COLLEGE OF SCIENCEs and Health Professions
Abstracts
Research Day 2013
001
RELATIONSHIP BETWEEN SUNDIAL TIME AND MECHANICAL CLOCK TIME
John Reinmann, M.S.1, 2; James A. Lock, Ph.D.3
1
Icing Technology Branch Chief, NASA-Glenn Research Center (retired);
2
Project 60 Program, Cleveland State University;
3
Physics Department, Cleveland State University
Sundials were the standard for telling time for many centuries before the development of reliable mechanical clocks in the
17th century. Many of these early sundials can still be seen today on medieval buildings throughout Europe, and several
modern working sundials in both private and public locations in cities throughout the world still hold a fascination for many.
Mechanical clocks are very precise instruments that run at the same rate throughout the year. But since the trajectory of the
sun in the sky changes from day to day, the difference between sundial time and clock time varies from day to day as well.
The daily correction to sundial noon with respect to clock noon is called the Equation of Time. Using a familiar and easily
understood model that includes both the earth’s elliptical orbit about the sun and the tilt of the earth’s axis of rotation with
respect to the plane of the ellipse, we obtain and numerically solve the exact equations for this daily correction. Then using a
Taylor series expansion of the equations, we not only obtain an approximation to the exact equations that is accurate to
within a few seconds, but also reveals the physical significance of each of its dominant terms.
002
APPLICATION OF NUMBER THEORY TO RSA CRYPTOGRAPHY
Shane S. Poloha; John Walsh, Ph.D.
Department of Mathematics, Cleveland State University
Perhaps one of the most useful and appreciated interactions of mathematics and computer science comes in the form of
cryptography. This field has become absolutely crucial in the modern world. Each day millions of people exchange financial
and personal information throughout the world over the internet and other electronic mediums. Without the necessary
protection, this information can easily be intercepted and abused by those with ill intentions. In simple terms, cryptography is
the study of how to protect this information and exchange it securely. Most of the techniques used in this field come from an
area of mathematics generalized as Number Theory. Thus, for this project, I will address many of these techniques and the
theory behind them. Furthermore, to apply these techniques to actual software development, I created several useful data
structures and a demonstrative application with the widely used Java programming language.
003
FEASABILITY AND METHODS OF LARGE SCALE BASELINE TESTING IN HIGH SCHOOL AND
COLLEGIATE ATHLETES USING THE CLEVELAND CLINIC CONCUSSION (C3) iPAD
APPLICATION
Jessica A. Jancewicz, BS1; Susan M. Linder, PT, DPT, MHS2; Karen O’Loughlin, PT, DPT 1; David D. Schindler, BS2;
Joshua R. Hirsch, BS2; Daniel E. Kana, BS2; Jay L. Alberts, PhD2
1
School of Health Sciences, Cleveland State University;
2
Department of Biomedical Engineering, Cleveland Clinic
1
Concussion, or mild traumatic brain injury (mTBI), is the result of traumatic biomechanical acceleration-deceleration and
rotational forces on the head causing declines in cognition, motor performance, vestibular function, or postural stability.
Concussion presents with a variety of symptoms and impairments, including deficits in postural stability, as well as cognitive,
motor, and vestibular function. To improve the detection and management of athletes with concussion, it is recommended
that a multi-dimensional assessment approach be used. The current model in concussion management does not include a
comprehensive baseline assessment to compare neurological function post-injury. The Cleveland Clinic Concussion (C3)
application was developed to help fill this gap and provide health care professionals with an on-site tool to comprehensively
evaluate the symptoms most prevalent in a concussion. The purpose of this project was to complete baseline performance
tests on athletes who participate in “at risk” fall sports including football, soccer, and volleyball. C3 baseline testing took
place at regional high schools and colleges whose teams are managed by a Cleveland Clinic Health System Athletic Trainer.
A volunteer sample of 4,921 male and female high school and collegiate athletes were included in this study. The C3 App
contains modules designed to evaluate baseline symptoms, cognitive function, motor function, and visual acuity. Specifically,
the C3 App contains: 1) Sport Concussion Assessment Tool 2 (SCAT2); 2) Balance Error Scoring System (BESS); 3) Trail
Making Test A&B; 4) Reaction Time; 5) Cognitive Assessment Profile (short-term memory, attention, cognitive ability) and;
6) Static & Dynamic Vision. Testers administered the SCAT 2, BESS, and Static & Dynamic Vision modules with the
athletes. The Trail Making Tests A&B, Simple Reaction Time, Choice Reaction Time and Cognitive Assessment Profile
modules were completed by the athletes independently. C3 was able to provide a comprehensive assessment with optimal
validity as well as cost efficiency.
004
NOMATIVE DATA FOR ARABIC SPEAKING OMANI ADULTS ON COMMONLY USED
NEUROPSYCHOLOGICAL MEASURES
Chelsea K Kociuba, B.A.; Amir Poreh, Ph.D.; Alya Sultan, M.A,; Kelly Martincin, B.A.; Ashley K. Miller, B.A.
Department of Psychology, Cleveland State University
Normative data for Arabic speaking adults is limited. This study provides normative data for commonly used
neuropsychological measures using a sample of 200 Arabic speaking Omani adults aged from 18-50. Included are the norms
for six neuropsychological measures assessing attention, memory, and verbal and nonverbal fluency. Similar to previous
cross-cultural studies the age and education appear to have a strong effect on the subjects’ test performance. Although the
verbal memory measure in the present study produced identical results to those that have been published in other countries,
most of the other measures did not produce equivalent scores. These results support the important role linguistic, cultural, and
educational experiences play in the performance of subjects on neuropsychological tests.
005
CENTRAL PATTERN GENERATORS FOR QUADRUPEDAL ARBOREAL LOCOMOTION OF
CHIPMUNKS TAMIAS SIBIRICUS
Alexandra Powers,1;Ulrich Zurcher, Ph.D.2; Andrew Lammers, Ph.D.3
1
Department of Biological, Geological and Environmental Sciences, Cleveland State University;
2
Department of Physics, Cleveland State University;
3
School of Health Sciences, Cleveland State University
Locomotion requires to coordinated periodic movements of muscles and ligaments, which in turn requires periodic firings of
neurons. The patterns are self-generated and do not depend on external stimulus. The corresponding oscillators are known as
central pattern generators [CPG]. The CPGs for fore- and hind limb movement have been studied quite extensively in the
literature. We focus on CPGs for the motion of the trunk, head, and tail of chipmunks. We observe wave-like behavior, which
generates the angular momentum about the mediolateral axis necessary for dynamic, rather than static, stability.
2
006
MOUSE TRACKING REVEALS FACILITATION IN A BILINGUAL STROOP TASK
Sara Incera, M.S.; Conor T. McLennan, Ph.D.
Department of Psychology, Cleveland State University
The purpose of this study is to analyze the continuous dynamics of facilitation in a bilingual Stroop task. Facilitation
(McLeod & McDonald, 2000) is observed when responses are faster or more accurate to congruent stimuli (e.g., red, rojo in
the color red) than to control stimuli (e.g., XXXX in the color red). Mouse-tracking data distinguish interference from
facilitation, providing new insights into whether color–word Stroop facilitation is caused by converging information
(coactivation model) or inadvertent reading (separate-activation model). Is the congruent word actually helping (converging
information) or are there reading errors that result in faster responses (inadvertent reading)? The current poster explains how
mouse-tracking data allow us to distinguish between these competing theories by observing the beginning of the mouse
movements. If reading errors occur, the trajectory of the mouse would show how participants ignore the color of the word at
the beginning of their movement. On the contrary, if converging is taking place, the trajectory should provide information
about when both paths (color and meaning) converge, making congruent conditions faster than control. Despite over 75 years
of research using the Stroop task, there remain important empirical questions in which this task can be useful. Mousetracking data can provide new insights contributing to theoretical developments, helping research to move forward.
007
THE IMPACT OF SELF-DEFENSE TRAINING ON FEMALE SURVIVORS OF SEXUAL ASSAULT
Caitlin M. Pinciotti, B.S.; Lisa Stines Doane, Ph.D.
Department of Psychology, Cleveland State University
207,754 people (age 12 and over) experience a sexual assault in the United States every year (U.S. Department of Justice).
While survivors are at a significantly higher risk for developing Posttraumatic Stress Disorder (PTSD) and maladaptive
coping skills, such as drug abuse (World Health Organization, 2002), the perceived control over their recovery decreases
these negative aftereffects (Frazier, 2003; Frazier, Steward, & Mortensen, 2004; Ullman, Filipas et al., 2007). Self-defense
programs which focus on female empowerment and strength may help to provide a sense of control both before and after an
assault. 200 female participants enrolled in the Rape Aggression Defense program throughout the country will take a brief
survey pre- and post-training to determine the impact of this formal self-defense training program on self-efficacy, coping,
and PTSD symptoms. The survey includes the following measures: The Self-Defense Self-Efficacy Scale (Ozer & Bandura,
1990), The National Violence Against Women Survey (NVAWS; Tjaden & Thoennes, 1998), PTSD Symptom Scale: SelfReport Version (PSS-SR; Foa, Riggs, Dancu, & Rothbaum, 1993), and the Brief COPE (Carver, 1997). It is predicted that
both groups, women with and without sexual assault history, will experience significant improvements in self-efficacy and
coping scores; women with sexual assault history will have greater changes in self-efficacy and coping scores than women
without sexual assault history; PTSD symptoms will decrease from the pre- to post-training among women who experienced
a sexual assault. The current study hopes to strengthen previous research on the positive benefits of self-defense training for
women, specifically female survivors of sexual assault.
008
USING SCHIZOTYPAL TRAITS TO DETERMINE THE EFFECT OF EFFERENT SIGNALS ON SELFMOTION SENSING
Lucinda V. Rohde; Naohide Yamamoto, Ph.D.
Department of Psychology, Cleveland State University
Self-motion sensitivity is comprised of input from two types of signals; afferent signals, based on proprioception and
vestibular senses and efferent signals, which are copies of motor commands issued by motor areas of the brain. These two
types of signals are present when body movements are willfully made, making it difficult to isolate the effects of efferent
signals on self-motion sensing. The present study addresses this issue by capitalizing on the theory that efferent signals are
dysfunctional in schizophrenia. That is, by investigating self-motioning sensing of schizophrenia patients, unique
contributions of efferent signals to self-motion sensing may be identified. To validate this idea, we compared performance of
individuals with high schizotypal traits and those with low schizotypal traits in distance perception tasks that involved
sensing self-motion-related signals. It was hypothesized that individuals with high schizotypal traits would perform less
3
optimally in tasks that require more use of efferent signals than the individuals with low schizotypal traits. In one task,
participants viewed a target that was 4-8 m away and walked to it without vision (real walking; efferent and afferent signals
both used). In the other task, they only imagined walking to the target (imagined walking; primarily using efferent signals). In
both tasks, time required to reach the target (either physically or in imagination) was measured for comparison. Results
showed that low schizotypal participants tended to “walk” more quickly to the target in imagined walking than in real
walking, which represents a normal pattern of performance in these tasks shown in the literature. By contrast, high
schizotypal participants exhibited smaller differences between real and imagined walking, indicating that they performed
imagined walking differently from low schizotypal participants. These findings were consistent with the hypothesis and
suggest that efferent signals do play a role in self-motion sensing.
009
DEFICIENCY OF LARGE RIBOSOMAL SUBUNIT PROTEIN L13A IN MACROPHAGES PROMOTE
ATHEROSCLEROSIS IN MICE
Abhijit Basu, Ph.D.1; Darshana Poddar, M.S.1; William Baldwin, Ph.D.2; Jonathan D. Smith, Ph.D.3; Maria Febbraio,
Ph.D.4; Barsanjit Mazumder, Ph.D.1
1
Department of Biological, Geological and Environmental Sciences; Center for gene Regulation in Health and Disease;
Cleveland State University; Cleveland, Ohio, USA;
2
Department of Immunology, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, USA;
3
Department of Cellular and Molecular Medicine, Cleveland Clinic Lerner Research Institute, Cleveland, Ohio, USA;
4
Department of Dentistry, University of Alberta, Edmonton, AB, Canada.
Inflammation is a part of host’s immune response aimed to remove infectious stimuli. Though it is beneficial initially,
uncontrolled syntheses of inflammatory molecules can result in chronic persistent inflammation and eventually leads to
development of diseases such as atherosclerosis. Pro inflammatory molecules such as chemokines selectively recruit
monocytes, neutrophils and leukocytes to sites of vascular injury, inflammation and developing atherosclerosis. We have
identified one cellular mechanism that prevent overexpression of inflammatory molecules and thereby protect cells from
developing chronic inflammation. This pathway, which depends on ribosomal protein L13a, prevents expression of a group
of inflammatory molecules post transcriptionally. In this study, we explored the role of L13a in atherosclerosis using a
macrophage-specific L13a knock-out (KO) mice. Using hyperlipidemic apoE-/- mice, we showed, upon feeding with high fat
diet, L13a-KO mice exhibit an increased atherosclerotic lesion with an elevated levels of plasma cytokines/chemokines. This
study provides evidences for an atheroprotective role of L13-dependent translational silencing.
010
THE TELOMERE DNA BINDING ACTIVITY OF TRYPANOSOMA BRUCEI TRF IS IMPORTANT FOR
SUPPRESSION OF SUB-TELOMERIC VSG SWITCHING
Imaan Benmerzouga, B.S.1; Xiaohua Li, Ph.D.2; Zhao Yanxiang, Ph.D.2; Bibo Li, Ph.D.1
1
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University, Cleveland, Ohio, USA.
2
Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Kowloon, Hong Kong,
Hung Hom, China.
Trypanosoma brucei, the causative agent of human African trypanosomiasis (HAT), effectively escapes the host immune
response by regulating the expression of its surface antigen, Variant Surface Glycoprotein (VSG), via monoallelic expression
and by regularly switching its expressed VSG via VSG switching.
VSGs are expressed exclusively from VSG expression sites located adjacent to telomeres, which are nucleoprotein complexes
at the natural chromosome ends. We have identified TbTRF as a telomere DNA binding factor, which is essential for
maintaining the telomere end structure. In this study we report that transient depletion of TbTRF leads to a significant
increase in the VSG switching frequency, and many switchers arose through the in-situ switching pathway that is seldom used
in WT cells. To investigate the underlining molecular mechanism, we focused on the telomere DNA binding activity of
TbTRF and solved the NMR structure of its myb domain, which harbors a helix-turn-helix motif conserved with the hTRF1
myb domain. The association of TbTRF with the telomere in T. brucei cells was not affected by the heavily modified J
-D-glucosyl-hydroxymethyluraci) usually found in T. brucei telomeres. However, several point mutations in the
4
TbTRF myb domain were identified to exhibit weakened DNA binding activity both in vitro and in vivo. The same mutants
also led to an elevated VSG switching frequency, although gene conversion is still the predominant switching mechanism in
these mutants as in WT cells. Our data indicate that the DNA binding activity of TbTRF is critical for VSG switching
regulation.
011
ACTIVATED NEUTROPHIL MEDIATE KIM-1 SHEDDING AND KIDNEY REMODELLING
Shreyas Vaman Lingadahalli, MBBS1, 2; Thomas McIntyre, PhD2
1.
Department of Biological, Geological and Environmental Sciences; Cleveland State University;
2.
Cellular and Molecular Medicine, Cleveland Clinic
Kidney performs a complex task of concentrating the urine, retention of salts and protein and excretion of metabolic toxins
and by the virtue of its function, is always under the chemical stress and subject to constant tissue damage. Post injury,
nephrons have the ability to regain their function by remodeling including, clearing of apoptotic and necrotic debris. Kidney
Injury Molecule-1(KIM-1/TIM-1/HAVCR-1) is a phosphatidylserine receptor that recognizes the apoptotic bodies and
directs them to the lysosomal degradation. KIM-1 a type I transmembrane glycoprotein, although not constitutively
expressed, is expressed in injured epithelial cells. It is known that the extracellular domain is cleaved by, MMP-9 and MMP3, and the cleaved protein can be detected in the urine, making it a sensitive and non-invasive biomarker of renal injury. In rat
renal injury model we observed neutrophil infiltration at the site of injury. To understand the role of neutrophis and its
relation to KIM-1, we hypothesized, activated neutrophils increases shedding of the KIM-1 extracellular domain via
MMP activity and thus reducing the uptake and clearing of apoptotic bodies. The purpose of the study is to determine if
activated neutrophils can cause KIM-1 shedding and identify the MMP involved in this cleavage. Immortalized human
proximal tubular (HK2) cells were treated with supernatant from PAF (pro-inflammatory lipid) activated neutrophils. We
observed an increase shedding of KIM-1 on treatment with activated neutrophil supernatant and that shedding was blocked
after pretreatment of neutrophils with an MMP-9 inhibitor. Pretreatment with MMP-3 inhibitor did not show any change. We
also found KIM-1 mediated uptake of apoptotic bodies is reduced after the KIM-1 ectodomain shedding .Our study suggests
that activated neutrophils can cause KIM-1 shedding via MMP-9 that consequentially reduces the phagocytic activity of the
epithelial cells and thus affect the kidney remodeling.
012
MYOD DIRECTLY INCREASES TRANSCRIPTION OF THE PRO-APOPTOTIC BCL2 FAMILY
MEMBER PUMA
Terri J. Harford, Ph.D. and Crystal M. Weyman, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
We have previously reported that the muscle regulatory transcription factor MyoD plays a role in the elevated expression of
the pro-apoptotic Bcl2 family member PUMA, and thus the ensuing apoptosis that occurs in response to various apoptotic
stimuli. Herein, we report that MyoD directly increases the transcription of PUMA. In silico analysis of the PUMA extended
promoter revealed three potential MyoD binding sites, of which only one is conserved. We, therefore, have created luciferase
reporter constructs containing segments of the PUMA extended promoter. Luciferase expression from the vector containing
the -2000 to +225 region of the PUMA promoter transfected into 23A2 myoblasts lead to a 500-fold induction when
compared to control vector alone. When this construct is transfected into myoblasts silenced for MyoD expression, luciferase
expression is significantly reduced. Additionally, when this construct is transfected into fibroblasts, which do not express
MyoD, luciferase expression is also significantly reduced compared to myoblasts. However, when fibroblasts are cotransfected with a construct constitutively expressing MyoD, the 500-fold increase in luciferase expression is restored.
Through a series of truncations of this reporter construct, we have determined that the region from -1170 to +225,
containing the conserved MyoD binding site, is sufficient for this MyoD induced increase. Additionally, when this conserved
binding site is deleted in the reporter construct, induction is again significantly decreased. Chromatin immunoprecipitation
(ChIP) analysis using anti-MyoD antibodies confirmed MyoD binding to this region encompassing the site identified through
reporter analysis within the PUMA extended promoter. Binding of MyoD to this region increased in myoblasts cultured in
differentiation medium (DM, no serum) when compared to myoblasts cultured in growth medium (GM, 10% serum). Finally,
in myoblasts silenced for MyoD expression, the increased binding of MyoD at this region when cultured in DM is abolished.
5
013
MODEL OF CILIARY MOTION IN PHYSIOLOGICAL CONDITIONS
Albert R. Sundberg1, B.A.; Andrew H. Resnick1, Ph.D.; Partha Srinivasan2, Ph.D.
1
Department of Physics, Cleveland State University
2
Department of Mathematics, Cleveland State University.
The cilia is a structure that is ubiquitous in mammalian biological systems. The study of this organelle has the potential to
alleviate or cure many common diseases such as immotile-cilia syndrome, primary ciliary dyskinesia, situs inversus,
hydrocephalus, infertility, and retinitis pigmentosa. The mechanisms of cilia as a sensor are not completely understood in
light of the know physiology. This study is intended to elucidate the details of ciliary movement using computational
methods based on classical mathematical models.
014
EXAMINING THE ROLE EFFERENT SIGNALS PLAY IN THE SELF-MOTION SENSING
COMPONENT OF SPACE PERCEPTION IN INDIVIDUALS WITH VERY HIGH SCHIZOTYPAL
TRAITS
Saliha Qadir; Naohide Yamamoto, Ph.D.
Department of Psychology, Cleveland State University
Spatial navigation is an important aspect of an individual’s daily life. An important constituent in spatial navigation is selfmotion sensing that detects motion and position of body parts via efferent and afferent signals. Afferent signals are present
when the body is moved, regardless of whether the movement is made intentionally or by external forces. On the other hand,
efferent signals are only present when one intends to move one’s body. Thus, in most circumstances when efferent signals are
present afferent signals are also present. As a result, it is difficult to identify unique roles of efferent signals in self-motion
sensing. In this particular research we aim to examine whether efferent signals have a distinctive part in self-motion sensing.
Participants with varying degrees of schizotypal (i.e., premorbid schizophrenia) traits will be selected using a questionnaire;
they will then carry out tasks in which they will estimate the distance traveled or the time required for traveling that distance.
These tasks will vary in their involvement of afferent and efferent signals (e.g., actively walking to a previewed target vs.
being passively moved while sitting in a wheelchair). Participants with schizotypal traits are part of this research, because
schizophrenia is a disorder which can be theorized as having dysfunctional efferent signals. It is predicted that participants
with very high schizotypal traits will estimate the distances and the time less optimally than those with low or no schizotypal
traits, but only when the tasks involve efferent signals. These results will allow us to analyze whether efferent signals have a
distinguishing role in self-motion sensing. Moreover these results will also expand our knowledge in the theory which
constitutes schizophrenia as a disorder of efferent signals.
015
COMPARATIVE STUDY OF POLYMER CHAIN DYNAMICS IN AQUEOUS SOLUTIONS BY FPR AND
DLS
Kiril A. Streletzky1, Ph.D.; Ryan McDonough1; Rafael Cueto2, Ph.D.; Paul Russo2, Ph.D.
1
Department of Physics, Cleveland State University
2
Department of Chemistry, Louisiana State University
Self-diffusion of tagged polymer chains in aqueous solutions of hydroxypropylcellulose (HPC) was measured by
Fluorescence Photo-bleaching Recovery (FPR) and compared to mutual diffusion of scattering species in the same solutions
measured by Dynamic Light Scattering (DLS). The effect of the dye presence on thermodynamic concentration fluctuations
observed by DLS was also studied. The observed multimodal spectra in DLS and FPR were analyzed with CONTIN and
stretched exponential fits. A set of consistent dissimilarities in the modal distributions of FPR and DLS spectra was found.
This indicates a comparative limitation or sensitivity in range of detectable diffusive processes between FPR and DLS in this
complex system. In addition, it was found that the fluorescent tag and/or tagging process seem to alter the mutual diffusion
processes seen by DLS. In particular, a slower mode which is apparent in the non-tagged sample does not appear in the
tagged sample. It seems likely that the dye chemically affects the polymer chains keeping them from clustering with each
6
other, altering the solvent environment preventing formation of polymer clusters responsible for the slow mode usually seen
in HPC.
016
APPARENT SPHERE TO ELONGATED PARTICLE TRANSITION OF ELASTIN-LIKE POLYPEPTIDE
THERMIREVERSIBLE MICELLES
Kaitlin W. Vandemark1; Ali Ghoorchian2, Ph.D.; Nolan B. Holland2, Ph.D.; Kiril A. Streletzky1, Ph.D.
1
Department of Physics, Cleveland State University
2
Department of Chemical and Biomedical Engineering, Cleveland State University
Biosynthesized polymers can be designed to assemble into environmentally responsive nanoparticles. Such a system
consisting of an oligomerization domain connecting three elastin-like polypeptide (ELP) chains has been developed. These
polypeptides reversibly transition from aqueous soluble polymers to amphiphiles when the temperature is raised above the
ELP transition temperature. As amphiphiles these can assemble into micelles under appropriate solution conditions. A
particular system has been designed to reversibly form micelles at a neutral pH. However, the shape and size of micelles was
found to depend strongly on salt concentration. We used polarized and depolarized dynamic light scattering to study
temperature-driven formation of micelles of various geometries under different solvent conditions. We also monitored the
sphere-to-elongated particle transition of the ELP micelles with addition of salt. The apparent radius, shape, and dynamics of
micelles strongly depend on salt concentration, with two distinct salt regimes and a broad transition region. At low salt
concentration (0-15 mM), largely spherical micelles were found with a hydrodynamic radius of 10-13 nm. At intermediate
salt concentration (15-35 mM) the transition from spherical micelles to elongated particles is observed. At high salt
concentrations (above 35 mM), the micelles again reach a stable structure consisting of highly anisotropic particles with
aspect ratio of about 10-15:1.
017
CHARACTERIZING FeOOH NANORICE IN SOLUTION USING POLARIZED AND DEPOLARIZED
DYNAMIC LIGHT SCATTERING
Phil Dee1; Olga V. Dement'eva2, Ph.D.; Victor M. Rudoy2, Ph.D.; Kiril A. Streletzky1, Ph.D.
1
Department of Physics, Cleveland State University, Cleveland, Ohio, USA;
2
Frumkin Institute of Physical Chemistry and Electrochemistry, Moscow, Russia
Spindle-shaped (nanorice) particles of iron(III) oxyhydroxide (FeOOH) are used for targeted synthesis of core/metal nanoshell structures known as complex composite nanoparticles (CNPs). Characterization of FeOOH nanorice within native
solution environment is essential for understanding their properties and successful synthesis of CNPs. The combination of
Dynamic Light Scattering (DLS) and carefully designed Depolarized Dynamic Light Scattering (DDLS) allows to measure
translational and rotational dynamics, structure, and size distribution of nanorice in situ. DLS/DDLS provides sampling of a
large number of nanorice particles as opposed to a few particles typically probed by the imaging techniques. In addition,
FeOOH provide a nice prolate ellipsoid and solid cylinder models were used to deduce FeOOH nanorice dimensions from
measured diffusion properties. For orthodox methods based on Perrin model, prolate ellipsoidal model produced nanoparticle
lengths and aspect ratios within 10-20% of the transmission electron microscopy (TEM) results. The cylinder model based on
Tirado/de la Torre approach performed significantly worse. For an alternate method after Glidden, et.al (M. Glidden, M.
Muschol, J. Phys. Chem. C, 116, 8128 (2012)), the opposite was true and straight cylinder dimensions surprisingly proved to
be more accurate, though the model itself was found to be not fully self-consistent. The effects of number concentration of
FeOOH nanoparticles in solution on their dynamics were also studied in detail in order to understand the degree of coupling
between the rotational and translation diffusion under different conditions.
7
018
APPLICATION OF THE TIME COURSE OF VARIABILITY EFFECTS IN SPOKEN WORD
RECOGNITION
Laura Elias; Conor McLennan, Ph.D.
Department of Psychology, Cleveland State University
There are two types of models of spoken word recognition that aim to describe how speech is represented and processed. The
abstractionist model explains talker variability as acting as noise that does not affect performance in recognizing spoken
language; in contrast, the episodic model explains that talker variability does influence the performance in terms of accuracy
and speed. The central question is no longer which model explains how spoken words are recognized; it is accepted that both
are used at different times during processing. Evidence has been found that the abstractionist model explains early
processing, whereas the episodic model describes how later processing occurs consistent with the time-course hypothesis.
The time-course hypothesis has a lot of supporting evidence obtained through basic research. In the current study, I examined
if the time-course hypothesis can be applied to real world tasks that involve processing spoken words, particularly in a
dispatcher task. Participants were shown two maps and primed the locations of several images. All participants were shown a
map for the easy condition and a different map for the hard condition. During the hard condition there was background noise
presented with the image words. In the target task participants were asked to identify the locations primed in the target block.
My predictions are consistent with the time-course hypothesis. In the easy condition, I predict participants’ reaction times and
accuracy of image location will be the same for repeated words regardless of whether or not they are spoken by the same
talker; however, in the hard condition, I expect that the same talker condition will have faster reaction times and greater
accuracy than the different talker condition.
019
STRUCTURED LEISURE PARTICIPATION OF AN ADULT MALE WITH DOWN SYNDROME
Brittany M. Holmes, B.S.; Lisa M. Hooker, BSHS; Lyndsey A. Mitchell, BSHS; Kathryn E. Toledo, BSHS; and Rebecca J.
Winne, BSHS; Susan Bazyk, Ph.D., MHS, B.S., OTR/L, FAOTA
School of Health Sciences, Master's of Occupational Therapy Program, Cleveland State University
The purpose of this case study is to explore the experience of an individual with Down syndrome during leisure participation
at a local YMCA. Data were collected using observations during one personal training session and one karate class, and indepth interviews with the participant, his mother and trainer. Qualitative data analysis resulted in four major themes
including: somewhere he belongs, benefits of structure leisure participation, parental roles, and components needs for
success. Study findings indicate there are many physical and psychosocial benefits to both exercise and structure leisure
participation, including increased social participation, improved confidence, and improved balance and muscle strength.
