea of learning
about a new culture through firsthand interactions and research.
This research experience abroad
has made her want to continue
her studies in this direction.
KEY TERMS
• Supra
• Tamada
• Mugham
A B S T R AC T
This study seeks to show the utility of metaphors as a tool for understanding a new culture. Many travelers find cultures very different from
their own difficult to understand due to a lack of sufficient means to
navigate the culture. Psychological research suggests that metaphor
can be very useful when trying to understand new and complicated
concepts. The authors traveled to two countries infrequently visited by
Americans, the Republic of Georgia and Azerbaijan, and developed a
metaphor for each: the Georgian feast and Azeri carpet. Metaphors
were investigated through literary research before and after the trip, as
well as from interviews conducted with Georgian and Azeri citizens
during the trip. This paper will summarize research on and theories of
metaphor, provide a brief overview of the Georgian feast and Azeri carpet, and give examples of how these metaphors can be used to understand the politics and society of the two countries. The interviews and
historical research show that understanding the feast and carpet can
facilitate one’s understanding of the people and customs of Georgia
and Azerbaijan. Rather than referring to the rules of one’s own culture
or using the typical travel guide, travelers can rely on the flexibility of a
cultural metaphor to guide their exploration of a new culture.
FA C U LT Y M E N T O R S : A L A N L A M B E R T, P H . D. ,
A S S O C I AT E P R O F E S S O R O F P S YC H O L O G Y &
J OA C H I M FAU S T, P H . D.
Professor Lambert conducts research on the expression versus suppression of stereotypes. A primary goal of his research is to understand how
aspects of the “situation,” the perceiver’s personality, as well as transitory mood states (e.g., happiness versus sadness) might lead people to
either use-or avoid using-stereotypic knowledge as a basis for responding to others. In addition to his work on stereotyping, he is interested in
the processes underlying perceptions of risk and self-vulnerability.
Dr. Faust teaches a freshman FOCUS seminar course called “Global
Culture and the Individual”. The class teaches students how to interact with different cultures, and how to dialogue about difficult topics.
AC K N OW L E D G E M E N T S
We would like to thank Gerald Early, Kathleen Fields, and the Center for
Programs for helping to make this project possible by funding our
research in the Republic of Georgia and Azerbaijan. In addition to our
faculty mentors Dr. Joachim Faust, Dr. Alan Lambert, and Professor
Jim Wertsch, as well as Dean Mary Laurita, who helped guide us
before, during, and after our trip. We would like to thank all of the people we met in both the Republic of Georgia and Azerbaijan who were so
generous and willing to take us into their culture, share their thoughts
and ideas, and educate us with their perspectives and experiences.
53
hen we arrived at the airport in Tbilisi, Georgia, I walked up to the airport officials at the counter with a big American smile on my face and said, “Hello.” Even
having been warned that smiling at strangers was an American phenomenon, I have to
admit that I was slightly disappointed that all I got in response was a grimace and a palm
waiting for my passport, and I said to myself, “Andrea, you’re not in Kansas anymore.”
Many travelers have this sort of experience in a new country. They rely on social
schemas from their own culture or guide books that attempt to educate them on proper etiquette for a given culture only to find themselves startled when the rules they
arrived with will not do. Cultures and the people who employ them are unique and
mutable; as such, a more flexible means for understanding a culture is necessary.
Metaphor’s flexibility and ability to produce new and unique meaning for every reader (or traveler) make it a useful tool for exploring a new culture. This study will summarize research on and theories of metaphor, provide a brief overview of the
metaphors chosen for the Republic of Georgia and Azerbaijan, and give examples of
how our metaphors can be used to understand the culture of these two countries.
Metaphor has been a source of intrigue in psycholinguistics for several decades.
Initially, metaphors were thought to have a merely ornamental function and were useful in “coining terms for new concepts, such as the leg of a triangle.”1 However, these
thoughts have evolved to allow for a more complex analysis of metaphor and its role
in conveying concepts. The two primary theories most relevant to this research are
comparison theory and interaction theory.
Comparison theory suggests that metaphor is a comparison between objects
whose literal meanings are different. Aristotle believed that metaphors were used for
stylistic purposes only, and to make language “more elegant and beautiful rather than
more meaningful.”2 Bréal took this theory a few steps further, suggesting that
metaphors are not simply ornamental but are a basic component of language use:
“metaphors teach us little save what we knew already; they demonstrate only the universal intelligence, which does not vary from one nation to another.”3 According to
Campbell, some metaphors can have multiple mutable meanings for different individuals at different times.4
Proponents of interaction theory typically agree that metaphors are sometimes
used to replace literal statements, but “good metaphors actually relate the topic [the
subject] and the vehicle5 [the term being used metaphorically] to produce a resulting
meaning that is new and transcends both.”6 There is an interaction between the subject and the vehicle that can produce a completely new meaning. These types of
metaphors are psychologically important because “they require readers to make inferences and to draw implications rather than merely to react.”7 Readers can be creative
and reflect on previous knowledge to make new meaning.
Theoretically, it could be argued that cultural metaphors would be a useful tool for
exploring a new culture: “it is common sense that it is easier to understand new things
if they are cast in terms of old.”8 Furthermore, research by Verbugge and McCarell
shows that adults can easily and consistently comprehend metaphors.9 Metaphors can
be used to make complex behavior and cultures more understandable to people outside the cultural context.
W
54
This research aims to explain the Georgian and Azeri culture through the use of
cultural metaphors. Cultural metaphors can be utilized as a tool to engage with and
better understand a new culture. Each metaphor is an “activity, phenomenon, or institution, which members of a given culture consider important and with which they
identify emotionally and/or cognitively.”10 While a cultural metaphor does need to
generalize information, “Metaphors are not stereotypes. Rather, they rely on features
of one critical phenomenon in a society to describe the entire society.”11 Initial
research showed the Georgian feast and Azeri traditional carpet to be appropriate cultural metaphors for Georgia and Azerbaijan.12 13
THE GEORGIAN FEAST
Khachapuri (Georgian cheese bread), mtsvadi (shish-kebabs), badridzhani (stuffed
eggplant), tqemali (plum sauce), sliced tomatoes and cucumbers are just a few of the
things one would find at the table of a supra, or Georgian feast. The feast is a tradition
in which many Georgians participate. Friends and family come together for hours of
eating, drinking, singing, and dancing. “The atmosphere at the Georgian table is so
friendly and candid that even the enemies are likely to make up.”14 The feast emerged
as an expression of Georgia’s cultural independence during a time of political dependence under the Russians.15 As the country stayed under Russian influence for almost
two hundred years, the supra persisted as a mark of their identity.
There are a few key players in a Georgian feast. The tamada, or toastmaster, makes
toasts throughout the feast. Research conducted at a feast in Georgia totaled over 30
toasts. The topics of the toast range from friendship and love, to wine and its special
qualities, to the landscape and poetry.16 It is up to the toastmaster to be aware of how
intoxicated the others around the table are and to pace his toasts to keep intoxication
relatively low. Being a tamada is truly an art and a great honor to the man assigned the
position, as traditionally women were not present at feasts. The feast emphasizes
developing relationships and provides a setting for friends to reflect and dialogue. It is
the tamada’s task to guide the reflection through his toasts.
The tamada is not the only person to give toasts; fellow Georgians around the table
are expected to respond to the tamada’s toast. The responses must be on the same
topic as the tamada’s latest toast. “The tamada acts like a director of the party
announcing traditional toasts…Guests at the party are like actors responding to the
toasts and often with their own interpretation.”17 This sort of exchange allows the
tamada to retain control of the flow and pace of the feast, while still allowing anyone
who wishes to make a toast.
The food served at the feast is prepared by women including the hostess, who is the
woman of the house where the feast is being held. The women spend the day baking
bread, grilling meat, and slicing vegetables. While the table is where the men usually
socialize, the women use their time in the kitchen to reflect and socialize with their
friends. As mentioned, women were traditionally not present at feasts. When they initially began to join men at the table, they did not make toasts or drink alcohol. Now,
women do attend feasts and can make toasts and drink, but many prefer to not to sit
at the table, viewing feasts as “a guy thing.”
55
Generally, the host of the house serves the food to the guests. He makes sure their
wine bowls stay full, and that there is enough food for everyone at the table.
Throughout the entire feast, the host continues to bring out food from the kitchen. At
the feast attended for this research, the host even brought out roses for all of the
women sitting at the table. Similar manifestations of hospitality were apparent
throughout the feast: a male singing group performed for the guests and several of the
toasts were directed to the Americans sitting at the table.
SUPR A AND GEORGIAN CULTURE
As a cultural institution distinctive to the Republic of Georgia, the Georgian Feast can
be used as a lens to help understand this unique country. The longevity of the feast is
a testament to the extensive history of the Georgian people with a culture that dates
back to the sixth century B.C. Archeologists believe that inhabitants of this region have
been producing wine since the forth millennium B.C. and may even be home to the
first wine grape.18 Wine is one of Georgia’s greatest exports and acts as the foundation
of the supra, bringing together people from different backgrounds for the sake of hospitality. During this research, several toasts were made in honor of wine as the force
that binds humans together and strengthens relationships.
Historically, Georgia is a land that values tolerance. Despite invasions of Persians,
Mongols, Turks, and most recently Russians, Georgia has always tried to adapt to and
tolerate other cultures while still retaining the essence of its own culture. Although
these aggressors would temporarily control the region, the Georgian people maintained their language, religion, and sense of Georgian identity. Similarly, the origins of
some foods present in the Georgian feast are the result of these past invaders. For
example a type of meat dumplings called khinkali is seen as the variation of similar
dumplings found in Mongolian cuisine. Several recipes have a Turkish influence, especially along the coast of the Black Sea, such as a cheese and noodle dish called achmamakarina which resembles makarna firinda found in Turkey.19 This tradition of tolerance is also demonstrated in the high level of Georgian hospitality. Guests and foreigners are welcomed and greatly respected at the supra. Whether described as “guest
lovers,” “friendship,” “love,” or “helping people,” this notion of hospitality was a clear
theme in all the interviews of this study.
While the values expressed in the feast are enduring, the supra is not the same as it
was two centuries ago or even a generation ago, a result of the change this culture has
recently undergone. Several Georgians mentioned that feasts have become more infrequent, shorter, more gender equal, and even more “European”. Another participant
noted that originally men sat around the table and ate with their hands in a process
that connected people with food, drink, and friends. Once a very male dominated culture, gender roles in both Georgian society and the feast have begun to change as well.
Women can now be included in the table and the tamada no longer has as great a level
of power and status associated with it. The apparent modernization seen in the
Georgian feast reaches past this cultural institution.
The changing feast over the past few decades is evidence of Georgia as a changing
nation. Since the fall of the Soviet Union in 1991, Georgia has gone through considerable political and social change.20 In 2003, the Georgian people undertook the peace-
56
ful Rose Revolution with the shared desire to rid their nation of past corruption and
rapidly move it forward as an active world player. Following this political transformation, several changes have taken place over the past few years, such as improvements
in roads, and an increase in freedom of speech, the military force, and international
alliances. By using new outlets like Non-Governmental Organizations and a less censored media, Georgians are able to express their ideas in a more modern and global
environment. Small oppositional groups once allied with the current ruling party have
split away and are trying to make adjustments that will help Georgia sustain and build
its democracy.
The interviews and feast experiences indicated that the transformation of the supra
is in part due to Georgia’s younger generation. Young Georgians said that they often
found feasts “boring,” “too long,” and “hard with the generation gap.” Similarly, adults
made reference to “the devaluing,” “the change in gender roles,” and how “it used to be
more serious.” Likewise Georgian youth have begun to look beyond their borders for
a sense of identity. There is a growing connection to European culture. While most
participants saw themselves as Georgian, the youth alluded to a certain pull they feel
away from Georgia and towards Europe.
This cultural shift towards Europe also coincides with a push towards European
politics. With the European Union’s recent growth in size and power, many countries,
including Georgia, are now looking to the EU in hopes of partnership and future
membership. Traveling throughout the country one can see the European Union flag
flying next to the Georgian flag even in small, rural towns. Furthermore, some see
Georgia’s current membership in the Council of Europe as a testament to their
“Europeanness”. In addition to European support, in recent years, Georgia has relied
on the United States as a key component in its current efforts to reform and rebuild.
In an attempt to strengthen international relations, President Bush made a visit to
Tbilisi in the summer of 2005 and then welcomed Georgian President Mikheil
Saakashvili in Washington in the fall of 2006. There is even a main road in Tbilisi bearing President Bush’s name and picture as evidence of this new relationship.
