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Project Manager

Bellevue, Washington, United States
February 24, 2018

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Manjula Yeleswarapu, Ph.D.

Seattle, WA

Cell: 617-***-****


VA-Puget Sound, Seattle July 2014-March 2015


Fat Clearance role of APOE 2 isoform as in protection from various diseases such as Alzheimer’s is the outcome of my project. Technology standardized by me is Transfections, Reporter Assays as Alkaline Phosphatase (AP) and Luciferase Reporter Assay and also Chromatin Immunoprecipitation (CHIP) and PCR to establish Alzheimer’s disease and cancer model in Human Cell lines as UII8 and HAPG2.

University of Washington, Seattle Sep 2008-March 2009

Research Scientist,


Electrophysiological and Behavioral studies in mouse models of cortico-striatal plasticity is the outcome of my project. Generated cortico-striatal plasticity in mice by injecting them with psycostimulants such as cocaine and amphetamine. I focused my studies on gestational effect of psycostimulants, which showed symptom’s like Parkinson’s disease. Psycostimulants are known to increase the dopamine neurotransmitter levels in-vivo, but the gestational effect is unknown. The mechanism of synaptic plasticity is studied in striatal slices and dissociated striatal cells by patch clamping experiments. Patched cells were characterized by biocytin immunohistochemistry. Patch clamping data is correlated with behavior and multiphoton imaging studies done in those mice. I observed very interesting differences in synaptic plasticity at different age groups of mice and also observed significant differences in male and female mice. These studies help’s in understanding the effects of alterations made in dopaminergic neurotransmission in neurons of striatum at different age groups and sex of mice. Impaired dopaminergic neurotransmission in striatum is the leading cause for various neurological disorders hence this research is very important to find cures for movement and learning disorders which include both neurodegenerative and neurodevelopmental diseases.

Seattle Children’s Hospital & Regional Nov 2007-June 2008

Medical Center, Seattle


Identification of possible role of microchimeric cells in Human and Mouse models of Lupus and Multiple Sclerosis is the outcome of my project. Microchimeric cells are stem cells which pass from mother to offspring during pregnancy. The hypothesis is that these cells loose tolerance to body immune system due to some unknown reason and triggers autoimmune diseases such as lupus and multiple sclerosis. This is a novel field of research and needs lots of studies to conclude anything. Lupus is devastating autoimmune blood disorder with lot of damaging effects on brain. Multiple sclerosis is also an autoimmune brain disorder. Interestingly, immunohistochemical, DNA Fluorescent Insitu Hybridization and quantitative PCR studies done on the postmortem human brain tissue, lymphocytes from patient blood samples and mouse tissue showed that there is an increase in microchimeric cells in these diseases. In order to understand the gene expression in microchimeric cells in disease verses normal conditions, I isolated microchimeric cells using LASER capture microdissection and studied gene expression pattern in them using microarray experiments and quantitative RT-PCR technique. I still need to repeat these experiments many times to reach to a conclusion. This is a very important research because if microchimeric cells are triggering autoimmune diseases then we can develop strategies to make them tolerate body immune system or specifically eliminate them.

Harvard Medical School, Beth Israel-Deaconess, Apr 2004-Aug 2007

VA Medical Center, Boston

Research Fellow and Lab Manager,

Molecular Physiology

Active and inactive Neurotransmitter Nitric oxide isoform expression in neurons and Adenocarcinoma is the outcome of my project. Nitric oxide is a gaseous neurotransmitter extensively studied in gut. It is known to relax smooth muscle cells in gut to open the pyloric sphincter for passage of food from one part of stomach to next part. The function of nitric oxide in adeno- carcinoma and striatum part of brain, specially regulating the dopaminergic neurotransmission is unknown. I focused my research in identifying novel active isoforms of neuronal nitric oxide synthase enzyme expressed in nerve terminals by using western blot and immunoprecipitation in gut which I will compare with the ones expressed in brain during normal and pathological conditions. This type of research is very important because brain disorders severely affect neuronal cells in brain not much to the gastrointestinal neurons. So, if we can find the differences between these neurons, especially protection mechanisms, these can be used to protect brain cells as well.

