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Sample Applications
UNIVERSITY OF CINCINNATI COLLEGE
OF MEDICINE
MEDICAL STUDENT SUMMER RESEARCH FELLOWSHIP APPLICATION
Application Receipt Deadline is Friday, March ##, 2###
I. STUDENT PORTION OF APPLICATION:
Name: Sample
Application I
Social Security #:
Address: |
Academic Year:
Birth Date:
Phone #:
Email Address: |
Education (begin with baccalaureate):
| Institution and Location |
Degree & Year |
Scientific Field |
|
UC Berkeley, Berkeley, CA
MCP-Hahnemann U, Philadelphia, PA
UCCOM, Cincinnati, OH
|
BA, 1995
MMS, 1999
MD, 2003 |
Molecular & Cell Biology
Medical Sciences
Medicine |
Honors:
Outstanding Volunteer Award, Summit Medical
Center, Oakland CA
Honorable Mention, Colgate Essay Contest
Certificate of Recognition, People's Emergency Center, Philadelphia,
PA
Major Career Interests:
Academic medicine in Neurology or Internal
Medicine
Have you received a previous University of Cincinnati Summer Research
Fellowship? NO
If YES, indicate date.
Describe any research and/or professional experience,
including publications.
(Use only this page.)
6/98-7/99 MCP<>Hahnemann
University, Philadelphia, PA Center for NeuroVirology and
NeuroOncology, Laboratory for Molecular NeuroVirology, GRADUATE
STUDENT , PI: Kamel Khalili
Working under the direction of principal investigator and postdoctoral
fellow: participating in the studies of the correlation between
HIV and Progressive Multifocal Leukoencephalopathy through its etiological
agent JC virus. Projects: 1) Study the effect of HIV
protein Tat and Vpr on JC virus at the level of DNA replication
by method of Dpn 1 assay; at the transcriptional level by examining
the HIV proteins' effect on JCV late promoter via luciferase activity
assay; and at the translational level by western hybridization technique.
2) Investigate the effect of pur alpha on the cell cycle of neoplastic
cells (astrocytoma) via FACS analysis. Results are compared
between extracellularly administer of pur alpha and stable transfection
of pur alpha-GFP construct. 3) collaborating with postdoctoral
fellow on the efforts to isolate GBP-i and its associated proteins
in PMA induced U87-MG cells by the method of Differential Display
and various screening techniques.
1/97-3/97 University of Southern California,
Los Angeles, CA Hoffman Medical Research Center
Department of Microbiology, LABORATORY TECHNICIAN, PI:
Lucio Comai
Working under the direction of principal investigator: participating
in the study of RNA polymerase I transcription associated factor
interactions. Project: elucidating the RNA Pol I transcription
associated factors expression differences in undifferentiated and
TPA induced differentiated U937 cells.
3/95-6/96 UCSF, San Francisco, CA, Department
of Biochemistry/ Biophysics, Genetics/Development lab, LABORATORY
ASSISTANT, PI: Didier Stainier
Working under the direction of principal investigator: participating
in zebrafish heart development project. Project: isolating and characterizing
zebrafish homologue gene, XEGR (angiotensin receptor II) and VEGF
(vascular endothelial growth factor) by methods of PCR and library
screening.
RESEARCH PROPOSAL: (Use three pages maximum, single-spaced.)
Using the following format, type in the shaded spaces.
The space will expand to fit your need
Title
Chromium Mesoporphyrin Induced Heme Oxgenase-1 Gene Expression as
a Protective Mechanism Against Stroke.
Hypothesis to be tested
Induction of the Heme Oxygenase-1 (HO-1) heat shock gene
with chromium mesoporphyrin (CRPP) will protect brain against
ischemia.
Specific Aims
1) Describe the induction of the HO-1 heme oxygenase heat
shock gene in brain following (a) intraperitoneal
and (b) intraventricular injection of different doses of chromium
mesoporphyrin .
2) Determine whether HO-1 induction by chromium mesoporphyrin
protects the brain against stroke.
Background & Significance
Cells have evolved a number of mechanisms to help them
survive in various hazardous conditions. Often an extreme
environmental insult activates a battery of genes whose products
protect the cell from its effect. One category of these
protective proteins, Heat-Shock proteins (HSPs), is induced,
in part, by denatured proteins produced during heat shock, ischemia,
low glucose level, and many other stresses. Overproduction
of HSP70, for instance, protects brain cells in vitro against
injuries that produce necrosis and some types of apoptosis.