Despite their current movement towards change, Georgians still possess very
strong ties to their cultural roots and practices. The data in this study demonstrated
that Georgians are first and foremost Georgian. Bound by a collective memory of foreign invasion and occupation, as well as prosperity and strength, they have been tested throughout the centuries. No matter how boring a Georgian teenager may find a
supra, he or she still knows all the songs and traditional toasts by heart. As the feast
changes along with their political and social identity, Georgians still retain what they
value most – hospitality, a collective society, the arts, food, the Georgian language, and
interpersonal relationships.
AZERI CARPET
Similar to the Republic of Georgia, Azerbaijan is also a newly independent country
(from Russia), struggling to become a member of the European Union. At the same
time, there are a lot of differences between Georgian culture and Azeri culture. By
understanding Azeri carpet and mugham, music that is often played while sitting on
the carpet, a traveler can better understand Azerbaijan.
57
Carpets are one of Azerbaijan’s top exports; Azeris make special smaller rugs just
for tourists to purchase. One could not walk far in downtown Baku, Azerbaijan’s capital, without spotting someone selling carpets. In 1992, Azerbaijan joined UNESCO
and began to assist UNESCO in the preservation of the heritage of the Silk Roads. This
heritage is very important to Azeris, as they use silk in many of their carpets.
Azerbaijan is very well known for its carpet production.21
Many Azeri pastimes (including mugham) are performed while sitting on these
carpets. When traveling to Sheki, a farm town in the mountains of Azerbaijan, it is not
uncommon to sit, drink tea, and eat baklava in a traditional tea house with carpets
hanging on the walls. Every museum features carpets, such as Shirvanshah’s palace in
Baku and Xan Saray, the khan’s palace, in Sheki. The walls of Xan Saray are painted
with a paisley-like design that is often seen in Azeri carpets.
There is an entire museum in Baku dedicated to carpets. The museum holds carpets
made in different regions in Azerbaijan, from different time periods, and for different
occasions. Each region in Azerbaijan has its own different style of carpet. Azeris employ
different technique for weaving the carpets, different materials, different patterns and a
different feel to their carpets. Carpets often tell a story and contain many details.22 One
would have to study some carpets for hours to see everything depicted in them.
AZERI CARPET AND CULTURE
The origins of Azeri carpet making date back centuries, and it continues to be a craft
that is honored and perfected today. As a practice, carpet making unites all Azeris
including those living across national borders in Iran, Georgia, and Armenia as an art
of “symmetry, bold design, and juxtaposition of color.”23 Azeri carpets are divided into
four distinct groups based on region, each one unique in its use of color and design.24
Less than one hundred years ago, the Azerbaijan of today consisted of several smaller
kingdoms25 which led the different schools of carpet making. The Tabriz style of
Southern Azerbaijan uses a weave of bold dark colors in a thick pile that often depicts
nature or hunting scenes, while the Gobu-Shirvan school is known for pieces used in
weddings, such as saddle bags or cloths in patterns of vivid colors.26 Carpets are usually named after the village in which they were created, and thus show this sense of
regional pride that is still expressed throughout the nation.
Azeris are very proud of their landscape, and Azerbaijan is home to nine out of the
world’s eleven climatic zones.27 Carpets demonstrate an attention to the surroundings
by reflecting the natural environment in their design. The rich colors that weave in
and out of each carpet are all made naturally from available plants such as pomegranate, grapes, quince, and saffron.28
In more recent years, the financial position and landscape of Azerbaijan has been
dramatically changed by the country’s oil wealth. The capital city of Baku has been
especially affected. Old, small stone buildings and store fronts can be seen along with
clumps of new, modern office and apartment buildings. This architectural discontinuity is a manifestation of Azerbaijan’s changing economy. As the Baku oil supply has
helped facilitate its transition to a newly independent country, Azerbaijan has gained
much international attention, especially with the new Baku-Tbilisi-Ceyhan oil pipeline.
58
The tradition of carpet making has also recently been changing. In the middle of the
twentieth century, internationally known carpet maker Latif Karimov began a modern
movement in this longstanding craft. In addition to researching and classifying Azeri
carpet into one hundred forty four different styles, he is most famous for his carpet
entitled “Song of the Centuries,” which brings together the varying traditions of each
region in one masterpiece that pays tribute to this national art form.29 This work reflects
a modernizing and changing mentality that extends beyond carpet making.
The modernization of carpet making has also inspired politically motivated pieces.
Several have been created to honor former president and national symbol, Heydar
Aliyev. Following two unstable periods of leadership after the fall of the Soviet Union,
Heydar Aliyev became the second elected president of the communist-free government. He is attributed with moving Azerbaijan to the more advanced, prosperous, and
developed state that it is in now. Aliyev was replaced by his son Ilham in 2003 in an
election that received much criticism from the opposition and the international community. Many claim that methods were used in the election process that gave Ilham
Aliyev an unfair advantage which led to his victory.30 Today, the legacy of Aliyev is still
very prominent. There are large billboard sized pictures of Aliyev placed throughout
Baku and small town centers promoting different Azeri institutions, such as the train
system or telephone service. There are also several images showing Aliyev with his son,
which represent the transfer of wisdom and power between generations. By depicting
Aliyev in carpets, his importance to the Azeri people is forever celebrated.
AZERI MUSIC AND CULTURE
Although a separate form of art, the traditional Azeri music of mugham is often presented in the same contexts as carpets. Musicians play mugham while surrounded by
and seated on carpets and use the intricate designs to guide the progression of the
music. The practice of mugham has also seen a shift in recent years. While it is a folk
tradition that goes back several centuries, lately mugham has been merged with jazz to
form a more modern sound that is gaining popularity. It is even speculated that it will
be performed in 2008 at Eurovision, a widely televised international pop music competition. This evolution in Azerbaijan’s textile and oral tradition is accompanied by a
general shift towards change.
As a young democracy, Azerbaijan is still in the process of changing from a dependent Soviet republic to an independent democratic nation. The support for independent
political movements is growing as well as the opportunity for women to gain candidacy. Azerbaijan’s economy has been growing at a steady pace over the past decade. With
recent membership to the Council of Europe in 2001, Azerbaijan has gained a greater
international status and is becoming a bridge between the East and the West.
Like Georgia, however, Azerbaijan has still made it a priority to retain its distinctive culture. Certain traditions, such as the Azeri carpet making and mugham, have
remained integral parts of their society and education. Some of the youth indicated
that a growing European identity is not mutually exclusive with being Azeri. Their
strong feelings of patriotism along with the desire to understand and preserve their
rich culture are what make them Azeri.
59
CONCLUSION
Metaphors offer a different way of engaging with a culture. By providing a framework
to more profoundly understand experiences, encounters, and practices, they help to
organize and make sense of all that accompanies entering a new culture. Research on
interaction theory of metaphor, in particular, shows that metaphors can help create
new meanings from previous knowledge. In the case of this study, metaphors created a
starting point which aided in building a comprehension of two national cultures. By
understanding an important cultural institution as well as, how it can provide insight
into the society in which it is found, an individual can become equipped with a matchless tool to enter a new culture. Like stereotypes, it can be dangerous and nearsighted
to take cultural metaphors too definitively; there will always be people, situations, and
practices that do not seem to fit. Metaphors can be one of many tools used in cultural
exploration and can be an aid that accompanies experiences within different cultural
contexts. Ultimately, for this investigation, cultural metaphors provided the opportunity to more deeply explore and appreciate these two unique and remarkable nations.
Notes
1
Andrew Ortony et. al., “Metaphor: Theoretical and Empirical Research,”
Psychological Bulletin 85 (1978): 921.
2
Ibid.
3
Ibid., 922.
4
P.N. Campbell, “Metaphor and linguistic theory,” Quarterly Journal of Speech 61(1975): 1-12
5
The ‘topic’ and ‘vehicle’ are terms metaphor researchers use to describe a metaphorical statement. “For example, in the metaphor The question of federal aid to parochial schools is a bramble
patch, the topic is federal aid to parochial schools, the vehicle is bramble patch” (p. 921).
6
Andrew Ortony et. al., “Metaphor: Theoretical and Empirical Research,”
Psychological Bulletin 85 (1978): 923.
7
Ibid.
8
Ibid., 937.
9
R. R. Verbrugge and N. S. McCarrell, “Metaphoric comprehension: Studies in reminding
and resembling,” Cognitive Psychology 9 (1977): 494-533.
10 Martin J. Gannon, Understanding Global Cultures, 3rd ed. (Thousand Oaks:
SAGE Publications, 2004).
11 Ibid., 17.
12 Inna Naroditskaya, “Azerbaijani Mugham and Carpet: Cross-Domain Mapping,”
Ethnomusicology Forum 14 (2005): 25-55.
13 Darra Goldstein. The Georgian Feast: the vibrant culture and savory food of
the Republic of Georgia (Los Angeles: University of California Press, 1993).
14 Besiki Sesauri and Matuta Bjalava, “Georgian Cuisine: The Tradition of Georgian Feast
or Supra,” (Besiki Sesauri), http://sisauri.tripod.com/ref/cuisine/cuisine.html#intro
15 Zurab Karumidze, interviewed by the author, St. Louis, MO, May, 9 2006.
60
16 Darra Goldstein. The Georgian Feast: the vibrant culture and savory food of
the Republic of Georgia (Los Angeles: University of California Press, 1993), 27.
17 Besiki Sesauri and Matuta Bjalava, “Georgian Cuisine: The Tradition of Georgian Feast
or Supra,” (Besiki Sesauri), http://sisauri.tripod.com/ref/cuisine/cuisine.html#intro.
18 Darra Goldstein. The Georgian Feast: the vibrant culture and savory food of
the Republic of Georgia (Los Angeles: University of California Press, 1993), xiv, xix.
19 Ibid., 6.
20 For a general discussion of Georgia and the Rose Revolution, see: Zurab Karumidze and James V.
Wertsch, eds., Enough: The Rose Revolution in the Republic of Georgia 2003 (New York: Nova
Science Publisher, Inc., 2005).
21 The Ministry of Culture of the Republic of Azerbaijan, Azerbaijan Carpets, 1.
22 Ibid.
23 Farida Sadikhova, “Carpets made to last: a walk through Baku’s National Carpet Museum,”
(Azerbaijan International, 2000), http://www.azer.com/aiweb/categories/magazine/82_
folder/82_articles/82_carpet_museum.html.
24 Ibid.
25 Mark Elliott. Azerbaijan with excursions to Georgia (Surrey, UK: Trailblazer
Publications, 2004), 36.
26 Farida Sadikhova, “Carpets made to last: a walk through Baku’s National Carpet Museum,”
(Azerbaijan International, 2000), http://www.azer.com/aiweb/categories/magazine/82_
folder/82_articles/82_carpet_museum.html.
27 Fariz Ismailzade, interviewed by the author, Baku, Azerbaijan, 6 June, 2006.
28 “Azerbaijan Art of Carpet-Making,” http://www.media-az.com/azerbaijan/art.html.
29 Farida Sadikhova, “Carpets made to last: a walk through Baku’s National Carpet Museum,”
(Azerbaijan International, 2000), http://www.azer.com/aiweb/categories/magazine/82_
folder/82_articles/82_carpet_museum.html
30 Mark Elliot, Azerbaijan with excursions to Georgia (Surrey, UK: Trailblazer
Publications, 2004), 47–48.
61
Summaries of Student Work
Toward a Better Understanding of...
External Connections of the
Dorsolateral Prefrontal Cortex
Abhisek C. Khandai,
Mentor: Dr. Joseph L. Price
Executive functions such as decision-making, planning, and goal-directed behavior
have been found to take place in the prefrontal cortex (the cortex anterior to the motor
cortex) area of the brain. The prefrontal cortex of monkeys and humans consists of at
least two different regions, the orbitomedial (OMPFC) and dorsolateral prefrontal
cortex (DLPFC). These areas can be further divided into areas based on cellular (architectonic) differences and can be categorized into regions based on intrinsic connections, as well as differing external axonal connections to other parts of the brain, providing insight into the region’s function. Previous studies of the Price lab have analyzed the connections of the orbitomedial region in depth. These studies have shown
that there are two distinct and complementary “networks” within the orbital (ventral)
and medial regions of the OMPFC, the former being linked to food sensation and
reward, the latter involved in visceral awareness. However, the analysis of the DLPFC
has been incomplete, although previous studies have identified a number of architectonic areas and examined external axonal connections. These earlier findings suggest
the presence of several distinct regions within the DLPFC, each having different connections and functions. Three regions have been hypothesized in the ventral, caudal,
and dorsal areas of the DLPFC.