University of California, Davis Sep 2001-Mar 2004

School of Medicine, Post-Doctoral Fellow,

Human Molecular Genetics, Medical Microbiology & Immunology

Role of Polyadenylation in Rett Syndrome is the outcome of my project. Lower levels or mutations in Mecp2 gene in brain causes neurodevelopmental disorder Rett syndrome. Mecp2 protein is expressed primarily by two types of transcripts short transcript and long transcript. Longer transcript has a long 3’ non coding region (3’UTR). Switch from short to long form alters the mRNA stability and subcellular localization in neurons. I identified and quantified these transcripts at various developmental stages in mouse brain by using Northern blot, RT-PCR and quantitative PCR. I also developed a cell culture model for Rett Syndrome and quantified Mecp2 gene and protein levels with development using LASER Scanning Cytometry. Transfected these cells with siRNA (Decoy approach) to knock down Mecp2 expression. Future aim is to knock down 3’UTR and study the effect on neurotransmission in cells. It is a very important research because this will give us an idea that which transcript is actually involved in neurotransmission and which transcript localizes itself or translate protein in nerve terminals.

Emory University, Atlanta Sep 2000-Aug 2001

School of Medicine, Post-Doctoral Fellow,

Human Genetics

Role of dynamin protein and transcriptional factors in Huntington’s disease is the outcome of my project

Huntington’s disease is a neurodegenerative disorder caused due to increase in glutamate

repeats on huntingtin’s gene. Developed a cell culture model for Huntington’s disease by overexpressing huntingtin gene with different number of glutamate repeat’s. The severity of disease depends on increasing number of glutamate repeat’s. I co-transfected these cells with dynamin construct to study the effect of dynamin along with the severity of disease. I found very interesting results which indicate that dynamin protein in involved in mitochondrial fission and fusion, involved in formation of nerve terminals and also involved in internalization of D1 and D2 receptors in neurons which regulate the dopaminergic neurotransmission. Future studies are, to find out how dynamin levels effect neurotransmission in striatum. I also did preliminary studies to see how transcriptional factors affect Huntington’s disease. I made GST and His tag fusion proteins for Sp1 and Huntingtin in bacteria, purified the proteins and studied protein- protein interactions.

Emory University, Atlanta Nov1999-Aug 2000

Rollins Research Center, Post-Doctoral Fellow,

Molecular Biophysics

Novel slow intermediate free-radicals in Ischemia Reperfusion Injury is the outcome of my project. Free radicals generated during ischemia reperfusion injury causes severe damage to brain cells. I developed a liposomal membrane model to study Ischemia reperfusion injury and also synthesized cobalt complexes to amplify free radical damage. I increased the levels of fast acting radicals by filling the liposome’s with caged cobalt complexes and exposed them to LASER beam. Cobalt complexes released oxygen upon LASER exposure and damaged the membranes by forming free radicals. These radicals were trapped by spin traps and detected by Electron spin resonance spectrometry (ESR). My experiments proved that only oxy, hydroxy and peroxy radicals can be detected by regular ESR machine. It is not possible to detect other (hypothetical) fast generating radicals by using regular ESR machine. A cutting edge Pulsed electron spin resonance spectrometer can be used to detect free radicals generated at nanosecond time scale, may help us in identifying new free radicals. This research is very important because unknown fast acting free radicals damage organ’s during transplantation. This damage limits the time frame of organ transportation from one place to another and also use. We identified these damaging slow intermediate free radical’s as a marker this technology can save many lives.


Ph.D. in Biochemical-Engineering Thesis submitted: Nov 1999 (Indian Institute of Technology, Bombay, India) Degree awarded: May 2000

Used 1-3liter Bioreactor which is also a microbial fermentation sort of combined unique design by me during my Ph.D. as graduate studies. Bacterial fermentation to produce economically important products such as xanthan gum, ascorbic acid, pyocyanin etc. Analyzed these products using chromatographic techniques such as HPLC and GC. To understand the metabolism and molecular mechanism of product formation, expression of Sox RS regulon in presence and absence of Antioxidants and Oxidants were studied. This research is very important because this is the first time we showed the effect of optimum and excess free radicals generated in bacteria on product quality and quantity. This research has lots of applications in industry.

M.S. in Biochemistry, India July 1993-May1995

Summer project done on: “Effects of pesticides on mice”, the results showed that pesticides interfere with electron transport system in mitochondria and generate free radicals. These mice showed symptom’s like Parkinson’s disease. This is a very important study to show how environmental pollutant’s leads to devastating neurological diseases.