Finally, the model of “Occupational spin-off” is discussed as it relates to this research. This model focuses on the
precipitating factors that lead to initial occupational participation, continued participation, and eventually seeking out new
experiences.
020
A STUDENT-LED COMMUNITY COLLABORATION: A PARTNERSHIP WITH A NATIONALLYRECOGNIZED HARDWARE STORE TO PROMOTE OCCUPATIONAL THERAPY
Jaime E. Bass, B.S.; Christine M. Cecil, M.Ed.; Robin L. Chilton, M.B.A.,OTR/L; Jocelyn R. Freyman, B.L.S.; Richard L.
Morris, B.S.H.S.; Emily N. Swogger, M.A.H.G.
School of Health Sciences, Occupational Therapy Program, Cleveland State University
The American Occupational Therapy Association (AOTA) Centennial Vision was designed to be a road map for the future of
the profession to commemorate the Association’s 100th anniversary in 2017. It envisions occupational therapy as a powerful,
widely recognized, science-driven, and evidence-based profession with a globally connected and diverse workforce meeting
society’s occupational needs. The Centennial Vision encourages AOTA members to become strong advocates for the
occupational therapy profession. Five occupational therapy students (OTS) at Cleveland State University sought to
implement this call to action by collaborating with a local Home Depot store in an underserved community for a Fire Safety
Event. The purpose of this collaboration was to develop a partnership with a local business that already had an established
8
presence in the community and to educate participants about the role of occupational therapy and fire safety through
interactive activities, handouts, and displays. Demographics and feedback were collected with a voluntary survey, an
interview with a Home Depot staff member, and student reflections. The results of the demographic survey indicated an
average respondent age of 35.5 years old, 47% ranked high school as their highest level of education and 48% reported a
family income less than $20,000. In addition, the Home Depot staff reported an increase in storewide product sales and fire
safety products during and directly following the event and participation was estimated to be the highest among all Home
Depot stores in the district. Three themes emerged from the OTS reflections: 1) Home Depot staff enthusiasm, 2) the need for
occupational therapy in the community, and 3) the importance of advocating for and providing education about the
occupational therapy profession. It is recommended that occupational therapy programs facilitate similar types of community
outreach programs by developing community-collaboration courses and providing opportunities to perform similar public
events.
021
WHAT FACTORS BEST PREDICT FALLS IN COGNITIVELY IMPAIRED OLDER ADULTS?
Nicole T. Dawson, P.T., M.A.; Katherine S. Judge, Ph.D.
Department of Psychology, Cleveland State University
Falls accounted for 2 million non-fatal injuries to adults over the age of 65 in 2007. Most studies include healthy,
community-dwelling adults; with few studies examining older adults with cognitive impairment, who are twice as likely to
fall. Several theories attempt to explain this increased risk including decreased executive function, decreased visual attention,
and/or difficulty with dual-task resource allocation. The current study examined falls in forty-seven cognitively impaired
older adults. Participants completed a cognitive assessment: MMSE, clock-drawing test, working memory task (WM), verbal
fluency task (VF), and visual attention task (VA), as well a dual-task paradigm using the 10-meter Walk Test during
performance of each cognitive task. The 10-meter Walk Test also measured gait velocity. The number of falls incurred over
the past 12-months was used as the outcome. Mean age of participants was 84.9 (SD 9.5) with MMSE score of 19.1 (SD 3.1)
indicating mild to moderate cognitive impairment. Twenty-two participants (47%) reported recurrent falls over the past 12months with the number of falls across all participants averaging 2.2 (SD 3.0). Gait velocity (GV) was .55 meters/second
(m/s: SD .17) in the single task condition which significantly decreased in dual-task conditions of WM (p<.001), VF
(p<.001), and VA (p<.001) to .35 m/s (SD .15), .33 m/s (SD .13), .34 m/s (SD .12), respectively. VA in the single task
condition was significantly correlated with falls (r=-.33, p=.037). Significant group differences also were identified in
VAsingle (F1,38=4.7, p=.037). These results provide preliminary evidence that the cognitive construct of visual attention is
important in assessing fall risk in older adults with cognitive impairment. Measures of visual attention may assist clinicians in
identifying older adults with cognitive impairment who may be at the highest risk for falls.
022
TRANSITION TO COLLEGE PROGRAM FOR HIGH SCHOOL STUDENTS WITH DISABILITIES: A
GRANT PROPOSAL
Kathryn A. Staraitis, B.S.; Lauren E. Baird, B.A.; Mary C. McHale, B.S., B.A.; William J. Williams, B.S.; Glenn D.
Goodman, Ph.D.
School of Health Sciences, Master of Occupational Therapy Program, Cleveland State University
We are seeking funding to provide a college transition program for high school students with disabilities in Cuyahoga
County. In high school, students have the resources of IEPs or 504 plans allowing them to take a passive role in modification
and adaptations for personal educational success. When students with disabilities begin a post-secondary education, there is a
role shift that occurs simultaneously with the transition. Implementing an effective transition program for these students will
provide the education and training necessary to master self-advocacy concerning their disability. Students with disabilities
need an effective transition program to receive the best college education possible. The overall goal of the program is to
address several pertinent areas found within current research to compose a successful transition program. These include
appropriate guidance and counseling, self-determination and advocacy, early intervention, parental support, assistive
technology, knowledge of laws and awareness of rights, and financial support. In 2008, CSU was awarded a grant from the
State Library of Ohio, to fund a pilot transition program for high school students with disabilities transitioning to college. It
was evaluated and suggestions for improvements were made. We are seeking to receive funding to re-implement this
transition program, making necessary improvements. Grant funding would total $244,914 over a two year period. Outcomes
9
will be measured through post-tests composed of measured typing speed, accuracy and other computer skills, a selfdetermination survey, exit interview, and a focus group. Long-term outcome measures include tracking the students that
apply to post-secondary institutions, of those, how many are accepted, and finally how many attend. The transition program
will therefore address a community-wide unmet need for transition services in the Cleveland area for students with
disabilities, and the low attendance of young adults with disabilities at college and universities.
023
STRUCTURE-ACTIVITY STUDIES OF INDOMETHACIN DERIVATIVES AS TUBULIN INHIBITORS
AND COMPUTATIONAL PREDICTION OF BINDING MODE
Snigdha Chennamaneni1, B.S.P.S.; Bin Su1, 2, Ph.D.
1
Department of Chemistry, Cleveland State University;
2
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Microtubules are the basic components of cell structure, which take part in a wide number of pivotal cellular functions. Drugs
that are able to modulate the microtubule assembly either by inhibition of tubulin polymerization or by blocking microtubule
assembly are of great interest in anti-cancer therapy. In the past few years, several small synthetic molecules that have indole
nucleus as a core structure have been identified as tubulin inhibitors. Indomethacin, a well-known NSAID, with an indole
nucleus was chosen for our study. In order to investigate the structure activity relationship (SAR) between the anti-cancer
effect and some potentially relevant chemical properties, we prepared a series of indole derivatives, where the structure of
indomethacin was gradually modulated, varying the carboxylic group nature by substitution with different amide groups and
the amide hydrogen nature by introduction of different benzoyl substituents. Interestingly, these modifications resulted to a
set of compounds exhibiting higher potency than the parent compound. One of the Indomethacin analogues showed stronger
antiproliferative activity against multiple cancer cell lines via interfering with tubulin polymerization. Tubulin polymerization
assay indicated that this compound inhibited tubulin assembly at high concentrations, but promoted this process at low
concentrations. To rationalize our experimental results, ligand docking was carried out. These analyses suggested a molecular
basis for the interaction of the synthesized compound with tubulin. This compound was selected as the lead compound for
further SAR study, devoted to the optimization of the cytotoxic effect and other mechanism investigation.
024
BEST METHODS IN EDUCATING CLIENTS IN TODAY’S WORLD
Benjamin J. Alger, B.A.; Leigh A. Farr, B.S.; Christine J. Fink, B.S.; Mallory L. McCarthy, B.S.; Carmin L. Milazzotto,
B.A.; Glenn D. Goodman, Ph.D., OTR/L; Joanna DeMarco, DHSc, CHES
School of Health Sciences, Master of Occupational Therapy Program, Cleveland State University
Client education is a major concern in healthcare today. As healthcare continues to evolve, client education needs to be a
widely discussed topic. Three aspects enhancing the delivery of client education include: using technology, ensuring clientcentered care, and facilitating empowerment. Utilizing popular technology - such as Facebook, Twitter, and YouTube - helps
to educate clients through readily available and cost effective means. Being client-centered, by understanding how each client
learns best, can lead to improved retention and follow-through. Empowering clients through education provides them the
tools to be active participants in treatment options, better cope with their illness, have an improved quality of life, and be
empowered to seek the care and resources they need. As healthcare professionals, being aware of the best strategies for
delivering client education is essential in providing the best care.
025
THERMODYNAMIC STABILITY OF PROTECTIVE ANTIGEN MEASURED BY HIS-HDX-MS
Vennela Mullangi, B.Pharm.1, 2; James Bann, Ph.D.3; David Anderson, Ph.D.1; Masaru Miyagi, Ph.D.2
1
Department of Chemistry, Cleveland State University;
2
Center for Proteomics and Bioinformatics, Case Western Reserve University;
3
Department of Chemistry, Wichita State University
10
Histidine hydrogen deuterium exchange mass spectrometry (His-HDX-MS) has been successfully used to investigate the
structural changes of several proteins. We anticipated that protein stability analysis by His-HDX-MS has capability to
determine the thermodynamic stability of individual domains in a muti-domain protein, therefore would be a useful tool for
proteins that have multiple domains. The commonly used circular dichroism and tryptophane fluorescence method do not
provide such spatial resolution. We analyzed protective antigen (PA) produced by Bacillus anthracis, which has four distinct
domains, and binds to cellular receptor capillary morphogenesis protein 2 (CMG2) through domain 2 and 4. PA and PACMG2 complex were incubated in D2O buffers (pH 7.5) containing different guanidine hydrochloride (Gdn-HCl)
concentrations at 37 °C for 48 hrs. After the incubation the protein samples were equally divided into three tubes, and
digested by Lys-C, Lys-C/chymotrypsin and Lys-C/Glu C, respectively. The resulting digests were pooled and analyzed by
LC-MS using quadrupole/TOF mass spectrometer. The experimentally obtained rate constants (kφ) for HDX of individual
histidine residues were corrected by considering the changes of water concentration associated with the volume effect of
Gdn-HCl. The correction factor was experimentally determined and used it to correct the experimentally obtained kφ values.
Out of 10 histidine residues in PA four residues were found to be well protected from the solvent in its native structure. The
kφ values for the four histidine residues [His211 (domain 1), His253 (domain 1), His336 (domain 2), and His616 (domain 4)]
were therefore monitored to determine the thermodynamic stability of individual domains. The denaturation curves reflected
by the kφ values of these residues in different Gdn-HCl concentrations clearly indicated that the three domains monitored in
PA were stabilized upon receptor binding. This data supports the capability of His-HDX-MS method for determining the
thermodynamic stability of different domains in a protein.
026
PROTEASOME PROTEOLYSIS PROMOTES PLATELET FUNCTION AND THROMBOSIS
Nilaksh Gupta, M.Sc1,2; Wei Li, Ph.D.1; Belinda Willard, Ph.D.1; Roy L. Silverstein, M.D, Ph.D.1,3; and Thomas M.
McIntyre, Ph.D.1
1
Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH;
2
Department of Biological Geological and Environmental Sciences, Cleveland State University; Cleveland, OH;
3
Department of Medicine, Medical College of Wisconsin, and Blood Research Institute, Blood Center of Wisconsin,
Milwaukee, WI.
Treatment of hematologic cancers by proteasome inhibition causes thrombocytopenia and reduces thrombosis. We tested the
hypothesis that platelets modify their proteome by ubiquitination and proteasome proteolysis, and this contributes to the
function of these terminally differentiated cells. We found platelets expressed proteasome hydrolytic activities, and
proteasome inhibition by MG132 strongly suppressed formation of occlusive thrombi in damaged carotid arteries. The gpIbV-IX complex is the high affinity thrombin receptor, and proteasome inhibition reduced platelet aggregation by low thrombin
concentrations. Spreading and clot retraction after MG132 exposure also were aberrant, and MG132 suppressed microparticle
shedding stimulated by several agonists. Proteasome substrates are marked by ubiquitin, and platelets modified their
proteome by mono- and poly-ubiquitination. Stimulation increased this decoration. Platelet cytosol contained fragments of
the cytoskeletal proteins Filamin A and Talin-1 decorated with ubiquitin. MG132 both increased this decoration and
prevented their proteolysis. Mass spectrometry revealed a single MG132-sensitive tryptic cleavage after R1745 in an
extended Filamin A loop, which would separate its actin binding domain from its carboxy terminal gpIbα binding domain.
Platelets therefore contain a ubiqitin/proteasome system that marks cytoskeletal proteins for proteolytic modification to
promote productive platelet-platelet and platelet-wall interactions.
027
MIXING ENTROPY IN DEAN FLOWS
Petru S. Fodor, Ph.D.; Miron Kaufman, Ph.D.
Department of Physics, Cleveland State University
We investigate mixing in Dean flows by solving numerically the Navier-Stokes equation for a circular channel. Tracers of
two chemical species are carried by the fluid. The centrifugal forces, experienced as the fluid travels along a curved
trajectory, coupled with the fluid incompressibility induce cross-sectional rotating flows (Dean vortices). These transversal
flows promote the mixing of the chemical species. We generate images for different cross sections along the trajectory. The
mixing efficiency is evaluated using the Shannon entropy. Previously we have found, P. S. Fodor and M. Kaufman, Modern
11
Physics Letters B 25, 1111 (2011), this measure to be useful in understanding mixing in the staggered herringbone mixer.
The mixing entropy is determined as function of the Reynolds number, the angle of the cross section and the observation
scale (number of bins). Quantitative comparison of the mixing in the Dean micromixer and in the staggered herringbone
mixer is attempted.
028
THE INHERENT EFFECT OF THE B DOMAIN OF COAGULATION FACTOR V ON ACTIVATED
PROTEIN C INACTIVATION OF FACTOR VA
Jasmine Manouchehri; Michael Kalafatis, Ph.D.; Joseph Wiencek
Department of Chemistry, Cleveland State University
The purpose of this study was to determine if the carboxyl-terminal within the B domain of factor V accelerates the rate of
inactivation of factor Va by activated protein C (APC). By experimentation, it was determined that increasing the
concentration of APC increases the rate of inactivation of factor Va by APC. There were two recombinant factor V species
applied to the experiments. One recombinant factor V species had a change in the amino acid from an arginine to glutamine
at 709, 1018, and 1545 of factor V, and another recombinant factor V species had a deletion of amino acids from 1489 to
1532 of factor V which represents the deletion of the B domain of factor V. Through experimentation with a control, it was
determined that the recombinant factor V species with the change in the amino acid from arginine to glutamine at 709, 1018,
and 1545 resulted in a greater rate of inactivation of factor Va by APC while the recombinant factor V species with the large
deletion of amino acids of factor V from 1489 to 1532 had the lowest rate of inactivation of factor Va by APC. An increased
amount of the B domain of factor V showed an increase in the rate of inactivation of factor Va by APC. Therefore, these
results continue to support the hypothesis that the carboxyl-terminal of the B domain of factor V is a factor in the increased
rate of inactivation of factor Va by APC and that the carboxyl-terminal of the B domain of factor is also responsible for the
enhanced cleavage of factor Va by APC at Arg306.
029
GLYCO-MEMBRANE MIMETIC SYSTEM BASED ON TETHERED BILAYER LIPID MEMBRANE
Valentinas Gruzdys, B.S.; Yong Ma, Ph.D.; Jacob Weingart, Ph.D.; and Xue-Long Sun, Ph.D.
Department of Chemistry, Cleveland State University
Cell surface carbohydrates existing as glycoconjugates such as glycoprotein, glycolipids, and proteoglycans play significant
roles in many physiological and pathological events, such as cell-cell signaling, immune recognition events, pathogen host
interaction, etc. Therefore, cell surface carbohydrate-mediated interactions provide a starting point to develop essential
approaches for understanding carbohydrate-protein interactions and for developing carbohydrate-based biosensors for
biomedical research and applications. Cell membrane mimetic systems play fundamental roles in understanding biomolecular
interactions on the cell surface and provide enormous opportunities in developing products for biomedical research and
applications. Among them, tethered bilayer lipid membrane (tBLM) has emerged as the most attractive platform because of
its quasi-natural setting and that its size, geometry and composition can be tailored with great precision to the complexity of
biological membrane. Herein, we report glyco-membrane mimetic system based on tBLM fabrication. Briefly, introduction
of dibenzylcyclooctyne (DBCO)-PEG-lipid into liposome allows the vesicle/bilayer to immobilize onto an azide surface
through a copper-free click cycloaddition reaction. Subsequent rupture of the immobilized liposome followed by a second
liposome fusion leads to the tBLM formation, which contains DBCO for further chemically selective modifications on its
surface. The tBLM was covalently functionalized with azide-containing glycans under biocompatible condition, and thus
provides a straightforward approach for glyco-membrane mimetic system fabrication. The lectin-binding activity of the
glyco-membrane mimetic system was confirmed by fluorescence microscopy. The glyco-membrane mimetic system is very
versatile as it can be adapted easily to different types of support for a variety of biological and biomedical research and
applications.
12
030
GR1HIGHCD11B+ CELLS SUPPRESS B CELL DIFFERENTIATION AND LUPUS-LIKE DISEASE IN
LUPUS-PRONE MALE MICE
Abhishek Trigunaite, M.Sc.1,2; Ayesha Khan, B.S.2; Evan Der2, Anne Song2, Sanjay Varikuti, M.S.2; Trine N. Jorgensen,
Ph.D.1,2.
1
Department of Biological, Geological and Environmental Sciences; Cleveland State University;
2
Department of Immunology, Lerner Research Institute, Cleveland Clinic
Objective: Systemic lupus erythematosus (SLE) develops much more readily in females than in males. In the past, research
has focused primarily on identifying mechanisms pertinent to the pathology in females, while little effort has been made to
delineate active protective mechanisms in males. In the current study we present evidence for a new male-associated
mechanism of protection from lupus-like disease development in lupus-prone (NZB x NZW)F1 mice.
Methods: We identified previously uncharacterized cellular and functional differences in myeloid cells between male and
female (NZB x NZW)F1 mice using flow cytometry, confocal imaging, in vivo antibody-mediated depletion and in vitro cell
co-culture assays.
Results: A population of Gr1highLy6G+CD11b+ myeloid cells was found to be constitutively elevated in male (NZB x
NZW)F1 mice as compared with females, and regulated by testosterone. The cells were located adjacent to spleen B cell
follicles in vivo and found to directly inhibit cytokine-induced differentiation of naïve B cells into antibody-secreting cells in
vitro. Most noticeable, treatment with anti-Gr1 depleting antibodies elevated spontaneous anti-nuclear autoantibody
production in male (NZB x NZW)F1 mice, while a similar approach in female mice had no effect on disease development.
Conclusion: Lupus-prone male (NZB x NZW)F1 mice harbor elevated levels of a population of myeloid cells with
pronounced immunosuppressive capacities specifically targeting B cells and antibody production in vivo. We suggest these
cells represent a male-driven inhibitory mechanism involved in the control of B cell pathogenesis, delaying (or preventing)
lupus-like disease development in otherwise genetically predisposed male (NZB x NZW)F1 mice.
031
SYNTHESIS AND ANTI-CANCER MECHANISM INVESTIGATION OF DUAL HSP27 AND TUBULIN
INHIBITORS
Bo Zhong, Ph.D.1; Snigdha Chennamaneni, B.S.P.S1; Rati Lama, B.S.1; Bin Su, Ph.D.1,2*
1
Department of Chemistry, Cleveland State University
2
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Heat shock protein 27 (Hsp27) is a powerful chaperone protein and its expression is increased in response to various stress
stimuli including anti-cancer chemotherapy, which allows the cells to survive in the lethal conditions and causes drug
resistance. Hsp27 is an attractive molecular target for anti-cancer drug development. We previously identified lead
compounds that bound to Hsp27 and tubulin through biotinylated small molecular probe, affinity chromatography and
proteomic approaches. Systematic ligand based optimization in the current study significantly increased the cell growth
inhibition and apoptosis inducing activities of the compounds. Furthermore, these compounds were examined with tubulin
polymerization assay and Hsp27 chaperone activity assay. Compared to the lead compounds, one of the new derivatives 20
exhibited much better potency to inhibit tubulin polymerization, but a decreased activity to inhibit Hsp27 chaperone function,
suggesting that the structural modification dissected the dual targeting effects of the compound. The most potent compounds
20 and 22 exhibited strong cell proliferation inhibitory activities at subnanomolar concentration against 60 human cancer cell
lines conducted by Developmental Therapeutic Program at the National Cancer Institute and represented promising
candidates for anti-cancer drug development.
032
ENCAPSULATION OF NITRIC OXIDE SYNTHASE OXYGENASE DOMAIN (NOS OXY) IN
ELECTROSPUN FIBERS: FABRICATING NITRIC OXIDE (NO) RELEASING ANTITHROMBORIC
MEMBRANES
Joseph B. Apisdorf, B.A.; BhagyaGunasekera, M.S.; Mekki Bayachou*, Ph.D.
Department of Chemistry, Cleveland State University
13
Nitric oxide synthase enzymes (NOSs) are responsible for the catalytic conversion of the amino acid L-arginine to Lcitrulline with the release of one equivalent of NO. Among other effects on the cardiovascular system, NO is known to
counteract platelet aggregation, and therefore has the ability to prevent the thrombosis cascade on the surface of bloodcontacting medical implants and vascular grafts. NO-releasing surfaces have been proposed to counteract thrombosis on the
surface of implanted devices. In the past, we proposed the use of NOS enzymes, in alternating layer-by-layer thin film
architecture, as the catalytic source of NO released using substrates of the NOS reaction in blood or plasma.
The objective of this work is to prepare and characterize membranes of electrospun fibers embedding NOSoxy domain as a
catalytic source of NO release, using polymer matrices, polycaprolactone (PCL) and Tecophilic®. The electrospinning
process uses a syringe containing the material to be electrospun; the latter is driven towards a collector drum by an electric
field.
The resulting material is characterized by a number of imaging methods, optical microscopy, and atomic force microscopy
(AFM). In addition, the NOS-modified electrospun membranes are further characterized electrochemically to evaluate the
presence and activity of iNOSoxy. To probe function, we analyze our iNOSoxy-containing material for enzymatic activity
under physiologic conditions by monitoring actual levels of NO released over time.
We will discuss our overall results, which, among other aspects, show that the native structure of the encapsulated iNOSoxy
is conserved in the pockets within the electrospun fibers. Also, we show that the entrapped iNOSoxy retains enzymatic
function and, under the right conditions, can sustain the release of nitric oxide. We will compare and contrast the results for
the various matrices and under the different conditions.
033
MOTIVATING FACTORS FOR PARTICIPATION IN EXERCISE IN AGED INDIVIDUALS
Philip D. Simon1, Bette R. Bonder, Ph.D.1,2
1
School of Health Sciences, Cleveland State University;
2
Department of Psychology, Cleveland State University
Introduction and Overview: The many benefits of physical activity for aged individuals have been reaffirmed by a large
amount of research. Unfortunately, there is a disparity between the awareness of these benefits and the proportion of aged
individuals who participate in the physical activity necessary to achieve these advantages. The existing literature has
adequately identified the motivation for aged individuals participating in exercise in a broad sense, but little is currently
known regarding their motivation for specific exercise decisions and actions.
Purpose: The aim of this study was to investigate factors that aged individuals identify as contributing to their decisions
about exercise—specifically frequency, type, and intensity.
Method: To investigate these issues, interviews were conducted that sought to discover factors that aged individuals identify
as contributing to their general decisions about whether or not to exercise, as well as about type, intensity, and frequency of
their exercise. Interviews were conducted with 15 adults of 65 years of age or older in whom no serious physical or cognitive
impairments were found. Recorded interviews were then transcribed, analyzed, and interpreted.
Findings: We found that most well aged individuals had positive outlooks on exercise and were primarily motivated to
exercise in general by a holistic outlook view of their health. Older adults in this sample cited factors such as abundant free
time and enjoyment of the activity as primary factors influencing their decisions regarding frequency, type, and intensity of
exercise. The findings of this study offer insights that may guide health care practitioners in developing more successful
interventions for the health education of aged individuals. It may also allow for physical activity programs to be designed and
executed that are consistent with the expressed motivations of older adults in order to maximize program effectiveness.
034
USING CHANGE DEAFNESS TO INVESTIGATE ATTENTIONAL ALLOCATION DURING SPOKEN
WORD RECOGNITION
Matt Wetzel; Samantha E. Tuft, B.A.; Maura L. Krestar, M.A.; and Conor T. McLennan, Ph.D.
Department of Psychology, Cleveland State University
14
Change deafness is a phenomenon in which listeners fail to detect auditory changes. This phenomenon can be used to analyze
the allocation of cognitive resources during spoken word processing. One theory suggests that when hearing spoken words
listeners allocate attention to different types of information (phonology or talker). The proposed study will consist of two
experiments. In Experiment 1, participants will perform a lexical decision task in which they will hear words and nonwords
and decide on each trial whether they are hearing a real word or a nonword. One group will hear taboo words, another will
hear neutral words. In Experiment 2, participants will perform a shadowing task in which they will repeat the word they are
hearing on each trial. One group will hear famous talkers, another will hear non-famous talkers. In both experiments, the
talker will change to a new but distinguishable talker half way through the experiment. In Experiment 1, we predict listeners’
attention will be pulled to the linguistic information for the group listening to taboo words. In Experiment 2, we predict
listeners’ attention will be pulled to the indexical information (the talkers) for the group hearing words spoken by famous
talkers. The degree of change deafness (the number of participants who fail to detect the change in talkers) will be analyzed
as a function of condition (taboo versus neutral; famous versus non-famous). We predict significantly more change deafness
in the taboo and non-famous conditions. That is, more participants should notice the talker change (less change deafness)
when hearing neutral (compared to taboo) words or words spoken by famous (compared to non-famous) talkers. The change
deafness paradigm is a novel way to examine listeners’ attentional allocation during lexical processing, and the current study
has implications for theories of spoken word recognition.
035
MOTOR SYSTEM EXPRESSIONS OF ANXIETY AND DEPRESSION
Ashley Chafin, B.S1., Andrew Slifkin, Ph.D.1
1
Department of Psychology, Cleveland State University
Physiologically healthy individuals have “complex interactions between multiple regulatory processes that function over
different time scales” (Lipsitz, 1994, p. 102). Such complexity of internal processes is reflected in the complexity of system
output, and loss of complexity in system output is a strong indicator of physical illness and aging (e.g., Lipsitz & Goldberger,
1992; Slifkin & Newell, 1998). However, research on health and complexity is limited to the study of physiological systems.
Few studies have addressed the relations between mental health and complexity (e.g., Bain et al., 2011; Balon, et al., 1992;
Thorton & Gilden, 2005). Here, in the current proposed research, we would examine the relations between depression and
anxiety and how each may be expressed as changes in complexity of motor output: The SCL-90-R would be used to assess
depressive and anxious intensity, and motor performance would be assessed by requiring participants to generate 500
consecutive aiming movements. Time series analysis -- in particular, spectral analysis -- would be used to quantify time
series complexity. We would predict that as anxiety and depression intensity increase across participants, there would be
correlated decreases in the complexity of motor output.
036
DECREASE OF STEADY-STATE LEVELS OF STAT BY NONSTRUCTURAL PROTEINS NS1 AND NS2
OF PNEUMONIA VIRUS OF MICE (PVM)
Jayeeta Dhar, MS; Tanmay Majumdar, Ph.D.; Valentina Verbovetskaya, MD; and Sailen Barik, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Pneumonia virus of mice (PVM) and human respiratory syncytial virus (hRSV) are enveloped, negative sense, singlestranded RNA viruses of the family Paramyxoviridae, subfamily Pneumovirinae, genus Pneumovirus. RSV is the leading
cause of pediatric illness; every year it causes more than 85,000 hospitalizations in USA alone. Antiviral therapy is not very
effective for hRSV. PVM is the only other virus related to RSV in Paramyxoviridae family. We are using the PVM infection
model because it reproduces many of the clinical and pathologic features of the more severe forms of RSV infection in
human infants. The main aim is to study the viral infection process and explore vaccination and immunomodulatory
strategies. Previous studies have shown that two non-structural (NS) proteins of RSV, namely NS1 and NS2 inhibit interferon
(IFN) α/β. Previously, our laboratory has shown that NS1 and NS2 of RSV, working singly as well as, together strongly
suppress the host cell's type I IFN-mediated innate immunity by degrading or inhibiting multiple cellular factors needed for
either IFN induction or response, including RIG-I, IRF3, IRF7, TBK1, and STAT2. Based on these results we hypothesized
that NS proteins of PVM may have a similar effect in IFN pathway. We created recombinant clones of these two NS proteins
of PVM. In our preliminary experiments, we have observed that the both NS1 and NS2 of PVM lower the steady-state level
15
of mouse STAT2 (signal transducer and activator of transcription), a necessary and essential transcription factor for IFN
inducible antiviral genes, in addition they affect human STAT2 and STAT1. In an attempt to understand the mechanism of
this loss we employed a series of recombinant STAT deletion mutants and STAT1-STAT2 chimera. Our preliminary results
offer a glimpse of STAT sequence regions, particularly the N-terminal sequence, that are important for this loss.