To study these cortico-cortical connections, retrograde and anterograde axonal
tracers were injected into the various hypothesized regions of the DLPFC in macaque
monkeys (macaca mulatta), valuable animal models in their high structural homology to the human brain. After sacrifice, the brains were prepared for histological staining and demonstration of the tracers. The brains were then serially sectioned via
microtome to create coronal brain slices to easily visualize labeling of cells and axons
by tracers in various parts of the brain. These slices were mapped, rostrally to caudally from the premotor cortex to the posterior temporal cortex, using the Accustage
MDPlot 5.2.0 computer and microscope system. The maps of successive sections were
elaborated with grey-white matter boundaries, and collated to produce composite
images of areas of the cortex, allowing patterns of connections to be readily visualized
across the brain.
62
The resulting patterns of cortico-cortical connections exhibited clearly distinct
systems of labeling from each injected region, leading to differing functional areas.
This supports the hypothesis that geographically close areas in the DLPFC belong to
different functional systems. However, more injections must be made into other parts
of the DLPFC, and continuing analysis of labeling needs to be done before the hypothetical ventral, caudal, and dorsal areas can be sufficiently delineated or disproved.
Further study will lead to better discrimination of separate functional areas within the
DLPFC, as well as the understanding of its overall functional role in the brain. The
importance of such “neuro-cartography” cannot be understated. Apart from an elucidation of the executive functions of the brain, the DLPFC has also been implicated in
various psychiatric diseases, from depression to schizophrenia. Understanding the
connections and subdivisions within this region can hopefully lead to better diagnoses
of such brain disorders, and perhaps better treatment as a result of increased knowledge of the neuroanatomy.
63
Toward a Better Understanding of...
Ecological Trap for Ovipositing Mosquitoes
Mediated Through Preferences for
Contaminated Habitat
Julia Buck
Mentor: Dr. James Vonesh
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
Although the toxic effects of pesticides on non-target species in aquatic systems have
received considerable attention in the recent ecotoxicology literature, we know almost
nothing about whether pesticides can alter the habitat selection behavior of organisms
with complex life cycles. In a previous experiment, I explored the oviposition response
of several aquatic taxa to a common pesticide, carbaryl, and found opposite effects on
mosquito and treefrog oviposition: treefrogs avoided contaminated pools, while mosquitoes were attracted to them. Because toxic effects of the pesticide reduced survival
of mosquitoes, this scenario represents an ecological trap. The pattern of oviposition
detected could reflect 1) preference for carbaryl, 2) avoidance of potential competition
with treefrog tadpoles, or 3) both. We tested these alternatives using a mosquito oviposition site selection experiment in which we manipulated the presence of carbaryl and
treefrog eggs. Mosquito egg rafts were removed daily, placed in uncontaminated water,
and allowed to hatch so that mosquitoes could be identified to species. More than six
times as many Culex restuans egg rafts were laid in the carbaryl treatment as in either
of the other two treatments combined (p<0.0001). There was no difference in the
number of mosquito egg rafts in the control and treefrog egg treatments. This provides evidence that ovipositing mosquitoes detect carbaryl directly and that the ecological trap is mediated through an apparent preference for contaminated habitat.
64
Toward a Better Understanding of...
Optimization of Design Parameters
for Future Gamma-Ray Telescopes
Christopher Weaver
Mentor: James Buckley
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
The goal of this research was to simulate the behavior of a high resolution camera for
a gamma-ray telescope in order to determine the extent of the benefits of improving
current telescope designs. Higher resolution designs could allow IACT (Imaging Air
Cherenkov Technique) telescopes to detect gamma-rays at lower energies and better
differentiate between gamma-rays and hadronic cosmic rays. IACT telescopes work by
detecting Chrenkov light emitted by charged particles traveling at more than the speed
of light in air which are created in cascades when high energy gamma-rays interact
with the atmosphere via pair production of electrons and positrons.
To determine the capabilities of a high-resolution camera, we used a combination of
software Monte Carlo simulations and analysis programs. The simulation software generates random gamma-rays according to a realistic spectrum and, for each gamma-ray,
generates a particle cascade and the accompanying shower of Cherenkov light. Then, the
collection of the light by the telescope and the output of the camera’s electronics is simulated. Because of background light in the sky, IACT telescopes only record data (trigger) if signals in a number of photomultiplier tubes exceed some predefined trigger conditions. If triggering conditions are met, the simulated data are recorded. The finished
data are analyzed by ‘cleaning’ the image to determine which pixels contain useful data
and ‘parameterized’ to numerically describe its characteristics.
For the high resolution camera, we found that the existing triggering and image
cleaning algorithms were not applicable, thus we are working to develop general
methods that will be scalable to any size or resolution camera. However, we do not
know the optimum parameter values to use with these new algorithms, therefore we
are using simulations to determine which combinations give the best results. Once we
have determined how these algorithms should be applied to the existing and hypothetical cameras, we will be able to compare the performance of cameras with different
angular resolution, and hence, make an informed decision about the optimum camera design for a new gamma-ray telescope.
65
Toward a Better Understanding of...
Regulation of the Urea Transporter
UT-A in Rats with Adriamycin Induced
Nephrotic Syndrome
Benjamin Nanes
Mentors: Dr. Janet Klein and Dr. Jeff Sands
Urea, the major waste product of amino acid metabolism in the body, is a small, polar
compound that generally cannot pass directly through cell membranes. Several families of urea transporters allow the facilitated diffusion of urea across cell membranes in
many tissues. UT-A, one of the urea transporters most commonly found in kidneys,
plays an important role in urine concentration, through the maintenance of an osmolality gradient in the kidney medulla. Previous studies have shown that conditions, such
as diabetes mellitus, which are associated with large amounts of solute entering the kidneys or a decrease in urine concentrating ability, result in the increased expression of
UT-A in the kidneys. Rats with adriamycin (Doxorubicin) induced nephrotic syndrome, a condition characterized by damage to kidney glomerular cells, production of
protein containing urine, and progressive renal failure, were tested for similar effects.
Fourteen rats weighing approximately 200g were injected with about 7.6mg adriamycin per kg body weight; six additional rats were not treated. The rats were placed in
individual cages for 21 days, and their food consumption, weight, and urine were monitored periodically. Urine samples were tested for volume, osmolality, protein content,
leukocytes, blood, and ketones. Three adriamycin treated rats were removed at day 19
for a separate experiment, and the remaining seventeen rats were sacrificed on day 21.
At sacrifice, blood samples were collected from each rat, and kidneys, livers, and hearts
were removed and weighed. Kidneys were dissected into cortex, outer medulla (OM),
inner medulla tip (IM tip), and inner medulla base (IM base). Abundance of UT-A in
the tissue lysates was measured using western blot analysis.
Throughout the course of the study, the adriamycin-treated rats developed
increasing levels of urine protein, suggesting that adriamycin did induce nephrotic
syndrome as expected. Urine osmolality tended to be higher and urine volume lower
in the adriamycin-treated rats, though exceptionally high variability in the treatment
group made these comparisons uncertain. Expression of UT-A in any of the kidney
sections and in liver was not significantly changed, though expression of AQP2, a
water transport protein, was increased in kidney IM tip. These results differ significantly from those observed in rats with uncontrolled diabetes mellitus, where expression of UT-A is clearly increased. Though both conditions involve an unusually large
66
amount of solute entering the kidney, the adriamycin-treated rats did not exhibit
decreased urine concentration. This suggests a direct relationship between UT-A
expression and urine concentrating ability, not the osmolality of the filtrate entering
the kidney. In the heart, UT-A expression was increased with adriamycin treatment. In
addition, expression of UT-A in the hearts of the untreated rats was strongly correlated with blood urea levels. Adriamycin treatment disrupted this relationship. It is possible that UT-A expression may be regulated by urea concentration gradients, and that
adriamycin may disrupt this mechanism.
67
Toward a Better Understanding of...
The Limitations of a Mixed Phase
in Strange Star Crust
David Eby
Mentor: Dr. Mark G. Alford
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
It has been suggested that the form of matter seen in atomic nuclei, containing only
up and down quarks bound into protons and neutrons, is metastable and that the fully
stable state is a mixture of up, down, and strange quarks. This state is known as
“strange matter”, and there may exist “strange stars” made entirely of strange matter.
It has recently been hypothesized that the crust of a strange star may not be a sharp
surface, but a mixed phase of vacuum and small pieces of strange matter
(“strangelets”). At the outermost edge there would be pure vacuum followed by
nuggets of quark matter, which by moving deeper into the star crust, would increase
in density until they became rods and finally slabs of quark matter. The purpose of
this research has been to evaluate the stability of different strangelet structures.
The core of this research has been a computerized numeric model based on the
Poisson equation, commonly used in potential theory. We modeled strangelets by
using Wigner-Seitz cells that are formed similarly to exotic atoms with a nucleus of
strange matter surrounded by a vacuum populated by electrons. The results of this
model were then used to construct enthalpy diagrams that allowed us to determine the
relative stability of these structures under a variety of conditions. These models
enabled us to evaluate the free energy of the Wigner-Seitz cell as a function of its pressure corresponding to greater depth in the crust.
We found that the nugget-shaped strangelets were always the dominant structure
despite variances in several parameters of both the strange matter and the cell configuration. This indicates that if a mixed phase occurs, it will be very limited in terms of
structures. The results of this work will help inform experimentalists where and in what
form strangelets may be detected in addition to clarifying this form of quark matter.
68
Toward a Better Understanding of...
Guanxi: A Reemerging or Disappearing Key
to China’s Labor Markets?
Linda Zhou
Mentor: Dr. Dottie Petersen
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
This research focuses on guanxi, a uniquely Chinese cultural phenomenon. It can be
loosely translated as personal ties, but it not only means human connections, it also
refers to a Chinese phenomenon that denotes implicit, long-term, utilitarian business
relationships that are based on reciprocal obligations, assurances, and understanding.
It is complex, personal, and represents the totality of a bond between two parties. As
such, it is crucial to business and therefore economics. Beginning in the 1990s, guanxi
networks have operated as informal mechanisms that facilitate employment processes in Chinese cities by connecting job seekers and prospective employers. My research
examines the role of guanxi in the rapidly developing and changing Chinese economy.
In the context of China’s economic history, current trends, economic policies, and culture, my research asks: what is the future nature of guanxi as China changes institutionally? I seek to know whether guanxi’s importance will increase or decrease as a
major method for people during the employment search process., or will its function
change? This topic is interesting because China’s economic system is changing significantly, and capitalist-style labor markets are emerging. I am interested in how globalization and free-marketization will affect traditional employment practices, especially
that of hiring potential employees. For example, how should employers and potential
employees act, and what resources should they rely on? I have read many authors with
various points of view as well as interviewed several experienced business people. I
have not yet come up with an “answer” to my question. At its present stage, my
research project is simply analyzing the different scholars’ views and how they are similar and different from one another, and the significance of these similarities and differences. This research may eventually lead into the future role of guanxi in Chinese
business practices by analyzing what part guanxi plays in labor economics. For example: how business people hire employees, how workers locate and secure jobs, how
employees transfer jobs and/or industries, etc. I would try to develop a cultural model
of the most efficient and successful way for Chinese people in China to find employment. Finally, I would also like to explore what the changing role of guanxi means for
China’s economic public policy, especially with regard to the open economy: international trade, international negotiations, and globalization.
69
Toward a Better Understanding of...
Individualism, Economics & Revolution:
Socioeconomic Ideology in
Revolutionary France and America
Andrew Schupanitz
Mentor: Dr. Gerry Izenberg
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
Much has been made of the supposed similarities between the American and French
Revolutions. Many historians have cited parallel ideologies of liberty and equality at
work in both events. R. R. Palmer, for example, in his Age of the Democratic Revolution
melts both revolutions into the broader, more sweeping emergence of liberal democratic forms of government in the Western world. But if this is true, how does one account
for the wildly divergent outcomes of the French and American Revolutions? If the two
events are closely related ideologically, why is it that the American Revolution resulted
in relative peace and prosperity while the French Revolution collapsed under the weight
of a Jacobin dictatorship and brought France once again under the control of an authoritarian ruler? This research challenges the notion that the French and American
Revolutions were born of the same broadly historical movement.
The research first looks at the American debates over ratification of the Constitution.