Overproduction of HSP70 in vivo protects the brain against injury
produced by ischemia and prolonged seizure. Previous studies
have also found that viral transfection with a HSP-70 construct
in rat brain confers significant protection to the striatum
and hippocampus following a stroke produced by a one-hour middle-cerebral-artery
occlusion. These findings suggest the potential therapeutic
application of heat shock proteins in protecting brain against
variety of acute destructive or chronic degenerative processes.
Heme oxygenase 1, also known as HSP32, is synthesized mainly
in microglia and is one of several related heme oxygenase proteins
that metabolize heme to carbon monoxide, iron and biliverdin.
HO-1 is synthesized in response to heat shock, heme, oxidative
stress and ischemia. Furthermore, HO-1 also increases
its gene expression following cisterna magna injections of blood
or hemoglobin almost exclusively in microglia throughout brain.
Over production of HO-1 protects vessels against heme and hemoglobin-mediated
injury. In light of this observation, our study attempts
to investigate the efficacy of chromium mesoporphyrin, a heme
analog, on HO-1 induction and subsequent potential in brain
protection against ischemic injury.
The finding of Heme Oxygenase I protection on brain against
blood and hemoglobin-mediated is clinically significant in that
it elucidates a significant intrinsic physiological protective
mechanism against stresses. Should CRPP demonstrate inductive
effect on HO-1 gene and whose
products confer protective effect on brain cells, it would open
a new avenue for potential treatment modality in acute destructive
or chronic degenerative processes. Drugs designed specifically
for this purpose could be offered to patients with high risks
of stroke so that the deleterious effect of cerebral infarction
can be minimized following a stroke. Even the situation used
in this animal study could be used clinically, where patients
undergoing high risk procedures like CABG surgery could be pre-treated
to protect the brain, heart and other organs against ischemia.
Methods
Chromium mesoporphyrin will be injected intraperitoneally
into adult Sprague Dawley rats in doses ranging from 0.01mg/kg
to 100mg/kg. The drug will be injected into a second group
of ketamine (100mg/kg) and xylazine (20mg/kg) anesthetized
animals via the cisterna magna,
doses ranging from 0.1 micrograms to 20 micrograms dissolved
in 5 microliters of sterile saline.
One day after intraperitoneal or citernal injections, animals
are anesthetized and the brains removed as rapidly as possible.
The brains will be processed for (a) HO-1 mRNA using Northern
blots (b) HO-1 protein using Western blots (c) and HO-1 immunocytochemistry.
This part of the study will describe the dose response induction
of HO-1.
In the second part
of the study animals will be injected with the dose of chromium
mesoporphyrin that produces the maximal HO-1 induction, either
i.p. or icv. Control animals will be injected with the same
volumes of vehicle used for the drug. One day after the injections
animals are subjected to a stroke. This is produced by anesthetizing
animals, and inserting a suture into the internal carotid artery
and advancing it to the middle cerebral artery. The suture is
left in place for 2 hours to produce a middle cerebral artery
infarction. The suture is then removed and the wound cleaned
and sutured. Animals are allowed to survive for 24h at which
time they are re-anesthetized, and perfused with 4% paraformaldehyde.
The brains are removed, sectioned and stained.
Plans for Data Analysis and Interpretation
In the first part of the study we will describe the dose
response for HO-1 induction in brain by chromium mesoporphyrin.
This drug induces HO-1 by inhibiting the enzyme activity. We
will also determine which cells express HO-1. It is predicted
that i.p. drug will induce HO-1 in blood vessel endothelial
cells, since the drug may not cross the blood brain barrier
very well. Following injection into the cisterna magna, the
drug may induce HO-1 in cells that can transport porphyrins
into CNS cells ¡V which is unknown, but may include microglia
(brain macrophages) and neurons.
The second part
of the study will measure the areas of infarction using a computer
system in the lab through the entire infarct in each animal.
Knowing the areas of infarction and the distances between sections,
the volumes of infarction are calculated for each animal. It
is anticipated that if HO-1 induction protects the brain, that
the infarction volumes in the chromium mesoporphyrin treated
animals will be smaller than in the vehicle injected control
animals.
II. MENTOR PORTION OF APPLICATION
Research Sponsor Jack Qqqqq,
Dept of Science
GENERAL PLAN for student training program (To be completed
by mentor). Include information about the nature and frequency
of direct interaction with the student. Identify individuals
who will participate in student training, and briefly describe
conferences and lab meetings the student will attend.
The student will be involved
in an active stroke research program that is getting underway
in the Vontz. He will participate in a weekly basic science
journal club and research seminar. In addition, I will meet
with xxxxx once per week to review how the work is going, and
to go over some aspect of the science that is being studied.
He will present his ideas and work at least twice to the research
group over the course of the summer.
He will be helped in his studies by yyyyy
who is my technician who will help with the animal handling,
injections, brain removals, and histology and immunocytochemistry.