037
COORDINATION OF CONTINUOUS AND DISCRETE COMPONENTS OF ACTION
Stephanie L. Kilian, B.A.; Andrew B. Slifkin, Ph.D.; Ashley Chafin, B.S.; Jasmine M. Manista, B.A.; Patrick Byrne
Department of Psychology, Cleveland State University
In studies on human factors, human-computer interface, and ergonomics, a button press is used to indicate the end position of
a movement. In contrast, studies on movement control typically rely on the peak position of a movement trajectory to
indicate movement end, without any additional response. The main purpose of the current study was to determine whether the
same movement endpoint position is reflected by both the button press and the peak movement position. An experiment was
designed to examine changes in the relation between button press position and peak movement position as a function of
increases in movement difficulty, i.e., index of difficulty (ID). Participants within each condition made 1000 consecutive
targeted movements and were instructed to press a mouse button when they arrived within a target area. We recorded both
button press position and peak movement position. The conditions included two levels of ID (2 and 5 bits), and within each
ID level there was a small and large scale version of the target display. Analyses were based on the last 25 movements of
each condition. We hypothesized that the button press position would be coincident with the peak movement position across
ID levels. However, the button press occurred before the peak at ID 2, whereas the button press occurred after the peak at ID
5. Thus, the coordination between continuous hand (mouse) movement and a discrete finger (button) press can vary. As will
be discussed, the results suggest parallel processing of hand movement and button press information at ID 2, but serial
processing at ID 5.
038
ROLE OF CORE CIRCADIAN CLOCK PROTEIN BMAL1 IN CALORIE RESTRICTION MEDIATED
INCREASE IN LONGEVITY
Sonal A. Patel, B.S.; Yuliya V. Debrosky, B.S.; Rohini V. Khapre, B.S.; William E. Samsa, B.S.; Roman V. Kondratov,
Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease
(GRHD); Cleveland State University
Organisms from unicellular to complex multicellular are shown to exhibit behavioral and physiological rhythms (circadian
rhythms) that persist with the period of approximately 24 hour. These rhythms are generated by the internal system known as
the circadian clock. The clock is entrained by light and by feeding regimens. We recently found that the circadian clock
regulates longevity. Dietary restriction and especially CR (calorie restriction) has been shown to extend the lifespan of
diverse species and prevents various age related disorders. We hypothesized that the clock and clock genes play an important
role in calorie restriction mediated longevity. To test this hypothesis we performed an analysis of expression of clock
controlled genes by real time PCR in calorie restricted and ad libitum fed mice. We found that calorie restriction significantly
affects amplitude and phase of clock gene expression in wild type mice. Interestingly CR has little effect on the expression in
Bmal1 deficient mice (mice with disrupted circadian clock). These results suggest that CR does not extend longevity of
Bmal1 deficient mice.
039
CIRCADIAN REGULATION OF mTOR SIGNALING PATHWAY
Rohini V. Khapre, B.S.; Yuliya V. Dubrovsky, M.S.; Sonal A. Patel, B.S.; William E. Samsa, B.S.; Roman V. Kondratov,
Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease
(GRHD); Cleveland State University
16
Understanding mechanisms of aging is important for the treatment and prevention of age-associated pathologies. However
these mechanisms are not well understood. Recently we have demonstrated that the circadian clock (an internal time keeping
system) regulates longevity in mammals but molecular mechanisms are not known. The aim of our current study is to
investigate possible interconnection between the circadian clock and mTOR (mammalian target of rapamycin) signaling
pathway. mTOR pathway is nutrient response pathway involved in cell growth and proliferation; many recent studies indicate
role of mTOR as important regulator of aging. Here we demonstrate that circadian system regulates mTOR signaling in vivo.
Analysis of liver and heart from WT mice reveals circadian rhythms in phosphorylation of known mTOR targets such as
S6K1, 4EBP and others. These rhythms are disrupted in the tissues of Bmal1 knockout mice providing potential mechanistic
explanation of reduced longevity of these mice. Further analysis of expression of mTOR complex components and upstream
regulators demonstrated that some of them have circadian rhythms on protein and mRNA levels. Taken together, these results
suggest that circadian clock controls aging by regulating mTOR signaling pathway through BMAL1 dependent control of
gene expression.
040
THE EXPERIENCES OF PHYSICAL THERAPY STUDENTS DURING AN INTERNATIONAL SERVICE
LEARNING PROGRAM: A PHENOMENOLOGICAL STUDY
Karen E. Vitak, D.P.T.; Beth A. Ekelman, Ph.D., D,OTR/L; Deborah D. Espy, Ph.D., PT; Nada S. Salem; Stephanie L.
Wisniewski
School of Health Sciences, Cleveland State University
The purpose of this phenomenological qualitative study was to explore the experiences of 6 entry-level Doctor of Physical
Therapy students as they participated in a service learning program in Belize, Central America. Participants wrote daily
reflections in personal journals which were transcribed verbatim. Researchers completed open, axial, and selective coding to
capture the essence of the participants’ experience. Results were triangulated using multiple investigators. Three themes
emerged from the data: cultural immersion and enlightenment; establishment of meaningful relationships; and personal and
professional growth and reflection. The participants were able to gain an appreciation for another culture besides their own in
which the connections they made with their patients aided to their overall experience. Findings reinforce the notion that
implementing a service learning immersion experience promotes cultural awareness, overall enrichment, and further
development of a more well-rounded healthcare provider.
041
SURFACE GLYCO-FUNCTIONALIZED LIPOSOMES BASED ON DIFFERENT ANCHORING LIPIDS
Pratima Vabbilisetty, M.S. and Xue-Long Sun, PhD*
Department of Chemistry, Cleveland State University
Liposomes have been widely used as biomimetic models of cell membranes and as drug/gene delivery carrier systems.
Surface modification of liposomes facilitates for many enormous potential applications, such as enhanced stability, bioactive
liposome conjugates, and targeted drug/gene delivery. Anchoring lipids are needed for grafting ligand of interest onto the
liposome surface by direct liposome formation, post-insertion, or post-coupling of the preformed vesicles. In addition, the
anchoring lipids play important roles in liposome chemical and physical characteristics and liposome stability as well. In this
report, glyco-functionalized liposome systems based on two kinds of anchoring lipid, phosphatidylethonalamine (PE) and
cholesterol (Chol) were prepared by post chemically selective functionalization via Staudinger ligation. The size and stability
of the liposomes were confirmed by dynamic light scattering (DLS) and fluorescent dye release kinetics. Particularly, anchor
lipid effects on the stability of glyco-functionalized liposomes were investigated comparing two different anchor lipids,
namely CholPEG2000-TP and DSPE-PEG2000-TP. Furthermore, the density and accessibility of grafted carbohydrate residues
on the liposome surface were evaluated with Lectin binding assay and the encapsulation efficiencies of the different anchor
lipids were also determined.
17
042
THE EFFECT OF BILINGUALISM ON SPATIAL MEMORY
Jayleen A. Meléndez; Naohide Yamamoto, Ph.D.
Department of Psychology, Cleveland State University
Prior research on spatial memory has suggested that when individuals learn a spatial layout of objects from more than one
perspective, they tend to remember object locations more accurately or quickly from one particular preferred perspective than
from other learned perspectives. The objective of the present study is to explore conditions in which individuals can establish
more than one preferred perspective. To this end, we will examine whether English-Spanish bilinguals can remember two
perspectives of a spatial layout equally well when they learn them in different languages. The bilingual participants will be
given two sets of verbal descriptions that describe the same object locations from two different viewpoints. These
descriptions will be written either in the same language (English or Spanish; monolingual conditions) or different languages
(one in English and one in Spanish; bilingual condition). After learning the two different perspectives, participants' memories
for the object locations will be tested without using language. It is predicted that participants in the bilingual condition will be
able to maintain two preferred perspectives in memory, one learned in Spanish and another learned in English. On the other
hand, participants in the monolingual conditions are expected to have only one preferred perspective, as has been the case in
the literature. These results would broaden our understanding of how spatial memory is affected by language in general and
by bilingualism in particular.
043
INTERPROFESSIONAL EDUCATION IN REHABILITATION: A SYSTEMATIC REVIEW
Lori D. Chandar, B.H.S.; Kelly M. Mangan, B.A.; Kim M. Mangan, B.A.; Catherine M. Parrino, B.F.A.; Karen M. Keptner,
MS, OTR/L
School of Health Sciences, Occupational Therapy Program, Cleveland State University
A review of literature reveals that the majority of errors in medicine are due to poor communication amongst healthcare
providers. In response, inter-professional education (IPE) has gained popularity at universities in an effort to prepare
professional healthcare students to engage in quality collaboration once they are clinicians. Various obstacles that impede the
acceptance and implementation of IPE include: a poor understanding of the roles of other disciplines, confusion in what IPE
entails, lack of funding and staff, and scheduling conflicts amongst university departments for the purpose of training. The
aim of this systematic review was to determine if the research on IPE is strong enough to guide the creation of a model of IPE
for professional rehabilitation programs. A keyword search yielded 2,239 articles. After titles and abstracts were screened for
relevance, the remaining articles were screened for rigor. Results and implications of this systematic review look promising.
044
NOVEL ROLE FOR MYOD AS A SENSOR OF DNA DAMAGE
Gregory M. Kliment, B.S. and Crystal M. Weyman, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Apoptosis (programmed cell death) is induced at the same time as differentiation (specialization) in skeletal myoblasts as
well as other systems. In skeletal myoblasts, these two processes are both initiated by culture in differentiation media (DM:
absence of growth factors) and result in mutually exclusive physiological endpoints. However, the simultaneous regulation of
these two processes is not understood. While the muscle regulatory transcription factor MyoD is known to control the process
of differentiation in skeletal myoblasts, our lab has recently discovered that MyoD is also controlling the apoptotic process in
response in culture to DM by direct up-regulation of the pro-apoptotic Bcl2 family member PUMA. We have also discovered
that, through direct up-regulation of PUMA, MyoD plays a role in the apoptotic process initiated by the DNA damaging
agent, etoposide. Our hypothesis is that a novel role for MyoD as a sensor of DNA damage could correlate both of these
findings. We are testing our hypothesis by comparing the activation of key checkpoint kinases (Chk1, p38, c-abl and ATM)
in response to culture in DM for various time-points with the activation of these kinases in response to culture in Growth
Media (GM) supplemented with etoposide.
18
045
THE YELLOW BRICK ROAD: AN OCCUPATIONAL JOURNEY INTO RETIREMENT
Mary K. Molnar, B.A.; Adina A. Morgan, B.S.H.S.; Vivian R. Atsu, B.A.; Samantha Wilds, B.S.H.S.; Stacey L. Nichols,
B.S.; John Bazyk, M.S.
School of Health Sciences, Occupational Therapy Program, Cleveland State University
Predictions pertaining to the growth in number of retired people and those transitioning to retirement, sparked by our nation’s
shifting demographics, acknowledge the inevitability of the social consequences. Individuals approaching the transition to
retirement, and ultimately retirement, face many challenges. Among those challenges are financial preparedness, physical
health, and psychosocial/emotional health. Psychosocial/emotional well-being presents a major concern for those making the
transition, yet receives less emphasis than its counterparts. To better understand the barriers and facilitators of well-being
during retirement we asked the question: what factors are associated with wellbeing in retirement and what are the lived
experiences of those transitioning from worker to retiree? After conducting an extensive literature review five themes
emerged that strongly correlated to positive well-being after retirement; they were: volunteering, planning and choice
regarding the time and circumstances of retirement, having a positive attitude, the importance of peer support and
continuity/occupational identity. With the intention to increase the breadth and depth to the current body of knowledge
regarding the lived experience of those transitioning from worker to retiree we conducted a qualitative research study using a
grounded theory approach. We recruited and interviewed a convenient sample of five community dwelling retirees.
Interviewers conducted one-on-one, semi-structured interviews lasting approximately 60 minutes. Interviews were recorded,
transcribed, and analyzed for common themes. Through this process we identified three common themes: continuity,
engagement, and financial preparedness. Retirees reporting high levels of continuity, engagement, and financial preparedness
experienced smooth, swift transition into retirement and subsequent well-being throughout retirement. Occupational therapy
practitioners play a major role in bolstering engagement in meaningful occupations and promoting continuity across the
lifespan despite age or disease related deficits. In this way, occupational therapists can facilitate productive, healthy lifestyles
that increase life satisfaction and well-being for those embarking on their journey into retirement.
046
CHAIN-END FUNCTIONALIZED GAGS FOR BIOORTHOGONAL CONJUGATION AND GLYCOREENGINEERING OF RECOMBINANT THROMBOMODULIN
Jacob Weingart, Ph.D.1, Rui Jiang, Ph.D.1, Nikola Paulic, B.S.1, and Xue-Long Sun, Ph.D.1,2
1
Department of Chemistry, Cleveland State University
2Departmetn of Chemical and Biomedical Engineering, Cleveland State University
Glycosaminoglycans (GAGs) are linear polysaccharides composed of two alternating sugars, usually an uronic sugar and an
amino sugar, with varying patterns of sulfation determining the type. Found throughout the body displaying high degrees of
heterogeneity, GAGs are involved in many complex biological and chemical signaling events such as CNS development,
coagulation, and cellular signal transduction. Thrombomodulin (TM) is a glycoprotein that contains a GAG domain which
contributes TM’s thrombin binding and stability and plasminogen activation activities. Previous research has concluded that
this domain, containing chondroitin sulfate (CS), can inhibit the coagulation process by enhancing the binding affinity of the
integral membrane protein thrombomodulin to thrombin 10- to 20-fold, subsequently activating Protein C and
downregulating clotting Factors Va and VIIIa. We hypothesized that incorporation of CS or heparin GAGs into the
recombinant thrombomodulin (rTM), the active domains 4-6 of the native protein, should enhance the binding affinity to
thrombin creating a more effective antithrombotic. This presentation reports the synthesis and characterization of chain-end
functionalized CS and heparin derivatives, which will be used to investigate various bio-orthogonal conjugation reactions in
an attempt to create novel GAG-rTM conjugates to test their effects on rTM’s binding affinity to thrombin.
19
047
OCCUPATIONAL THERAPY PROGRAM SERVICE LEARNING
Beth Ekelman, Ph.D., JD, OTR/L.; Robert Griffiths, B.S.; Beth Iorio, B.A.; Olga Wood, B.S.;Shannon Yarborough, B.S.
School of Health Sciences, Master of Occupational Therapy Program, Cleveland State University
Currently, students from disadvantaged socioeconomic and/or educational backgrounds are not well-represented as healthcare
professionals. These students face many barriers that make it difficult for them to transition to college and complete their
studies successfully. Students from the Cleveland Public Schools and inner ring suburbs face similar obstacles to obtaining an
undergraduate education. Mentoring programs have been shown to promote competence, relatedness, and autonomy for
participants (Britner, et al., 2006). To help entering freshman interested in healthcare professional programs to succeed at
CSU, Beth Ekelman, Associate Director of the School of Health Sciences and Associate Professor of the university’s Master
of Occupational Therapy Program, is developing a service learning mentoring program which will target incoming freshman
students from Cleveland Public Schools and inner ring suburbs who are interested in pursuing a health career. This mentoring
program will pair incoming freshman from disadvantaged backgrounds with occupational therapy students enrolled in a
service learning course. The occupational therapy student mentors will help and encourage their mentees to: develop time
management and life balance skills, attend campus extracurricular programs, establish relationships with peers, and engage in
a community service activity. OBJECTIVES: 1.Participants will apply time management strategies to create schedules. 2.
Participants will participate in campus activities and meet peers. 3. Participants will complete a community service activity.
OUTCOME MEASURES: The success of the mentoring program for the mentee participants will be measured by individual
interviews and focus groups, as well as by the Academic Attitudes Scale, (Wong, 1998) the Sense of Belonging
Questionnaire, (Hoffman et al., 2003) and the Student Adaptation to College Questionnaire (Bake & Siryk, 1999).
048
THE EFFECTS OF INTER-TALKER VARIABILITY ON LISTENERS’ PERCEPTION OF SPOKEN
TABOO WORDS
Samantha E. Tuft, B.A.; Conor T. McLennan, Ph.D.; and Maura L. Krestar, M.A.
Department of Psychology, Cleveland State University
In the current experiment, we examined the effects of inter-talker variability on the listeners’ perception of spoken taboo
words. Previous spoken word recognition research using the long-term repetition-priming paradigm, in which listeners
respond to two separate blocks of spoken words, found performance costs for stimuli mismatching in talker identity. That is,
when words were repeated across the two blocks and the identity of the talker remained the same (e.g., male to male) reaction
times (RTs) were faster relative to when the repeated words were spoken by two different talkers (e.g., male to female). Such
performance costs, or talker effects, followed a time course, occurring only when processing was relatively slow. More recent
research has found that explicit and implicit attention towards the talker led to talker effects (even during relatively fast
processing). The purpose of the current study was to examine how word meaning could affect the pattern of talker effects.
Participants completed an easy lexical decision task and participants’ mean accuracy rates and RTs were analyzed. We
hypothesized that hearing taboo words would surprise the listeners and grab their attention, such that talker effects are
obtained even when processing is relatively fast. The results are consistent with the attention-based hypothesis that talker
effects emerge when participants hear both spoken taboo and neutral words. In addition, taboo words were responded to
faster than neutral words, suggesting that spoken word recognition can be affected by word meaning. The results of the
current study have important implications for theoretical models of spoken word recognition and how attention plays a role.
049
THE EFFECTS OF STRESSFUL LIFE EVENTS, THEIR RELATIONSHIPS AMOUNG
DEMOGRAPHICS, AND THE IMPLICATIONS THEY HAVE ON MENTAL HEALTH
Katie M. Gonzalez and Boaz Kahana, Ph.D.
Department of Psychology, Cleveland State University
Stress had been a concern in human health for as long as humanity has existed, but it is more of a concern in today’s society
for various reasons. The purpose of this study is to examine stress on many different levels and its role in the lives of
different individuals. Our sample will consist of college students with a variety of backgrounds and demographics. Past
20
studies have shown differences in stress and stress coping methods between men and women and have shown differences in
behaviors following stress. This study will examine the relationship of stressful life events and trauma to mental health
indicators such as depression and anxiety. The data will then be compared based on different demographics. Managing stress
and learning the most effective ways to cope with stress seem to be important factors in the overall health of individuals
within our society. Educating these individuals on the effects of stress and beneficial ways of coping with stress would be
expected to improve health overall.
050
TWO MOUSE STRAINS REGULATES CHOLESTEROL ESTER METABOLISM
Brian M. Ritchey, B.S.1,2; Peggy Robinet, Ph.D.2; Jonathan D. Smith, Ph.D.2,3
1
Department of Chemistry, Cleveland State University
2
Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic
3
Department of Molecular Medicine, Cleveland Clinic Lerner College of Medicine of Case Western Reserve University
Atherosclerosis is a complex, multi-factorial disease characterized by the accumulation of lipids in large arteries.
Macrophages are key drivers of atherosclerotic plaque formation, acting to activate inflammation and increase lipid burden in
the artery wall. Advanced plaques can rupture and trigger life-threatening ischemic events such as heart attack and stroke. In
fact, atherosclerosis is the leading cause of cardiovascular disease and death in the world.
Previous work in the Smith laboratory has demonstrated that genetic background is a major determinant of atherosclerosis
susceptibility in mice, where DBA/2 apoE-/- mice develop ten times larger aortic root lesions than AKR apoE -/- mice. My
research focuses on the study of foam cells generated in culture from these two strains.
Foam cells were generated utilizing bone marrow derived macrophages derived from AKR and DBA/2 mice using a range of
acetylated LDL concentrations. A striking finding from this study was that DBA/2 foam cells accumulated about 2-3 times
more cholesterol ester than AKR foam cells, while AKR foam cells accumulated roughly twice as much free cholesterol.
Investigation into this phenotype revealed that the rate of cholesterol ester hydrolysis was more than two-fold higher in AKR
foam cells vs. DBA/2. A subsequent study, utilizing an inhibitor of lysosomal acid lipase, demonstrated that macrophage
cholesterol ester hydrolysis is mediated primarily through this enzyme. It was also determined that autophagic flux was
significantly faster in AKR foam cells, mirroring the cholesterol ester hydrolysis findings.
The role of autophagy in macrophage cholesterol ester hydrolysis will be investigated with greater precision going forward.
Gene mapping, using the hybrid mouse diversity panel, and lipid droplet proteomic studies will be conducted with the goal of
eventually elucidating genes and/or cellular processes that contribute to the cholesterol ester storage phenotype, as well as the
difference in atherosclerosis susceptibility.
051
CAN VIEWING IMAGES OF THIN WOMEN NEGATIVELY AFFECT EMOTION RECOGNITION
PERFORMANCE?
Kim D. Thomas, B.A., Teresa A. Markis, M.A. and Conor T. McLennan, Ph.D.
Department of Psychology, Cleveland State University
Images depicting thin women can lead to body dissatisfaction. Social Comparison Theory explains this with two processes.
They are known as “upward comparison,” which is defined as comparing oneself to a more attractive image and “downward
comparison,” which is defined as comparing oneself to a less attractive image. Upward comparison can result in depression
and body dissatisfaction. Downward comparison can result in an increased body image. Extreme body dissatisfaction is a
core feature of eating disorders. Previous research has shown that individuals with eating disorders, such as anorexia, have a
deficit in emotion recognition and processing. Building upon this theory, this study is designed to examine the effect of
priming normal individuals (those without an eating disorder) with pictures depicting thin women on their performance on an
emotion recognition task. Prior to coming to the lab, participants completed The Eating Disorder Inventory. Once in the lab,
participants were placed in one of three priming groups: thin-ideal prime, overweight prime, or a neutral prime. After
priming, all participants performed a facial emotion recognition task. In this task, participants viewed 40 faces on a computer
screen and were asked to choose one of four emotions (sad, angry, surprise, or happy) to describe the face. Based on previous
21
findings, it was hypothesized that the individuals primed with the thin images would take longer to respond, and make more
errors in recognizing the emotions than both the neutral and overweight prime groups. Results of this experiment will be
discussed.
052
SIALIC ACID QUANTIFICATION WITH LC-MS/MS METHOD
Dan Wang, M.A., Xiang Zhou, Ph.D., Xue-Long Sun, Ph.D.
Department of Chemistry, Cleveland State University
Sialic acids (SAs), a family of 9-carbon containing acidic monosaccharides, often terminate the glycan structures of cell
surface and secreted glycoconjugates such as glycoproteins and glycolipids. SAs play crucial roles in many physiological and
biological processes. It has been demonstrated that cancers and cancer staging may be associated with a significant
overrepresentation of SA on the surface glycoproteins of cancer cells compared with normal cells. Also it is well known that
the amount of free SA and lipid- and protein-bound SA is elevated in plasma from cancer patients compared with healthy
individuals. Therefore, SA has been considered as a biomarker for certain types of cancers and some other diseases. Due to
the great significance of SA in physiological and clinical applications, several analytical methods have been developed for
SA measurements. These include colorimetric, fluorometric, and chromatographic methods. However, all of these methods
suffer from some limitations, such as less sensitive and interference when dealing with biological samples. In this work, a 3,
4-diaminotoluene derivatization of SA combined with LC-MS/MS was developed for sensitive and robust quantification of
SA. The method had a linear calibration range of 1.0 to 1.0 × 10 3 ng/mL and a lower limit of quantification of 1.0 ng/ml. The
derivatization product was stable for at least 3 weeks in -20 oC. The repeatability of derivatization was carried out in 7
consecutive days. The relative error (RE) was from -7% to 7%. This method could be used for SA quantification of biological
samples.
053
PREFERENCE FOR COLORED NOISE
Katrina B. Wolfgang and Andrew B. Slifkin, Ph.D.
Department of Psychology, Cleveland State University
Pink noise output is found in a range of biological and physical systems. Previous studies have shown that speech and many
popular forms of music have a pink (1/f) noise structure (Voss and Clarke, 1978; Ro and Kwon, 2009).
Because composers have composed music with a pink noise structure, it follows that people should prefer sound with that
structure. Otherwise, it would have been expected that music would have been composed with a different structure. The
current study would investigate peoples’ preferences for sound with three different types of colored noise structures: white
noise, pink noise, and brown noise. By using noise sound samples rather than real music samples, sound would be stripped of
associations that might bias preference, namely, musical artist and musical genre. Here, participants would be presented with
the three different types of colored noise samples, and each type would be presented at three different volumes. We would
predict that pink noise would receive the highest preference scores. Knowing which type of noise is most preferred has
potential applications. It has become increasingly popular to use noise to aid sleep, to enhance performance, to reduce outside
noise, and even for enjoyment. White noise has mostly been used for those purposes. However, it could be that it would be
better to use the most preferred type of noise instead.
054
PLATELET IL-1: AUTOCRINE SIGNALING IN LIPOPOLYSACCHARIDE ACTIVATION AND
THROMBOSIS
Padmini Narayanan, MSc.1,2; G.Thomas Brown, MD.Ph.D.1; Roy L. Silverstein, M.D.,Ph.D.3; Thomas M. McIntyre, Ph.D.1
1
Department of Cellular and Molecular Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH
2
Department of Biological, Geological and Environmental Sciences, Cleveland State University,
Cleveland, OH
3
Medical College of Wisconsin, Milwaukee, Wisconsin.
22
Platelets are critical components of the septic response to Lipopolysaccharide (LPS) through MyD88 coupled –TLR4
receptor signaling. This results in stimulated splicing of stored intron containing IL-1 hetronuclear (hn-RNA) RNA,
translation of newly formed mRNA, pro-IL-1 cleavage to active cytokine and its release along with microparticles and
exosomes. Thrombin also stimulates IL-1 hnRNA splicing and microparticle sedding but results in poor pro-IL-1
translation. We report that platelets express functional IL-1R1 receptor and responded to recombinant IL-1β or IL-1b
associated with particles shed by LPS stimulated platelets. Specific blockade of IL1R1 with IL1Ra (Anakinra ®) suppressed
this, so IL-1 stimulated its own synthesis in an autocrine signaling loop. Platelet microvesicles shed upon LPS stimulation
were capable of activating endothelial cells (HUVECs) and HEK-BLUE – IL-1 sensor cells (for detection of bioactive IL1 ) and the activity was abrogated with the pre-incubation of IL-1Ra. IL-1 rapidly accumulated within occlusive thrombi
formed in intact, and sterile, murine carotid arteries after transmural oxidative damage initiated by ectopic FeCl 3. Thrombus
IL-1 associated with platelets, and later with infiltrating monocytes. We infer that IL-1 produced upon stimulation is
different from the short-lived secretory form of the protein as accumulation of IL-1 is observed upon thrombus maturation.
Results suggest IL-1 protein newly synthesized upon LPS activation, are associated with the shed particles are biologically
active, and induce a stimulatory loop in platelets through IL1R1, this IL-1 autocrine loop greatly amplifies LPS activation.
055
ANALYZING THE WAYS IN WHICH PARENTS DISCUSS THE DIAGNOSIS OF AUTISM SPECTRUM
DISORDER WITH THEIR DIAGNOSED CHILDREN
Arielle N. Bowers1; Christopher M. France, Psy.D.1
1
Department of Psychology, Cleveland State University
The prevalence of autism spectrum disorders (ASDs) such as Autistic Disorder and Asperger’s Disorder has increased in
industrialized nations in recent decades. According to a recent study released by the U.S. Department of Health and Human
Services, 1 in 50 school-aged children have been diagnosed with ASD in the United States (Blumberg et al., 2013). One issue
faced by parents of children with an ASD is when, how, and (in some cases) whether to discuss the diagnosis with the
affected child. Little systematic research has been conducted to date regarding the above issues. The primary purpose of this
study is to gather information from parents of children with ASDs regarding whether, when, and how the parents discussed
the child’s ASD diagnosis with the child. We further hope to understand whether the parents view such diagnosis-related
discussions as helpful, are satisfied with their chosen timing of the discussion or other aspects of the information provided to
the child as part of the discussion, and how the information appears to have affected the diagnosed child. A survey design
incorporating closed and open-ended questions and Likert-type ratings will be utilized. Parents of children with ASD
diagnoses will be recruited both on the Cleveland State University Campus and at the Cleveland Clinic Center for Autism. It
is hoped that the results of this study will be useful in considerations of how best to discuss an ASD diagnosis with a
diagnosed child.