Focusing specifically on the Federalist papers, as well as Federalist responses to antiFederalist writings, the research finds strong parallels between the socioeconomic/political philosophies of the leading Federalists (notably Hamilton and Madison) and those
of the major Scottish Enlightenment thinkers – specifically David Hume and Adam
Smith. Like Hume and Smith, Hamilton and Madison take human selfishness as given,
and seek to structure government in such a way that this self-interest will contribute to
the overall well being of society. Just as Smith’s Invisible Hand explains how self-interested individuals can unknowingly work toward the benefit of society at large, so too
does Hamilton and Madison’s elucidation of the U.S. Constitution explain how selfinterested individuals in government will be guided to work for the good of the nation
at large. As Madison explains, rather than trying to change human nature, the United
States government will be the “greatest of all reflections on human nature.”
This research next looks at the socioeconomic and political philosophy of the French
Revolution. Unlike their American counterparts, the French revolutionaries often refuse
to take selfishness as given or self-interest as a morally legitimate motivation to action –
economic or otherwise. Instead, Jacobin leaders such as Robespierre and Saint-Just –
inspired largely by the writings of Rousseau – seek to impose a Republic of Virtue and
become increasingly intolerant of any loyal opposition or faction within the Assembly.
This growing intolerance finally results in the Reign of Terror. The final part of the
research tries to account for these differences in ideology – particularly the moral standing of self-interest – and offers a mix of institutional and past-ideological explanations.
70
Toward a Better Understanding of...
Efficacy of the Ketogenic Diet in Children
with Surgically Approachable Epilepsy
Rebecca Stainman
Mentor: Dr. Eric H. Kossoff
Epilepsy is a neurological disorder where neurons fire abnormally, causing epileptic
seizures. After the failure of multiple antiepileptic medications, the ketogenic diet
and/or surgery are considered for seizure control. The ketogenic diet is a high-fat and
low-carbohydrate diet that creates a ketotic state believed to reduce seizures. Surgery
involves localizing the seizure focus using MRIs and EEGs, followed by a resection.
Many parents are hesitant to commit to surgery and wonder if the ketogenic diet
would be an appropriate treatment. This research tests if the ketogenic diet was as
effective as surgery, and if children with surgically approachable epilepsy responded to
the diet as well as other children on the diet with multifocal or generalized seizures.
We conducted a retrospective study with patients on the ketogenic diet at Johns
Hopkins University. These children were divided into separate groups, those who had
surgically approachable epilepsy and those who did not. This comparison allowed us
to see whether children with surgically approachable epilepsy were at a disadvantage
on the diet. The group with surgically approachable epilepsy was further divided into
those who went on to have surgery and those who did not. This allowed us to compare the efficacy of the treatments. For all of the patients, seizure reduction was measured at 6 months.
We expected that children with surgically approachable epilepsy would not
respond as well to the diet as they would to surgery or compared to other children
with epilepsy that were on the diet. Our results showed that children with surgically
approachable epilepsy did respond well to the diet; however they were more likely to
have a >90% seizure reduction with surgery. Furthermore, these children did not
respond as well to the diet as those with multifocal or generalized seizures.
71
Toward a Better Understanding of...
Effects of Management on Garlic Mustard
(Allaria Petolata) Fitness
Brittany Teller
Mentor: Dr. Tiffany Knight
This research was supported by a Washington University/Howard Hughes
Medical Institute Summer Undergraduate Research Fellowship Award.
Garlic Mustard (Alliaria petiolata), an invasive plant species, increasingly threatens
forest and woodland understories where it out competes native species in Missouri
and in many similar areas of the Midwest and plains states. The most non-toxic and
reliable way to remove garlic mustard is by manually removing the plant and its root
system from its stand. However, this process is often subject to human error that
affects both the survival and reproductive success of the target garlic mustard to an
extent that was previously unknown.
In this study, six similar garlic mustard plants were subjected to one of six treatments inspired by human error. The treatments were applied in early April and midMay which corresponds to early and late in the reproductive growth season of this
population of garlic mustard. Fifty replications of the treatments were created. Plants
were either: pulled completely and left in the original stand, clipped at 50% early and
late, clipped at base early and late, or subjected to a control treatment. The treated
plants were monitored throughout the rest of the growing season and were finally harvested to measure average seed weight and number of seeds produced.
Garlic mustard plants that were pulled and left lying above their original stand
produced just as many fruits as control plants at half the weight. Plants that were
clipped at 50% early in the growing season continued to grow and produced fewer
seeds at lower seed weight than control plants. All other monitored plants, such as
those that were clipped at the base or clipped at 50% late in the season had no fecundity following treatment assuming that the reproductive parts of those particular
plants were disposed. Future research will examine and compare germination rates of
the aforementioned seeds.
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Functional Domains of
RNA Binding Protein CUGBP2
Pavel Rodriguez
Mentor: Dr. Shrikant Anant
This research was supported by a Washington University/Howard Hughes
Medical Institute Summer Undergraduate Research Fellowship Award.
CUGBP2 (also called NAPOR, ETR-3, or BRUNOL3) is a ubiquitously expressed RNA
binding protein with highest expression observed in muscle and brain. It was originally believed to play a role in regulating alternative splicing. We identified the protein
based on its ability to interact with apobec-1, the catalytic component of the
apolipoprotein B (apoB) mRNA complex in a yeast two-hybrid screen of liver cDNA
library. Furthermore, we have determined that CUGBP2 binds to AU-rich sequences
immediately upstream of the edited cytidine in apoB mRNA, and represses C to U
editing of apoB transcripts. In addition, we have determined that CUGBP2 is a nuclear
protein that is sometimes found in the cytoplasm where it can regulate stability and
translation of Cyclooxygenase-2 mRNA. However, the mechanism of CUGBP2 function is currently unknown. As a first step to understanding CUGBP2 function, we proposed to perform a structure: function study by generating deletion mutants.
CUGBP2 has significant structural homology to HuR, a member of the ELAV RNA
binding protein family. Both CUGBP2 and HuR encode three RNA recognition motifs
(RRMs) with the first two RRMs (RRM1, RRM2) in tandem, and the last one (RRM3)
separated from the first two by a linker region. A major difference between the proteins is the presence of an A/Q rich linker region that is present only in CUGBP2. The
hypothesis being tested is that RRM1 and RRM2 are involved in mRNA binding and
stabilization, while the A/Q rich linker region and possibly RRM3 are engaged in the
inhibition of COX-2 mRNA translation. To understand the role of the different
domains, we have generated a series of deletion mutants, including deletion of the
individual RRMs and the A/Q-rich bridge region, as well as deletion of both RRM1
and RRM2. In addition, we generated constructs that encoded just the individual
domains, cloned into plasmids pCMV-Tag2B (for expression in mammalian cells) and
pGEX-4T3 (for expression in bacteria). Further studies were performed with these
deletion mutants. We have recently identified that the full length CUGBP2 localizes to
the nucleus and shuttles between the nucleus and cytoplasm. Using immunocytochemistry, we have determined the localization of the individual proteins in two
human cell lines, HCT116 colon adenocarcinomas cells and HeLa cervical carcinoma
cells. We are also determining the effects of the mutants on stability and translation of
COX-2 mRNA. We have also begun bacterial expression and protein purification using
affinity chromatography for further studies. These studies will delineate the role of the
different domains of CUGBP2 in stability and translation.
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Toward a Better Understanding of...
What Would Sex Education
from the Left Look Like?
Kristi Nigh
Mentor: Dr. Susan Stiritz
Helping students navigate and understand the sexual culture of their society should be
a major goal of sexuality education curriculums for high school and college students.
Rather than treating sexuality as simply personal, we position it as the core issue of our
times. Our progressive high school and college-level curricula use literature as a lens
to explore how the current American sexual culture supports and legitimizes gender
inequalities, including a social, political, and economic supremacy. By featuring stories that present themes of sexuality and characters that students can relate to on a
personal level, our curricula provide opportunities for students to reflect upon the
lives and societies of the characters they read about as well as examine, critique, and
begin to understand their own sexualities as they fit in within the context of their own
sexual cultures. Because we believe that respect for difference, pleasure, reciprocity,
and dialogue would positively transform our society, we also expose students to the
power of mutuality. This presentation details how we assess needs, craft a mission
statement, set learning objectives, construct activities, select instructional materials,
and evaluate students’ success on both the high school and college levels.
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Toward a Better Understanding of...
The Role of JIP-1 in Axon Transport
Pavan Bhat
Mentor: Dr. Aaron DiAntonio
This study investigates the process of axon transport, which is the movement of molecules between cell body and the synapses of the neuron. In some motoneurons this
distance can be quite large, almost a meter. The focus of this research is the Drosophila
protein JIP-1. In vertebrates, JIP-1 has been shown to interact with Kinesin, a motor
protein that carries cargo down axons, and with JNK a signaling molecule. In
Drosophila, our lab has found that JNK signaling, regulated by Wallenda, functions in
axon transport. This investigation addresses whether Wallenda functions through JIP1 to regulate transport.
Two approaches are used to address this question. The first approach is to develop
an assay to determine whether Wallenda, Kinesin, and/ or Dynein motors associate
with JIP-1 in Drosophila embryo extracts. Antibodies that bind to JIP-1 and to beads
to purify JIP-1 and associated proteins will be used. This technique is called a coimmunoprecipitation. Another set of antibodies specific to different proteins of interest will be used, in order to detect which proteins have bound to JIP-1. The second
approach is to knockdown expression of JIP-1 using a RNA interference technique and
look for genetic interactions with Wallenda. The technique of RNA interference or
RNAi involves creating a special double stranded RNA that directs destruction of JIP1 RNA transcripts, thus effectively silencing the JIP-1 gene.
My hypothesis is that JIP1 will bind with Kinesin and/or Dynein. I also expect to
see axon transport defects in the JIP1 RNAi larvae. These results are important
because JIP1’s role in axon transport is not completely understood. Understanding
axon transport is important because many neurological disorders are associated with
a defect in axon transport.
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Toward a Better Understanding of...
Hopping Rates in Scandium Hydride
and Magnesium Hydride
Erik Carl
Mentor: Dr. Mark Conradi
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
The study of metal hydrides and their capabilities to store and transport hydrogen has
played a vital role in developing hydrogen as a viable alternative fuel source. My
research under the Conradi group focuses on measuring hopping rates of hydrogen
gas through metal hydrides, specifically magnesium hydride and scandium hydride, by
means of nuclear magnetic resonance.
Nuclear magnetic resonance exploits the fact that nuclei posses magnetic moments
due to their intrinsic angular momentum as predicted by quantum mechanics. When
placed in a magnetic field, solids, such as metal hydrides, obtain a macroscopic magnetic moment due to the addition of all of the individual magnetic moments of their
nuclei. By applying radio frequency pulses to metal hydrides, the macroscopic magnetic moment can be perturbed and manipulated in such a way as to yield information on the hopping rates of hydrogen gas through the lattice of the metal hydride.
My research has found that the hopping rates of hydrogen are significantly faster
in scandium hydride than they are in magnesium hydride. This would seem to indicate that scandium hydride would prove more advantageous in hydrogen fuel cells
because the hydrogen can travel faster within the scandium hydride lattice than the in
magnesium hydride lattice. However, magnesium hydride can store significantly more
hydrogen than scandium hydride. Research under the Conradi group has shown that
magnesium scandium hydride actually possesses faster hopping rates of hydrogen gas
than both scandium hydride and magnesium hydride and can also store more hydrogen than scandium hydride. Therefore of the three compounds, magnesium scandium
hydride would prove to be more advantageous in hydrogen fuel cells.
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Toward a Better Understanding of...
Noise Reduction by Spectral Contrast Shaping
Nathan J. Killian
Mentor: Dr. Dennis Barbour
Some neurons in the auditory cortex exhibit a substantial dynamic response range at
intermediate spectral contrast values. Spectral contrast represents variance in sound
energy distribution across frequency. Previous studies using spectral contrast
enhancement to improve the intelligibility of noisy speech have demonstrated some
benefit for normal listeners and cochlear implantees. We reasoned that filtering or
shaping the contrast of noisy speech to match the contrast tuning of these auditory
cortex neurons may prove to be a useful strategy for improving speech intelligibility
in noise. Such a strategy may by particularly useful for cochlear implantees, whose
native contrast filtering circuitry is likely to be nonfunctional. We decomposed noisy
speech into frequency and contrast components, applied an exponential weighting to
randomly selected contrast values, and finally reconstructed a time-domain signal by
summing across the individual filters. Additionally, it may be possible to perform this
weighting based on the value of a single, continuously updated contrast parameter.
This would lend the operation quite readily to use in cochlear implants.