He will also be helped by Dr. zzzzzz who will teach him how
to do Northerns and Westerns, and who will do the strokes that
are a part of the study. A portion of his time will involve
learning how to do the surgeries required to do the strokes,
doing the brain sectioning, and the analysis required to determine
stroke sizes in individual groups of animals .The project is
intentionally meant to be self-limited so that he can
participate in all aspects of the study from examining protein
induction, to performing the stroke animal studies, to examine
the stroke infarct data. It is unlikely that he will able to
complete the entire study himself, but there will be sufficient
data by the end of the summer so that he would be able to state
in preliminary paper the doses of protoporphyrin required to
induce HO-1 in brain, and whether a given dose of drug would
protect. We have chosen this project with the specific goal
of making it clinically relevant, and making it possible to
participate in all portions of the study in the course of a
single summer.
If the proposed project will involve use of RADIOACTIVE ISOTOPES,
list:
Authorized User: Jack Qqqqq
Authorization Number: 99-S63-06-1
If the proposed project will involve the use of ANIMALS, list:
Title of Approved IACUC Protocol:**
Gene Regulation in Cerebral
Ischemia and Hemorrhage
angry
Principal Investigator: Jack Qqqqq
IACUC Protocol Number: 99-12-09-02
** (Must be currently approved or have evidence of approval
by June 1, 2000)
UNIVERSITY OF CINCINNATI COLLEGE
OF MEDICINE
MEDICAL STUDENT SUMMER RESEARCH FELLOWSHIP APPLICATION
Application Receipt Deadline is Friday, March ##, 2###
I. STUDENT PORTION OF APPLICATION:
Name: Sample
Application II
Academic Year: UCI
Social Security #:
Birth Date:
Address:
Phone #:
Email Address:
Education (begin with baccalaureate):
Institution and Location
Degree & Year
Scientific Field
University of Cincinnati Cincinnati, OH
BSN 1997
Nursing
Honors:
University of Cincinnati Presidential Scholarship Recipient
(1992)
Member of Sigma Theta Tau International Honor Society of Nursing
(1996 to present) University of Cincinnati College of
Nursing and Health Dean's List (1994 to1997)
Major Career Interests:
Surgery (unsure at this time of the exact subspecialty
I would pursue)
Have you received a previous University of Cincinnati Summer
Research Fellowship? No
If YES, indicate date.
Describe any research and/or professional experience,
including publications.
(Use only this page.)
I have worked in the surgical intensive care unit (SICU) at
the University Hospital since July 1997 as a registered nurse.
During that time, I have had the opportunity to collaborate
with multidisciplinary members of the health care team and learn
the intricacies of critical care of patients. Although
I have not been a formal member of the many research teams that
perform research in the SICU, I have aided many of the researchers
in the gathering of data, blood specimens, and consent.
This, however, will be my first experience with basic science
research, as well as my first opportunity for publication.
RESEARCH PROPOSAL: (Use three pages maximum, single-spaced.)
Using the following format, type in the shaded spaces.
The space will expand to fit your need
Title
Role of epinephrine in carbohydrate metabolism during subacute
sepsis
Hypothesis to be tested
Chronic propranolol infusion will normalize changes in carbohydrate
metabolism and Na, K pump activity seen in subacute sepsis.
Specific Aims
The aim of this study is to assess the impact of beta-receptor
blockade on the effects of subacute sepsis on plasma epinephrine
and norepinephrine, muscle glycogen and glucose-6-phosphate
(G6P), plasma glucose and lactate, muscle high-energy phosphates
(ATP and phosphocreatine-PCr) and muscle intracellular content
of Na and K ([Na]i and [K]i).
Background & Significance
Septic shock accounts for 100,000 deaths per year in the United
States and is the most common cause of death in intensive care
units (1). Sepsis results in a hypermetabolic state in
which many aspects of carbohydrate metabolism are abnormal,
including enhanced peripheral glucose uptake and utilization,
hyperlactacidemia, increased gluconeogenesis, depressed glycogen
synthesis, glucose intolerance and insulin resistance.
High blood lactate in critically ill septic patients is associated
with increased risk of organ failure and mortality (2).
Traditionally, stimulation of muscle glycolysis by epinephrine
(Epi) has been explained through increased activities of glycolytic
and glycogenolytic enzymes. However, recent studies from
this laboratory indicate that glycolysis and lactate production
increase in well-oxygenated muscle when Na-K pump activity is
stimulated (3,4). Thus, increased enzyme activities may
not be the primary cause of increased glycolysis but, rather,
the result of coordinated stimulation of the Na, K-ATPase and
of ATP-supplying glycolytic pathways. Although high circulating
Epi is associated with sepsis and its models, the major metabolic
changes in sepsis usually are not interpreted as effects of
Epi. Epi's role in the metabolic derangements of sepsis
needs to be reevaluated, as it may account for pathophysiologic
glycolysis through normal, but previously unknown, metabolic
mechanisms.