056
DOES DURATION AND DEGREE OF MINDFULNESS AFFECT IMPROVEMENT IN BALANCE
DURING THE IMPLEMENTATION OF A BALANCE EXERCISE INTERVENTION?
Kathleen J. Pantano, PT, PhD1; Jeremy E.C. Genovese, PhD2; David Schordock, SPT1; Umar Alabasi, BSc, MS, PT3
1
School of Health Sciences, Doctor of Physical Therapy Program, Cleveland State University;
2
College of Education and Human Services, Department of Curriculum and Foundations, Cleveland State University;
3
Department of Physical Therapy, King Abdulaziz University, Jeddah, Saudi Arabia
Mindfulness is described as the process of regulating one’s attention to bring out a quality of awareness to a current
experience, embracing aspects of curiosity, experiential openness, and acceptance (Bishop et al., 2004). Current evidence
suggests that mindfulness affects postural control, (Kee, et al., 2012, Mills & Allen, 2000), walking speed (Djikic et al.,
2008), and exercise intentions (Chatzisarantis & Hagger, 2007). The purpose of this study was to determine how mindfulness
affects balance improvement in physically active young adults during the implementation of a 3-week vs. 6-week balance
exercise intervention. Thirty-three physically active college students (24 female, 9 male) between the ages of 18-35 (24.8 ±
3.6) were recruited for the study after IRB approval. The Freiburg Mindfulness Inventory (FMI) (Walach et al., 2006) was
used to assess mindfulness at the initial and final stages of a balance exercise intervention. Balance outcome measures (the
BESS test and Y-balance test) were taken at initial, 3 and 6-week intervals. The balance exercise intervention consisted of a
23
warm-up and cool down period, and 8 progressive static and dynamic balance exercises. Subjects attended 2 exercises
sessions per week for 6-weeks. Paired t-tests were used to assess change in scores from baseline. In order to control for
multiple comparisons the significance level was set to p < 0.01. The results show statistically significant improvement in
balance on both the BESS test and the Y-balance test at 6-weeks, but not at 3-weeks. After 6-weeks of balance training,
participants showed a statistically significant increase in mindfulness as measured by the FMI. The results of this study are
important because they suggest that balance training might be an alternative to meditation as a means of increasing
mindfulness. Additionally, these results tell us that balance training must be of sufficient duration to have a positive effect on
balance.
057
A DEFICIENCY IN THE CIRCADIAN CLOCK PROTEIN BMAL1 RESULTS IN OSTEOPOROSIS
William Samsa1, BS; Anna Kondratova2, Ph.D.; Ronald Midura3, Ph.D.; and Roman Kondratov1, Ph.D.
1
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University;
2
Department of Molecular Genetics, Lerner Research Institute, Cleveland Clinic;
3
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic
Circadian Rhythms are approximately 24hr rhythms in physiology and behavior (Circa = Around; Diem = Day). The
circadian clock evolved as an adaptation to the Earth’s daily light/dark cycle. The molecular circadian clock is an
endogenously driven timekeeping device present in almost all organisms. At the core of the molecular circadian clock is
Brain and Muscle Arnt-like protein 1 (BMAL1). Recently, our lab has shown that a deficiency of BMAL1 in mice results in
progeria with accompanying age associated pathologies, including osteoporosis. Osteoporosis is a bone disease characterized
by decreased bone mass and bone mineral density, resulting in an increased risk of fractures. We have shown that along with
decreased bone mass, bones from BMAL1 deficient mice display an early closure of the epiphyseal growth plate, resulting in
shorter bones. Using micro-computed tomography scanning, we have shown that these bones have altered microarchitectural
parameters compared to wildtype bones, including decreased cortical and trabecular bone. To study the mechanism of bone
loss, we have examined the differentiation ability of mesenchymal stem cells (MSCs) into osteoblasts in wildtype and
BMAL1 deficient mice. We have found that the differentiation capacity of BMAL1 deficient MSCs is reduced, and that there
is a decreased ability of osteoblasts to form a mineralized extracellular matrix. Using the MC3T3 preosteoblastic cell line, we
have shown that osteoblastogenesis affects the expression of core circadian clock genes. Our data indicate that BMAL1 plays
an important role in bone homeostasis by positively regulating bone formation.
058
STAT6 TRAFFICKING AND ACTIVITY IN RENAL EPITHELIAL CELLS STIMULATED BY FLUID
FORCES, INTERLEUKIN-4 AND INTERLEUKIN-13
Brianna Boslett, B.S.1; Andrew Resnick, Ph.D.1,2
1
Department of Biological, Geological and Environmental Sciences; Cleveland State University
2
Department of Physics
The primary cilium is a solitary organelle comprised of microtubules that project from the apical surface of non-dividing
mammalian cells. In the nephrons of the kidney, epithelial cells line the tubule lumens with the primary cilium projecting into
the luminal space, exposed to flow of ultrafiltrate. In general, blood circulation and glomerular filtration generate a relatively
constant fluid flow through the tubules. Several pulses of flow are needed to bend the cilium, which then initiates numerous
signal transduction pathways such as calcium signaling, mTOR, Jak/STAT, Wnt, and Hedgehog signaling. Fluid flow, ciliary
bending and Ca2+ signaling are present in a healthy kidney and result in transcriptional inhibition of proliferative genes,
although the specific mechanisms are unknown. The relation between flow sensing by the primary cilium and cyst formation
is still unclear.
Current cell culture practices maintain cells in a stationary, or no-flow, environment. For cell types that are regularly in the
presence of flow in vivo, typical culture methods may limit the ability to gather applicable information about function. This
experiment will utilize a two-sided flow chamber to expose differentiated monolayers of renal epithelial cells to chronic fluid
flow. Using this technique, downstream Jak/Stat signaling initiated by primary cilium mechanosensation will be examined.
Specifically, this study will compare the STAT6 activity stimulated by fluid flow, to the STAT6 activity stimulated by IL4
24
and IL13, for which the mechanism has been well studied. STAT6 activity will be quantified in each condition by western
blotting using a phospho-STAT6 antibody.
Mccd 1296 (d) cells will be used to study the effects of renal fluid flow on Jak/STAT signaling by analyzing STAT6
activation in response to different physical and chemical stimuli normally present in healthy and/or disease states in vivo.
Future research will compare gene expression in each experimental condition.
059
EVAULATING THE EFFICACY AND POTENCY OF NON-NATURAL NUCLEOSIDES AS
THERAPEUTIC AGENTS AGAINST PEDIATRIC CANCERS
Neethika Yeruva, BS 1; Sravya Kanneganti, BS 1; Praveena Basireddy, BS 1; Lahari Komatireddy, BS 1; Anthony Berdis,
Ph.D. 1, 2
1
Department of Chemistry, Cleveland State University
2
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Approximately 10,000 children are diagnosed each year with leukemia or brain cancer. These malignancies are not only the
most common forms of pediatric cancer, they are also the most devastating as ~1,500 children will die within a year of
diagnosis. Although several treatments are available, chemotherapy is often the best choice against childhood cancers.
Indeed, nucleoside analogs such as Fludarabine and Gemcitabine can effectively treat pediatric blood cancers such as acute
lymphoblastic leukemia. However, these conventional nucleoside analogs are ineffective against most tumors including brain
cancers. In this report, we present data from a series of pharmacological studies testing the ability of non-natural nucleosides
to function as therapeutic agents against glioblastoma, the deadliest form of pediatric brain cancer, and acute lymphoblastic
leukemia (ALL), the most prevalent form of childhood leukemia. Our non-natural nucleosides lack conventional hydrogenbonding groups on the nuclobase yet retain a natural deoxyribose moiety. Both features are predicted to provide higher
potencies toward inhibiting enzymes involved in nucleic acid synthesis as well as increasing transport and metabolism. Our
results show that Fludarabine exhibits potent cytostatic and cytotoxic effects against a leukemia cell line (MOLT4) yet is
ineffective against the human brain cancer cell line (U87). Several of our 5-substituted indolyl-2’-deoxynucleosides display
potent anti-cancer effects against MOLT4 and U87 cells. Analogs containing large substituent groups (phenyl or cyclohexyl
moieties) show equal efficacy against leukemia and brain cancer cells. Likewise, analogs with smaller substituent groups
(ethyl and ethylene groups) show cytostatic and cytotoxic effects against both cancers. However, one analog designated 5ethylene-indolyl-2’-deoxynucleoside (5-EyIdR) is more potent against U87 cells compared to MOLT4 cells. This higher
potency suggests a unique mechanism of action toward killing these cells. Current studies are underway to define the
selectivity of 5-EyIdR against brain cancer cells and identifying the molecular target(s) associated with inducing cell death.
060
FROM START TO FINISH: STUDENT BELIEFS AS TO STRATAGIES FOR IMPROVING
UNDERGRADUATE RETENTION RATES
Peter M. Arian; Christopher M. France, Psy.D.
Department of Psychology, Cleveland State University
This study will investigate undergraduate retention from the perspective of current undergraduate students. We will use a
variety of quantitative measures (e.g., categorical questions, Likert-type ratings, etc.) as well as qualitative (e.g., open ended
questions). This study has been designed in two parts. One part looks into what factors may play a significant role in causes
and conditions linked to undergraduate dropout rates. The other part of this study explores several strategies that could
possibly be implemented by institutions as ways of combating decreasing undergraduate retention. In this study we will
attempt to not only identify factors contributing to undergraduate dropout rates, an area that has already received sizable
attention, but also to attempt to gather information exploring undergraduate beliefs as to strategies for improving college
retention rates. We believe that the information obtained from this survey will be useful for institutions and/or individuals in
finding effective strategies for improving retention at the undergraduate level.
25
061
THE EFFECTS OF EMOTIONAL STATES ON SELF-MOTION PERCEPTION
Kimberly Moran; Naohide Yamamoto, Ph.D.
Department of Psychology, Cleveland State University
As a reaction to developing a high state of physiological arousal, an individual would experience a fight or flight response in
hopes of increasing their likelihood of survival. During said response, the individual’s bodily system and functioning may be
adjusted in hopes of compensating for any inadequacies that may serve as an obstacle during their encounter with a potential
threat. Consequently, the individual’s senses and perceptions may change. The present study will investigate the effects of
emotional states on perception of walked distance. Blindfolded participants will be presented with aversive and neutral
auditory stimuli. Upon this exposure, the participants will either walk to a previewed target (target-directed walking) or be
guided along a linear path and verbally estimate the distance walked (experimenter-guided walking). We hypothesize that the
participants would underestimate their self-motion in the presence of the aversive sound because underestimation would help
them avoid the aversive stimulus; that is, if they thought they had walked shorter than they actually did, it would prompt
them to walk more, thereby helping them to move farther away from the aversive stimulus. Thus, we predict that in the
experimenter-guided walking, the participant would underestimate walked distance. On the contrary, in the target-directed
walking, the participant would overshoot the given target. These results would suggest that perceptual processes can be
adaptively modulated by internal states such as emotion.
062
SWAPPABLE RNA STRUCTURES IN THE MINOR SPLICEOSOME
Jagjit Singh, MS; Kavleen Sikand, Ph.D.; Girish C. Shukla, Ph.D.
Department of Biological, Geological and Environmental Sciences; Cleveland State University
In metazoans almost 1% of the total introns are U12 dependent and are spliced by distinct machinery known as minor class
spliceosome. snRNAs in U12 dependent spliceosome are functional analogs of U2 dependent spliceosome. U12 and U6atac
snRNAs are the important components of the catalytic core of the minor class splicing. U12 binds to the branch point
whereas, U6atac binds to the 5' splice site after the U11 interactions are destabilized. The 3' stem loop of the U6atac RNA is
considered as the guide element in minor class splicing. Various inter- and intramolecular RNA-RNA interactions are
facilitated by spliceosomal proteins and other splicing factors. p65 RNA binding protein, a component of U11/U12 di-snRNP
complex, has two RNA binding domains. Previous data show that the c-terminal domain of p65 interacts with terminal end of
U12 stem-loop (SL) III. Our data also show that the terminal end of the U12 SL III is functionally important for in-vivo
splicing. Structural and sequence similarity between the distal 3' SL of U6atac and the apical loop of U12 SL III give rise the
notion that p65 has potential to interact with U6atac. Our data show that the distal SL of 3' U6atac and terminal loop of U12
SL III are functionally swappable. In addition Electrophoretic Mobility Shift Assay (EMSA) data show that p65 c-terminal
domain interacts with the distal SL of 3' U6atac SL in a dose dependent manner. 3' distal U6atac SL mutagenesis analysis
revealed that point mutation A99C and C100G in the loop enhanced the splicing whereas U98G did not affect the splicing.
Abolishing the predicting stem by disrupting the base-pairing in the distal stem caused splicing defect. Restoration of the
entire base pairing in the stem by complementary mutations couldn't restore the WT splicing. In summary, our data show the
plasticity of RNA-RNA base-pairing interactions in U12-dependent spliceosome.
063
REGULATION OF ANDROGEN RECEPTOR THROUGH ITS 3’ UNTRANSLATED REGION
Jey Sabith Ebron, B.Tech; Kavleen Sikand, Ph.D.; Girish C. Shukla, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Androgen receptor (AR) is a member of the steroid receptor family, is a transcription factor that has an important role in the
regulation of both prostate cell proliferation and growth suppression. AR has been a target for Prostate Cancer treatment
through Androgen deprivation therapy (ADT). miRNAs, small noncoding RNA sequences which regulate gene expression by
post-transcriptional regulation are known to target many genes basepairing interactions with its 3’ untranslated region (UTR)
and in turn destabilize the message or repress protein synthesis. However, action of different miRNAs on the same gene is
not well understood. It is known that AR mRNA has evolved with a long 3’UTR and it is the target of many miRNAs which
26
leads to the differential regulation of the expression of AR. Our lab has shown earlier that AR is a target of miR 488* and
644. We have looked at the combinatorial action of these miRNAs on AR gene by cotransfection and we have found that the
repression of the AR gene is proportionally increased in cells treated with both the miRNAs in comparison with the cells
treated with miRNAs individually. Results from the apoptotic and cell viability studies confirm that AR gene is a target of
multiple miRNAs and confirm the additive effect of these miRNAs in AR gene regulation. Combinatorial regulation of
miRNAs may provide us a platform to understand the regulation of protein synthesis in different stages of cancer
pathogenesis and hence to identify therapeutic targets.
064
TOWARDS A TYPOLOGY OF L1 SPANISH L2 ENGLISH SPEAKERS’ PHONOLOGY USING THE
SPEECH ACCENT ARCHIVE CORPUS
Mark Selent, B.A.
School of Health Sciences, Speech and Hearing Program, Cleveland State University
This study makes a contribution towards a phonological typology of L1 Spanish L2 English speakers using a corpus
methodology. Speech samples gained from an elicitation paragraph from N=18 native Spanish Speakers from Latin America
who learned English were analyzed for phonological substitutions. This study offers a typology of the most common
phonological errors of these English learners. The use of a corpus methodology proved effective in rapidly gaining
knowledge about speakers’ phonological substitutions across a language group. The findings of this study have implications
for theoretical assumptions about foreign language acquisition and foreign accentedness.
065
VOICE ONSET TIME (VOT) ACROSS LIFE-SPAN, GENDER, AND ETHNICITY: A REVIEW
Mark Selent, B.A.
School of Health Sciences, Speech and Hearing Program, Cleveland State University
Voice onset time (VOT), the duration of the period of time between the release of a plosive consonant and the beginning of
vocal fold vibration, is a commonly discussed feature of speech production. Despite being a commonly studied phenomenon,
we lack accurate data about the role of age, gender, and ethnicity on VOT. The purpose of this investigation is to examine the
few studies that have been performed to gain insight into the effects of age, gender, and ethnicity on VOT. Since voicing is
language dependent, only studies examining North American native English speakers were reviewed. A survey of the
literature revealed these factors do not significantly affect VOT; however, there are several limitations of these studies and
there are more possibilities to further explore the way these variables interact with VOT.
066
THE HADRON BLIND RING IMAGING CERENKOV DETECTOR
Marie Blatnik1; Thomas Hemmick, Ph.D.2
1
Department of Physics, Cleveland State University;
2
Department of Physics, Stony Brook University
Heavy Ion Collisions in the Relativistic Heavy Ion Collider (RHIC) at Brookhaven Lab have hinted at the existence of a new
form of matter at high gluon density, the Color Glass Condensate or CGC. The existence of this new form of matter would
not only explain the initial state particle density at RHIC, but perhaps more importantly solve the issue of exploding gluon
density at low parton momentum fraction x inside the proton.
High energy electron scattering off of nuclei, focusing on the low-x components of the nuclear wave function, will
definitively measure this state of matter. However, when a nucleus contributes a low x parton, the reaction products are
highly focused in the electron-going direction and have large momentum in the lab system. High-momentum particle
identification is particularly challenging. A particle is identifiable by its mass, but tracking algorithms only yield a particle’s
momentum based on its track’s curvature. The particle’s velocity is needed to identify the particle. A ring-imaging Cerenkov
detector is being developed for the forward angle particle identification from the technological advancements of PHENIX's
Hadron-Blind Detector (HBD), which uses Gas Electron Multipliers (GEMs) and pixelated pad planes to detect Cerenkov
27
photons. The new HBD will focus the Cerenkov photons into a ring to determine the parent particle's velocity. Results from
the pad plane simulations and construction tests will be presented.
067
EVERYDAY BATTLES: ASSESSING THE SOURCES OF SOCIAL ALIENATION IN THE
WORKPLACE FOR VETERANS WITH POST-TRAUMATIC STRESS DISORDER
Mairi K. Mull
Department of Psychology, Cleveland State University
Previous research has indicated that PTSD is strongly correlated with low employability, and many veterans – even those
who have had apparently successful civilian careers – are eventually forced to rely on government support for their regular
income. A 2006 study also found that variables in workplace environment may contribute to this phenomenon, and in some
cases even exacerbate the symptoms of traumatized individuals. The proposed study would survey veterans diagnosed with
PTSD in order to identify particular aspects of a workplace environment which contribute to their alienation, with the
ultimate goal of providing data from which targeted interventions and specific coping strategies might be developed.
068
CLIMATIC AND ENVIRONMENTAL CHANGE FROM OXYGEN ISOTOPE OF PUNDERSON LAKE,
NORTHEAST OHIO
Ron VanBlarcum, B.S. and Fasong Yuan, Ph.D.
Department of Biological, Geological and Environmental Sciences, Cleveland State University
Ohio as the rest of the nation has experienced large changes in climate over the last century. The Great Flood of 1913 is a
premier example that demonstrated a large range of natural climatic variability. It is not clear how frequently such a
devastating event has occurred in the past. To put recent climatic changes into context, we explored the use of oxygen isotope
18
O) as an indicator of climatic and environmental change. A 39-cm-long surface sediment core was recovered from
a site with an ice cover of 10 cm and a water depth of 5.3 m in Punderson Lake in Northeast Ohio on February 23, 2013.
18
Porous water samples were extracted at every 1 cm interval with a centrifuge and measured
O with a liquid water
18
O record shows a 3.4‰ range of variability, from -8.1 to -4.7‰. There are several low-amplitude
18
(<0.5‰) decadalO values fluctuate greatly in the top 9 cm section.
18
O between 10-8 cm, followed by two large (2‰) positive
18
excursions between 8-5 cm and between 5O of lake-bottom water is 1‰ more negative than
that of the porous water extracted from the topmost 1 cm sediments. On the basis of six radiocarbon dates previously
18
O
remained relatively small until 1920. We suggest that the large isotopic fluctuations over the last 90 years be ascribed in part
to human activities (e.g., dam constructions), assuming that post-depositional isotopic alterations are minimal.
069
GC/MS ANALYSIS OF PHTHALATES PRESENT IN AIR FRESHENERS
Adel W. Aziz; Ekenedilichukwu N. Nnadi; Vishal Sagar; Anne O'Connor, Ph.D.; Bhagya Gunasekera; Nick Pallas.
Department of Chemistry, Cleveland State University
The purpose of this experiment was to discover Phthalates in Air Fresheners and fragrance oils. The reason we conducted this
research was because many American families use Air Fresheners in their daily lives. According to the research, it turns out
Phthalates present in such fresheners could lead to birth defects and other reproductive issues. Gas Chromatography by
Varian was used to detect phthalates in eight of the different samples that were used. Originally, a standard curve was created
appropriate for Dibutyl Phthalate. Samples were extracted and diluted using Hexane. After preparing the samples for the
GC/MS eight different samples were analyzed. Because of the limited resources, we could only detect Dibutyl Phthalate in
only four of the samples. However, there were several other phthalates present in all of the samples.
28
070
INVESTIGATION OF AN INTRON RETAINED SPLICE VARIANT OF CELF5
Ryan J. Coram; Andrea N. Ladd, Ph.D.; Twishasri Dasgupta, Ph.D.
Cleveland Clinic Lerner Research Institute, Department of Cellular and Molecular Medicine
The CELF protein family is a group of RNA binding proteins that have a variety of roles in cellular functions including
regulation of pre-mRNA alternative splicing, mRNA editing, and translation. A novel splice variant of CELF5 has been
discovered to be expressed in relatively high amounts in rat and mouse brain tissue and human cell lines. This variant retains
an intron containing a premature translation termination codon which is conserved in all three organisms studied. Translation
of this transcript would yield a truncated CELF5 protein. Nonsense-mediated mRNA decay (NMD) is a surveillance pathway
that is used by cells to eliminate transcripts that contain errors before translation initiation. This pathway degrades transcripts
that contain specific errors such as a premature stop codon. Since the CELF5 variant being studied contains a premature stop
codon, it may be a target of NMD however, there have been many cases reported of potential NMD targets able to escape the
system and create a protein. Studies have shown that some CELF proteins can function effectively when truncated at various
locations. It may be possible that the supposed truncated protein has productive biological activity and selection pressure has
driven the variant to escape NMD. The purpose of this work is to evaluate the expression pattern of this newly discovered
variant and gain information to hypothesize a function for the intron-retained variant. This work characterizes the expression
pattern of the intron-retained variant of CELF5 across tissues and stages of adult mice. Data is also gathered to gain evidence
as to whether or not a truncated protein is expressed by determining transcript stability. The results indicate that the
expression of the intron-retained variant of CELF5 is widely expressed in many tissues, but predominantly in nervous
system, skeletal muscle, and heart tissue. Transcript stability comparisons indicate that the intron-retained transcript is stable
and likely not an NMD substrate.. The presence of a protein product has yet to be determined and will be the focus of future
work.
071
PHOSPHOPROTEOMIC ANALYSIS OF THE SIGNALING TARGETS IN THYMOCTES DURING
PROLIFERATION AND DEVELOPMENT
Lin Zhang, B.Sc.; Xiang Zhou, Ph.D.; and Aimin Zhou, Ph.D.
Department of Chemistry, Clinical Chemistry Program, Cleveland State University
RNase L is one of the key enzymes involved in the function of interferons against viruses and other microbial pathogens.
Deficiency of RNase L contributes to the remarkable enlarged thymus glands containing significantly higher numbers of
thymocytes at the early stage than that from wild type mice, suggesting RNase L may play an important role in thymus
development and function. Therefore, the phosphorylation status in the thymoctes from RNase L null and wild type mice may
provide a better insight into RNase L mediated thymocytes proliferation and development. In this work, we performed the
systematic analysis of the phosphorylated protein profiles in the thymocytes from RNase L null and wild type mice. The
phosphorylated proteins were enriched by employing Immobilized Metal Affinity Chromatography (IMAC) and were
fractionated by 1-D gel electrophoresis. After excised from the gel, the phosphor-proteins were digested by trypsin in-gel.
The resulting peptide fragments were further separated by utilizing Fe-NTA phosphopeptide-enrichment columns, and then
subjected to liquid chromatography-tandem mass spectrometry (LC/MS) analysis. The results showed the phosphorylation
levels in the thymocytes from RNase L null and wild type mice were clearly distinct; a number of phosphor-proteins as the
potential signaling targets in thymocytes were identified. Further confirmation is in progress. Our findings may provide direct
evidence that RNase L is crucial for the thymus development through mediating unique protein phosphorylation in the
signaling pathways of thymoyctes.
072
A METEL-CONTAINING NUCLEOSIDE THAT FUNCTIONS AS A THERAPEUTIC AND
DIAGNOSTIC AGENT AGAINST CANCER
Jung-Suk Choi, Ph.D.1,2; Ayan Maity, M.S.3; Thomas Gray, Ph.D.3; and Anthony J. Berdis, Ph.D.1,2
1
Department of Chemistry, Cleveland State University;
2
Department of Biological, Geological and Environmental Sciences; Center for Gene regulation in Health and Disease;
Cleveland State University;
3
Department of Chemistry, Case Western Reserve University
29
Nucleoside transport and metabolism are important for the hyperproliferative nature of cancer cells. As such, they represent
important targets for therapeutic intervention. This report outlines the development of a novel metal-containing nucleoside,
designated Ir(III)-PPY, that possesses both therapeutic and diagnostic properties against cancers that overexpress nucleoside
transporters. The cytotoxic effects of this ‘theranostic’ nucleoside were evaluated against two human epidermal carcinoma
cell lines, KB3-1 (MDR-negative) and KB-V1 (MDR-positive). The cytotoxic effect of Ir(III)-PPY is both concentration and
time-dependent, exhibiting an IC50 value of 30 M after 48 hours of exposure. Flow cytometry analyses using propidium
iodide staining validates that the cytotoxic effects are caused by the ability of Ir(III)-PPY to block cellular entry into G2/M
phase. The cellular uptake of Ir(III)-PPY offers the possibility to visualize the intracellular distribution of this nucleoside
and/or its metabolites. Fluorescent microscopy shows that Ir(III)-PPY accumulates in the cytoplasm, nucleus, and
mitochondria of KB3-1 cells after 4 hours. However, significant nucleoside accumulation occurs in the nucleus and
mitochondria after 48 hours, and this localization is consistent with its ability to inhibit enzymes involved in cell-cycle
progression. In addition, pre-treatment with 10 M NBMPR, a known nucleoside transport inhibitor, causes reduced cellular
uptake and lower cytotoxicity of Ir(III)-PPY. These results indicate that uptake of the metal-containing nucleoside is
primarily mediated by a specific nucleoside transporter, human equilibrative nucleoside transporter 1 (hENT1). Collectively,
these data validate that metal containing nucleosides can function as unique theranostic agents against solid cancers.
073
DEVELOPMENT AND VALIDATION OF AN IN VITRO MULTI-CELLULAR SPHEROID MODEL AS A
TOOL FOR IDENTIFICATION OF POTENT ANTI-CANCER DRUG CANDIDATES
Rati Lama, B.S.1; Bo Zhong, Ph.D.1; Bin Su, Ph.D.1,2
1
Department of Chemistry, Cleveland State University
2
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Cancer is a leading cause of death worldwide. Two-dimensional (2D) monolayer cellular assays have been widely used as an
in vitro model for anti-cancer drug screening purposes. However, the conventional monolayer cell culture screening assays
fall short in predicting clinical response because they lack in vivo tumor characteristics. In contrast, three-dimensional (3D)
multicellular spheroid is a uniform symmetrical structure composed of self-aggregated cell clusters that closely mimic the
physiological environment of tumors in vivo. Studies have demonstrated that in vitro spheroids and in vivo tumors have
morphological similarities and exhibit resemblance in the penetration, binding and bioactivities of drugs. 3D spheroids
therefore serve as a bridge between in vitro monolayer and complex in vivo models. Despite numerous recognized advantages
of 3D cell culture models as drug screening tools, the complexity and lack of standardized protocols for 3D assays has
hindered their use in the mainstream pre-clinical drug screening processes. An easy-handling and cost-effective lung cancer
3D spheroid model suitable for small molecule anti-cancer compounds screening was developed, standardized and validated.
A compound library was generated with Nimesulide, an NSAID as a lead compound. The compound analogs were found to
be dual Hsp27 and tubulin inhibitors. A pilot screening was performed with 84 anti-cancer agents in spheroid and 2D models.
A correlational study based on the IC50 values from the 2D and 3D assays was conducted. There was no strong correlation
between the monolayer and spheroid assays, suggesting the two screening methods provide different information regarding
the potency of the tested drug candidates. This approach underlines the importance of 3D culture models as an excellent cellbased validation platform that contributes to the elimination of poor drug candidates at pre-clinical stage and to the
identification of the most promising drug candidates to enter in vivo animal studies.