B.S. in Biology and Chemistry, India July 1991-June 1993

Research project: “Effects of pesticides on benthic biota” was a voluntary work I did during my spare time to gain some experience in the field of free radical research. Fresh water invertebrate species which are very important to keep a healthy pond ecosystem, showed protein loss and mortality in geographical area’s (Madhya Pradesh) where excessive pesticides were used. Pesticides generated free radicals in these species which formed protein adducts and led to protein loss detected using spectrophotometer. Studies involve survey of vector born diseases due to imbalance of pond ecosystem. Water analysis for various pathological microbes is also done by me.

Research Interests

1. Normal and impaired molecular mechanisms of neurotransmission in striatum part of

brain using postmortem human brain, mouse brain tissue samples and cell culture


2. Synaptic plasticity studies in mouse models of neurodegenerative and

neurodevelopmental disorders

3. Identify different types of cells and their function in striatum of mouse brain.

4. Identify receptor distribution in striatal cells of mouse and human postmortem brain

tissue samples.

5. Identify novel protection mechanism’s which protects gut neurons from neuro-

degeneration and developmental disorders which are pretty common in CNS neurons

of mouse.

6. Role of Microchimerism in neurological autoimmune mouse brain diseases.

7. Studies to identify effects of natural products and drugs on neural circuits of mouse


8. Develop novel optical (Multiphoton) imaging techniques to understand

neurotransmission in mouse brain.

9. Identification of fast acting free radicals in biological membranes

10.Stem cell therapy for neurodegenerative disorders.

Awards / Honors

1. Best Research work contributed by a Young Scientist at International Congress on Sustainable Development of Environment, India, December 1997, for the paper on Environmentally induced mutations and generated free radicals in the plant pathogenic bacteria, Xanthomonas campestris.

2. Earned Fellowship to pursue Ph.D at Indian Institute of Technology, by scoring 98% in Graduate Aptitude Test in Engineering (GATE).

3. Competed and Qualified in All-India Entrance Examination to study Masters (M.S.) in Biochemistry at DAVV, Indore University, India.

4. Won State Level competition at undergraduate level for developing working models to harness renewable energy resources.

Technical Expertise

Cell Culture: Bioreactor design and use cell lines, Bacterial culture, Tissue Culture, Mammalian cell culture (ES cells such as P19 cells, cell lines and primary cultures) and lymphocyte isolation from normal and patient blood. Developed cell lines and screened them to obtain pure colonies

Animal Care and Handling: Genotyping, execution and interpretation of animal studies, behavior studies, dosing test articles via multiple routes.

Molecular Biology: Protein and Gene Expression studies (DNA, RNA and Protein isolation from Blood, Tissue and cells, PCR, Quantitative PCR, cDNA

synthesis, Multiplex PCR, In situ hybridization (ISH), Northern blot,

Western blot, SDS-PAGE, Immunoprecipitation, Primer design, Sequencing,

Mutagenesis, Transfections, siRNA technology, Decoy based inhibition of gene

expression and Cloning)

Imaging: Immuno-cytochemistry(ICC), Immuno-histochemistry (IHC) (Cutting, embedding and staining sections, Cartilage and bone decalcification), Confocal microscopy, Multiphoton Microscopy, Calcium ratio Imaging and LASER Capture Microdissection (LCM), Laser Scanning Cytometry, Automated Cellular Imaging System and Flow Cytometry.

High throughput Screening: Novostar Fluorescent Microplate reader

Biophysical and Electrophysiology Instruments operated: Electron spin Resonance Spectroscopy, Bioreactor, Ultracentrifugation, Ultrafiltration,enzyme linked immunosorbent assays (ELISA), HPLC, GC, Spectrophotometer and Patch Clamp set up.

Grant Writing

1. Grants and NIH progress reports

2. IACUC protocols (Animal use protocols), IRB (Human subject use protocols) and

Experimental protocols

3. Experimental Safety form writing

Selected Peer reviewed Articles

1. “Dopamine regulates prefrontal excitatory neurotransmission in the accumbens

nucleus” Dennis Dever, Janet Lowe, Wengang Wang, Anita Bhansali, Jessica Towne, Manjula Rao, Nigel S. Bamford (2009) Neuron, Manuscript under preparation.

2. “Prenatal Cocaine Exposure decreased nociception behavior and locomotor activity

but increased motor coordination in young adult Male mice” Y. Manjula Rao and

Nigel Bamford (2009), Brain Research, Manuscript under preparation.