As part of the "fight-or-flight" response, Epi has both
metabolic and physiologic effects that increase fuel availability.
The effects can be seen as contributing to muscle's ability
to sustain contractile activity. By a cyclic AMP-dependent
mechanism, Epi raises glycogen phosphorylase activity, increasing
availability of G6P. Epi also stimulates the Na, K-ATPase,
thereby reducing extracellular accumulation of K and maintaining
low intracellular Na during muscle activity (5). Stimulation
of glycolysis by Epi is largely blocked when the Na, K-ATPase
is inhibited, which strongly suggests that glycolytically-produced
ATP is preferentially used by the Na-K pump (4). These
new experimental results thus suggest that stimulation of glycogenolysis
and glycolysis by Epi is coordinated with stimulation of membrane
ion transport, and is not a generalized, non-specific increase
in glucose metabolism.
Knowledge of the causes of abnormal metabolism in sepsis
is a prerequisite for devising appropriate therapies.
Results from this laboratory have uncovered novel aspects of
skeletal muscle metabolism and function, i.e., stimulation of
glycolysis coupled to Na, K-ATPase activity. In septic
patients, persistently high Epi secretion would be expected
to cause increased Na-K pumping, glycogenolysis and glycolysis
that served no physiologic function related to muscle contractility.
To the contrary, the inappropriate hypermetabolism would become
pathologic. Changes in sepsis to carbohydrate metabolism
(depressed glycogen synthesis, glucose intolerance and insulin
resistance) are adaptive physiologic responses to exercise and
may be attributed to Epi's effects on skeletal muscle (6).
Therefore, blockade of adrenergic beta-receptors in sepsis would
be expected to counteract many of these pathologic effects.
Methods
The aim of this study is to assess the impact of beta-receptor
blockade on the effects of sepsis on (a) plasma Epi and norepinephrine,
(b) muscle glycogen and G6P, (c) plasma glucose and lactate,
(d) muscle ATP and PCr and (e) muscle [Na]i and [K]i.
We will study the extensor digitorum longus and soleus muscles,
which exemplify primarily white, fast-twitch and red, slow-twitch
fiber-types, respectively. Four groups (n=6-10) will be
used: (A) Septic-control, (B) Septic-propranolol, (C) Non-septic-control
and (D) Non-septic-propranolol.
Male Sprague-Dawley rats (70-90 g) will be used.
With rats under anesthesia [Na pento-barbital (50 mg/kg) and
atropine sulfate (0.04 mg/kg)], sepsis is induced by subcutaneous
injection of Escherichia coli. This model of non-fatal
sepsis has been extensively used by Lang et al (7) for studies
of whole body glucose metabolism. A PE50 catheter is passed
down a subcutaneous tunnel on the back for subsequent administration
of bacteria. This catheter is exteriorized on the back
of the neck and secured with tape. A suspension of E.
coli containing 2-5 x 10^10 CFU is injected through the catheter
twice on the day of surgery and once more on the next day.
Control animals will undergo all surgical procedures and anesthesia,
but receive saline instead of E. coli. This model results
in a reproducible, hypermetabolic, sub-acute sepsis, with increased
body temperature, elevated heart rate and altered whole-body
glucose metabolism [increased circulating lactate levels and
elevated glucose rate of appearance (Ra)] (7).
For beta-adrenergic blockade, Alzet osmotic minipumps
will be implanted in the abdomen to achieve sustained delivery
of propranolol. Pure L-propranolol is freely soluble in
water and, at a pump delivery rate of 0.024 ml/day, a dose of
15 mg/kg x day in a 100 g rat would be achieved by loading the
pump with propranolol at a concentration of 62.5 mg/ml.
Control animals will have pumps containing only normal saline.
Pumps will be implanted the day before sepsis to allow several
hours for the delivery of propranolol before the septic insult.
At 24 and 48 hours after induction of sepsis, hindlimb muscles
will be dissected with intact tendons from rats anesthetized
with sodium pentobarbital (50 mg/kg). For all procedures
except measurement of intracellular Na and K, muscles will be
rapidly frozen in liquid nitrogen.
Measurements of plasma Epi and norepinephrine, lactate
and glucose, muscle glycogen, G6P, lactate, ATP and PCr, and
Na and K will be made. Plasma norepinephrine and Epi levels
are determined by high performance liquid chromatography (HPLC)
with electrochemical detection (8). Blood for catecholamine
analysis is drawn from pentobarbital anesthetized animals.