074
CHARACHETERIZATION OF TRYPANOSOMA BRUCEI TELOMERIC PROTEINS IN ANTIGENIC
VARIATION
Vishal Nanavaty, M.S.; Bibo Li, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Trypanosoma brucei is a unicellular protozoan parasite that causes human African trypanosomiasis. T. brucei regularly
switches to expressa different surface antigen known as Variant Surface Glycoprotein (VSG), exclusively from subtelomeric
30
VSG expression sites (ESs) in a monoallelic fashion. This antigenic variation mechanism is the key for establishing a
persistent infection of T. brucei in its mammalian host. Two major pathways of VSG switching have been identified. One is
called in-situ switching, where the active ES promoter is silenced while a previously silent ES promoter is activated. Second,
homologous recombination (including gene conversion or crossover events) encompassing either the whole ES or a smaller
fragment including VSG can lead to the replacement of the original active VSG gene with any new VSG gene. A combination
of gene rearrangement and in-situ switch has also been observed in VSG switching events. Telomere is the nucleoprotein
complex located at the ends of linear chromosomes and is required forchromosome stability. We have recently shown that
TbRAP1, a telomere protein, is essential for complete silencing of ES-linked VSGs. Now we find that TbRAP1 also plays an
important role in VSG switching regulation. A temporary depletion of TbRAP1 led to an increased VSG switching frequency,
predominantly by VSG associated gene conversion. We have recently identified several potential candidates which may
interact with TbRAP1 via immunoprecipitation followed by mass-spectrometry analysis.
075
NON-CANONICAL ROLES OF THE MUSCLE REGUALTORY TRANSCRIPTION FACTOR MYOD
Divya Nair, M.Sc. and Crystal M. Weyman, Ph D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
A goal of our lab is to investigate novel roles for the muscle regulatory transcription factor MyoD. MyoD’s canonical role is
to regulate transcription of genes responsible for myoblast differentiation. Others have previously reported that when
myoblasts are cultured in differentiation media (DM), MyoD contributes to the increased expression of the cyclin dependent
kinase inhibitor p21, resulting in cell cycle exit. Using myoblasts silenced for MyoD expression, we have now determined
that MyoD also plays a role in cell cycle progression in GM. Experiments are underway to determine if this involves
regulation of p21. We have determined that MyoD positively regulates the expression of molecules critical to cap-dependent
translation. We have also determined that translation of PUMA is cap-independent and IRES mediated. We, therefore,
hypothesized that MyoD may play a role in the IRES mediated translation of PUMA. This is being investigated by analyzing
luciferase expression from constructs with or without the PUMA IRES element in both wild type and MyoD silenced
myoblasts. Finally, we have also previously reported that when myoblasts are cultured DM, MyoD regulates the expression
of the pro-apoptotic Bcl2 family member PUMA, causing a sub population of cells to undergo apoptosis. However, the
signaling responsible for determining when MyoD will increase the expression of myogenin and differentiation, or PUMA
and apoptosis, remains unknown. We now report that signaling by the p38 kinase appears to play a role in this decision.
076
IN VITRO ANALYSIS OF DESIASE CAUSING SYNONYMOUS MUTATION IN BLOOD
COAGUALTION FACTOR IX
Puja Nanavaty, M.S.; Sujata Jha, M.S.; Anton A. Komar, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Hemophilia B is a blood clotting disorder caused by mutations in the F9 gene, which encodes a serine protease in the blood
coagulation system known as factor IX (FIX). Mutations in F9 may lead to severe (FIX coagulant activity (CA) <1% of
normal), moderate (CA 1–5%) or mild (CA 5–30%) hemophilia B. Over 130 different non-synonymous mutations (which
change encoded amino acids) in F9 gene cause this disease. However, the effects of only a few have been analyzed to
determine the exact mechanism(s) by which these mutations contribute to FIX deficiency. A new paradigm that adds another
level of complexity to our understanding of the disease is effects of synonymous mutations. Synonymous mutations (which
do not change encoded amino acids) affect gene function mostly via perturbations of mRNA splicing and/or mRNA stability.
However, recent studies suggest that synonymous mutations also alter protein folding. It has been found that a mild form of
hemophilia B is caused by a synonymous GTG>GTA (Val107Val) mutation. All affected individuals had antigen and antiFIX antibody levels corresponded to mild hemophilia B. Studies of lymphocyte F9 mRNA in these individuals showed no
skipping or retention of introns and/or change in mRNA levels. To understand molecular mechanisms that alter FIX
coagulant activity due to changes in codon usage/translation rates, we explored how the GTG>GTA (Val107(153)Val) codon
substitution affects FIX mRNA translation rates. We found that it takes about 7 min to synthesize full-length (unprocessed)
FIX in a reconstituted in vitro translation system and that substitution of GTG encoding Val107(153) with GTA and/or 2
31
other synonymous codons encoding Val affects both global and local translation rates and the yield of the protein produced.
This project will add to our understanding of the relationship between genotype and phenotype.
077
KIDNEY EPITHELIAL CELL PROLIFERATION INDUCED BY CHANGES IN FLUID FLOW
VELOCITY
Vikram Neal Seetharaman, B.S.¹; and Andrew H. Resnick, PhD.¹,²
¹Department of Biological, Geological and Environmental Sciences; Cleveland State University
²Department of Physics, Cleveland State University
It was previously determined that, in mouse cortical collecting duct cells (MCCDC’s) bathed in culture media under
conditions of differentiation, changes in fluid flow velocity (Δvf) generated using an orbital shaker (o.s.) lead to increased
cell proliferation, a response thought to be transduced through the primary sensory cilia, organelles important for
mechanosensation. Since mature kidney epithelial cells (mKEC’s) of all types exist in a G0 non-dividing state, this
proliferation must be the result of cell cycle re-entry. Thus, I hypothesized that MCCDC’s subjected to Δvf de-differentiate
into migrating mesenchymal kidney cells before re-entering and continuing through the cell cycle, subsequently dividing.
To investigate this hypothesis, cells bathed in culture media will be allowed to differentiate while kept either under
conditions of no fluid flow (vf=0) or constant rapid fluid flow (vf=max) on an o.s. Cells will then be subjected to Δvf at 5 and
10 days post-differentiation, either from vf=0 to vf=max, or vice versa. At various time points after these Δvf, cells will be
harvested, DAPI will be used to stain nuclei for cell counting, and the RNA and protein will be extracted. qRT-PCR and
Western blot analyses will then be utilized to determine the concentrations of several mRNA transcripts/proteins known to be
markers of either KEC proliferation or differentiation. Markers of differentiation will include E-cadherin, important in
adherens junction formation in mKEC’s, and acetylated tubulin, critical for primary cilia outgrowth in mKEC’s. Markers of
proliferation will include phospho-Akt, phospho-mTOR, phospho-ERK1&2, cdk1, and c-Myc. If MCCDC’s do indeed dedifferentiate and actively divide in response to Δvf, then protein levels of both E-cadherin and acetylated tubulin should
decrease while protein levels of phospho-Akt/phospho-mTOR/phospho-ERK1&2 and mRNA levels of cdk1/c-Myc should
increase as compared to controls exposed to either vf=0 or vf=max only.
078
METAL CONTAINING NUCLEOSIDES THAT FUNCTION AS THERAPEUTIC AND DIAGNOSTIC
AGENTS AGAINST BRAIN CANCER
Jennifer N. Williams, M.S.1, 2; Jung-Suk Choi, Ph.D.1, 2; Anthony Berdis, Ph.D.1, 2
1
Department of Chemistry, Cleveland State University;
2
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Nucleoside transporters are essential components for the hyperproliferative capabilities of brain cancer cells. These
transporters play important roles to increase nucleoside metabolism which is necessary for higher levels of DNA and RNA
synthesis. The goal here is to use a series of metal containing nucleoside (MCN) analogues as novel chemical agents to study
how nucleosides are imported into cells. In addition, we expect that these MCNs will ultimately function as therapeutic
and/or diagnostic agents for brain cancer. Concentrative nucleoside transporters (CNTs) and equilibrative nucleoside
transporters (ENTs) represent the two classes of transporters that allow MCNs to travel in (and out) of brain cancer cells.
This study focuses on cyclometalated iridium nucleosides, designated Ir(III)-ppy, Ir(III)-bzq, and Ir(III)-pbo, that were tested
on a glioblastoma brain cancer cell line, U87. The therapeutic activities of these MCNs were tested against U87
glioblastomas using several biochemical methods. Cell viability experiments demonstrate that all compounds induce cell
death in a time- and dose-dependent manner. Ir(III)-bzq was identified as the most potent MCN, exhibiting an effective
concentration (EC50) of 10 M after 48 hours of exposure. The collective photophysical properties of our MCNs were also
used to visualize their intracellular distribution throughout localized regions of the cell. Fluorescent microscope images
reveal the accumulation of Ir(III)-bzq in the cytosol after 4 hours of exposure while significant nuclear localization is
detected at longer times (~24 hours). This timing corresponds with the onset of cell death and provides insight into the
mechanism of action of these MCNs. In addition, the cellular uptake and cytotoxicity of Ir(III)-bzq is significantly reduced
when U87 cells are pretreated with the natural nucleoside, adenosine. Collectively, the ability to measure the uptake of
32
MCNs coupled with their anti-cancer activities define these novel nucleoside analogues as “theranostic agents” – compounds
that possess both therapeutic and diagnostic activities.
079
DISPARITY IN TRAUMA AND POSTTRAUMTIC STRESS DISORDER IN AFRICAN-AMERICAN
PREGNANT WOMEN
Hannah E.Morris; Lisa Stines-Doane Ph.D.
Department of Psychology, Cleveland State University
Previous studies have found high rates of trauma and posttraumatic stress disorder (PTSD) among pregnant women, with
African American women being at highest risk for PTSD. Pregnant women are at an increased risk of PTSD with prevalence
of 6.8% versus 4.5% in non-pregnant women (Seng et al. 2010), while African American women are at greater risk for PTSD
compared to non-African American women, 13.4% versus 3.5% (Seng et al. 2008). Potential explanations for this racial
disparity in rates of PTSD include limited access to psychological interventions for African-American women, particularly
those who are low income and un-or underinsured, or that African-American women may be at greater risk of lifetime trauma
exposure, thus contributing to higher rates of PTSD at any point in the lifespan relative to Caucasian women. Further,
because African-American women have earlier average age at first pregnancy there may be less time between onset of the
traumatic event and pregnancy, thereby limiting the opportunity for natural recovery in these women. The purpose of this
study is to examine racial disparities in the prevalence of trauma and posttraumatic stress disorder (PTSD) in a diverse
community sample of pregnant women. Participants will be pregnant women recruited from a free inner-city obstetric clinic.
Eligibility criteria include being 18 years or older, and the ability to speak English. Participants will be interviewed using a
standardized, clinician-administered measure of DSM-IV PTSD severities. We expect to find that pregnant African-American
women will report higher rates of trauma history with more severe PTSD symptoms and lower rates of mental health
treatment when controlling for income and access to insurance. The U.S. Census Bureau states African Americans are 7.3
times as likely to live in high poverty neighborhoods with limited to no access to mental health services. If our hypotheses are
supported this will demonstrate a need for trauma and PTSD screenings in high risk areas such as free obstetric clinics, where
high risk pregnant African American women can be directed to appropriate care to improve their own mental health, which
has benefits for mothers and children.
080
TOWARD THE PURIFICATION OF ORNITHINE TRANSCARBAMOYLASE FROM
METHANOCOCCUS JANNASCHII
Elissar Daher1; Aditya Singh, Ph.D.2; and Jacqueline Vitali, Ph.D.2
1
Department of Biological, Geological and Environmental Sciences; Cleveland State University;
2
Department of Physics, Cleveland State University
Ornithine Transcarbamylase (OTCase) is an enzyme used to catalyze the reaction of ornithine and carbamoyl phosphate to
produce citrulline and phosphate. OTCase can be found in the process of arginine biosynthesis in plants and microbes and has
a role in the urea cycle of mammals. A genetic OTC deficiency in humans is a urea cycle disorder that results in
hyperammonemia. OTCase belongs to the transcarbamoylase family of proteins and has similarities in sequence and structure
with aspartate transcarbamoylase (ATCase). In both enzymes, the catalytic subunit contains two domains, the N-terminal or
CP-binding domain and the C-terminal or aspartate/ornithine binding domain.
We have been working with the OTCase from the hyperthermophilic archaeon Methanococcus jannaschii and are trying to
develop a successful strategy for its purification. We are working with epitubes. To this end, a heat step at 60 C and DEAE
anion exchange chromatography appear to be promising steps. In the poster, we will show our work up to this point. Using
the colorimetric assay, we have confirmed that our preparation contains the enzyme and that it follows Michaelis Menten
kinetics.
33
081
ARE INTRUSION RATES HIGHER IN CUED THAN IN FREE PHASES OF DIETARY RECALL TASKS
TASKS?
Albert F. Smith, Ph.D.; Kathryn Van Gunten; and Marvi Lalvani
Department of Psychology, Cleveland State University
We are comparing intrusion rates in free and cued parts of a dietary recall task. Twenty-four-hour dietary recalls were
collected by US Department of Agriculture scientists as part of a validation study in developing their five-step multiple-pass
method. Previous analyses of these data involved converting food items to a nutrients and focusing on the nutrient intake of
each participant.
For one day, each research participant ate all meals and obtained all food for between-meal snacks at the USDA Beltsville
Human Nutrition Research Center Human Study Facility. A record was made of all items eaten and taken away for snacks.
The next day, the participant was contacted and a 24-hour recall was obtained. First, the participant was to report all food
items eaten on the preceding day. Next, the participant was asked if he or she forgot to report anything in each of seven
different categories (e.g., coffee, tea, soft drinks, milk or juice).
We are examining reporting accuracy for actual food items. Every reported item is classified as either a match or as an
intrusion. Classification is being done by two coders to ascertain intercoder reliability. We will compare the rate of intrusions
in the free and cued phases of the procedure to determine whether higher intrusion rates are found in the cued phase.
082
ASSESSMENT OF RIVER MOUTHS AS UNIONID REFUGIA ALONG A LAURENTIAN GREAT LAKE
Trevor J. Prescott, B.S.1; Robert A. Krebs, Ph.D.1; Wentworth B. Clapham, Ph.D.1; David A. Klarer, Ph.D.2
1
Department of Biological, Geological and Environmental Sciences; Cleveland State University;
2
Old Woman Creek National Estuarine Research Reserve
Invasion of lakes and rivers by dreissenid mussels since the 1990’s have pushed out native species, particularly native
freshwater mussels in the Unionidae across the northern hemisphere, and perhaps most infamously, within the U.S. Great
Lakes. However, several small coastal areas along Lake Erie have been identified as refugia for native species, but the
conditions under which native species may persist are unknown. We identified that unionid species may frequently be using
river mouths (freshwater estuaries) even of small streams as refugia along the Lake Erie coastline. Comparing nine such
streams, we qualitatively assessed mussel diversity and compared species presence to land use based on remote sensing
techniques. Sampling focused on stream zones influenced by lake-water levels for three streams each in the western basin,
central basin and Sandusky Bay regions of Lake Erie. Eight of the nine streams possessed mussels: Pyganodon grandis (7
streams), Toxolasma parvum (5 streams), Quadrula quadrula (5 streams), Lasmigona complanata (5 streams), Leptodea
fragilis (4 streams), and Utterbackia imbecillus (2 streams), while Amblema plicata, Obliquaria reflexa and Uniomerus
tetralasmus were found each in only one stream. Diversity levels were not affected by any distinct bathymetric features,
although regional land use impacted diversity subtly. Importantly, river mouths are refugia for unionid mussels in regions
exposed to dreissenid infestation, and these areas must return to or come under regulatory control to monitor habitat
alteration, a process stopped in this region following the belief the dreissenid mussels had eradicated all species of interest.
083
YEAST 2-HYBRID SCREEN FOR T. BRUCEI RAP1- INTERACTING PROTEINS
Fan Wu, B.S.; Nicole Kresak, M.S.; Bibo Li, Ph.D.
Department of Biological, Geological and Environmental Sciences; Cleveland State University
Telomeres are specialized protein-DNA complexes located at the ends of eukaryotic chromosomes. They play an important
role in DNA stability. They act as caps on the end of chromosomes to preserve DNA from degrading and rearranging. In
Trypanosoma brucei, a parasite that causes sleeping sickness in humans and nagana in cattle, the Telomere complexes also
play important roles in allowing the parasite to evade the host immune responses by switching out its Variant Surface
Glycoproteins (VSG). VSGs are expressed exclusively from VSG expression sites, which are found at sub-telomeric regions.
Hence, understanding the VSG regulation by the telomere complex would help in developing means to eliminate this
34
parasite. Our previous studies indicate that TbRAP1 (Repressor Activator Protein 1) is a protein associated with the telomere
complex, and it plays an important role in regulation of VSG silencing and VSG switching frequency.
In order to better understand how VSG silencing and switching is regulated by TbRAP1, we explored more about the proteins
that interact with TbRAP1. By using TbRAP1 as bait, we identified a number of TbRAP1-interacting candidates in a yeast-2hybrid screen. We are currently validating the interactions between TbRAP1 and the newly identified candidates.
084
SELECTION OF IMPORTANT VARIABLES FOR DIAGNOSIS OF INFECTION ATTER TRAUMATIC
SPLENECTOMY VIA RANFOM FOREST
Xinwei Li, M.S.1; Ariadni Papana Dagiasis, Ph.D.1; Aman Banerjee, M.D.2
1
Department of Mathematics, Cleveland State University
2
Department of Surgery, MetroHealth Medical Center Campus, University Hospitals Case Medical Center
The study population consisted of 102 trauma patients who underwent total splenectomy at Level 1 Trauma Center from
January 1, 2005 through September 30, 2011. The data were collected on patient demographic, injury information, surgical
outcomes and clinical variables for post-operative days (POD) in 2 weeks. Infection is the most feared post-operative
complication associated with a high mortality. In this study, we utilized Random Forest (RF), a popular tree-based ensemble
method, to select variables of importance for recognizing infected patients after traumatic splenectomy. Clinical indicators
platelet count, white cell count, temperature, heart rate, and blood pressure out of 150 demographic and clinical variables,
were identified as important variables with significant RF score of importance. Other than primary signs of infection, i.e.
platelet count, white cell count and temperature, additional clinical predictors were extracted for diagnosis of infection for
traumatic splenectomy patients. The study helps clinicians differentiate infected and non-infected patients in early stage and
timely perform appropriate intervention to lower the mortality.
085 TESTS OF THE SUFFICIENCY OF A LETTER ROUTE FOR WORD IDENTIFICATION
Nevena Vasovic; Albert F. Smith, Ph.D.
Department of Psychology, Cleveland State University
Many models of visual word identification propose that identification is mediated exclusively by letter identification.
However, some empirical phenomena appear to require a route to word identification other than through letters. Two lexical
decision experiments, in which participants will decide whether a presented letter string is a word or a nonword, will be
described. The results of these experiments are expected to provide evidence concerning the plausibility of a route to word
identification that is not mediated exclusively by letters.
086
DEVELOPMENT OF A HIGHLY SENSITIVE LC-MS/MS METHOD FOR DETERMINATION OF NGP101 IN MOUSE SERUM
Harini Jogiraju, M.Sc.1; Xiang Zhou, Ph.D.1; Kiran K. Pedada, M.Pharmacy1; Werner Geldenhuys, Ph.D.2; Cornelis Van
der Schyf, Ph.D2; Samuel Crish,Ph.D.2; David Anderson, Ph.D.1
1
Department of Chemistry, Cleveland State University, Cleveland, OH;
2
Northeast Ohio Medical University, Rootstown, OH
An increasing focus of research in recent years is development of multifunctional drugs (in drug discovery), agents with more
than one therapeutic mechanism. Recent investigations have been aiming to develop neuroprotective agents to treat many
chronic neurodegnerative disorders like Parkinson’s, Alzheimer’s, and Huntington’s diseases. An aberrant calcium levels are
thought to play a large role in the pathogenesis of these disorders. NGP1-01 (8-benzylamino-8, 11oxapentacyclo[5.4.0.02,6.03,10.05,9]undecane) is a heterocyclic cage compound with multifunctional calcium channel
blocking activity that has been demonstrated to be neuroprotective in several neurodegenerative models. Our research team
has proposed that NGP1-01 as a potential intervention to produce neuroprotective effect. In the present work a quantitative
LC-MS/MS method has been developed to determine the pharmacokinetics and ability to cross the blood-brain and retinal
35
barrier for NGP1-01 in mouse serum, brain, and retina in-vivo at different time points. The LC-MS/MS system consisted of a
Shimadzu HPLC and an AB Sciex QTrap 5500 MS with positive electro spray ionization. The mass spec employed MRM
mode for sensitive and specific detection of the analyte and internal standard. The protonated molecular ions at m/z 266 and
268 for NGP1-01 and the internal standard revealed the predominant daughter ions at m/z 91 and 105 respectively upon
fragmentation in their product ion scan spectra. Therefore the MRM channels were set at m/z 266 → 91 for NGP1-01 and
280 → 105 for the internal standard. Based on the preliminary data, this method is reliable for a full length of
pharmacokinetic study of NGP1-01.
087
PHARMACOKINETIC STUDIES OF A NOVEL THIAZOLIDINEDIONE MITONEET LIGAND, NL-1,
USING A QUANTITATIVE LC-MS/MS METHOD IN DOSED MOUSE SERUM AND BRAIN
Kiran K. Pedada, M.Pharmacy1; Xiang Zhou, Ph.D.1; Harini Jogiraju, M.Sc.1; Richard Carroll, Ph.D.2; Werner J.
Geldenhuys, Ph.D.2, Li Lin, M.Sc.2; David J. Anderson, Ph.D.1
1
Department of Chemistry, Cleveland State University, 2121 Euclid Avenue, Cleveland, Ohio 44115;
2
Department of Pharmaceutical Sciences, Northeast Ohio Medical University, 4209 State Route 44, Rootstown, Ohio 44272
TZD compounds have shown promise as antidiabetics, antibiotics, antifungal and neuroprotective agents. Recently, a new
mitochondrial protein, mitoNEET, was identified as a target of several TZD compounds, altering mitochondrial respiration.
NL-1(5-[(3,5-di-tert-butyl-4-hydroxyphenyl)methyl]-1,3-thiazolidine-2,4-dione) is a novel ligand that binds to mitoNEET,
but does not activate PPAR-gamma. In previous work it has been shown that NL-1 prevents neurodegeneration in a
Parkinson's disease rodent model. In the present work a quantitative analytical pharmacokinetic study was employed to
measure NL-1 in serum and brain samples from dosed mice. Shimadzu HPLC system interfaced with an AB Sciex QTrap
5500 mass spectrometer employing negative electrospray ionization was used. The MS detection employed MRM mode set
at m/z 334 → 263 for NL-1 and 250 → 179 for NL-2. The developed method was applied for the measurement of NL-1
content in serum and brain samples from dosed mice at six different time points (0, 0.5, 1, 2, 6, and 24 hrs). Based on the
preliminary data, NL-1 drug reached the highest levels in the brain by 30 minutes, with a significant decrease in its serum and
brain levels by the 6 hours. Based on the preliminary data of these PK studies, the proposed method can be used to establish a
complete PK profile and for determining optimal dosing of NL-1 in future studies.
088
LASER TRAP STIFFNESS AND TRAPPING FORCE CHARACTERISATION OF MICRO SPHERES
Joseph P. Glaser; David M. Hoeprich, B.S.; Andrew H. Resnick, Ph.D.
Department of Physics, Cleveland State University
Optical traps have been in use in microbiological studies for the past 40 years to control particles on the micron scale. We are
interested in trapping and manipulating a primary cilium, a slender rod with aspect ratio l/r > 30, where ‘l’ is the cilium
length and ‘r’ the cilium diameter. In order to understand the trapping of the primary cilium, we first characterized the
mechanical stiffness and trapping force of the laser tweezers instrumentation. As a control, data from trapping micro spheres
was acquired, analyzed, and calibrated. The results obtained will inform the analysis of a trapped cilium.
089
ANALYSIS OF FLUID DYNAMICS AND REACTANT CONSUMPTION IN MICROCHANNEL BASED
FUEL CELLS
Joseph D’Alessandro; Petru S. Fodor, Ph.D.
Department of Physics, Cleveland State University
In this work, the fluid dynamics within a membrane-less microchannel fuel cell is analyzed computationally. The membraneless design is a result of the laminar nature of the fluid flow at small Reynolds numbers, restricting the fuel and oxidant to the
vicinity of the corresponding electrodes, without the need of a proton exchange membrane (PEM). The performance of such
cells is limited by the mass transport near the electrodes, with much of the reactants leaving the channel without coming in
contact with the catalytic surfaces. Various strategies to overcome this limitation are explored including separation streams
and channel surface modifications similar with the ones used in grooved micromixers. While the flow is still laminar in
36
nature, the addition of ridges to the top and bottom of the cell introduce a transverse element to the fluid flow, increasing
reactant consumption and overall cell efficiency. The characteristics of the cells are investigated as a function of the Péclet
number.
090
INCREASES IN TASK DIFFICULTY WHITENS THE SPATIAL STRUCTURE OF MOVEMENT
Patrick Byrne1; Andrew B. Slifkin, Ph.D.1; Jeffrey R. Eder, Ph.D.1; Ulrich Zurcher, Ph.D.2
1
Department of Psychology, Cleveland State University;
2
Department of Physics, Cleveland State University
Studies using a variety of experimental tasks have established that when humans repeatedly produce an action, the amount of
variability in system output is distributed across a range of time scales or frequencies. A finding of particular interest is that
fluctuations in the output of cognitive systems are the highest at the lowest frequencies with fluctuation magnitude (power)
systematically declining as frequency increases (e.g., for a review see Gilden, 2001). Such time-series structure—captured by
spectral analysis—is termed pink noise. (In contrast, white noise has equal amounts of power at all spectral frequencies.)
However, the appearance of pink noise is limited to tasks where action is executed in the absence of external, action-related
feedback (e.g., Gilden, Thornton, & Mallon, 1995; Gilden, 2001). A few studies have shown a white-noise structure for
action executed in the presence of sensory feedback (e.g., Miyazaki et al., 2004). Here, we sought to determine if movement
amplitude (MA) time-series structure would vary with variations of movement difficulty, viz., the index of difficulty (ID)
[log2(2A/W) (Fitts, 1954)]. The current task required young adults (n = 16) to generate long sequences of cyclical hand
movements over different movement amplitudes requirements (A) to targets that varied in their width (W). Levels of A and W
were combined to produce different levels of ID. Visual feedback of movement was always available. Given that increases in
ID are known to induce increased reliance on the available visual feedback (e.g., Flowers, 1976) we predicted an ID-induced
shift in time-series structure from pink to white noise. In other words, at low IDs movement should mainly be controlled by
internal information processes—with minimized visual feedback processing—and pink noise should result; however, as ID
requirements increase there should be increased reliance on visual feedback and time-series structure should shift toward
white noise. Indeed, as ID requirements increased there was shift in MA structure from pink to white noise.
091
COMPARISON OF JOB SATISFACTION IN HOSPITAL NURSES TO CORRECTIONAL NURSES
Megan M. Collingwood; Michael Horvath, Ph.D.
Department of Psychology, Cleveland State University
Typically studies on job satisfaction in nursing use hospital nurses as the sample population. There are, however, studies that
focus on industry-specific nurses and their job satisfaction but few compare their finding to the findings based on hospital
nurses. The present study compares the job satisfaction of hospital nurses and correctional nurses. The study is based on
findings from an online survey created to measure job satisfaction and its contributing variables (perceived organizational
support, level of stress, workplace safety, autonomy, and workload). A convenience sample of nurses was emailed a link to
the survey and they were told, upon completion, to forward the link to other qualified nurses. Predicted results are that
hospital nurses and correctional nurses will differ in their job satisfaction. More specifically, it is predicted that the
relationships between job satisfaction and its contributing variables (perceived organizational support, level of stress,
workplace safety, autonomy, and workload) will differ between the two groups and thus cannot be generalized across
different industries of nursing. This prediction is based on the idea that it is not the field of nursing that represents job
satisfaction but it is the specific industry of nursing that contributes the most to job satisfaction. If this is so, then
improvements in the workplace to increase job satisfaction among nurses should be based on research conducted for that
specific nursing industry and not on hospital nurses.