The normalized correlation metric (NCM)—verified previously to be an accurate
predictor of noisy speech intelligibility for cochlear implantees—provided quantitative estimates of intelligibility. Signal components of intermediate contrast (4–8 dB)
were found to be more important for increasing speech intelligibility than were high(8–20 dB) or low-contrast (0–4 dB) components. When preferentially emphasizing
contrast values in the intermediate range, we found NCM score increases of 10–20%
for speech and white noise having signal-to-noise ratios of –5, 0, and 5 dB.
Preferentially enhancing the highest and/or lowest contrast values resulted in substantially decreased NCM scores. Visual inspection of spectrograms indeed shows
dramatic noise reduction. These observations agree with neurophysiological expectations and imply that contrast shaping may be a useful noise-reduction algorithm
for normal-hearing listeners and/or hearing-impaired listeners such as cochlear
implantees. Listening studies in cochlear implant users are planned to test the efficacy of the algorithm.
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Toward a Better Understanding of...
Pictures of the Mind: the Pictorial
Foundation of Contemporary Neuroscience
Cara Dolan
Mentors: Dr. John Doris and Dr. Tony Jack
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
The purpose of our study is to investigate the effect that images presented in neuroscience articles have on the citation impact of the articles. We hypothesized a correlation between the number of pictorial images present in an article and the impact the
presented research has on its relevant field, as reflected through number of citations
received. We coded the number and type of figures for research articles published in
the year 2000 in two prestigious neuroscience journals, Nature, Neuroscience and
Neuron, and one of the top tiered general science journals, Science. Images were separated into three categories, basic, specialized and pictorial. Those categorized as basic
are recognizable quantitative representations of data, or standard forms of graph.
Specialized images might also represent quantitative data, but in a non-standard format. The final category was of pictorial images. The canonical case for pictorial images
would be an actual photographic picture, however we also included in this category all
figures that had a pictorial quality, such as the computer generated colored images that
are commonly used to present the result of brain imaging studies. After the initial data
were coded for, the number of citations each article received was recorded using the
Web of Science database. The type and number of figures present in each article were
then compared to the number of citations for each article. Both neuroscience journals
showed a positive correlation between number of pictorial images presented and
number of citations received. There was no such overall correlation found in the general science journal. These results suggest a unique importance of pictures to the field
of neuroscience.
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Toward a Better Understanding of...
Martin and Monradi:
An Ethics of the Ordinary
Maria Efimova
Mentor: Dr. Guinn Batten
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
As a junior in the Undergraduate Honors Fellowship program, I am in the early stage of
my project on theoretical approaches to the work of American minimalist painter Agnes
Martin and Italian still life painter Giorgio Morandi. My project asks how the artist’s aesthetic engagements with objects in the world become ethical engagements.
The central texts for my project are Jacques Lacan’s Seminar XX: On Feminine
Sexuality, the Limits of Love and Knowledge and Joan Copjec’s Imagine There’s No
Woman: Ethics and Sublimation. I am very interested in the relationship of these psychoanalytic, theoretical texts to the works of art. The artist and critic Mel Bochner has
said that for a critical text to make relevant commentary on art, “there has to be some
tension in the writing that reflects the conflict in the work.” I hypothesize that psychoanalytic theory tries to construct a kind of world or system around the individual in
the same way that a painting by Martin or Morandi strives to become inhabitable for
the viewer.
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Toward a Better Understanding of...
A New World Grain in an Old World:
A Study of Maize’s Impact on
Sustainability in Sub-Saharan Africa
Margot Danker
Mentor: Dr. Glenn Stone
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
Over the past century, maize cultivation and consumption in sub-Saharan Africa has
increased exponentially. While the crop was initially nothing more than a foreign garden crop grown primarily as a vegetable, the grain skyrocketed in popularity at the
turn of the twentieth century to become to a hugely popular, often monocropped
grain that has become the primary staple for millions of Africans. The reasons for this
explosion in maize cultivation are varied, but the efforts of seed companies combined
with new market forces that emerged during that time encouraged many farmers to
abandon more traditional indigenous crops, such as pearl millet and sorghum, in
exchange for the New World grain. This dramatic shift has had many consequences on
sub-Saharan economies, ecologies and cultures, particularly as new, higher-yielding
seeds that require more expensive and environmentally damaging inputs are introduced. Although some have benefited from this massive increase in maize cultivation,
namely governments, seed companies and larger scale farmers, others have not. As
agricultural practices become more industrialized and less sustainable, many smallholders, and especially female farmers, are experiencing insecurity, loss of indigenous
knowledge and debt as they become more dependent on maize. If more sustainable
practices are not introduced, these problems plaguing small-scale farmers will continue as food and economic insecurities persist.
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Toward a Better Understanding of...
Elucidating Preconditioning-Induced,
AKT-Mediated Signaling Mechanisms
Operative in Cerebral Endothelium that
Protect the Microvasculature from Stroke
Somalee Banerjee
Mentor: Dr. Jeffrey Gidday
This research was supported by a Washington University/Howard Hughes
Medical Institute Summer Undergraduate Research Fellowship Award.
Hypoxic preconditioning (HP) enhances cellular tolerance to ischemic injury in a
variety of organs; however the mechanisms underlying this protective effect have yet
to be fully elucidated. Since endothelial cells are the first to experience the detrimental effects of cerebral ischemia, maintaining their integrity is the body’s primary line
of defense to limit tissue injury. Protein kinase B (Akt) is an intracellular kinase that
regulates, after phosphorylation by PI3 kinase, the activity of a number of different
proteins that have protective roles after an ischemic injury. Thus, the focus of my project was to characterize the temporal expression of Akt and phosphorylated Akt (pAkt)
in the cerebral microcirculation and to identify some of its downstream effectors
mediating vascular protection in an adult mouse model of stroke. I hypothesized that
HP would increase pAkt levels in the microvessel endothelial cells, and that this would
lead to the phosphorylation of downstream molecular targets of Akt, which would
account for the vascular protection. A pro-survival effector examined was endothelial
nitric oxide synthase (eNOS), which, when activated by phosphorylation, produces
nitric oxide which exhibits protective effects within endothelial cells and at the bloodendothelial interface.
Groups of adult male Swiss Webster/ND4 mice were exposed to a HP stimulus (4
hours at 8% oxygen) after which they were recovered under normoxic conditions
prior to being sacrificed either immediately after preconditioning, or at 2, 6, 24, or 48
hours later. Cerebral microvessel-rich preparations from four individual mice were
pooled together and used for immunoblotting analysis. Results from this set of experiments demonstrated that Akt levels stayed relatively constant after HP. Expression
levels of pAkt, however, showed a maximum upregulation immediately after HP and
decreased to below naïve levels by 48 h after HP.
In order to facilitate the identification of downstream pAkt phosphorylation targets, the PI3-kinase inhibitor LY294002 (LY) was administered to separate groups of
mice 45 minutes prior to HP. Expression levels of eNOS and phosphorylated eNOS (peNOS) were measured to examine the necessity of pAkt in activating eNOS protein
levels. Protein profiles in cerebral microvessel fractions from mice treated with LY and
untreated control mice were examined at identical post-HP time points by
81
immunoblotting. Results from these experiments showed that LY markedly decreased
the expression of both Akt and pAkt normally observed to be elevated immediately
after HP. Also, there was a marked decrease in the expression levels of both eNOS and
p-eNOS immediately following hypoxic preconditioning in mice treated with LY as
compared to those without the inhibitor, suggesting that pAkt phosphorylates and
maintains p-eNOS levels in microvessel endothelial cells in response to HP.
Collectively, these results are the first to demonstrate the temporal profile of cerebral microvascular-specific expression of Akt and pAkt following hypoxic preconditioning. The lack of an increase in p-eNOS levels was unexpected and will be investigated further. Moreover, I will continue to leverage this experimental design to identify other phosphorylated proteins regulated by pAkt that are integral to HP-induced
protection of the endothelial cells that comprise the cerebral microcirculation.
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Toward a Better Understanding of...
Neural Evidence of Syntactic and
Referential Competition During
Sentence Processing
Amir Francois
Mentor: Dr. David January
This research was conducted as a part of the Washington University
Mellon Mays Undergraduate Research Fellowship Program.
This research investigates the neural evidence of syntactic and referential competition
during sentence processing in the left inferior frontal gyrus (LIFG; Brodmann areas 44
and 45), also known as Broca’s area. There are two main schools of thought on the role
of the LIFG: the first school believes that LIFG is integral to grammatical processing,
the seconds states that the LIFG is a major center in competition resolution no matter
the domain (including non-grammatical processing). Our experimental design uses
syntactic and referential competition as examples of grammatical and non-grammatical processing, respectively. We expect LIFG activation in both conditions, which
would support the non-domain specific competition resolution center hypothesis.
However, if the data show activation in only the syntactic condition, this would support the grammatical processing model. Functional magnetic resonance imaging was
used to acquire the brain activation data.
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Toward a Better Understanding of...
The Silent Victims of the AIDS Epidemic:
Rethinking the Illusion of Monogamy and
the Social Constructions of Marriage
Emily R. Hawkins
Mentor: Dr. Shanti Parikh
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
Earlier social analysis of HIV tended to emphasize the plight of the sex workers who
were exposed to AIDS as an occupational risk. Recently, however, there has been much
epidemiological and ethnographic literature that highlights the overwhelming evidence
that, instead, marriage and monogamy are what make a woman most susceptible to
contracting the HIV infection. Other than simply exposing this phenomenon and making broad generalizations about ineffective prevention in developing countries, less has
been done to question the structural forces that allow these patterns to occur.
Historically, the social commentary about people most frequently associated with
the disease has portrayed other groups, such as homosexual men, African-American
men, intravenous drug users and sex workers. Absent from these discussions are their
sexual partners, especially wives and lovers. Therefore, women are the crucial locus for
understanding the hidden dynamics of the cultural transmission of HIV because the
underlying social factors at work effectively silence their stories. Because dialogue
about issues such as sexual violence, marginalization of women, inequality, risk denial,
the illusion of monogamy in married relationships and the inability of women to
negotiate sexual protection are often considered socially unacceptable in many societies and therefore silenced—the victimization of women is often overlooked in much
of the current research and discussion. While married women are no longer an original category for analysis and it is clear that the central risk factor for HIV transmission is their husbands’ behavior, many unresolved questions remain.
This research challenges some of the fixed assumptions about the transmission of
HIV and redefines social categories by looking at the definitions of “sex worker” and
“married woman” while suggesting that perhaps these categories are not as stark as
they were in the past. Whereas a dominant way of looking at HIV spread has been
largely in terms of individual behaviors, I argue for a shift to a paradigm that would
consider the structural determinants that surround and facilitate those choices. By
examining the underlying implications of the cultural constructions of a married
woman living up to the standard of monogamy versus the unfaithful husband reality,
I will deconstruct the applications of cultural practices and reveal the factors which
allow married women to become victims of their social system.
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Toward a Better Understanding of...
Repetitious Element 1360 as a Target for
Heterochromatin Formation in
Drosophila Melanogaster
Alejandra Figueroa-Clarevega
Mentor: Dr. Sarah Elgin
This research was supported by a Washington University/Howard Hughes Medical Institute Summer
Undergraduate Research Fellowship Award and NIH grants GM68388 and GM73190 to SCRE.
Drosophila melanogaster has served as an excellent model for the study of genetics; it has
a fully sequenced genome, and many easily visible phenotypes. Like other eukaryotic
organisms, Drosophila displays highly effective mechanisms for DNA packaging. It has
been particularly useful in differentiating between the two basic types of chromatin
(defined as DNA and its associated proteins): euchromatin and heterochromatin, which
differ not only in their mechanism of packaging DNA, but also in the associated levels
of gene expression. Most actively transcribed genes are found in euchromatin; on the
other hand, heterochromatin is the form of packaging associated with gene silencing.
Proximity to heterochromatin has the consequence of inducing PEV (Position Effect
Variegation), in which a reporter gene is silenced in some of the cells where it is normally active. Earlier work has suggested that the repetitious element 1360, which is enriched
in heterochromatic domains, can serve as an initiation site for heterochromatin formation in the fourth chromosome. With this in mind, a P-element construct was designed
containing one or four copies of 1360 upstream of an hsp70-driven white reporter to
test whether this repetitious element is capable of initiating heterochromatin formation.