EGTA and reduced glutathione are added to the blood to prevent
auto-oxidation of the catechols. For plasma lactate and
glucose, fifty microliters of plasma will be deproteinized in
an equal volume of 0.4 N perchloric acid. After centrifugation
of the precipitated proteins, the supernate will be diluted
1:10 with distilled water before assay in triplicate.
Glycogen analysis is via amyloglucosidase digestion (9),
followed by fluorometric deter-mination of glucose. G6P
will be measured using a minor modification of the fluorometric
glycogen/glucose assay. Muscle lactate is assayed by a
standard microfluorometric enzymatic procedure (9) involving
reduction of NAD by lactate dehydrogenase (LDH) to produce NADH,
which is detected fluorometrically (excitation: 360 nm, emission:
530 nm) using a microplate reader (Bio-Tek). ATP and PCr
are assayed using an HPLC procedure (10). For the measurement
of intracellular Na and K, washout of ions from the muscle extracellular
space is accomplished immediately after dissection by a series
of four 15-min incubations at 0°C in ~3 ml of Na- and K-free
buffer containing (in mM) 263 sucrose and 10 tris (hydroxymethyl)
aminomethane, pH 7.4 (4). Thereafter, muscles are homogenized
in 5% trichloroacetic acid and ion content of the supernate
will be measured using an atomic absorption spectrometer (Shimadzu).
Plans for Data Analysis and Interpretation
A general purpose statistical package, Stata (Stata Corp., College
Station, TX) will be used for initital data analysis. Stata
combines speed, a comprehensive set of common routines [t-tests,
simple and multiple regression, ANOVA (with or without repeated-measures),
and follow-up comparison routines] with data management.
Final statistical analysis will be performed using the program
SAS.
1. Parrillo JE, Parker MM, Natanson C, et al.: Septic
shock in humans. Advances in the understanding of pathogenesis,
cardiovascular dysfunction, and therapy. Ann Intern Med 113:227-242,
1990.
2. Kirschenbaum LA, Astiz ME, Rackow EC: Interpretation
of blood lactate concentrations in patients with sepsis [Commentary]
. Lancet 352:922-923, 1998.
3. James JH, Fang CH, Schrantz SJ, et al.: Linkage of
aerobic glycolysis to sodium-potassium transport in rat skeletal
muscle. Implications for increased muscle lactate production
in sepsis. J Clin Invest 98:2388-2397, 1996.
4. James JH, Wagner KR, King JK, et al: Stimulation of
both aerobic glycolysis and Na+-K+-ATPase activity in skeletal
muscle by epinephrine or amylin. Am J Physiol E277:176-186,
1999.
5. Clausen T, Nielsen OB, Harrison AP, et al.: The Na
+,K+ pump and muscle excitability. Acta Physiol Scand 162:183-190,
1998.
6. Chiasson JL, Shikama H, Chu DT, et al.: Inhibitory
effect of epinephrine on insulin-stimulated glucose uptake by
rat skeletal muscle. J Clin Invest 68:706-713, 1981.
7. Hargrove DM, Bagby GJ, Lang CH, et al.: Adrenergic
blockade does not abolish elevated glucose turnover during bacterial
infection. Am J Physiol 254:E16-22, 1988.
8. Luchette FA, Robinson BRH, Friend LA, et al.: Adrenergic
antagonists reduce lactic acidosis in response to hemorrhagic
shock. J Trauma 46:873-880, 1999.
9. Lowry OH and Passonneau JV: A collection of metabolite
assays. 199-201, 1972.
10. Wiseman RW, Moerland TS, Chase PB, et al.: High-performance
liquid chromatographic assays for free and phosphorylated derivatives
of the creatine analogues beta- guanidopropionic acid and 1-carboxy-methyl-2-iminoimidazolidine
(cyclocreatine). Anal Biochem 204:383-389, 1992.
II. MENTOR PORTION OF APPLICATION
Research Sponsor Dr.
John Qqqqq, Dept. Surgery
GENERAL PLAN for student training program (To be completed
by mentor). Include information about the nature and frequency
of direct interaction with the student. Identify individuals
who will participate in student training, and briefly describe
conferences and lab meetings the student will attend.
Dr. Qqqqq will work with
the student on a daily basis. Ms. xxxxx will be trained
in all the methods to be used here (animal handling, dissection,
experimental set-up and performance, sample preparation, lactate
and glycogen assays, Na and K analyses) and will personally
perform each of them at least once. These procedures are
all currently in routine use in this laboratory. The group
works in close association on several projects simultaneously
and training will be by experienced personnel with additional
guidance from Dr. qqqqq, as needed.