37
092
THE BIO-MECHANICAL EFFECTS OF IL-5 ON THE CILIUM OF RESPIRATORY TRACT
EPITHELIAL CELLS
Briana M. McGinness, B.S.1, Andrew H. Resnick, Ph.D.2
1
Department of Biological, Geological and Environmental Sciences; Cleveland State University,
2
Department of Physics, Cleveland State University
Cilium plays an integral role in the health of the respiratory system. Dysfunction in cilia affects mucus and irritant clearance
in the respiratory tract. This is an element of the pathology of many respiratory disorders including asthma. The research
exploring the effects of cytokines expressed in the asthma response focus on IL-4 and IL-13. IL-13 is often co-expressed with
IL-4 and IL-5. The difference between IL-4 and IL-5 in their simultaneous expression in the presence of IL-13 is that IL-4
and IL-13 share a receptor (mediated by STAT-6) that IL-5 does not associate with (Laoukili et al, 2001). The activities of
IL-4 and IL-13 have been shown to effect the differentiation of respiratory tract epithelial cells, specifically inhibiting the
foxj1 promoter which promotes translation of ezrin, an actin binding protein that is necessary for ciliogenesis of motile (as
opposed to primary) cilium (Raksha et al, 2010). Ciliary beat frequency (CBF) was also reduced in the presence of these
cytokines. The research on the partnering activities of IL-13 and IL-4 does not explore the contribution of IL-5 in the
deficiency of cilium in the respiratory epithelium of asthma-affected cells. I have been trying to develop a line of Calu-3 cells
and differentiate them for the purpose of observing rate of ciliogenesis, CBF, and induced flow velocity under normal
conditions and with the addition of IL-5. After testing, the cells will be stained for alpha acetylated tubulin, actin, and ezrin.
Unfortunately, the cells currently available to me are not differentiating consistently and are not exhibiting the ciliated
phenotype that would be optimal for testing. Therefore, I will start growing BEAS-2B this spring. This is a human cell line of
bronchial epithelial cells that differentiate to ciliated squamous epithelium. The consistent ciliated phenotype of these cells
makes them more suited for the study.
093
THE ROLE OF RNASE L IN TYPE 1 DIABETES
Chun Zeng, B.S.; Lin Zhang, B.S.; Xin Yi, B.S.; and Aimin Zhou, Ph.D.
Department of Chemistry, Cleveland State University.
The cause of type I diabetes continues to be a focus of investigation. Studies have revealed that interferon (IFN)- in
pancreatic islets after viral infection or treatment with double-stranded RNA (dsRNA), a mimic of viral infection, is
associated with the onset of type I diabetes. However, how IFN- contributes to the onset of type I diabetes remains to be
obscure. In this study, we found that 2-5A dependent RNase L (RNase L), an IFN--inducible enzyme that functions in IFN
action against viruses and cell proliferation, played an important role in dsRNA-induced onset of type I diabetes. By using
RNase L deficient RIP-B7.1 mice which are more vulnerable to environmental harmful factors such as viral infection, we
demonstrated that deficiency of RNase L in mice resulted in a significant delay of diabetes onset induced by
polyinosinic:polycytidylic acid (poly I:C), a type of dsRNA, and streptozotocin (STZ), a drug which can artificially induce
type I-like diabetes in experimental animals. Immunohistostaining showed that the population of infiltrated CD8 + T-cells was
remarkably reduced in the islets of RNase L deficient mice, implicating RNase L may contribute to type I diabetes onset
through regulating immune responses. Furthermore, RNase L was responsible for the expression of certain proinflammatory
genes in the pancreas under the special condition. Our findings provide new insight into the molecular mechanism underlying
-cells destruction and may suggest novel therapeutic strategies for treatment and prevention of the disease based on the
selective regulation and inhibition of RNase L.
094
GENDER DIFFERENCES IN HELP-SEEKING BEHAVIORS OF INDIVIDUALS REFERRED FROM A
SUBSPECIALTY MEDICAL PRACTICE
Courtney N. Jones1; Katherine Lamparyk, Psy.D.2,
1
Department of Psychology, Cleveland State University;
2
Cleveland Clinic, Children's Hospital for Rehabilitation
Studies have shown it is more common for women to have a positive attitude associated with help-seeking behaviors,
resulting in an increased likelihood of engaging in the seeking of professional psychological help, compared to men (Nam,
38
2010). However, this has not readily been studied in pediatric medical populations. The study utilizes data of a larger medical
records project examining the referral patterns to psychology from a pediatric subspecialty medical practice. The process
includes a referral stage, where the patient is referred to a psychologist by their gastroenterology doctor, followed by the
consultation with the psychologist, who may then recommend treatment. Is there a significant difference between gender
when it comes to the stages that people reach in the overall process? Does the gender of the referring doctor, and whether or
not this matched the patient, make a difference in whether or not the patient follows through with the referral? Is this effect
moderated by the reason for the psychology referral (e.g. behavioral vs. anxiety, etc.)? These questions will be answered by
examining the medical charts of over 500 patients that were referred to psychology over a two year period.
095
WORKING MEMORY AND PROCESSING SPEED OF ADHD CHILDREN COMPARING TO THEIR
NON-ADHD PEERS AND THE ABILITY TO PREDICT THE LEVEL OF READING COMPREHENSION
IN ADHD CHILDREN BASED ON THESE ASPECTS OF INTELLIGENCE
Sara Al-Nimer, B.A.1; Katherine, Lamparyk, Ph.D.2
1
Department of Psychology, Cleveland State University;
2
Center for Pediatric Behavioral Health, Cleveland Clinic
Children with Attention-Deficit/Hyperactivity Disorder (ADHD) are vulnerable to a significant risk of poor outcomes
including healthy development of required knowledge and skills for academic achievement. A plethora of research in
Psychology has demonstrated Children diagnosed with Attention Deficit Hyper Activity Disorder suffer from weak Working
Memories in which Working Memory (WM) is believed to be a cognitive endophenotype of the disorder. Working memory
also plays a key role in the acquisition of other cognitive and motor skills that children who are diagnosed with ADHD are
prone to lack as a result. There is only a handful amount of research on the relation between ADHD and processing speed of
the brain. Therefore, in this research, we will compare scores from the Wechsler IQ tests - Working Memory and processing
speed subtests taken by children who are diagnosed with ADHD and non-ADHD children. These data was collected by a
Pediatric Psychologist in a hospital. Also, as there is an abundance of empirical evidence that IQ tests can be a predictor of
academic achievement by the acquisition of the necessary skills, we will use our results to investigate whether working
memory and processing speed would be a predictor of the reading comprehension skill in ADHD children. It is clear that if
an individual with ADHD has a disorder involving basic word reading, then he/she will most likely also have difficulty with
reading comprehension; however, studies of children with ADHD without reading basic word reading deficits have shown
that individuals with ADHD often have difficulties in reading comprehension even though they have intact word reading
accuracy (Ghelani, K., Sidhu, R., Jain, U., & Tannock, R. (2004), perhaps as a result of their ADHD-related executive
function deficits (Sesma, H.W., Mahone, E.M., Levine, T., Eason, S., & Cutting, L. (2009).
096
MANUAL ASYMMETRY, TASK DIFFICULTY, AND THE SPEED-ACCURACY TRADE-OFF
Saraha Martincak, B.A.; Andrew B. Slifkin, Ph.D.
Department of Psychology, Cleveland State University
A large literature has been devoted to understanding how handedness influences the control of movement. In general, righthand dominant individuals exhibit performance superiority with the right hand and weaker performance with the left hand.
Some evidence suggests that between-hand differences in performance should be amplified with increases in task demands.
In the current study, we examined whether dominant-hand performance superiority would be maintained at different levels of
task difficulty. To that end, under instructions for movement speed and accuracy, we examined performance of the left and
right hand under two levels of task difficulty. Under the low-difficulty condition, participants (n = 13) moved over a short
amplitude (A = 19.05 mm) to a target with a narrow width (W = 9.53 mm), whereas under the high-difficulty conditions,
participants moved over a larger amplitude (A = 152.40 mm) to a target with same narrow width (W = 9.53 mm). We
measured performance speed as movement time—shorter movement times would reflect higher speeds—and performance
accuracy was measured as the percentage of movements terminating within the target. The results showed that movement
time increased with increases in task difficulty, but the left- and right-hand movement times were the same at both the lowand high-difficulty conditions. Accuracy was high and equivalent for the left and right hands at the low-difficulty condition,
but the left hand was reliably less accurate than the right hand in the high-difficulty condition. Thus, handedness differences
39
were expressed as differences in accuracy but not differences in speed, and the differences in accuracy only emerged under
conditions of high difficulty. We will discuss potential explanations for those findings.
097
THE MOOD-ALTERING EFFECT OF MUSIC: IMMEDIATE LEVELS OF DEPRESSION AND
ANXIETY UPON THE PRESENTATION OF MUSIC AND THE EFFECT OF MUSIC GENRE ON
OUTCOMES
Lindsay E. Braun1; Paul D. Skalski, Ph.D.2
1
Department of Psychology, Cleveland State University;
2
School of Communication, Cleveland State University
With the quickly escalating presence of media in society, the number of studies on the effects of media on psychological
functioning has recently increased. In the present study, the effects of music on mood are examined. The purpose of this
study is to examine if the presentation of music has an immediate effect on levels of depression and anxiety and if the genre
of the song has any impact on the result. Using a sample of adult students at Cleveland State University, participants will be
randomly assigned into one of five conditions: control, jazz, classical, metal, or rap. The participants in the jazz, classical,
metal, and rap conditions will first be presented with a song selection that corresponds with the genre that they were initially
assigned. Those in the control group will not be presented with a song selection. All participants will then complete a survey,
which will consist of questions that indicate the participants' levels of depression and anxiety. Utilizing the data that are
collected, the outcomes of the control group and the music group will be compared. Also, the differences in the outcomes
among the genres will be analyzed. Due to the results of previous similar studies, the anticipated outcome is that the
presentation of music will cause a significant difference in levels of depression and anxiety in comparison to the control
group. It is also anticipated that the presentation of music will reduce depression and anxiety levels in genres that are
considered to be more soothing (i.e., classical and jazz) and increase depression and anxiety levels in genres that are
considered to be more abrasive (i.e., metal and rap). Significant findings can increase the knowledge of the effects of media
on psychological functioning, as well as provide further evidence of the mood-altering effect of music.
098
THE EFFECTS OF GOAL ORIENTATIONS AND COMPETITIVE ENVIRONMENTS ON LEARNING
AND TASK PERFORMANCE
Michael Horvath, Ph.D.; Nicole A. Celin, B.A.; Megan E. Crane, B.A.; Adam M. DeLong, B.A.; Kylie Paulina, B.A.;
Charles W. Benton; Sarah D. Borgos, B.A.; Jessica R. Burt, B.A.; Natalie J. DiNero, B.A.; Morris S. Edelstein, B.A.;
Katherine Elias; Laura J. Elias; Dana E. LoPresti; Gregory J. McKay, B.A.; Mark W. Murch, B.A.; Renee Popovic, B.A.;
Danielle R. Simpson, B.A.
Department of Psychology, Cleveland State University
In achievement situations, individuals are generally motivated in one of three ways: to learn as much as they can (mastery), to
demonstrate their competence (performance-approach), or to avoid demonstrating their incompetence (performanceavoidance). Previous research has shown that these orientations can facilitate learning and performance, but less is known
about how contextual factors such as a competitive environment affect these relationships. In this study, we predict that the
competitiveness of an achievement setting will have effects on both learning and performance, but that this effect will be
strongest for individuals with performance-approach orientations. To evaluate this hypothesis, we are randomly assigning
participants to one of six conditions (a combination of three levels of goal orientation crossed with two levels of
competitiveness). Participants engage in 50 trials of a complex and novel task, receiving feedback on their performance each
time. Afterwards, participants complete a test that measures their knowledge of the rules of the task. Finally, all participants
complete another 50 trials under the performance-approach / non-competitive condition in order to determine whether the
manipulations of goal orientation and competitiveness in the ‘learning’ phase of the task affected participants’ ultimate ability
to perform it.
40
099
THE EFFECT OF OTOLITH FUNCTION ON PERSPECTIVE TAKING
Jessica L. Willesch; Naohide Yamamoto, Ph.D.
Department of Psychology, Cleveland State University
The purpose of the proposed study is to gain insight into individual differences in spatial abilities, especially those observed
in perspective taking. It is hypothesized that people who perform perspective taking poorly have great difficulty in imagining
self-motion, which stems from perceptual deficiency in their self-motion sensing systems. In particular, we will focus on their
otolith function that measures linear components of body velocity and acceleration. In this study, participants’ otolith
function will be measured by a water level test, and those with good and poor otolith function will perform a judgment of
relative direction (JRD) test. This test requires participants to judge object locations from a novel perspective in a
remembered environment. The expected results are for participants who score poorly on the water level test (i.e., poor otolith
function) to also score poorly on the JRD test. These results would show that perceptual functioning of the otolith organ is
correlated with perspective taking ability, which suggests that increased difficulty in perspective taking originates from the
imagined movement deficit. The expected results can also help explain large individual differences that occur during JRD
tests.
100
RHOP-3 ANTIGEN AMPEROMETRIC IMMUNOSENSOR FOR THE DETECTION OF MALARIA
Ahmad Maarouf; Mekki Bayachou, Ph.D.
Department of Chemistry, Cleveland State University
In this work we present the groundwork to develop a sensitive and highly selective amperometric immunosensor for the
detection of Rhop-3, a malaria antigen protein. Rhop-3 protein has been recognized as a common signature among
different Plasmodium isolates from various geographical regions, and thus it is a potential marker for high throughput
screening for Malaria. The method of detection couples both the sensitive ability of immunosensing and fast response of
electrochemical detection. The method is based on an immunoassay with a response that is enzymatically amplified, and
whose product is electrochemically detected. To develop the method and provide a proof of principle, we used a generic
heme protein (NOS) since we have access to both NOS and NOS antibodies. First, a NOS antibody is immobilized on carbon
electrodes. The immobilized antibody is used to detect NOS protein when exposed to a solution containing NOS. Second,
exposure to NOS primary antibody and then to a horseradish peroxidase (HRP)-labeled secondary antibody will provide a
platform for amplification of the detection signal through the HRP reaction using thionine as an electron transfer mediator.
HRP catalyzes the reduction of H2O2 to H2O and the electrochemical signal is proportional to the amount of NOS initially
detected on the modified carbon electrode . We use cyclic voltammetry to monitor the catalytic activity, which is a direct
result of the formation of the immunocomplex between the immobilized antibody and its cognate antigen in the solution. This
immunosensing electrochemical technique is being optimized for the purpose to develop electrochemical immunosensors in
analyte samples containing target Rhop-3 antigen for malaria screening. Once optimized, the technique will enable the
development of cheap screen-printed electrodes for fast, accurate, sensitive, and selective diagnostic tools that can be adapted
for field devices.
101
SOURCE WATER COMPONENT DETERMINATION OF WEST CREEK, OHIO USING STABLE
ISOTOPE FRACTIONATION ANALYSIS
Daniel A. Gouch
Department of Biological, Geological and Environmental Sciences; Cleveland State University
Elements of low atomic number usually form several stable isotopes. When these isotopes react to form compounds, the
molecules produced differ from each other by the isotope they contain. The different atomic weights affect the way these
molecules respond to certain kinds of physical and biogeochemical processes that are mass dependent. Observed differences
in the isotope compositions of oxygen are caused by processes referred to as isotopic fractionation. Several methods were
used, during the summer of 2012, utilizing stable oxygen isotope fractionation to address questions of where the water comes
from in a stream and where the water goes once it reaches the ground.
41
The surface flow within a stream derives from a number of sources: direct precipitation falling on the stream, overland runoff
(including via anthopogenic modification, i.e. storm sewer discharges), and groundwater intrusion being the primary three
sources of water for surface flow. Each of these sources of has a distinct stable oxygen isotope chemical signature, which,
when combined together as surface flow will then have its own unique stable oxygen isotope chemical signature. By
determining the distinct chemical signatures from the primary source waters and surface flow, the relative volumes of input
from each primary source can be well estimated. Such information could be of use for determining groundwater retention
time, and useful to assess the potential long-term effectiveness of recently introduced projects designed to decrease the
amount of impervious surface runoff for a heavily modified small urban tributary to the Cuyahoga River - West Creek.
102
UNDERSTANDING THE ILLNESS EXPERIENCE OF INDIVIDUALS WITH DEMNTIA:
MODERATORS OF WELL-BEING
Lindsey M. Bisgrove, B.A.1, Katherine S. Judge, Ph.D.1
1
Department of Psychology, Cleveland State University
Until recently, few studies have investigated the psychosocial outcomes of individuals living with dementia. More
specifically, the relationship among depression, anxiety, and quality of life as well-being outcomes. The current study
examines the role of well-being outcomes and how key moderators (level of cognitive impairment, level of distress
experienced, relationship role strain) may influence these outcomes. It addresses how these specific moderators have an
impact, individually and mutually, on depression and anxiety symptoms and overall quality of life. This study utilizes the
stress process model for individuals with dementia as a framework to demonstrate the intricacies of the illness experience for
the individuals. Results indicate a significant positive correlation between symptoms of depression and symptoms of anxiety,
with higher symptoms of depression related to higher symptoms of anxiety. There is a significant negative correlation
between higher levels of symptoms of depression and QoL, with higher symptoms of depression related to a decreased
overall QoL. Results also show a significant negative correlation for higher levels of symptoms of anxiety and QoL, with
higher symptoms of anxiety related to a decreased overall QoL. Significant unique predictors of depression include PADL
distress, cognitive impairment, and relationship strain. Significant unique moderators of symptoms of anxiety include
cognitive impairment. Significant unique moderators of a lesser QoL included relationship strain, PADL distress, and
cognitive impairment. The knowledge that can be gained by examining previous and current research has the potential to
improve and develop future care practices and advance new intervention techniques.
103
EVALUATION OF CLIENT SATISFACTION WITH PRO BONO PHYSICAL THERAPY SERVICES AT
A FREE MEDICAL CLINIC
Mary Milidonis, PhD1; Scott Euype, DPT2; Karen O’Loughlin, DPT1; Karen Vitak, DPT1; Erin Duffy, BSHS1; Megan
Melchior, BSHS1
1
School of Health Sciences, Cleveland State University;
2
Cleveland Clinic
Little is known about client satisfaction with physical therapy services in a pro bono practice environment. The purpose of
this project was to evaluate client satisfaction with pro bono physical therapy services provided at the Free Clinic of Greater
Cleveland. Methods and Materials: A satisfaction survey was developed with research constructs related to treatment
outcomes, physical therapist attributes, processes and organization of care. The final survey was composed of 30 questions
and was administered by a trained interviewer who read the questions and recorded subjects’ responses immediately
following the first physical therapy visit. Chart abstraction was done to record subject co morbidities and physical therapy
diagnoses. Data Analysis: Completed paper surveys were scanned into Snap Survey Software to ensure reliable data entry.
SPSS 18 was used to define descriptive statistics. Results: Forty six patients with mean age 49.2 (9.9sd), 47.8% male, 61%
black, 59% unemployed were receiving pro bono physical therapy services and completed informed consent to complete the
satisfaction survey. The highest rated survey items included: ‘feeling the therapist listened’, ’was respectful’ and ‘explained
instructions well.’ A strong correlation (r = .731) between overall satisfaction with ‘returning to the clinic was found’.
Moderate correlations were found between satisfaction and ‘physical therapist gives detailed home instructions’ (r = .620)
and ‘following received instructions’ (r =.494). Lowest rated survey items after first visit were related to ‘complete
resolution of pain’, ‘restoration of full mobility’ and ‘making a complete recovery.’ Conclusions: As expected, optimal
42
treatment outcomes were not achieved in the first visit. However, patients still reported satisfaction with physical therapy
services and would return to the same office. Satisfaction was highest with physical therapist interactions and lowest with the
organization/structure of the clinic.
104
TRYPANOSOMA BRUCEI TIN2 INHIBITS VSG SWITCHING BY MAINTAINING THE SUBTELOMERE
INTEGRITY
Sanaa Jehi, M.S. and Bibo Li, PhD
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Trypanosoma brucei is a protozoan parasite and the causative agent of human African trypanosomiasis. T. brucei evades the
host immune responses by regularly switching its surface antigen – Variant Surface Glycoprotein (VSG), which is
exclusively expressed in a monoallelic fashion from VSG expression sites (ESs) located adjacent to the telomere. Telomeres
are specialized nucleoprotein complexes at the ends of linear chromosomes and are essential for chromosome stability and
genome integrity. Telomere functions are particularly important for antigenic variation in T. brucei. We have identified
TbTIN2 as an intrinsic component of the T. brucei telomere complex. Using a cell line carrying both positive and negative
selective markers in the active ES, we found that a temporary depletion of TbTIN2 led to a significant increase in the VSG
switching frequency. Most of the VSG switchers in TbTIN2-depleted cells appear to arise through gene conversion events
encompassing the entire active ES or loss of the active ES coupled with in-situ switching. Using ligation mediated-PCR, we
observed many more DNA double strand breaks (DSBs) in the subtelomeric region upon depletion of TbTIN2. This suggests
that wild-type TbTIN2 has an essential role in maintaining the subtelomere integrity. In addition, depletion of TbTIN2 led to
an increase in the association of RAD51 with the subtelomeric DNA. RAD51 homologs in other organisms are involved in
the strand invasion step during homologous recombination. Therefore, our observation suggests that DNA damage repair
mediated by TbRAD51 through homologous recombination led to subsequent increased VSG switching frequency.
105
A TRANS-SPLICED TELOMERASE RNA DICTATES TELOMERE SYNTHESIS IN TRYPANOSOMA
BRUCEI
Ranjodh Sandhu1, Samantha Sanford2, Shrabani Basu2, MinA Park2, Unnati M. Pandya1, Bibo Li1*, Kausik Chakrabarti2.*
1
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University, Cleveland, OH 44115.
2
Department of Chemistry, Center for Nucleic Acids Science and Technology, Carnegie Mellon University, Pittsburgh, PA
15213.
Telomerase is a ribonucleoprotein enzyme typically required for sustained cell proliferation. Although both telomerase
activity and the telomerase catalytic protein component, TbTERT, have been identified in the eukaryotic pathogen
Trypanosoma brucei, the RNA molecule that dictates telomere synthesis has remained unknown. Here we identify the RNA
component of T. brucei telomerase, TbTR, and provide phylogenetic and in vivo evidences for TbTR’s native folding and
activity. We show that TbTR is processed through trans-splicing, is a capped transcript that interacts and copurifies with
TbTERT in vivo. Deletion of TbTR caused progressive shortening of telomeres at a rate of 3–5 bp/PD, which can be rescued
by ectopic expression of a wild type allele of TbTR in an apparent dose-dependent manner. Remarkably, introduction of
mutations in the TbTR template domain resulted in corresponding mutant telomere sequences, demonstrating that telomere
synthesis in T. brucei is dependent on TbTR. We also propose a secondary structure model for TbTR based on phylogenetic
analysis and chemical probing experiments, thus defining TbTR domains that may have important functional implications in
telomere synthesis. Identification and characterization of TbTR not only provide important insights for investigation of T.
brucei telomere functions, which have been shown to play important roles in T. brucei pathogenesis, but also offer T. brucei
as an attractive model system for studying telomerase biology in pathogenic protozoa and for comparative analysis of
telomerase function with higher eukaryotes.
43
106
REALITY VERSUS PERCEPTION: A COMPARISON OF FOOD AVAILABILITY ON CLEVELAND
STATE UNIVERSITY’S CAMPUS COMPARED TO STUDENTS’ PERCEPTIONS
Samantha L. Bader, B.A.1; Albert F. Smith, Ph.D.2
1
Department of Health, Physical Education, Recreation and Dance, Cleveland State University;
2
Department of Psychology, Cleveland State University
We assessed the healthfulness of food available on Cleveland State University’s (CSU) campus, assessed students’ beliefs
about food availability, and will compare the two. To assess the healthfulness of available food, NEMS-R (Nutrition
Environment Measures Survey-Restaurants) and NEM-S (Nutrition Environment Measures Survey-Stores) evaluations were
conducted of CSU’s 13 food outlets. (NEMS was developed by researchers at Emory University and the University of
Pennsylvania to measure food availability within a specific environment.) To assess students’ perceptions, a brief paper
survey was distributed at >10 locations on CSU’s campus.
The average NEMS-R score of the 11 restaurants on CSU’s campus was 5.82 points on a scale of -27 to 63 points; the
average NEMS-S score of the 2 stores on CSU’s campus was 10.5 points on a scale of -9 to 54 points. Of these outlets, 18%
have nutrition information available on-site and 64% have nutrition information available online through the CSU campus
website. There were no price differences between healthy and unhealthy entrees.
A total of 605 individuals completed the brief survey; 59 surveys were excluded due to one or more missing responses,
leaving 546 surveys for analysis. Analyses of the survey data will examine students’ perceptions of healthy food availability
as a function of sex, residential status (on-campus resident vs. commuter), and frequency of dining on campus.
In conclusion, completed analyses show that, by NEMS criteria, CSU has low availability of healthy foods. Further analyses
of data will investigate the extent to which the beliefs of subgroups of students correspond to this assessment.
107
STRESS PROCESS MODEL OF CAREGIVER AND CARE RECIPIENT
Kelly Martincin, M.A.1 and Katherine Judge, Ph.D.2
1
Department of Counseling Psychology, Cleveland State University,
2
Department of Psychology, Cleveland State University
Demographic information such as race and education level, cognitive abilities, level of dependency, relationship strain, and
social isolation of the caregiver may create negative outcomes for the individual with dementia (IWD), including depression
and worse physical health outcomes. However, few studies have investigated this interaction between characteristics of
caregivers and psychosocial outcomes of IWDs. In addition, no conceptual model has been proposed that explicitly links both
caregiver and care recipient characteristics and the resulting interactions. The purpose of this poster is to provide a
comprehensive review of key caregiver variables that may contribute to outcomes of community dwelling individuals with
dementia. Drawing from the Stress Process Model (Pearlin, Megaghan, Lieberman, & Mullan, 1981) for Caregivers and the
Stress Process Model for Individuals with Dementia (Judge, Menne, & Whitlatch, 2009), this poster will present a theoretical
model for understanding and examining the stress process for caregiving dyads, including how primary stressors, secondary
stressors, and an individual’s environment can contribute to psychosocial outcomes. Discussion will focus on how this new
theoretical model can be used to examine the complex interactions between caregivers and IWDs, along with specific
interventions that may mitigate the negative symptoms of dementia for both care partners. Two measurable objectives of this
poster are first increased awareness of the current use of the Stress Process Model and second increased awareness of
potential new uses of the Stress Process Model, including use for the caregiving dyad of IWDs.
44
108
ROLE OF ZIP1 IN THE SYNAPTONEMAL COMPLEX IN MEIOSIS
Jasvinder Singh, M.S.; Hanna Morris and G. Valentin Borner, Ph.D.
Department of Biological, Geological and Environmental Sciences; Cleveland State University
My main research interest is the function of the synaptonemal complex and associated structures. Following a screen for
mutants with meiotic defects, I discovered a requirement for a subunit of the 20S proteasome in synaptonemal complex
assembly and crossover formation. Chemical inhibition of the proteasome also results in defective recombination and
synaptonemal complex morphogenesis. Strikingly, the proteasome is recruited to distinct positions along meiotic
chromosome axes in a DSB-dependent manner. Together, these results suggest that locally tethered proteasome particles
directly control key events along meiotic chromosomes. Presently, I am examining the relationship between the proteasome,
synaptonemal complex formation and distribution of crossover markers along the meiotic chromosomes.
109
LOCUS OF CONTROL AND ASSIGNED VERSUS SELF-SET GOALS IN OLDER ADULTS
Makara N. Scruggs; Katherine S. Judge, Ph.D.
Department of Psychology, Cleveland State University
It has been recognized that goals produce higher performance when compared to having no goals in a variety of settings.
Along with goals, locus of control also has been commonly associated with performance outcomes and looked at as a
mediator in goal setting and achievement. What research has not investigated, however, is the role of locus of control in
performance with self-set goals versus assigned goals. This could have important implications when it comes to rehabilitation
and physical therapy settings. Older adults often constitute a large portion of individuals using physical therapy. When
working with older adults in this setting it is very important to understand their motivation dynamic and the factors that affect
their motivation to increase progress and success within their treatment protocol. The present study will investigate whether
older adults with an internal locus of control perform better on physical therapy tasks when given assigned goals versus
setting their own goals. It is predicted that older adults who have an internal locus of control will perform better on given
tasks when using self-set goals versus assigned goals.
110
ENDOTHELIAL NITRIC OXIDE SYNTHASE (eNOS) ON LIPID NANODISCS BASED ON MEMBRANE
SCAFFOLD PROTEINS: TOWARDS A SOLUBLE ASSEMBLY REFLECTING NATIVE-LIKE
FUNCTION OF eNOS.