This construct was injected into Drosophila embryos and transformants carrying the
transgene were recovered. To generate additional lines, an X-chromosome insertion line
was used in a cross to mobilize the P-element. A total of 5,000 males were screened for
mobilizations. Without the 1360 element upstream of hsp70-white, only 1% of the lines
recovered from previous P-element mobilization screens were variegating. In other
words, this 1% indicates the percentage of P-element insertions in domains able to
induce heterochromatin formation, phenotypically visible as a variegating eye. We have
observed that with one copy of 1360, 4% of the recovered lines are variegating; with four
copies of 1360, 12% of the recovered lines are variegating. These results suggest that
increasing the number of copies of 1360 increases the likelihood of initiating or maintaining heterochromatin formation. Furthermore, when looking at the distribution of
variegating lines in terms of their chromosomal location, we observed that the majority
of inserts were in the second and third chromosomes, which unlike the fourth chromosome have a large portion of euchromatin. This leads us to believe that we are recovering variegating lines with P-element insertion sites outside regions that are known to be
heterochromatic by the addition of the repetitious element 1360.
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Toward a Better Understanding of...
Continuing the Technology Debate:
The Effects of Interactivity and
Visual Stimulation on Learning
Andrea Young
Mentor: Dr. Desiree White
This research was conducted as a part of the Washington University
Mellon Mays Undergraduate Research Fellowship Program.
This research focuses on the efficacy of interactive, visually stimulating technology
(such as video and computer games) on quality of learning. Interactivity seems to have
a powerful influence on how well people learn information and tasks. As such, interactive media has become a growing target of psychological investigation. Much of the
research conducted has focused on the effects of video games, computer programs,
and television on aggression, however, interactivity can be a very powerful teaching
tool and very productive when used in an educational context.
Only in the past decade or so have experimenters been interested in whether or not
interactive media can have an effect on cognitive ability. The majority of the research
does indeed show that use of interactive technology and skill and knowledge acquisition at least correlate. Keeping in mind some of the flaws of past experiments, we are
in the process of designing an experiment to test the efficacy of an interactive, computerized biology textbook. Subjects (from Washington University’s subject pool) will
be placed in one of three conditions. In the first condition, subjects study a lesson that
is interactive and visually stimulating. In the second condition, the lesson is visually
stimulating but not interactive. The final condition is the passive condition; while the
lesson is still on the computer, it is neither visually stimulating (in comparison to the
other two) nor interactive.
Participants return to the lab 48 hours after studying the lesson to complete a
paper and pencil test on the lesson they studied as well as a questionnaire. The test
includes three sections: definitions, multiple choice, and short answer. The questionnaire assesses learning styles by placing subjects on four different scales: measuring
whether the subject is (1) a verbal or visual, (2) active or reflective, (3) sequential or
global, or (4) sensing or intuitive learner. We expect that subjects in the first condition will perform better on the test than subjects in the other two conditions due to
interactivity and that subjects in the second condition will perform better than those
in the third, because we do expect some effects for visual stimulation (even if only
keeping the participants attention). We expect that there will be little variation based
on learning styles.
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Toward a Better Understanding of...
Design and Construction of
a Formula-Style Race Car
Melissa Holtmeyer
Mentor: Dr. Jeremy Neubauer
A project from the Mechanical Engineering department that provides a wealth of
applied engineering experience is the Formula SAE (Society of Automotive Engineers)
racecar, designed and built by students for students. The goal of building such a car is
to compete against over 140 universities from around the world at the annual Formula
SAE Student competition held in Detroit, Michigan every May. In addition, a total of
seven Formula SAE student competitions exist throughout the year around the world.
The car used for these challenges is a small formula style, open wheel vehicle weighing around 450 pounds, powered by a 600-cc motorcycle engine. This car shares many
of the same features found at the highest level of motor sports, where speed, handling,
and reliability are key components of success. A very different feature, however, is that
all of the components are designed, analyzed, and manufactured by college students.
While these students have almost endless freedom in completing these tasks, they
must work within the rules of competition and the financial and manufacturing capabilities of the team. The most important of these rules states that no second year cars
may return to international competition. It is mine and the team’s responsibility to
design and provide documentation for a brand new car each competition year.
I have been involved with this project for the past two and one-half years and serve
as lead designer for the 2007 competition car. My designs incorporate the ease of manufacturability and practicality, and include the following components for the 2007
entry: Drive train, Driver Controls (Gas Pedal, Brake Pedal, and braking system central
location), Braking system, Exhaust. Additionally, the following projects in progress,
including the above: Chassis complete, Engine selected, Machining beginning on
Suspension, corners, and steering, Fuel system design, Impact attenuator design. It is
also a near term goal for the team to become environmentally friendly by fueling with
E85, instead of gasoline, and possibly designing for the Formula Hybrid challenge.
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Toward a Better Understanding of...
Difference-Blind Politics
in a Multicultural State:
The Political Implications of
Identity in Democratic Societies
Marquita James
Mentor: Dr. Gerald Izenberg
This research was conducted as a part of the Washington University
Mellon Mays Undergraduate Research Fellowship Program.
America has long been proud of its diversity as evidenced by the motto on the country’s state seal- out of many, one. The many were initially the thirteen states that combined to form the United States. As America became increasingly more diverse, and
especially in the 19th and 20th centuries as that diversity gained more political recognition, more questions arose about who the many are and what their relationship
should be to the one. In democratic societies, which are by definition comprised of
individuals working together to accomplish common goals, the kinds of people who
choose to cooperate politically are important.
According to modern democratic principle, those political associations should
change depending on the issue at hand. Along the same line of thought, people should
be treated equally—without any consideration of arbitrarily assigned characteristics
such as race, sex, or even disability. This difference-blind application of the laws
should be uniformly applied, to prevent both privileging and disadvantaging any
member or segment of the unified political body. Identity as an American should provide a strong enough foundation for individuals to develop political associations.
However, the fallacy of invoking difference-blind principles in the United States is that
these principles have not always been applied. Hence, what seems a novel, human
rights grounded, difference-blind approach can become a reinforcement of institutionalized American historical inequalities.
Here, I will showcase a segment of my ongoing research project that attempts to
flesh out the political implications of racial and cultural identity. How are we to justify recognizing people as members of collective groups in democratic societies that
tout the merits of equality and individualism? Do racial and identity groups have
legitimate claims to special political consideration? In the future, I will assess what the
political recognition of identity groups might mean in practice by examining historical and contemporary case studies.
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Toward a Better Understanding of...
Bronze Age Steppe Dynamics and
Stable Strontium Isotopes
Taylor Hermes
Mentor: Dr. Michael Frachetti
During the Bronze Age (c. 2500 – 1000 BC), populations practiced the socio-economic strategy of mobile pastoralism across Central Asia. This region and time period is
particularly noted for the proliferation of horseback riding, wheeled transport, metallurgy, and Indo-European language. This environment enabled a rich and diverse cultural community to develop that challenges frameworks of complexity. What was once
thought of as a singular “Andronovo” cultural community, is now better understood
as an adaptive and diverse society, reflecting a somewhat common but varied material culture. Moreover, recent interpretations present models of oscillating levels of
complexity that illustrate a non-uniform socio-political landscape. This study investigated transitions between consolidations and fractionalizations of society through stable strontium isotopic analysis on human archaeological remains.
The geographic locality of this study was the Koksu Valley of the seven-river system region of eastern Kazakhstan known as Semirech’ye. This area fits into the larger
cultural landscape of the Central Asian steppe zone as a microcosmic model to test
theories of social dynamics. The presence or absence of non-local individuals at the
archaeological pastoral camp complexes of Begash created a clearer picture of the
mechanisms that drove social, political, and economic organization within the Koksu
Valley and farther across the Central Asian steppe.
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Toward a Better Understanding of...
Constructing Various Mutants of
Pathogenic Bioluminescent Salmonella for
a Murine Model of Infection
Aasish Manglik
Mentor: Dr. Jeffrey McKinney
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
Salmonella infections can lead to gastroenteritis, osteomyelitis, bacteremia and sepsis
in many hosts, including humans. In developed countries, Salmonella typhimurium
continues to compromise food safety; in less developed countries Salmonella typhi
causes typhoid fever and significant mortality. To study how Salmonella causes disease,
the McKinney lab works with a fully virulent enteric subspecies Salmonella typhimurium, which we are sequentially altering by genetic modifications for gain or loss of
functions of interest.
Our lab has integrated the P. luminescens luxCDABE operon into the chromosome
of S. typhimurium. The resulting Salmonella exhibits constitutive bioluminescence,
emitting photons that can be measured using a specially optimized camera. Using this
system, we can spatially and temporally track Salmonella dissemination in real time,
in living mice, by observing the bioluminescent signal given off by the bacteria.
To study the events involved in Salmonella pathogenesis, I have made otherwise isogenic derivatives of our bioluminescent Salmonella strain. Thus far, I have created precise in-frame deletions of three genes thought to be critical to Salmonella induced disease. These include invC, a Salmonella Pathogenicity Island I (SPI-1) gene, sseB, a SPI2 gene, and msbB, a gene involved in the final synthetic steps of the endotoxin
lipopolysaccharide (LPS). Classically, SPI-1 genes are thought to mediate initial invasion into the host cell, which are the epithelial cells lining the intestines. SPI-2 genes are
often cited as important for Salmonella survival inside the host cell, after the invasion
process. The endotoxin LPS triggers the so-called septic shock response in the host.
Although the biologic functions of these genes have been previously explored, we
can now assess their influence over the spatiotemporal details of pathogenesis in our
whole animal model. By comparing the spatiotemporal patterns of the bioluminescent
signals of our respective invC-, sseB-, msbB- Salmonellae strains, we hope to gain further insights into the in vivo relevance of the SPI-1, SPI-2, and LPS mediated disease
pathways during infection. In time, this may also help us create precisely attenuated
Salmonellae strains for use in other contexts.
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Toward a Better Understanding of...
Role of the Transcription Factor HIF-1a
in Epithelial Inflammation
Rebecca Martin
Mentor: Dr. Jeffrey M. Arbeit
The primary question of this work is: What is the role of the transcription factor HIF-1a
in epithelial inflammation? We hypothesize that over-expression of HIF-1a in keratinocytes (basal cells of the epidermis) will lead to enhanced inflammation. Experiments
were performed in vivo, hoping that a better understanding of the role of HIF-1a in skin
inflammation could lead to better treatment for inflammatory skin conditions.
To evaluate the effects of over-expression of HIF-1a, we created a transgenic line of
mice called K14-HIF-1a Double Proline Mutant (DPM), which constitutively expressed
HIF-1a under the Keratin-14 promoter in keratinocytes. Comparing DPM mice with
non-transgenic (NTG) mice, we examined inflammatory cells, and blood and lymphatic vessels in untreated and phorbol myristyl acetate (PMA) treated ears. We used tissue
staining techniques such as immunohistochemistry and immunofluorescence in evaluating levels of inflammation. Stains were performed on 5 mm sections of treated and
untreated NTG and DPM ears. Pictures were then taken using a bright field and fluorescence microscope. When quantification of data was needed, positive inflammatory
cells in a 20X field were counted and statistical analysis was performed.
The PMA-treated DPM cohorts demonstrated increased T-cells, macrophages,
neutrophils and mast cells, and greater dilation of blood and lymphatic vessels. The
number of inflammatory cells observed in untreated DPM ears was approximately
twice the number observed in untreated NTG ears. Only six hours after PMA treatment, DPM mice demonstrated dilated blood vessels and epidermal micro-abscesses
filled with neutrophils. After 10-days from the initial PMA treatment, the inflammation in DPM ears was not resolved as it was in the NTG; instead, it progressed with
increased inflammatory cells, lymph-vessel and blood-vessel growth in the dermis,
and thickening of the epidermis with rete ridge-like projections. These experiments
confirmed our hypothesis that over-expression of HIF-1a in PMA-challenged keratinocytes leads to inflammatory conditions resembling human psoriasis.
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Biodiversity and Energy Availability:
Testing the Evolutionary Rates Hypothesis
Anne K. Greenberg
Mentor: Dr. Barbara Schaal
This research was supported by a Washington University/Howard Hughes
Medical Institute Summer Undergraduate Research Fellowship Award.
The inverse relationship between latitude and biodiversity is one of the strongest patterns in biology. Among many competing mechanisms to explain this pattern, the evolutionary rates hypothesis predicts that higher levels of environmental energy (i.e. temperature and ultraviolet radiation) in tropical, low latitude regions lead to elevated rates
of molecular evolution, which in turn leads to increased rates of speciation and thus
more biodiversity. We are using the biogeography and evolution of a creeping evergreen
vine, Mitchella repens (Rubiaceae), to test the hypothesis that heightened levels of environmental energy lead to increased rates of molecular evolution and divergence.