In addition, as results
are obtained, Ms. xxxxx will be trained in data analysis, graphics
and statistical procedures required for presentation of her
work. She will obtain practice in presenting her results
and defending/explaining them to the group at regular small
meetings with Dr. yyyyy and Dr. qqqqq as well as in the bimonthly
lab meetings of Dr. zzzzz's group. The latter is attended
by Dr. zzzzz, typically involves 10-14 people and requires a
formal presentation with visual materials (slides or overheads).
It is anticipated that the results will be incorporated in a
paper or other communication from the laboratory in which Ms.
xxxxx will assist in preparation. Assistance will be provided
in the preparation of slides and posters for Ms. xxxxx's presentations
to the Medical Student Research Fellowship group.
If the proposed project will involve use of RADIOACTIVE
ISOTOPES, list:
Authorized User:
Authorization Number:
If the proposed project will involve the use of ANIMALS,
list:
Title of Approved IACUC Protocol:**
Abdominal Sepsis and
Metabolism (latest approval date: 10-26-99)
Principal Investigator: Qqqqqqq
IACUC Protocol Number: 89-09-05-02
** (Must be currently approved or have evidence of approval
by June 1, 2000)
UNIVERSITY
OF CINCINNATI COLLEGE OF MEDICINE
MEDICAL STUDENT SUMMER RESEARCH FELLOWSHIP APPLICATION
Application Receipt Deadline is Friday, March ##, 2###
I. STUDENT PORTION OF APPLICATION:
Name: Sample
Application III
Academic Year: UCI
Social Security #:
Birth Date:
Address:
Phone #:
Email Address:
Education (begin with baccalaureate):
Institution and Location
Degree & Year
Scientific Field
University of Illinois -Urbana Champaign
B.S., 1999
Biochemistry
University of Illinois -Urbana Champaign
B.S., 1999
Biology Honors
Honors:
Cum Laude, Biochemistry and Honors Biology
Chancellor's Scholar at the University of Illinois
Edmund W. James Scholar at the University of Illinois
Golden Key National Honor Society
Phi Eta Sigma Honor Society
Red Cross Youth Recognition Award
Major Career Interests:
Clinical Pediatrics with a focus in research
This summer will provide me a foundation in clinical research
techniques and experimental design which should be useful throughout
my career. It should also provide me an opportunity to
work with top clinical investigators from in an interdisciplinary
study and to directly coordinate a research study.
Have you received a previous University of Cincinnati Summer
Research Fellowship? NO
If YES, indicate date.
Describe any research and/or professional experience,
including publications.
(Use only this page.)
June 1999-August 1999
Science Policy Internship at the National Institute for Child
Health and Development at the National Institutes of Health
-Conducted a longitudinal resource evaluation of NICHD directed
research and centers programs under Mona Rowe at the Office
of Science Policy. Results were formally presented to
Dr. Duane Alexander, the NICHD Institute Director.
June 1997-August 1997
Pathology Summer Internship at the University of Cincinnati
College of Medicine
-Studied the mechanisms of antiatherogenic surfactants using
chemical and physiological models under Professor Patrick Tso
using peroxidation assays.
June 1996-May 1997
Protein engineering laboratory at the University of Illinois
-Engineering a Cytochrome P450 functional mimic from Cytochrome
c Peroxidase using site directed mutagenesis under Professor
Yi Lu
June 1997-August 1999
Performed gel electrophoresis, PCR, primer design, protein expression,
protein purification, ligand titrations, and UV-Vis Spectroscopy
June 1995-August 1995
Howard Hughes Biology Summer Internship at the University of
Cincinnati
-Studied the serotonin-induced spawning of Ohio River
Zebra mussels under Professor Daniel Gist.
-Performed fieldwork along the Ohio River, made slides
using microtones designed serotonin delivery protocols, and
performed microscopic analyses of gametes.
June 1994 -August 1994
Medical Summer Intern at the Children's Hospital Medical Center
in Cincinnati
-Researched the respiratory condition Stridor under Dr.
Frederick James in Cardiology.
-Compiled and analyzed data from patient reports, observed
subjects, and conducted library research. Proficient use
of spreadsheets.
RESEARCH PROPOSAL: (Use three pages maximum, single-spaced.)
Using the following format, type in the shaded spaces.
The space will expand to fit your need
Title
Prevalence of reduced bone mass is non-ambulatory institutionalized
children and adults
Hypothesis to be tested
Children and adults who are non-ambulatory will have significantly
lower bone mineral density/mass of selected bones and higher
frequency of fracture than sex and age matched controls.
Specific Aims
The specific aim of this proposal is to evaluate the baseline
bone mineral mass/density and plain film x-ray findings to assess
the frequency of osteopenia in institutionalized non-ambulatory
patients.