Tunde S. Veres, B.A.; Mona Patel, M.A.; Ghaith AlTawallbeh, M.A.; Mekki Bayachou, Ph.D.*
Department of Chemistry, Cleveland State University.
Nitric oxide (NO) is an important signaling molecule involved in neurotransmission, immune responses, and in vasodilation
among other roles in the cardiovascular system. NO is synthesized by a family of enzymes called NO synthases (NOSs),
which catalyze the oxidation of L-arginine to citrulline and NO. Endothelial nitric oxide synthase (eNOS) is one member of
this family, and is primarily associated with plasma membranes. Understanding the details of the molecular function of this
enzyme is of critical importance to understand eNOS dysfunction and its pathological implications. All the work published
on the mechanisms eNOS function is based on recombinant soluble forms, and the possible effect of phospholipid
membranes present in the enzyme’s native microenvironment is not known.
To mimic the enzyme’s microenvironment and characterize its native-like kinetic and thermodynamic parameters, we are
preparing eNOS assembled on self-contained phospholipid nanodiscs formed by lipoproteins scaffolds. Nanodiscs have
recently been used as models of membranes for membrane-bound proteins in order to study the protein’s native function. In
this project, we are preparing and characterizing eNOS/nanodiscs assemblies for the first time. Homogenous eNOS/nanodiscs
samples are purified using nickel nitriloacetate affinity column followed by size exclusion chromatography to target
assemblies of one protein per nanodisc. Griess and hemoglobin-dioxy assays are used to measure activity of free and
nanodisc-bound enzymes. Redox titration of eNOS/nanodisc assemblies is used to determine the thermodynamic parameters
45
of the heme catalytic center in the assembly versus free enzyme. Results on the characterization of activity of eNOS/nanodisc
assemblies will be presented and discussed.
111
NITRIC OXIDE SYNTHASE (NOS) IN THIN FILMS AS A NITRIC OXIDE GENERATING COATINGS
TO COUNTERACT THROMBOSIS ON MEDICAL DEVICES
Bhagya Gunasekera, M.S.; Mekki Bayachou, Ph.D.*
Department of Chemistry, Cleveland State University
Thrombosis is a major problem on blood-contacting medical devices. Like the native endothelium, artificial coatings
releasing small amounts of nitric oxide (NO) are known to counteract platelet adhesion, and thus the thrombosis cascade.
Various sources capable of releasing NO have been proposed as coatings for blood-contacting devices. We proposed a novel
approach using recombinant NOS in thin layers, which generate NO using substrates from the bathing medium. We first
proposed and introduced the Layer-by-layer (LbL) adsorption of inducible NOS (iNOS) on polyethylenimine (PEI) matrix on
representative surfaces. In the current work, we study the effect of pH on NOS loading during the fabrication of layer-bylayer coatings. We also study the effect on the catalytic efficiency of immobilized NOS and the resulting NO fluxes under
various conditions.
We examined how the pH of the protein solution and the charge density of PEI matrix can modulate enzyme loading. We
used buffered iNOSoxy solutions and both linear and branched PEI solutions to investigate the charge-driven LbL
immobilization process.
We used Fourier Transform infrared spectroscopy (FTIR) to characterize immobilized iNOSoxy layers. Atomic force
microscopy (AFM) suggests more iNOSoxy units immobilized on films formed at pH 8.6 and/or using the branched version
of the PEI matrix. We used catalytic reduction of exogenous NO mediated by immobilized iNOSoxy in the LbL films as a
measure of film activity. Results show higher activity for films immobilized at pH 8.6 compared to pH 7. Monitored NO
release using the Griess assay show higher levels of NO released from films constructed with iNOSoxy solution at pH 8.6
compared to pH 7. Also, a higher average NO flux is observed from PEI/iNOSoxy LbL films having the branched version of
PEI. We will discuss these findings in light of platelet adhesion on surfaces coated by the various LbL films.
112
PROFILING OF LIPIDS IN ALGAE, SCENEDESMUS DIMORPHUS BY MASS SPECTROMETRY
Satya Girish Avula1 M.S.; Joanne Belovich2 Ph.D.; Yan Xu1 Ph.D.,*
1
Department of Chemistry, Cleveland State University
2
Department of Chemical Engineering, Cleveland State University
As concern grew over the supply of fossil fuels, other sources of energy are sought. Algae has significant promise as a
feedstock for biodiesel production. Microalgae have the ability to synthesize, accumulate, and store large amounts of lipids
(up to 40% of dry biomass) in its cells, which are precursors for biodiesel. In order to optimize growing conditions of various
microalgae species in the engineered gravity settlers for maximizing biodiesel production, qualitative analytical methods are
needed for lipid profiling.
In this work, we reported a qualitative analytical method for profiling neutral and polar lipids from algae species
Scenedesmus dimorphus extracted by Bligh-Dyer method. A total of 116 lipids including phosphocholines (PC),
phosphotidylethanolamines (PE), phosphatidylinositols (PI), phosphatidylglycerols (PG), phosphotidic acids (PA), monoand tri-acylglycerides (MAG and TAG), as well as a series of fatty acids (FA), were identified using multiple mass
spectrometric platforms including quadrupole time-of-flight mass spectrometer (Q-TOF), quadrupole linear-ion-trap mass
spectrometer (Q-Trap) and multiplexed precursor ion scanning (MPIS), and gas chromatograph and mass spectrometer (GCMS). These lipids and fatty acids were annotated by tandem mass spectral (MS/MS) database matching with Lipid Maps
database and LipidViewTM software.
46
113
NON-CANONICAL ROLE FOR THE DR5/FADD/CASPASE PATHWAY IN THE REGULATION OF
EXPRESSION OF THE MUSCLE REGULATORY TRANSCRIPTION FACTOR MYOD
Jason A. Ross, B.S.; Crystal M. Weyman, PhD.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Our lab has previously reported that the stable expression of a dominant negative Death Receptor 5 (dnDR5) in proliferating
myoblasts inhibits differentiation by decreasing mRNA and protein expression of the master muscle regulatory factor MyoD,
and without altering MyoD mRNA stability. Further, we have shown that the acetylation of histones associated with the
MyoD promoter in dnDR5 myoblasts is decreased and that treatment with a histone deacetylase (HDAC) inhibitor allows
partial recovery of MyoD expression and differentiation in differentiation-defective, dnDR5-expressing myoblasts,
suggesting that an increase in histone acetylation is sufficient for MyoD expression. The expression of MyoD is
transcriptionally regulated by a distal regulatory region (DRR), which includes a non-canonical serum response element
(SRE) that is required for MyoD expression in both differentiated and proliferating myoblasts. Contained in this cis-element
is a CArG box required for MyoD expression; it has been demonstrated that two trans-acting factors, SRF and Mef-2, bind to
this CArG element in proliferating and terminally differentiated myoblasts respectively. Myoblasts stably expressing dnDR5
exhibit an increase in the amount of Mef-2 bound at this CArG element when compared to Mef-2 bound to this CArG
element in wild-type (WT) cells. We infer that there is an inverse relationship between MyoD expression and Mef-2
recruitment, and that recruitment of HDAC(s) by Mef-2 bound to the CArG element in the MyoD DRR may at least partially
explain the reduction in MyoD protein in proliferating and differentiating myoblasts and the concomitant delay in
differentiation observed when dnDR5-expressing cells are induced to differentiate. The goal of the present project is to
elucidate the mechanism(s) by which dnDR5 expression affects the level of Mef-2 at the CArG element.
114
GEOGRAPHIC VARIATION IN SONGS OF THE VEERY (CATHARUS FUSCESCENS) ALONG THE
APPALACHIAN MOUNTIANS
Courtney L. Brennan, B.S.1; Andrew W. Jones, Ph.D.2
1
Department of Biological, Geological, and Environmental Sciences; Cleveland State University;
2
Cleveland Museum of Natural History
The Veery (Catharus fuscescens) is a Neotropical migrant that breeds in secondary growth and woodland habitats subject to
fragmentation and loss (Bevier et al. 2005). The Veery breeding distribution spans from southeastern British Columbia across
southern Canada and south into the northeastern United States through the higher elevations of the Appalachian Mountains
from western Maryland to northern Georgia (Bevier et al., 2005). Veery song may vary regionally in the Appalachians
(Bevier et al., 2005) but these variations have not been studied. During May-June 2012 I recorded over 1,500 songs from 95
individual Veeries from four locations along the Appalachian Mountains. By comparing various features of their song, this
study investigates geographic variation among breeding Veeries along the Appalachian Mountain range. Song divergence
among bird populations is recognized as a potential mechanism of premating isolation and may be a step towards speciation
(Ruegg et al., 2006).
115
THE UPTAKE OF HEAVY METALS AND DIFFERENTIAL EXPRESSION OF HEAVY METAL
REGULATED PROTEINS IN CHRISTMAS FERN
Paul Ilkanich; Pratheek Koneru; Anne O’Connor, PhD; Chamari Walliwalagedara, PhD; Robert Wei, PhD
Department of Chemistry, Cleveland State University
Plants are considered the most cost effective and environmentally sound way to clean up soils and water contaminated with
toxic metals. The Chinese fern, Pteris vittata, is currently used to remediate arsenic-contaminated soils. A major
disadvantage of this plant species is that it has proven difficult to grow, especially under cold climate conditions. We
therefore tested another fern species, Polystichum acrostichoides (Christmas fern). One major aim of this project was to
determine whether Christmas fern was capable of hyperaccumulating arsenic (As), lead (Pb) and cadmium (Cd). Another aim
was to learn important biochemical changes that occur when the plant is subjected to stress by the toxic metals. Six-eight
47
week old plants were purchased from a local green house and were grown hydroponically in nutritive water that contained the
respective metals for seven days. The plants were ground in liquid nitrogen and stored at -80C upon harvesting. Proteomic
analysis and metal uptake measurements were performed using two different concentrations (150 & 300 µM) of metal ions.
The determination of metal uptake was performed using HNO3 digestion (USEPA method 3050B) followed by Inductively
Coupled Plasma spectroscopy (ICP). The proteomic analysis was performed using polyethylene-glycol fractionation (PEG)
followed by one (SDS-PAGE) and two dimensional gel electrophoresis (2-DE) technique. A total of 17 protein spots were up
regulated and 3 proteins were down regulated upon various metal exposures. Metal uptake and proteomic analysis results will
be discussed.
116
INFLUENCE OF LATE TRANSITION METAL DOPING ON METHANE SELECTIVITY IN FISCHERTROPSCH CATALYSIS
Peter C. Psarras, B.A.; David W. Ball, Ph.D.
Department of Chemistry, Cleveland State University
The effect of late transition metal substitution into Fe(100), Ni(111), and Co(0001) surface analogs has been investigated
with density functional methods. The surface is modeled using a 7-atom cluster, with perimeter atoms saturated with
hydrogen to approximate surface coordination. All calculations were performed at the B3PW91 level of theory with the
LANL2DZ basis. Eight surface adsorbates were studied: C, CH, CH 2, and CH3 represented the hydrogenating steps on
surface carbide, while C + CH, CH + CH, C + CH3, and CH2 + CH2 represented four competitive coupling pathways. It has
been shown that a BrØnsted-Evans-Polanyi relationship exists between the effective barrier to methane formation and the
chemisorption of carbon. A review of the effect of Cu, Ag, Au, and Pd on adsorbate binding energies together with a
comprehensive view of the surface stability of other reactionary intermediates can help to create a model for catalytic
improvement.
117
BORONIC ACID-BASED GLYCO-AFFINITY LIGAND FOR GLYCOMICSAPPLICATIONS
Vinay Kumar Kodithyala and Xue-Long Sun*
Department of Chemistry, Cleveland State University
Cell surface carbohydrates existing majorly as glycoproteins and glycolipids are involved in many biological processes such
as cell communication, and the molecular recognition and cell targeting. However, the mechanisms of most processes at the
molecular level are still unclear. It is therefore very important to develop novel tools for efficient analysis of complex
carbohydrates structures and functions, known as glycomics. In addition, certain carbohydrate molecules can be used for bioanalytical, pharmaceutical and targeted drug delivery applications. Herein, we report synthesis and characterization of
boronic acid (BA)-silica gel conjugates as glyco-affinity ligand for carbohydrate capturing and modification applications. The
BA-silica gel conjugates were synthesized by amidation of carboxylic acid-functionalized silica gel with amino phenyl
boronic acid in the presence of EDC, and were characterized IR spectroscopy. The sugar capturing capacity and specificity
were confirmed by Alizarin Red displacement assay. Further, on-resin glycosylamine formation from the captured sugar was
investigated. Overall, the boronic acid (BA)-silica gel conjugates serve as both glyco-capturing ligand and solid phase for
carbohydrate synthesis and thus will provide an important tool for glycomics research and applications.
118
COMPARISON OF GENETIC DIVERSITY IN THE FRESHWATER MUSSEL PYGANODON GRANDIS
BETWEEN LAKE ERIE AND LAKE ONTARIO
Matthew T. Begley, B.S.1 and Robert A. Krebs, Ph.D.1
1
Department of Biological, Geological and Environmental Sciences, Cleveland State University
Pyganodon grandis is a freshwater mussel (Family Unionidae) common in the Great Lakes. Its mode of dispersal, like other
Unionid mussels, is through larval glochidia which are parasitic on fish gills or scales. P. grandis glochidia have the ability to
parasitize the scales of a wide array of host fish and as a result are more widely distributed throughout the Great Lakes. In
light of this, it still may not be expected that female mitochondrial DNA would pass from Lake Erie to Lake Ontario, over
48
Niagara Falls, through glochidia attached to fish. Sequences of the mitochondrial cytochrome oxidase C subunit 1 (CO1)
gene from 260 samples of P. grandis show one dominant haplotype across Lake Erie, from the western basin to the Niagara
River. Our results from collections of P. grandis from inlets along the eastern end of Lake Ontario suggest that the same CO1
haplotype is the most common there as well. Lake Erie began to flow through the Niagara River into the Ontario Basin
~12,400 years ago with fluctuating flow rates occurring since that period. This flow is responsible for the formation of
Niagara Falls. Our results imply that there may have been more recent gene flow between these two large lake populations to
maintain the common haplotype in the two lakes.
119
mTOR IN BECN1-INDEPENDENT AND AMPK-DEPENDENT AUTOPHAGY OF FLUDARABINERESISTANT LEUKEMIC B CELLS
Arishya Sharma, M.Sc. (Hons.)1,3; Allison Janocha , B.S.E.4; BT Hill, M.D., Ph.D2; M Kalaycio, M.D., Ph.D2; Serpil C.
Erzurum, M.D.4 A Almasan, Ph.D.1;
1
Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic;
2
Department of Hematologic Oncology and Blood Disorders, Taussig Cancer Institute, Cleveland Clinic;
3
Deparment of Biological, Geological and Environmental Sciences; Cleveland State University;
4
Department of Pathobiology, Lerner Research Institute, Cleveland Clinic
Fludarabine (Fd), a purine analog, is a frontline therapeutic for chronic lymphocytic leukemia. Fd resistance is a significant
clinical problem, however, the molecular determinants are incompletely understood. Unlike apoptosis, autophagy, a cellular
catabolic degradation process, causes both cell death and survival. The regulation cell death and acquired Fd-resistance by
autophagy was examined in leukemic cell lines and their Fd-resistant (FdR) derivatives. In sensitive cells, Fd induced
apoptosis and autophagy. MCL-1 degradation following Fd treatment freed pro-apoptotic BIM and autophagy regulator
BECN1, leading to cell death-associated autophagy in Fd-sensitive cells. However, in FdR cells low levels of BIM and
BECN1 sequestration by MCL-1 prevented cell death. Inhibition of apoptosis using siBIM and autophagy using siBECN1,
shATG7, or LAMP2 significantly reduced Fd-induced cell death. Paradoxically, FdR cells were addicted to basal autophagy,
which was independent of BECN1, but it was dependent on AMPK. Interestingly, p70S6 kinase phosphorylation, a marker of
mTOR activation, could predict Fd response in a large cell line panel. FdR cells were addicted to basal mTOR activation and
mTOR inhibition synergized with Fd to cause cell death. AMPK is known to inhibit mTORC1, which, in turn, is known to
inhibit autophagy by ULK-1 phosphorylation. Further investigation unraveled the antinomy of the coexistence of all the three
pathways in FdR cells. Our data suggest that mTORC1 hyperactivation resulted in a prominent metabolic reprogramming in
FdR cells as demonstrated by significantly higher extracellular acidification (ECAR) and oxygen consumption rates; and
higher sensitivity to 2-deoxyglucose and serum starvation of FdR compared to parental Nalm-6 cells. Importantly, mTOR
inhibition could significantly reduce ECAR in FdR cells. Additionally, the mTORC1 and the ULK-1 complexes co-existed in
FdR cells due to their spatial uncoupling. Thus, we establish that both autophagy and mTOR pathway activation are critical
determinants of Fd-resistance and delineate the responsible molecular mechanism.
120
CYTOTOXIC EFFECTS OF TRAIL ALONE AND IN COMBIATNION WITH ELLAGIC ACID IN
VITRO AND IN VIVO
Katherine A. Turner, B.S.1, 2; Daniel Lindner, MD, Ph.D.2; and Michael Kalafatis, Ph.D.1, 2, 3
1
Department of Chemistry, Cleveland State University;
2
Taussig Cancer Institute, The Cleveland Clinic;
3
Department of Biological, Geological and Environmental Sciences; Cleveland State University
Cancer is a widespread disease characterized by unregulated cell growth and evasion of apoptotic stimuli. A direct and
selective cancer therapy is the application of Tumor necrosis factor-Related Apoptosis-Inducing Ligand (TRAIL).
Recombinant human TRAIL (rhTRAIL) can induce apoptosis (programmed cell death) in a broad range of transformed
human cells. However, some cancers remain resistant to rhTRAIL-induced apoptosis. Casein kinase 2 (CK2) is a ubiquitous
kinase that is found to promote cancer growth. CK2 is up-regulated in the nucleus of cancer cells and its activity can induce
the activity of inhibitors of apoptosis causing resistance to cell death. By inhibiting CK2, resistant cells may become sensitive
to TRAIL-induced apoptosis. Therefore, co-treatment with the natural CK2 inhibitor, ellagic acid (EA), is proposed. In vitro
analysis of malignant melanoma cell line A375 and prostate carcinoma cell line DU145LUC shows that both cell lines are
49
sensitive to TRAIL and EA alone as single-agent treatments, compared to the untreated control. Combination treatment of
TRAIL plus EA results in a greater decrease of cell viability compared to the single-agent treatments in both cell lines. Chou
and Talalay median effect analysis shows that the combination treatment of TRAIL and EA works in a synergic mechanism
to inhibit cancer cell proliferation. Athymic nude mice bearing A375 xenografts show tumor growth inhibition and prolonged
survival rate following treatment with TRAIL alone and in combination with EA compared to the untreated control group
and the group treated with EA alone. In contrast, in mice bearing xenografts with DU145LUC, EA alone and TRAIL plus EA
were the most effective in inhibiting tumor growth and increasing survival rate compared to the untreated control mice and
the mice treated with TRAIL alone. These data demonstrate that EA can potentiate the apoptotic effect of TRAIL in a
synergistic mechanism in vitro and in vivo.
121
COMBINED HETEROZYGOCITY IN TWO PREVIOUSLY-UNIDENTIFIED CARBOXYLASE
MUTATIONS CAUSES SEVERE VITAMIN K-DEPENDENT CLOTTING DEFICIENCY
Aisulu Usubalieva, B.S.1; Kevin W Hallgren, B.S.1; Mark A Rishavy, PhD1; Dhouha Darghouth, PhD2; Jean-Philippe Rosa,
MD2; and Kathleen L Berkner, PhD1
1
Department of Molecular Cardiology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH
2
U770, INSERM, Le Kremlin-Bicetre, France
The gamma-glutamyl carboxylase activates vitamin K-dependent proteins, which are essential for functions that include
hemostasis, arterial calcification, signal transduction, bone development. Naturally-occurring carboxylase mutations result in
two distinct diseases: severe clotting deficiency or pseudoxanthoma elasticum that is associated with mild bleeding and
mineralization of soft connective tissue. The reason why naturally-occurring mutations cause two diseases is unknown. The
gene for the carboxylase is autosomal, and we identified a patient with severe bleeding who is compound heterozygous for
two mutations: Trp493Cys and Arg704stop. The parents of the patient with either the Trp493Cys or the Arg704stop mutation
and one wild type carboxylase gene are unaffected, indicating that combination of these two mutations results in the disease.
We have expressed the mutants in insect cells, which lack endogenous carboxylase but express fully active recombinant
enzyme. Functional analysis indicated that the Trp493Cys had poor carboxylase activity, which might be the reason for the
defective hemostasis in the patient. In contrast, the Arg704stop mutant that is truncated in the C-terminal 55 amino acids had
wild type carboxylase activity. However, the Arg704stop mutation must contribute to defective carboxylation because the
parent with one copy of Trp493Cys and one copy of wild type carboxylase is unaffected. This defect may be decreased
protein stability, as the expression level of the Arg704stop mutant was lower in insect cells than that of wild type
carboxylase. We are currently testing this possibility using immortalized lymphocytes derived from the patient and his
parents’ blood. By characterizing these mutants we hope to better understand the carboxylation mechanism and find an
approach for the treatment of patients with hemostatic disorders and pseudoxanthoma elasticum.
122
IMPROVING CLASSIFICATION ACCURACY IN SPECTRAL ANGLE-BASED HYPERSPECTRAL
IMAGE ANALYSES
Leanna N. Ergin, B.S. and John F. Turner II, Ph.D.
Department of Chemistry, Cleveland State University
The efficacy of hyperspectral image segmentation is strongly reliant on the limitations of pixel-wise classification methods,
most of which are based on chemometric strategies that do not account for spatially dependent variations across the image
field of view. Spatial inhomogeneity arises from a variety of causes including non-uniform sample illumination and unequal
optical coupling efficiencies between axial and off-axis rays in the image path. These inconsistencies are often manifested in
the data as variations in the signal to noise ratio across the image field. In the work presented here, we describe a novel
method of hyperspectral image classification based on spectral angle and spectral vector pointing stability that reduces the
misclassification frequency. We provide a theoretical framework upon which the accuracy of the classified image can be
estimated without a priori information about the spatially dependent variations. In addition, we demonstrate the utility of the
method on both modeled hyperspectral image data and Raman hyperspectral data obtained for cold-drawn poly-L-lactide.
50
123
A NOVEL APPROACH TO INTEGRATING LOW-LIGHT SCATTERING SPECTROSCOPY
Jonathan R. Damsel, B.S., and John F. Turner II, Ph.D.
Department of Chemistry
The need for non-destructive analytical tools has motivated recent instrumental and methodological progress. Spectroscopic
techniques provide a non-contact approach to physical and chemical analysis. Brillouin inelastic scattering spectroscopy can
be used to determine mechanical properties such as the Young’s modulus of a sample by measuring the frequency shift of the
light scattered by the sample’s acoustic phonons. Another inelastic scattering spectroscopy, Raman spectroscopy is widely
used to provide chemical identity and to investigate conformational changes of a sample. For the first time, these
complimentary techniques are combined to simultaneously provide in depth analysis of the elastic properties and chemical
composition of a sample non-destructively. Tandem multi-pass fabry-perot etalons are conventionally employed to provide
spectral resolution of the Brillouin scattered light
cm-1 from incident light. We employ an
alternative method using the virtually imaged phased array (VIPA) to provide spectral resolution. We have developed a
numerical model of the VIPA optical response and characterized the performance of the VIPA-based spectrograph. We have
obtained Brillouin and Raman spectra, and discuss these results in the work presented here. Additionally, we compare the
numerically modeled Brillouin spectrum to the spectrum obtained by the VIPA based spectrograph. The benefits of this
method to future applications will be discussed.
124
EXAMINING THE EFFECTS OF TIME, AGE, VALENCE, AND VARIATION IN EMOTIONAL
INTONATION ON SPOKEN WORD RECOGNITION
Maura L. Krestar, M.A.; Conor T. McLennan, Ph.D.
Department of Psychology, Cleveland State University
Emotional intonation is essential for optimal verbal communication between humans. Recent research has found that
variation in emotional intonation and other nonlinguistic features of speech has consequences for spoken word recognition,
and that the occurrence of these consequences depends largely on the time course of perceptual processing. Furthermore,
while specificity effects often emerge in implicit memory tasks, they are not always evident in older adults or on tests of
explicit memory. The proposed study will attempt to extend previous time course findings to emotional intonations varying in
valence (i.e., sad, happy, and neutral) in younger and older adults. In a long-term repetition-priming paradigm, intra-talker
variability in emotional intonation is expected to reduce magnitudes of priming when processing is relatively slow, but not
when processing is relatively fast. The proposed study will also provide information regarding negative and positive
attentional biases in younger and older adults, respectively. Younger adults are predicted to be biased toward words in a
negative emotional intonation (sad) and older adults are predicted to be biased toward words in a positive emotional
intonation (happy). The results of the proposed study will make important contributions to theoretical views on the
representation of words in the mental lexicon, including how the issue of representational specificity may depend on factors
such as age, attentional biases, and type of emotional intonation.
125
AMINO ACID SEQUENCE 473-487 OF PROTHROMBIN IS REQUIRED FOR INHERENT
COAGULANT ACTIVITY
Joesph R. Wiencek, BA1; Jamila Hirbawi, Ph.D.2; Michael Kalafatis Ph.D1, 2
1
Department of Chemistry, Cleveland State University;
2
Molecular Cardiology, Cleveland Clinic
The prothrombinase complex is the significant enzymatic complex that activates prothrombin (Pro) to thrombin with rates
compatible with survival. Prothrombinase is composed of the enzyme factor Xa (fXa), and the cofactor factor Va (fVa)
associated on a procoagulant membrane surface in the presence of calcium ions. Previous studies have suggested that fXa
interacts with Pro within amino acid region 473-487 in a fVa-dependent manner. Thus we further investigated the functional
importance of amino acid region 473-487 of Pro in coagulation. Site-directed mutagenesis was used to construct a
recombinant Pro molecule with the region 473-487 deleted (rProΔ473-487). The deletion and wild type Pro (rPro WT) were stably
transfected in BHK-21 cells. The two recombinant molecules were purified according to a well-established protocol and, at
51
the last step, a FPLC equipped with a strong anionic Mono-Q 5/50 column and a calcium gradient were used to isolate
properly carboxylated rProΔ473-487 and rProWT. Both recombinant molecules were assessed, by gel electrophoresis, to examine
their ability to become activated into innate thrombin by fully assembled prothrombinase or fXa alone. The recombinant
molecules were also investigated for clotting and chromogenic activity. Gel electrophoresis revealed that consumption of
rProΔ473-487 by prothrombinase and thrombin formation was considerably impeded when compared with thrombin formation
following cleavage of rProWT by prothrombinase. In contrast, membrane-bound fXa alone, in the absence of fVa, exhibited a
marked increase in the rate of cleavage at Arg271 and activation of rProΔ473-487 as compared to cleavage at Arg271 and
activation of rProWT. Additionally, clotting assays revealed rProΔ473-487 was devoid of clotting activity and severely impaired
in its amidolytic activity while rProWT had clotting and chromogenic activities comparable to human plasma-derived Pro. Our
data demonstrate that amino acid sequence 473-487 of Pro is necessary for optimal rates of activation by prothrombinase.
Also these investigations suggest that amino acid region 473-487 is required for innate thrombin activity.
126
REGULATORY SEQUENCE 1000-1008 OF HUMAN COAGULATION FACTOR V MAINTAINS THE
PROCOFACTOR IN A QUIESCENT STATE
Joesph R. Wiencek, B.A.1; Jamila Hirbawi, Ph.D.2; Michael Kalafatis, Ph.D.1,2.