Mitchella repens has an ideal distribution for testing the evolutionary rates hypothesis.
This species exists in both temperate eastern North America and tropical Central America,
and has a sister species (M. undulata) that occurs in temperate Asia. Temperate populations of these species are exposed to similar amounts of environmental energy, while the
tropical populations of M. repens experience much higher levels of environmental energy.
To determine whether increased levels of environmental energy lead to an
increased rate of molecular evolution, we collected ten samples from ten populations
of Mitchella repens across 100-mile transects in Mexico and the eastern United States.
We sequenced multiple regions of the chloroplast genome to find regions that are variable enough to allow us to create a haplotype network for this species. We have successfully identified two sufficiently variable regions of the chloroplast genome, psbDtrnT and rpL32-trnL, and have sequenced the psbD-trnT region in every Mexican
sample, allowing us to create a haplotype network for the Mexican individuals.
Another haplotype network will be constructed for the eastern North American samples once sequencing of these samples is completed. Analysis of these networks will
allow us to measure the rates of divergence between local populations as estimated by
Fst, a measure of the distribution of variation between local populations.
Higher levels of divergence between the Central American populations of Mitchella
repens compared to their North American counterparts will support the evolutionary rates
hypothesis. If our data support the evolutionary rates hypothesis, more work will be
required to fully determine whether higher levels of available energy are the precise cause
of these observations. Among other possibilities, an increased rate of speciation may be the
consequence of more diverse habitats in the tropics, which leads to a decrease in gene flow
between individual populations and an increase in speciation. Higher levels of divergence
could also be related to the increased amount of available land or decreased seasonal variation at lower latitudes. These questions can be investigated in future related studies.
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VERITAS Observation of GRB060501
Rashied Amini
Mentor: Dr. Henric Krawczynski
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
Gamma Ray Bursts (GRBs) are nature’s most powerful phenomena. Spurred by the
collapse of large stars, or supernovas, GRBs project gamma rays with very high energies at or above an order of 1 TeV. Following a GRB, there is an observed “afterglow”
that consists of electromagnetic radiation at radio, IR, visible, UV, and X-ray energies.
The Very Energetic Radiation Imaging Telescope Array System (VERITAS) is a telescope array built to detect the Cherenkov radiation that emanates from the shockwave
of air showers caused by gamma rays entering earth’s atmosphere.
On the evening of May 1st, 2006, a GRB was detected by NASA’s SWIFT Burst Alert
Telescope. The burst’s location was then radioed across the GRB Coordinates Network
and picked up by numerous observatories. In the observation of GRB060501, there
were numerous difficulties in pinpointing an afterglow due to hardware, weather, and
background problems experienced by many observatories. In order to find an ever
elusive afterglow in the gamma ray energy range, the VERITAS Cherenkov telescope
system observed post-burst. This research focused on the VERITAS observation of the
burst and the associated methods of discovering gamma ray afterglow. We reported on
a search of an excess of events on time scales between one and several runs, and on
very short (30 s) time scales. This preliminary analysis does not show strong evidence
for TeV gamma-ray emission associated with the GRB. The data should be studied
again after further optimization of the gamma-hadron separation cuts.
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Lead Adsorption to
Titanium Dioxide Nanoparticles
Carolyn Maus
Mentor: Dr. Daniel Giammar
Heavy metals such as lead are toxic and non-biodegradable, making them persistent environmental contaminants. Lead is usually introduced into bodies of water by human activity; electric utilities, metal smelters, and lead pipes are several sources. Lead poses an especially acute danger to children, in whom even low levels can cause severe developmental
defects. One method of removing lead from water is adsorption, whereby a sorbent is
added to the water. The contaminant adheres to the sorbent surface through chemical or
electrostatic bonds, and the solid mass formed can be removed. Since adsorption occurs
proportional to the surface area of the sorbent, nanoparticles are the subject of current
study. Nanoparticles, or those particles with sizes under 100 nanometers, have very high
surface area to mass ratios, making them efficient sorbents. Furthermore, it is hypothesized that nanoparticles’ structural properties may affect their reactivity with contaminants. Previous experimentation has also shown that oxides are effective for heavy metal
removal. For these reasons, titanium dioxide nanoparticles are the focus of this research.
This project studied lead adsorption to various sizes of titanium dioxide nanoparticles over a pH range of 2-8. Both commercially produced and laboratory-synthesized
particles were used. Experiments were carried out in batch reactors with known concentrations of lead and titanium dioxide. Ionic strength and pH were also controlled.
Samples were taken from each reactor after equilibration and were filtered, centrifuged,
and acidified. These samples were analyzed for lead content using inductively coupled
plasma mass spectrometry. The amount of lead remaining in solution was compared to
the initial amount of lead added to determine how much adsorption had occurred.
The experiments confirmed that more sorption occurred at higher pH values, which
was expected for a metal cation like lead. The results also showed that the nanoparticles
adsorbed more lead per unit mass than the coarser particles that were also studied, supporting the idea that nanoparticles are more efficient sorbents. However, when adsorption was normalized to surface area, the coarser particles adsorbed more lead than any
of the nanoparticles. This indicates that the nanoparticle crystalline structure may not
provide the same density of highly reactive surface sites for adsorption. Adsorption was
also greater to anatase particles than particles composed of both anatase and rutile titanium dioxide, suggesting that even between particles of similar size, crystalline composition makes a difference. Further experiments are being conducted to determine if
adsorption trends are observable for materials that only differ in size, and if so, at what
size are the effects apparent. Studying materials that have the same crystalline phase and
were synthesized by the same method will clarify this size effect. An additional research
question is whether the trends seen so far are applicable to adsorbates other than lead.
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Study of Decays from Excited States of 10C
Kevin Mercurio
Mentor: Dr. Lee Sobotka
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
The structure of nuclei with about 10 nucleons (A~10) is interesting both because
these nuclei exhibit a great variety of interesting phenomena and because they are at
the crest of what is possible with theories starting with realistic 2 (and 3-body) nucleon-nucleon interactions yet deal with the interactions between all A objects. The interesting phenomena include: a) the generation of “neutron skins” in neutron rich nuclei,
where the neutron density far exceeds the proton density, b) so called Borromean
nuclei, which are particle bound but with the removal of a nucleon the remainder falls
apart into individual nucleons and alpha particles and finally c) these nuclei exhibit
well defined molecular-like structures. Calculations show that the later is true for
10
4Be6, which can be described by a molecular orbital picture of two valence neutrons
surrounding a two-centered a-particle core.
Our study of 106C4 is also motivated by the relative scarcity of data on this nucleus
as compared to its mirror 10Be. The later has 6 particle-bound levels (including the
ground state) and many more known resonances. 10C has only two bound levels and
three reported resonances. This work has two main goals. First is an attempt to confirm the reported unbound levels as well as to search for additional unbound structures. Our second aim is to study the decay chain of all observed resonances.
10
C becomes unbound to 2p’s and 2a’s at 3.7 MeV of excitation. As only the first
excited state is below this threshold, the study of all other excited states requires the
detection of all four of these particles and the study of the correlations between them.
Doing so affords the possibility to determine the structure of these excited states that
have been described as two “valence” protons around an 8Be core. (Just as some excited states in 10Be are well described as valence neutrons around an 8Be core.)
In order to gather these data, we performed a secondary beam experiment in
which a 10C beam was generated by 10B +p -> 10C +n reactions. The 10C nuclei were then
directed to a secondary target of 9Be by a spectrometer system. The 2a + 2p products
were detected with a high resolution silicon-strip detector array. We verified two of the
three previously reported excited states in 10C above the 2p + 2a threshold and determined that most decay via 10C -> p + 9B -> 2p + 8Be -> 2p + 2a, through the ground
states of both 9B and 8Be.
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Toward a Better Understanding of...
Rate of Hydrolysis of
4-(p-Nitrophenoxy)-2-Butanone
by Amine Base
Dafang Zhang
Mentor: Dr. John-Stephen Taylor
Certain setbacks have plagued modern chemotherapeutic advances on the treatment
of cancers and viral infections. Chemotherapeutic drugs often rely upon the higher
reproductive rates of cancer cells over normal cells to target the disease. Such a mechanism affords slight selectivity and detrimentally affects normal body cells. Moreover,
a given cancer cell, through random mutation, may possibly be immune to a certain
chemotherapeutic drug. A more selective, universal approach to cancer treatment is
needed. We have recently proposed the new approach of a PNA-based RNA-triggered
prodrug activation system. In such a system, a cytotoxic drug is released only in those
circumstances where the target cancerous nucleic acid is attacked simultaneously by
two halves of complementary sequence, to which are affixed a prodrug and a substrate.
Formerly, we employed a coumarin ester as the substrate that made up the prodrug
component. For the catalytic component, we attached imidazole to PNA. Presently we
utilize 4-(p-nitrophenoxy)-2-butanone as the substrate. In order to attain higher rates
of substrate hydrolysis, we have tested said rates for a variety of amine bases. A phosphate buffer was used to maintain a pH 7 environment. A 30% methanol-70% distilled water solvent system was chosen because such a system dissolves the substrate
and is miscible in the phosphate buffer. The amine base that demonstrated the fastest
rate of hydrolysis was 2-dimethylaminoethylamine.
We have shown that the contribution of the phosphate buffer to substrate hydrolysis is negligible since it is exceedingly slow. However, at pH 7.00, substrate hydrolysis
by hydroxide is more than 3610-fold faster than that by 2-dimethylaminoethyamine.
Nevertheless, since we have previously demonstrated that the hydrolysis of a coumarin
ester by free imidazole is accelerated more than 60000-fold when put under a PNAbased RNA-triggered drug-releasing system, there is reason to expect that 2-dimethylaminoethylamine may prove to be a feasible catalyst in a biological system.
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Toward a Better Understanding of...
Guatemalan Border Control and Implications
for the Trafficking of Persons
Matthew T. Miller
Mentor: Dr. Jim Wertsch
The issue of border control is very important in the study of human trafficking
because trafficking victims often find themselves trafficked across international borders by their modern-day slave traders. This trafficking occurs by a variety of means
and in innumerable locations throughout the world. This summer, my research
sought to examine the border regions of Guatemala—particularly, official border
crossing points—in order to understand the dynamics at borders and in order to
determine the level of border control that exists in these vitally important, but often
neglected, loci of trafficking of persons.
The study that I undertook this summer relied primarily on observational data
generated by multiple expeditions to four different border crossing points in
Guatemala—El Florido, El Corinto, Tecún Umán, and Agua Caliente. In each trip,
detailed notes and extensive photographs were taken to document important findings.
Further, when possible, different types of transportation and scenarios were utilized in
order to more fully understand the dynamics at the border.
During this field study, significant problems at both the procedural and individual
level of Guatemalan border control were evident. Glaring examples of professional
misconduct (including, bribery) and extraordinarily lax oversight were observed, in
addition to larger procedural inadequacies that all but guarantee that a person could
cross the border without ever being examined. Definitive conclusions based upon this
cursory study of the border crossing points of Guatemala would be premature. This
field study was limited in numerous ways—by time, resources, number of observed
sites, etc.—and, therefore, its significance to the study of human trafficking and the
study of borders may be that it raises important questions and, hopefully, spurs future,
far more comprehensive direct research of borders and border crossings.
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Toward a Better Understanding of...
Mapping the MIA1 Gene in
Chlamydomonas Reinhardtii
Michelle L. Miller
Mentor: Dr. Susan Dutcher
This research was supported by a Washington University/Howard Hughes
Medical Institute Summer Undergraduate Research Fellowship Award.
Chlamydomonas reinhardtii is a unicellular biflagellate green alga that has the ability
to phototax, or respond to light stimulus, both through environmental factors and
genetic expression. It is important in medical research, particularly for studying cellular motility, as many cells in the human body have cilia. Mutations affecting motile
cilia can cause primary cilia dyskinesia, sterility, increased respiratory infections, and
reversal of asymmetrical organs, such as the heart and stomach. These patients often
have defects in the dynein arms of their cilia.
A screen for phototactic mutants conducted by King and Dutcher (1997) identified a strain with a modified inner dynein arm (mia1) in its flagella, which caused it
to swim slowly, and also found that one of the subunits of the I1 inner dynein arm
required for responding to directional light showed hyperphosphorylation. To continue studying the gene, its precise location needs to be found. It was known that MIA1
was linked to another gene, ACT1, which was used as a phenotypic marker. The mia1
mutants were then crossed with the polymorphic lab strain, S1D2, so that there would
be polymorphisms to score where recombination events might have occurred.