Once data is available regarding the frequency of reduced bone
mass, a companion protocol will be developed to determine whether
use of a bisphosphonate, Aledronate sodium, will increase bone
mineral density and potentially prevent future fractures.
Background & Significance
Cerebral Palsy (CP) is a condition caused by damage to the brain,
usually occurring before, during or shortly after birth, and
is characterized by an inability to fully control motor function.
CP affects 112,000 children, and is prevalent among 0.2% of
the entire population. Within the combined child and adult
CP population, it is estimated that 1/3, or 150,000 individuals
are severely affected and non-ambulatory, meaning immobile.
Though few of these individuals exhibit primary diseases of
the bone, immobilization predisposes bone resorption.
This may result in bone mineral density reduction and increased
susceptibility to bone fractures. Non-ambulatory CP patients
are particularly susceptible among the pediatric population
due to their relatively long lifespans as many survive well
into adulthood. Though this study is not limited to CP
patients, their condition is representative of other neural
and neuromuscular diseases which immobilize young people and
consequently subject them to conditions favoring increased fractures.
Few studies have investigated the quantitative relationship
between the frequency of fractures in the nonambulatory young
population and their associated bone mineral density, and none
have systematically evaluated potential interventions that might
result in bone accretion and decrease the risk of fractures
and its associated morbidity, mortality, and medical costs.
It is estimated that 10% of the patients at one of the institutions
where subjects will be recruited have sustained fractures during
routine care in the last year. If this is representative
to the population as a whole, as many as 15,000 children and
adults within this non-ambulatory population will have non-traumatic
fractures at home or chronic care facilities in the US during
the next year. A minimum cost for fractures per individual
in this population may be as high as $5000-10,000; a total figure
in the U.S. alone might be as high as $75-150 million/year.
The long term goal for this study is to identify means to prevent
fractures in this population which would provide savings for
the care of these patients as well as reduce their discomfort
and risk of other medical complications.
Using multivariate analysis, the best predictor of bone
mineral density was determined to be ambulatory status followed
by nutritional status.(1) As reported by Wilmhurst et
al, children with CP presented with increasing lumbar spine
bone mineral density with increased mobility.(2) Lee et
al studied 50 patients over two years who had severe handicaps
with radiological evidence of osteopenia ranging in age of 1-22,
and reported that during the study period 28% of the patients
sustained 29 fractures, mainly in the long bones of the lower
extremities.(3) Anticonvulsant usage was inversely related
to bone mineral density at both the femur and the lumbar spine.
Though nutritional status correlated with bone density, certain
studies reported that no relationships were observed between
bone density and vitamin D metabolite levels, osteocalcin, Ca,
P, or alkaline phosphatase levels.
Methods
Study subjects will be recruited from three area nursing homes
(Brookside, Camelot Lake, Fairfield Center) which provide care
for children and adults with neuromuscular disease beginning
in childhood. Subjects who are institutionalized and non-ambulatory
between 6-40 years of age will be considered eligible for participation
in this study (n=50). Subjects will be excluded if they
have intercurrent chronic medical conditions that may affect
bone metabolism independent of the effects of immobilization
and thereby confound the results of the study. Patients
must be in sufficient good health to tolerate the moderate amount
of blood drawing as well as travel.
All subjects qualified for this study will travel to the Clinical
Research Center at the Children's Hospital Medical Center for
bone density measurements (lumbar spine, total body, and hip).
Bone density will be measured by dual energy X-ray absorptionometry
using the Hologic Dual Energy X-Ray absorptiometer (DEXA, Hologic
QDR-2000, Waltham, MA). All study subjects will have plain
X-rays taken to examine for the presence of old and new fractures.
The view will include a single view of the long bones (femurs,
humerii, radia, ulnai, tibiae, fibulae), and a view of the pelvis
and the chest. Fractures [defined as the radiologic evidence
of a break in the cortex or presence of callus] will be counted
for each patient. In cases where a fracture if found or
suspected, another view may be taken to confirm the finding.
A bone age will also be assessed in subjects less than age 20.
Measurements of Ca, P, 25 OH vitamin D, and 1,25 (OH)D2 vitamin
D and four biochemical markers of bone resorption or turnover
will be used as well. Serum osteocalcin and the bone fraction
of alkaline phosphatase will be used as turnover markers and
free deoxypridinoline and N-telopeptide crosslinks will be used
as resorption markers.
Osteocalcin will be measured by radioimmunoassay using a kit
from INCSTAR (Stillwater, MN) which has a sensitivity of 0.2
ng/ml. The RIA employs simultaneous addition of sample
from serum, rabbit anti-bovine antibody and [125I] bovine osteocalcin
followed by an overnight incubation at 2-8 degrees C.