1
Department of Chemistry, Cleveland State University
2
Molecular Cardiology, Cleveland Clinic
The proteolytic conversion of prothrombin to thrombin is catalyzed by the prothrombinase complex composed of the
enzyme, factor Xa (fXa), the cofactor, factor Va (fVa), assembled on a membrane surface in the presence of divalent metal
ions. Factor V (fV) is synthesized as a procofactor with nominal procoagulant activity. Following three catalytic cleavages by
thrombin (Arg709, Arg1018 and Arg1545) amino acids 710-1545 of the B domain of factor V are removed and the protein
becomes activated. Without the proper removal of the B domain the fV protein will not show cofactor activity. A
comparative sequence analysis of fV among mammals revealed that amino acid sequence 1000-1008 of the B domain of fV
contains 7 conserved basic amino acids and could play a major part in inhibiting the interaction of fVa with fXa. Thus we
investigated the functional importance of the basic amino acid region 1000-1008 within fV. We constructed three mutant
recombinant fV molecules as follows: fVQ3 all activation sites (R709/R1018/R1545) were mutated to glutamine, fV ΔB9 had
region 1000-1008 deleted and fVΔB9/Q3 was a combination of both mutations. Kinetic analyses revealed that fVQ3 was
severely impaired in its interaction with fXa before and after incubation with thrombin. In contrast, fVΔB9 and fVΔB9/Q3 had
similar affinity for fXa when compared to wild type (fVa WT) before and after activation by thrombin. Two-stage clotting
assays revealed that while fVQ3 was devoid in clotting, fV ΔB9/Q3 had clotting activity comparable to fVaWT. Under similar
experimental conditions prothrombinase assembled with recombinant fVΔB9/Q3 had a Km that was comparable to the Km of
prothrombinase assembled with fVaWT while the kcat was decreased by ~20-fold. Increasing the concentration of fVΔB9/Q3
within the mixture resulted in a 57-fold increase in the Km and a 4-fold increase in the kcat of prothrombinase. Overall, our
data demonstrate that amino acid sequence 1000KTRKKKKEK1008 controls spontaneous binding of fV to fXa.
127
STRUCTURAL CHARACTERIZATION OF GLYCOPROTEIN BY MASS SPECTROMETRY
Sujatha Chilakala, MS; Yan Xu, PhD*
Department of Chemistry, Cleveland State University
A common post-translational modification of proteins is glycosylation. By glycosylation, a given protein may contain
multiple forms of glycan moieties at different locations. Glycan moieties are involved in signal transduction, extracellular and
cell- cell recognition, and molecular trafficking etc. Structural characterization of proteins and their post-translational
modifications is particularly important for the study of human diseases such as heart disease, cancer and diabetes. To
correlate the functional features of a glycoprotein with its structure (both temporally and spatially), the detailed structural
analysis of glycans is required.
Human IgG is a glycoprotein. Its physiologic significance and function are mainly regulated by the N-glycans attached to
each heavy chain. In this work, structural characterization of human IgG was achieved using trypsin digestion, enzyme
deglycosylation, permethylation, PMAA (partially methylated aditol acetates) reactions with LC-MS, LC-MS/MS, GC-MS
and bioinformatics. The detailed glycan composition, linkage and configuration of human IgG were elucidated.
52
128
OSTEOACTIVIN INDUCES TRANSDIFFERENTIATION OF C2C12 MYOBLASTS INTO
OSTEOBLASTS
Gregory R. Sondag1,2, Sibel Salihoglu1, Suzanne Lababidi1, Cale Crowder1, Fouad Moussa1,2, Samir Abdelmagid1, Fayez
Safadi1,2
1
Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH
2
School of Biomedical Sciences, Kent State University, Kent, OH
Osteoactivin (OA), is a novel osteogenic factor important for osteoblast differentiation and function. OA has been shown to
stimulate matrix mineralization and transcription of osteoblast specific genes required for differentiation. OA has shown to
play a role in wound healing as OA expression was shown to increase in post -fracture calluses. OA has also been shown to
be present in muscle as OA is upregulated in cases of denervation and unloading stress. The regulatory mechanisms of OA in
muscle and bone have not yet been determined. In this study, we infected C2C12 myoblasts with a retroviral vector
overexpressing OA under the CMV promoter and these cells were able to transdifferentiate from myoblasts into osteoblasts.
Immunofluorescence showed that skeletal muscle marker MF-20 was severely downregulated in cells overexpressing OA and
contained significantly less myotubes compared to uninfected control. C2C12 myoblasts overexpressing OA showed an
increase in bone specific mineralization such as alkaline phosphatase and alizarian red staining, and also showed an increase
in Runx2 protein expression. Overexpression of OA in C2C12 myoblasts reduced cell proliferation and protein expression of
cyclinD1. We detected increased levels of phosphorylated Focal Adhesion Kinase (FAK) in C2C12 myoblasts
overexpressing OA compared to control. Taken together, our results suggest that OA is able to induce transdifferentiation of
myoblasts into osteoblasts by increasing levels of phosphorylated FAK.
129
IDENTIFICATION OF TRYPANOSOMA BRUCEI TELOMERE SPECIFIC PROTEIN COMPLEX
COMPNENTS
Ali A. Lattif, Ph.D.; Vishal Nanavaty, M.S.; and Bibo Li, Ph.D.
Department of Biological, Geological and Environmental Sciences; Center for Gene Regulation in Health and Disease;
Cleveland State University
Trypanosoma brucei is a protozoan parasite that causes African sleeping sickness in humans and nagana in cattle. This
parasite is able to evade the host immune system because it switches the variant surface glycoprotein gene (VSG) expressed
when it is in a mammalian host. Telomere-specific proteins are essential for telomere functions. We have shown that
telomere proteins play important roles in VSG expression and switching regulation. Six proteins have been identified in
mammalian telomere complex, but only three telomere protein homologues in Trypanosoma brucei, TbRAP1, TbTRF, and
TbTIN2, have been identified by our laboratory. In order to identify additional protein components of the T. brucei telomere
complex, we have established a cell line with one endogenous allele of TbTRF and TbTIN2 deleted and the other allele
tagged with a FLAG-HA-HA epitope. We aim to use two-step immunoprecipitation to specifically pull-down the
TbTRF/TbTIN2 protein complex and to identify its components by mass spectrometry analysis.
130
MODIFIED CARBON FIBER MICROELECTRODES WITH RUTHENIUM OXIDE NANOPARTICLES
FOR SENSITIVE DETECTION OF NITRIC OXIDE IN BIOLOGICAL SAMPLES
Tiyash Bose, M.Sc.; Thomas Bomberger, B.S.; Mekki Bayachou*, Ph.D.
Department of Chemistry, Cleveland State University
Nitric oxide is an important biological molecule that has diverse functions in human physiology. The concentration of NO in
tissues and cells are of vital importance and the presence of too high or too low concentration of this reactive metabolite is
the source of a variety of disease states. The major challenges with NO measurement are its low nano-molar concentrations in
tissues and short half-life. Out of the available analytical methods, electrochemical tools are most promising because they
allow for miniaturization of probes as well as direct and accurate detection.
We fabricated the microelectrodes in house using single carbon fibers of 7μm diameter mounted on copper wires, sealed in
glass capillaries with 2-mm of the tip exposed. We have developed a method based on electrodeposited ruthenium oxide
53
nanoparticles on the surface of bare carbon fiber microelectrode as a platform for catalytic NO detection. The
electrodeposition of ruthenium oxide nanoparticles is performed in perchloric acid solution containing RuCl3 precursor with
constant cycling of the potential at 100 V/s scan rate for 20 minutes. Under optimum conditions, the nucleation of ruthenium
oxide occurs on the surface of the carbon fiber, with somewhat aligned growth of the nuclei that are in the 100-nm range in
diameter as characterized by field emission scanning electron microscopy (FESEM).
Electrocatalytic oxidation of the NO is assessed by cyclic voltammetry and amperometry in standing solutions. The
ruthenium-modified microelectrodes exhibit rapid and reproducible response to NO at low applied potential (+0.5V vs.
Ag/AgCl). The modified carbon microelectrodes typically show a five-fold increase in sensitivity compared to bare carbon
microfibers. We also show that this modified NO sensor has excellent linearity in relatively a wide range, including very low
concentrations of NO.
131
SITE-SPECIFIC FUNCTIONALIZATIONS OF RECOMBINANT THROMBOMODULIN BY
ENZYMATIC AND CHEMICAL METHODS
Lin Wang, Rui Jiang, Jacob Weingart, Xue-long Sun*
Department of Chemistry, Cleveland State University
Thrombomodulin (TM), an endothelial cell surface membrane glycoprotein, is a cofactor for protein C activation via
thrombin, and therefore a crucial regulator in protein C pathway and coagulation cascade. TM contains six epidermal growth
factor (EGF)–like structures, in which the forth to sixth EGF-like region (TM456) is the minimum functional domain
responsible for the activity of protein C activation. Therefore, TM456 is attractive anticoagulant drug candidate. Herein, we
report site-specific functionalization of recombinant thrombomodulin (rTM) with poly(ethylene glycol) (PEG) and lipid via
sortaseA A (SrtA)-mediated ligation (SML) and copper-free click chemistry (CFCC). SrtA is a transpeptidase from S. aureus
that recognizes and cleaves LPXTG motif on the target protein, and then catalyzes the linkage of the target protein with the
compound containing glycine. CFCC is a bioorthogonal reaction of dibenzocyclooctyne group (DBCO) with azide linker to
form a stable triazole. A truncated rTM456 derivative with azidohomoalanine for CFCC modification and LPETG tag at the
C-terminal for the recognition of SrtA was expressed in E. coli. and purified with Nickel affinity column. Site-specific
PEGylation and lipidation of rTM456 were performed by SML as well as CFCC. The conjugation products of PEGylation
and lipidation were confirmed by SDS-PAGE and western blot. Site-specific functionalizations of rTM leads to molecules
being arranged in a definitively ordered fashion and facilitating the preservation of the protein’s biological activity and
pharmacokinetic properties.
132
THE RELATIONSHIP BETWEEN THE CONSUMPTION OF BREAKFAST, CONSUMPTION OF
CAFFEINE, STRESS, AND EXAM SCORES
Malgorzata K. Bernas
School of Health Sciences, Cleveland State University
The following study was performed to see if relationships exist between the consumption of breakfast, consumption of
caffeine, stress, and exam scores could be found. A cross-sectional study design was conducted in which the surveys
completed by the students were collected at one time point. The sample population consisted of 71 neuroscience students
who attend Cleveland State University. The results show that students who consume breakfast the morning of the exam have
a higher level of alertness. However, the level of alertness does not differ between students who consume caffeine and
students who do not consume caffeine the morning of the exam. The results also show that there is a significant difference
between the amount of stress in a student’s life and their level of alertness. A borderline statistical significance (pvalue=.052) was found between how stressed a student felt before taking the exam and their final exam score. However, due
to the small sample size, a correlation between stress and exam scores could not be determined. In conclusion, although a
direct relationship was not found between the consumption of breakfast, consumption of caffeine, stress, and exam scores,
the findings show that consuming breakfast and avoiding stress are essential to staying alert while taking an exam.
54
133
CLEAVAGE AT ARG1018 IS NOT REQUIRED FOR FACTOR V ACTIVATION
Mahesheema Na, MSc1*; Joesph R Wiencek, B.A.1*; Jamila Hirbawi, PhD2*; and Michael Kalafatis, PhD1,2,3*
1
Department of Chemistry, Cleveland State University;
2
Molecular Cardiology, Cleveland Clinic;
3
Department of Biological, Geological and Environmental Sciences; Cleveland State University
The coagulation cascade is initiated following vascular injury which results in the exposure of endothelial tissue factor to the
blood flow and that in turn results in robust thrombin generation. The proteolytic conversion of prothrombin to thrombin is
catalyzed by the prothrombinase complex. This enzymatic complex is composed of the cofactor factor Va (fVa), the enzyme
factor Xa (fXa) associated on a procoagulant membrane surface in the presence of divalent metal ions. Factor V circulates as
a single chain inactive precursor consisting of three domains A1-A2-B-A3-C1-C2. The molecule must be activated by
thrombin following three sequential cleavages at Arg709, Arg1018, and Arg1545 to generate the active cofactor. It was previously
reported that cleavage at Arg1545 by thrombin requires prior cleavage at Arg1018.We used site directed mutagenesis to create
several recombinant fV molecules with the activation sites mutated to Glutamine (RtoQ). We have also used a recombinant
mutant factor V molecule with the region 1000-1008 from the B region deleted (fV∆B9). We have created recombinant fV
molecules fVWT (wild type), fVQQR (only cleavage at Arg1545 is available), fVRQQ ( Arg709 available), fVQRQ, ( Arg1018
available), fVRQR ( Arg709 and Arg1545 available), fVQQQ (no cleavage available), fV∆B9/RQR, and fV∆B9/QRQ. The recombinant
molecules were expressed in COS-7 cells, purified to homogeneity and assayed for clotting activity. Kinetic analyses
demonstrated that fVaRQR and fVaΔB9/RQR have similar affinities for fXa, while fVa QRQ, and fVaΔB9/QRQ were impaired in their
interaction with fXa. The kcat values for prothrombinase assembled with fVa RQR andfVaΔB9/RQR were similar to the kcat
obtained with prothrombinase assembled with fVaWT, while prothrombinase assembled with fVaQRQ and fVaΔB9/QRQ had 2fold and 7-fold reduced catalytic efficiency respectively, when compared to the kcat values obtained with prothrombinase
assembled with fVaWT. Our data demonstrates that cleavage at Arg1018 is neither essential nor required for activation of factor
V.
134
CLEAVAGE OF FACTOR V BY THROMBIN AT ARG709 IS DEPENDENT ON THE INTEGRITY OF
AMINO ACID REGION 1000-1008 OF THE PROCOFACTOR
Mahesheema Na, MSc1*; Joesph R Wiencek, B.A.1*; and Michael Kalafatis, PhD1,2,3 *
1
Department of Chemistry, Cleveland State University;
2
Molecular Cardiology, Cleveland Clinic;
3
Department of Biological, Geological and Environmental Sciences, Cleveland State University
Thrombin generation is a key step in the hemostasis mediated by prothrombinase complex. Prothrombinase is an enzymatic
complex between factor Va (fVa) and factor Xa (fXa) assembled on a membrane surface in the presence of divalent metal
ions. To study the effect of region 1000-1008 from the B-domain on thrombin cleavage at Arg709/Arg1545. We generated
recombinant fV mutant molecules by deleting the highly conserved region 1000-1008 (∆B9) and by mutating one or two of
the activating cleavage sites from arginine to glutamine. We have generated recombinant factor V molecules as follows:
fV∆B9/QQR, fVQQR , fV∆B9/RRQ , fVRRQ . The functional activity of the recombinant mutants was assessed by prothrombinase
assays .The appearance of the heavy and light chain of fV following incubation with thrombin was visualized following SDSPAGE and staining with silver. SDS-PAGE demonstrates that thrombin activation of fVRRQ resulted in the appearance of the
heavy chain only, fV∆B9/RRQ shows resistance to thrombin and no heavy chain formation. In contrast, SDS-PAGE
demonstrates that thrombin activation of both fVQQR and fV∆B9/QQR resulted in the appearance of the light chain. Kinetic
analyses revealed that the KD of fVa∆B9/QQR for fXa is 0.5nM which is similar to that of fVa PLASMA ,while prothrombinase
assembled with fVaRRQ and fVaQQR have values similar to the kcat of prothrombinase assembled with fVa WT, the kcat values
for prothrombinase assembled either fVa∆B9/QQR or fVa∆B9/ RRQ were decreased by 5-fold.Our data clearly shows that amino
acid region 1000-1008 from the B domain is essential for thrombin cleavage at Arg709
55
135
FUNCTIONAL MORPHOLOGY OF RAT HANDS AND FEET: CORRELATION WITH THE ABILITY
TO GRIP TREE BRANCHES DURING LOCOMOTION
Jessica E. Fonce, B.S.; Andrew R. Lammers, Ph.D.
School of Health Sciences, Cleveland State University
Anatomy and function are usually closely related. Since locomotion on tree branches is common among mammals, we expect
to find that the anatomy is well adapted for gripping narrow cylindrical branch-like substrates. We hypothesize that the
ability of rats to grip arboreal supports relies on musculature responsible for adducting the first digit (thumb and big toe) and
opposing medial-most and lateral-most digits. We dissected the hands and feet of four rat cadavers. There is a substantial
muscle believed to be responsible for the flexion/adduction of the thumb in the hands. We believe that this muscle is
equivalent to the opponens pollicis and/or adductor pollicis in the human hand. We also found lumbricals as well as dorsal
and palmar interossei. These lumbricals are probably responsible for flexing metacarpophalangeal joints as necessary for
gripping. The palmar interossei are believed to be responsible for the adduction of the rat digits just as they are responsible
for adduction of fingers in humans. Foot anatomy looked very similar to that of the hands. Based on the anatomy alone, rats
are built for locomotion across cylindrical branches as well as terrestrial substrates.
136
THE EFFECTS OF PRIMING A MUSCULAR IDEAL ON THE SUBSEQUENT PERCEPTION OF BODY
IMAGE WORDS IN MALES
Stephanie A. Henley, B.A.; Teresa A. Markis, M.A.; Conor T. McLennan, Ph.D.
Department of Psychology, Cleveland State University
The majority of body image research has focused on females, and limited research has been conducted with males. However,
research has shown that body dissatisfaction (BD) is prevalent among males and associated with unhealthy outcomes
(Karazsia & Crowther 2009). Muscular ideal internalization and social comparison are predictors of BD in males. Little
research has been conducted to understand the internal mechanisms associated with male BD. In the present study, the effect
of priming a muscular ideal on the subsequent perception of male body image words is examined. Half of the participants
viewed muscular male models, and half viewed plants (control group). All participants participated in a Stroop task including
three categories of word stimuli: body (BICEPS), plant (FERNS), and neutral (BOOK). Participants were asked to ignore the
meaning of the word and to name the color each word appeared in as quickly and accurately as possible. They were also
asked to complete a questionnaire to determine overall satisfaction with their body. We hypothesized that those primed by the
muscular ideal, especially those with a high level of body dissatisfaction, would respond significantly slower to body words
versus neutral and plant words. Inconsistent with our hypothesis, the results showed, regardless of which pictures they
viewed (e.g. muscular male models or plant pictures), the males that had a higher level of body dissatisfaction were
significantly faster at responding to the color of the body words. It appears that males, contrary to females, are avoiding the
body words. Results and practical implications are discussed.
137
ASSESSMENT OF INCORPORATION OF DNA ANTIMETABOLITE (TERMINAL VS. INTERNAL) BY
LC-MS/MS
Ye Feng1, Lan Li1, Lili Liu2, Stanton L. Gerson2, Yan Xu1, 2,*
1
Department of Chemistry, Cleveland State University
2
Case Comprehensive Cancer Center, Case Western Reserve University
Fludarabine (9-β-D-arabino-furanosyl-2-fluoradenine or F-ara-AMP) is an adenosine analogue, which has been used as
chemotherapeutic agent for the treatment of hematological malignancies. This drug can be phosphorylated to its
triphosphorylated form (F-ara-ATP) in cells, and then incorporated into DNA strands. It is generally accepted that
fludarabine is overwhelmingly incorporated to the terminal site of DNA strands and acts as DNA chain terminator. However,
the recent studies revealed that fludarabine could also be incorporated into internal positions of DNA strands, and base
excision repair pathway may be one of the mechanisms attributed to the drug resistance.
56
To better understand the toxicity mechanism of fludarabine, we have developed LC-MS/MS methods in conjunction with
nuclease digestion procedures to quantitatively determine the amount of fludarabine incorporated at DNA terminal and
internal positions. In this work, the incorporated drug molecules were first released from the DNA backbone in the form of
either F-ara-A (from terminal positions) or 3’-F-ara-AMP (from internal positions). Then, the released drug molecules were
identified and quantified by LC-MS/MS. Our data showed that the majority (> 95%) of incorporated fludarabine molecules
were at internal positions of DNA rather than at terminal positions as previously reported. This work provided direct
evidence that fludarabine incorporation in DNA is mainly taken place at internal positions of DNA strands. This new toxicity
mechanism of fludarabine will certainly lead to new a therapeutic strategy to overcome tumor resistance to fludarabine.
138
A NOVEL LC/MS/MS METHOD FOR QUANTIFICATION OF CANGLIOSIDES IN HUMAN PLASMA
Qianyang Huang, M.S.1; Xiang Zhou, Ph.D.1; Heng Wang, Ph.D.2; and Aimin Zhou, Ph.D.1
1
Department of Chemistry, Clinical Chemistry Program, Cleveland State University, Cleveland, Ohio;
2
DDC Clinic, Center for Special Needs Children, Middlefield, Ohio
Gangliosides belong to a family of glycosphingolipids characterized by mono- or poly-sialic acid-containing oligosaccharides
linked by 1, 3- and 1, 4- β glycosidic bonds with subtle differences in structure, which are abundantly present in the central
nervous system of many living organisms. Their abnormal cellular surface expression and physiological functions are
generally believed to pathologically contribute to considerable neurological disorders including Alzheimer and Parkinson
diseases. In this study, we developed and validated a sensitive LC/MS/MS/ method for determination of GM2, GM3, GD2,
and GD3 in human plasma. The analytes and internal standard were extracted from plasma using a one-step protein
precipitation procedure with the addition of methanol. Then the samples were analyzed by LC/MS/MS with a Multiple
Reaction Monitoring (MRM) mode in MS detection to obtain superior sensitivity and specificity. The assessment of
extraction recovery, calibration linearity, precision, and accuracy were also established. The method was successfully applied
to measure the ganglioside levels in plasma from normal human subjects.
139
DETERMINATION OF BISPHENOL A β-D-GLUCURONIDE IN HUMAN URINE SAMPLES BY LCMS/MS
Simuli L. Wabuyele, M.Sc.1; Dan Wang, M.Sc.1; Shuming Yang, Ph.D.2; Li Li, Ph.D.2; Yan Xu, Ph.D.1, 2
1
Department of Chemistry, Cleveland State University;
2
Case Comprehensive Cancer Center, Case Western Reserve University
Bisphenol A (BPA) is a synthetic estrogen and is widely used to make polycarbonate plastics and epoxy resins. The prevalent
use of BPA-containing plastics in consumer products frequently exposes humans to this compound. Since BPA is an
endocrine-disrupting compound, the adverse physiological effects of BPA are subjects of ongoing clinical studies.
The measurement of BPA in biological samples often produces false results due to the fact that BPA is readily available in
the laboratory environment and reagents. Since bisphenol A β-D-glucuronide (BPA-G) is a metabolite of BPA which does
not exist in the environment without metabolism, it can therefore be a biomarker to assess human exposure to BPA. The
existing analytical methods of BPA-G use either solvent matrix for calibration or laborious enzymatic hydrolysis for sample
preparation: they often result in bias results and are susceptible to environmental contamination.
In this work, we have developed an isocratic liquid chromatography tandem mass spectrometry (LC-MS/MS) method for
quantitative measurement of BPA-G in human urine using d6-BPA-G as the internal standard (IS). A simple liquid-liquid
extraction protocol was used for efficient urine samples preparation. Chromatographic separation of BPA-G and the IS from
urine matrix was achieved on a Waters X-select HSS T3 column at a flow rate of 0.2 mL/min in less than 10 min.
Quantification was performed by AB Sciex QTRAP 5500 mass spectrometer using negative turbo-ion-spray ionization in
multiple-reaction-monitoring (MRM) mode with mass transitions m/z 403.2 → 227.1 for BPA-G, and m/z 409.0 → 233.1for
the IS, respectively. The method has a linear calibration range of 0.500-1000 ng/mL (r = 1.00) using a weighted 1/x linear
regression, relative recoveries were > 97%, and matrix factor ranged from 0.94-1.01. The method developed was successfully
applied to the measurements of BPA-G in 39 patients’ urine samples.
57
140
FXR, A NOVEL RECEPTOR IN BONE CELL DIFFERENTIATION AND FUNCTION
Fouad M. Moussa1,2, Suzanne L. Lababidi1, Afif S. Sanyurah1, Lana N. El-Shaar1, Kim M. Novak1, Samir Abdelmagid1,
Yanqiao Zhang1, Fayez F. Safadi1
1
Department of Anatomy and Neurobiology, Northeast Ohio Medical University, Rootstown, OH
2
School of Biomedical Sciences, Kent State University, Kent, OH
Farnesoid X receptor (FXR) is a receptor key in maintaining bile acids and cholesterol homeostasis. While it has been known
that FXR is expressed in liver, and adipose tissue, its expression in bone are not well understood. In this study, we examined
the expression of FXR in bone by qPCR analysis and demonstrated FXR in normal long bone and calvaria. Next, we
examined bone mass of FXR KO mice using micro-CT and showed increased bone volume and trabecular thickness in FXR
KO compared to WT mice. Survival and proliferation of primary osteoblasts were significantly less in the FXR KO
compared to WT. We next examined the effects of FXR agonist (GW4064) on bone cell differentiation. Primary osteoblasts
and MC3T3-E1 osteoblast-like cells were treated with GW4064 and differentiated, they demonstrated less alkaline
phosphatase staining at day 14 and less matrix mineralization staining at day 21 compared to untreated culture. Osteoclast
differentiation of hematopoietic stem cells showed less number of osteoclasts coupled with less TRAP activity in the FXR
KO compared to the WT mice. Taken together, these results suggest a functional role of FXR in regulating bone homeostasis.
Further studies are warranted to know the mechanism of FXR in bone cell differentiation.
141
THE EFFECT OF STEREOTYPE THREAT ON REPORTED MEDICAL HISTORY
Lisa P. Gaynier, M.A.1; Steve Slane, Ph.D.1; Leslie Fisher, Ph.D.1; Kyle Znamenak, M.A.1; Swagata Banik, Ph.D.2
1
Department of Psychology, Cleveland State University;
2
Baldwin Wallace College
An unexplored process that may contribute to poor quality healthcare is stereotype threat, the phenomenon of being at risk of
confirming a negative stereotype about one's identity group when the stereotype is made salient (Aronson & Steele, 1995).
While over 100 studies have confirmed stereotype threat effects in performance settings, this project investigates the
phenomenon in everyday interactions between patients and healthcare providers. We hypothesize that this form of crossracial/ethnic and cross-gender anxiety contributes to health disparities by making patients reluctant to accurately disclose
health behaviors that may be detrimental to their health. Our findings indicate clear cross-gender correlations that support the
hypothesis. Race/ethnic samples were too small to support reliable conclusions.
142
OPTIMIZATION OF SITE-SPECIFIC DOUBLE FLUORESCENT LABELING OF RECOMBINANT
THROMBOMODULIN
Rui Jiang, Lin Wang, Jacob Weingart and Xue-Long Sun*
Department of Chemistry, Cleveland State University
Thrombomodulin (TM), an endothelial thrombin receptor, acts as a major cofactor in the protein C anticoagulant pathway.
TM contains six epidermal growth factor (EGF)–like structures, in which the forth to sixth EGF-like region (TM456) is
responsible and the minimum functional domain for the activity of Protein C activation. Recombinant TM456 has been
proposed as potential antithrombotic agents. In order to modify TM456 in a controlled way, new functionalities were
introduced in a defined manner and attentions have focused on methods that enable the efficient and site-specific covalent
conjugation of recombinant TM456. In this study, one LPETG tag for sortaseA-mediated ligation (SML) and one unnatural
amino acid, azidohomoalanine for copper-free click chemistry (CFCC) were genetically introduced at the C terminal of
TM456. After recombinant expression by E. coli B834 cells, enzyme cleavage and purification, the target protein TM456Azide-LPETG (TM456AL) was obtained. Double fluorescent labeling of TM456AL was performed by SML and CFCC at
the C-terminal in one-pot reaction and the double labeled protein was confirmed by fluorescent scanner. Different reaction
conditions were evaluated by varying pH, reaction time, concentrations of substrates and sortaseA. In conclusion, we have
demonstrated a protein engineering technique that permits site-selectively double labeling of the recombinant TM456AL
which allows further double modification of protein in one-pot reaction such as adding two molecules bearing diverse
functions.
58
143
RAMAN CHEMICAL IMAGING OF POLY-L-LACTIDE BIOIMPLANTS USING AN ACOUSTO-OPTIC
TUNABLE FILTER
Venkata N K Rao, Bobba; and John F, Turner II, PhD
Department of Chemistry, Cleveland State University
Poly-L-lactide (PLLA) is a biodegradable and biocompatible implant polymer that can be endowed with varying degrees of
elasticity and piezoelectricity depending on the extent of its long-range molecular order. In its amorphous state, PLLA
exhibits no net piezoelectric response. Cold-drawing PLLA at temperatures above the glass-transition temperature and below
the cold crystallization temperature induces molecular ordering. As the draw ratio increases, corresponding increases in the
piezoelectric properties and degree of crystallinity are observed. At draw ratios greater than 4.5, fibrillization begins to
dominate and these molecular order-induced properties diminish. By tailoring the crystallinity of PLLA, its degradation rate,
Young’s modulus, and piezoelectricity can be enhanced for a particular application in vivo.
In the work presented here, we have developed a wide-field Raman imaging system based on a narrow passband acoustooptic tunable filter. Our implementation overcomes some of the current limitations of AOTF imaging systems and combines
novel multivariate strategies with AOTF-based acquisition to provide high resolution images of crystalline content. The
multivariate method employs a new Gram-Schmidt orthogonalization method to establish a reference spectrum for automated
image analysis based on spectral correlation. A full theoretical treatment of the multivariate approach and the design of the
novel wide-field AOTF Raman imaging system are presented.
59