Progeny exhibiting the mia1 phenotype were scored with the approximately 25 CAPS
markers designed around the gene. Recombinants flanking mia1 were identified, and
the region has been narrowed from one linkage group to one 3 Mb contig to just 70
kb, which contains eleven possible gene candidates and is covered by two BACs.
Transformations are currently being performed to see if the mutants can be rescued with one of the BACs. The goal is to identify a single gene and to sequence in
order to look for mutations. Then it can be further studied to aid in understanding the
role of MIA1 in the control of the phosphorylation of the inner dynein arm and the
processes of gene regulation, flagellar motility, and phototaxis.
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Toward a Better Understanding of...
Public Policy and Poverty in
East St. Louis, Illinois
Elizabeth Williams
Mentor: Dr. Leslie Brown
This research was conducted as a part of the Washington University
Mellon Mays Undergraduate Research Fellowship Program.
During the summer of 2006, I worked to (1) locate and understand working definitions
for the term poverty and the phrase public policy; (2) familiarize myself with the history of East St. Louis from 1920-1990; (3) place that history in the context of United
States history during that period (with an emphasis on African-American history); and
(4) begin the process of finding and understanding the relationship between poverty
and public policy. In defining poverty and public policy, I found that by simply comparing these definitions in their broadest sense, a relationship begins to emerge which
provides the foundation for the building of a relationship, more specific in nature,
between poverty in East St. Louis and public policy.
Poverty may be briefly defined as the condition of lacking predictable and steady
means of meeting basic life needs especially in the face of limited options for securing
those means. Public policy is policy that is made in the name of the public, initiated by
government, and is both what the government intends to do and what the government
chooses not to do. Since all actions of government affect the public, public policy is in
short whatever government chooses to do or not to do. When there is poverty, insufficient means for meeting basic life needs, it is either a result of absolute lack of choice
or limited options. Slavery is an example of poverty resulting from lack of choice. When
looking at poverty that results from limited options, specifically citywide poverty, government becomes accountable for passing or not having passed laws that have allowed
for a community’s decline into poverty. Alongside personal choice, three of the greatest causes of poverty are 1) lack of employment opportunities, 2) lack of education, and
3) state discrimination. In studying the history of African Americans in the United
States, all three of these causes become highly visible. This is no less true when studying the history of specific black communities within the larger community. In East St.
Louis, for decades citizens have been denied job opportunities, quality education, and
access to fair housing and transportation practices.
At this point, I am in the process of locating and analyzing instances of state discrimination with regard to East St. Louis. I am also still working on gaining knowledge
of African- American history and a familiarity with the various policies that have had
the most impact upon them. While I began my project not wanting to focus mainly on
race, research has shown that because of various historical occurrences, in East St. Louis
race and poverty have long gone hand in hand. I would like to continue analyzing the
ways in which the causes of poverty, which have been visible on a national level, have
affected East St. Louis, Illinois.
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The Body Strikes Back:
Media Theory and AIDS Activism
Ian Schatzberg
Mentor: Dr. Jami Ake
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
The influx of media imagery that resulted from the proliferation of home video produced a great degree of hesitation within media critics. Individuals such as Jean
Baudrillard and Fredric Jameson paused and asked, where had the “real” gone in a
society dominated by generated imagery? Although contemporary discussions within
media and cultural studies may have developed beyond these initial concerns, such
criticisms have not lost their theoretical strength. Examining these respective theorist’s
bodies of work in terms of their rhetoric helps us engage in a type of critical thinking
that occurred at the onset of the nineteen-eighties. Such uses of rhetoric function as
an undeveloped site from which we may begin to understand the psychic implications
of living in a mass-media society.
Between 1983 and 1984, we witness within media theory a conception of the body
that evacuates its materiality. Such a discourse envisions the body as a vessel that
serves simply as a receptacle for spectacular imagery. As Guy Debord articulates, the
rate of image distribution in postmodern society has the effect of mediating our relationship with surrounding individuals and ourselves. Existing beside this conversation
in the streets of New York City and throughout the nation, a medical phenomenon
silently presents itself as a force to be reckoned with: the growing number of AIDS
cases. My research suggests that the presence of AIDS, and the tactics activists used to
combat its spread between 1986 and 1988, effectively challenge and make problematic our understanding of the postmodern and its influx of media imagery in the early
1980s. AIDS media activism foregrounds the body in a way that reengages its corporeality within spectacular media.
While theorists envisioned an erasure of the body, one determined by the markings of race, class and gender, AIDS video activists utilize their media in order to bring
marked bodies into the public sphere. We witness queer subjects engaged in a contestation of their visual portrayals and we encounter the tension that arises when marked
bodies express themselves within a medium capable of unmarking them.
Consequently, the impetus behind my research lies in examining issues of representation and identifying who marked individuals, specifically queer subjects, re-inscribe
the body into the mass media landscape.
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Binding Characteristics of the
PAT Protein Family - Perilipin,
ADRP TIP47, S3-12, and OXPAT
Michelle Mo
Mentor: Dr. Perry Bickel
This research was supported by the Washington University Office of
Undergraduate Research through a Hoopes Undergraduate Research Award.
Little is known about the way proteins bind onto lipid droplets and transport them
throughout cells. By creating pieces of proteins from two specific lipid binding regions
– known as the 11mer repeats and the four helix bundle – of the PAT (PerilipinAdipophilin-TIP47) protein family, more can be understood about how lipids are
bound. The PAT protein family consists of Perilipin, ADRP (adipophilin or Adipose
Differentiation Related Protein), TIP47 (Tail Interacting Protein of 47 Daltons), S312, and OXPAT. Each PAT protein shares many similarities with others in the family
and some differences as well. Based on their differences they are divided into two categories, CPATS (constitutively TAG-associated PAT proteins) and EPATs (exchangeable TAG-associated PAT proteins). The four helix bundle region and the 11mer
repeat region are hypothesized to act differently for different proteins, and consequently contain different lipid binding abilities.
In this work, ten novel DNA constructs were created and expressed in cells so that
the lipid binding characteristics of the protein they encode could be observed under
immunofluorescence microscopy. It is hypothesized that the 11mer repeats and the
four-helix bundle regions are crucial areas for binding to lipid. Without them, proteins
most likely do not know when or where to bind to lipid droplets, thus causing problems within cells. It is highly possible then, that certain metabolic disorders may be
caused by lower or higher amounts of various PAT proteins or defects in certain PAT
proteins. If a cell fails to effectively store intracellular lipid, insulin production is either
greatly reduced or does not occur at all. Roger Unger’s theory of lipid overflow
addresses the condition in which excess lipid no longer can be stored or partitioned in
adipocyte, and ‘overflows’ into muscle tissues and organs. This condition can lead to
lipid insensitivity and other disorders. Such an understanding is potentially significant, considering that failure to effectively partition lipids into storage is implicated in
cardiomyopathy and Type II Diabetes.
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Toward a Better Understanding of...
Reading Islam in the Italian Renaissance:
Representations of Islam and the East in
16th Century Travel Narrative as seen in the
Itinerary of Ludovico di Varthema
Jessica Pryde
Mentor: Dr. Michael Sherberg
This research was conducted as a part of the Washington University Undergraduate Honors Fellowship Program.
The end of the fifteenth century brought about a new desire to explore lands unknown
to Western Europeans. Columbus, de Gama, and numerous other explorers and
adventurers sought many ways to reach “India” and later “the Indies” and opened a
new world to many in the process. Once travelers, settlers, and adventurers began to
direct their attention to this new world, exploration of the exotic, mysterious old
world did not cease. Not ten years after Columbus had left on his first trip to the West
Indies in 1492, a Bolognese man, Ludovico di Varthema, set about in the opposite
direction, returning six years later after having become the first Christian known to
have entered the holy Islamic cities of Mecca and Medina and claiming to have past
the Malaysian Peninsula and Borneo before returning to Italy. When reading his travel narrative, the Itinerary, many places are familiar, yet exotic, and Varthema, always
present in the happenings story, shows himself to be superior to those around him
time and again. He manages to awe readers with vignettes of his fantastic adventures
while maintaining the portrait of the mundane aspects in the lives of Arabs, Persians,
and Indians. This research seeks to answer the following questions: How does
Varthema represent Islam and the Eastern people? How does this coincide with the
mindset and occurrences of his fellow Continental Europeans? How does Varthema’s
narrative differ from other works of the time?
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Exploring the Meanings of the Huipil
Erendira Quintana Morales
Mentor: Dr. David Browman
This research was conducted as a part of the Washington University
Mellon Mays Undergraduate Research Fellowship Program.
In academic literature, the huipil is often described as a square-shaped blouse made of
one to three panels of woven cloth whose length can vary from the ankles to the feet.
Visual examples show that the cloth is typically white and decorated along the collar
and the sleeve openings. In the past century, the huipil has undergone many changes
in its colors and designs that challenge us to think about what makes a huipil different from other clothing.
A closer look at how this garment is made illustrates two principal styles of decoration: brocading and embroidery. As a result of this distinction, I believe that the use
of each technique leads to a particular design type that is influenced by commercialization in a different way. Because much of the planning of the designs and colors
takes place during the preparation for weaving, in many instances the weaver is aware
of the intended use for the garment she is creating before she begins to weave.
Knowing that the garment is intended for sale to a tourist allows her to emphasize different aspects of the huipil through the designs and colors she chooses.
The ways in which this garment is created and used shows that there is an important relationship between women and the huipil. Weaving is one of the activities used
to differentiate between genders since it is an activity that even in mythology is associated with the feminine role. Women use the huipil in many ways; for example, to
wear during religious ceremonies or for daily use, to provide clothing for their families, to express their personal and community identity in public, to establish ties with
other women and across generations (particularly in the learning process) and to
obtain income in the tourist market.
In understanding the transformation of patterns in the huipil, I see the Maya
weavers as active agents of change in their culture. Through the sale of the huipil to
tourists, Maya women are connected to an international market and are thus affected
by the changing trends and tastes of their clients and the images of Maya culture
around the world. Their participation in this international market influences their roles
within their communities. In my research, I hope to understand some of these changes
and to portray Maya people as dynamic members of the international community.
103
Wahington University
Spring 07 Peer Review Board
NATURAL SCIENCES COMMITTEE
Cassie Sadinski, junior
Christopher Rinaldi, sophomore
David Kurland, junior
Sarah Frazier, sophomore
Amanda Adeleye, junior
Srikar Rao, senior
SOCIAL SCIENCE/HUMANITIES COMMITTEE
Andrew Schupanitz, senior
Julie Kling, senior
Olga Lozovskaya, freshman
Amelia Clark, senior
Morgan Grossman-McKee, freshman
Kelly MacArthur, senior
Nick Niles, senior
104
Humanities Research
Review Committee
Faculty Oversight Committee
For Undergraduate Research
Barbara Baumgartner
Lecturer in Women’s Studies
Joseph Ackerman
William Greenleaf Eliot Professor of Chemistry
Eric Brown
Associate Professor of Philosophy
Steven Fazzari
Professor of Economics
Susan Rotroff
Jarvis Thurston & Mona Van Duyn Professor
Wayne D. Fields
Lynne Cooper Harvey Distinguished Professor
in English
William J. Whitaker
Senior Artist in Residence in Drama
Natural Science Research
Review Committee
Beata Grant
Professor of Chinese Language and
Religious Studies
Jonathan M. Chase
Associate Professor of Biology
Hillel J. Kieval
Gloria M. Goldstein Professor of Jewish History
and Thought
T. Joseph Kappock
Assistant Professor of Chemistry
Robert Lamberton
Professor of Classics
Kathryn Miller
Professor of Biology
Maria Lander
Assistant Professor of Romance Languages
and Literature
Social Science Research
Review Committee
Joseph Loewenstein
Professor of English
David L. Browman
Professor of Anthropology
Linda Nicholson
Stiritz Distinguished Professor of Women’s Studies
Robert L. Canfield
Professor of Anthropology
Itai Sened
Professor of Political Science
Peter J. Kastor
Assistant Professor of History
Richard Jay Smith
Ralph E. Morrow Distinguished University Professor
Mark A. McDaniel
Professor of Psychology
Gary J. Miller
Professor of Political Science
105
OFFICE OF UNDERGRADUATE RESEARCH
Umrath 100
Campus Box 1026
Phone: (314) 935-7342
Fax: (314) 935-4384
E-mail: [email protected]
Web: http://ur.wustl.edu