Phase separation is accomplished by the addition of a complex
of goat antirabbit serum, carrier rabbit serum, and polyethylene
glycol.
Bone
Alkaline Phosphatase will be measured by an ELISA (Alkaphase-B-Metra
Biosystems) which has low crossreactivity with liver isoforms
(5%) and activity with intestinal and placental forms.
Specimens should be taken from nonhemolyzed blood. The
captured BAP will also be assayed for activity by using a pNPP
substrate. The detection limit is 0.7 U/L. BAP levels
can be influenced by estrogen status, calcium intake, or immobilization.
Deoxypridinoline,
found in collagen type I and isolated in this study from urine,
will be assayed by ELISA using a kit provided also provided
by Metra Biosystems. Values will be corrected for urinary
dilution by urinary creatinine analysis.
Crosslinked
N-telopeptides of Type I collagen, also will be assayed by ELISA
using kit provided by Osteomark. Assay values are also
corrected for urinary dilution by urinary creatinine analysis
and expressed as nanomoles bone collagen equivalents per liter
per millimole creatinine per liter. Higher values should
be anticipated for postmenopausal women as compared to premenopausal
women, 35 (SD:15) versus 57 (SD:39) nm CE/BCE/mM creatinine
respectively.
Plans for Data Analysis and Interpretation
This pilot study will allow sample size calculations for the
planned followup intervention study. Descriptive statistics
will be used to express results (bone density, fractures, biochemical
tests). DEXA scan results will be normalized to Z-scores
based upon comparisons to normative data previously collected
for children and adults for children and adults from CHMC or
from literature controls using similar densinometry equipment.
Statistics between the ambulatory and nonambulatory populations
will also be tested, as will correlations between scored variables
within both populations.
References
(1) Henderson RC, Lin PP, Green WB. Bone mineral
density in children and adolescents who
have spastic cerebral palsy. J Bone Joint Surg 1995; 77-A: 1671-1681.
(2) Wilmhurst S, Ward K, Adams JE et al. Mobility status
and bone density in cerebral palsy.
Arch Dis Child 1996; 75:164-5.
(3) Lee JJK, Lyne ED, Kleerekoper K et al. Disorders
of bone metabolism in severely handicapped
children and young adults. Clin Ortho Related Res 1989;
245: 297-302.
II. MENTOR PORTION OF APPLICATION
Research Sponsor
James Qqqqq, M.D. Dept. Pediatrics
GENERAL PLAN for student training program (To be completed
by mentor). Include information about the nature and frequency
of direct interaction with the student. Identify individuals
who will participate in student training, and briefly describe
conferences and lab meetings the student will attend.
Xxxxx (student) will
serve as a research coordinator for the proposed project.
The project has been approved by the IRB at the Children's Hospital
and the Scientific Advisory Committee of the General Clinical
Research Center. Xxxxx and I will meet with staff at 3
area nursing homes regarding the recruitment and logistics of
bringing subjects to the GCRC for study. He will coordinate
all of these activities with the assistance of myself and staff
of the GCRC. He will interact with me on a daily basis
to work out procedural problems related to the project.
He will develop a database with the assistance of staff of the
GCRC for collection of data and will enter and check all data
for accuracy and completeness. He will learn the techniques
of dual energy x-ray absorptiometry (DEXA) including methods
to download data directly into spreadsheets. He will learn about
radiation safety measures as it relates to this portion of the
study. He will spend time in the laboratory, as time permits,
assisting in the assays for the minerals, bone markers
and vitamin D metabolites to be measured as a portion of this
study. We will have a defined weekly one hour meeting
to review progress on this project. At this time, Xxxxx, myself
and a nurse representative from the GCRC will discuss progress
and problems that need to be addressed to ensure completion
in a timely fashion.
In addition, Xxxxx
will be encouraged to attend Drs. Yyyy and Zzzzz weekly lab
meeting where progress on their projects will be discussed.
He will be encouraged to make clinical rounds with me when I
am on call in June and he will be encouraged to attend all conferences
related to the GI Division (3-1 hour conferences/week) which
include a clinical care conferences, a topic conference and
a combined Pediatric/Adult GI conference alternating with a
Pathology Conference. He will also have the opportunity to observe
endoscopies that I perform as time permits.
If the proposed project will involve use of RADIOACTIVE ISOTOPES,
list:
Authorized User: Not applicable
Authorization Number:
If the proposed project will involve the use of ANIMALS, list:
Title of Approved IACUC Protocol:**
Not applicable
Principal Investigator:
IACUC Protocol Number:
** (Must be currently approved or have evidence of approval
by June 1, 2000)
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