3. “Sex specific differences in behavior of young adult mice after prenatal exposure to

cocaine”Y Manjula Rao and Nigel Bamford (2009) Brain Research, Manuscript

under preparation.

4. “Calmodulin bound and Calmodulin lacking forms of nNOS in the nerve terminals in

the gut: implications for nitrergic neurotransmission” Y. Manjula Rao, Arun

Chaudhury and Raj K Goyal AJP gastrointest.Liver physiol. (2008) 294,G 627-634.

5. “PIN is associated with cytosolic but not membrane-bound nNOS in the nitrergic

varicosities in mice gut: implications for nitrergic neurotransmission”. Arun

Chaudhury, Y.Manjula Rao and Raj K Goyal AJP gastrointest.Liver physiol. (2008)

In Press

6. “Telomere Maintenance In LCM Purified Barrett’s Adenocarcinoma Cells And

Impact Of Telomerase Inhibition In Vivo” Masood A Shammas, Aamer Qazi,

Ramesh B Batchu, Robert C Bertheau, Jason Wong, Manjula Y Rao,

Madhu Prasad, Selvarangan Ponnazhagan, Diptiman Chanda, Christopher P

Steffes, Nikhil C Munshi, Immaculata DeVivo, David G. Beer, Sergei

Gryaznov, Donald W Weaver, & Raj K Goyal, Cli.Can. Res (2008) 14. 4971-


7. “Elevated methyl-CpG-binding protein 2 expression is acquired during postnatal

human brain development and is correlated with alternative polyadenylation”

Damina Balmer, Jared Goldstine, Y. Manjula Rao and Janine M. LaSalle

J Mol Med (2003) Volume 81, Number 1 pp: 61- 68

8. “Free radical aspects of Xanthomonas campestris cultivation with liquid phase oxygen

supply strategy”. Y. Manjula Rao, A. K. Suresh and G. K. Suraishkumar,

Process Biochemistry (2003) Volume 38/9 pp: 1301 – 1310

9. “Interaction of Huntington Disease Protein with Transcriptional Activator Sp1”

Shi-Hua Li, Anna L Cheng, Hui Zhou, Suzanne Lam, Manjula Rao, He Li, and

Xiao-Jiang Li, Molecular and Cellular Biology, Mar.(2002) Volume 22, No. 5

pp: 1277-1287, 2002

10. “Improvement in bioreactor productivities using free-radicals: HOCl induced

overproduction of xanthan gum from Xanthomonas campestris and its mechanism

Y.Manjula Rao and G.K.Sureshkumar, Biotec. Bioeng. Jan 5 (2001) Volume 72,1,

pp: 62-68

11. “Direct biosynthesis of ascorbic acid from glucose by Xanthomonas campestris

through induced free radicals”. Y. Manjula Rao and G.K. Suresh Kumar.

Biotechnol. Letters 22(5): 407-411; Mar 2000

12. “Oxidative-stress-induced production of pyocyanin by Xanthomonas campestris and

its effect on the indicator target organism, YMRao and G.K.Sureshkumar.

J. Ind. Micro. Biotechnol, Nov. (2000) Volume 25, No.5, 266-*** (****)

13. “Oxygen supply without gas-liquid film resistance to Xanthomonas campestris

cultivation” G.Sriram, Y.Manjula Rao, A.K.Suresh, and G.K.Sureshkumar.

Biotechnol.Bioeng.(1998) 59, 714-723

14. “Nicotinamide and alpha-tocopherol combination partially protects t-butyl hydro

peroxide-induced neurotoxicity: Implication for neurodegenerative disease”

M.S.Parihar and Y.Manjula et al. Current science (1997), vol.73, 3, 290-292

Selected Presentations

1.“Maternal Cells Respond to Chronic Renal Fibrosis” Manjula Rao, PhD, Jesus Lopez-Guisa, PhD, Kendall Blair, Debra George, Anne Stevens, MD, PhD, Fifth Annual Fellows’ Research Day on Friday, April 25th at Seattle Children’s Hospital Research Institute 2008.

2. “Telomerase Activity and Telomere Length in LCM Purified Barrett’s Esophageal

Adenocarcinoma Cells.” Masood A Shammas, Aamer Qazi, Ramesh B Batchu, Jason

YY Wong, Manjula Y Rao, Christopher S Bryant, Sanjeev Kumar, Madhu Prasad,

Christopher P Saeffes, Immaculata De Vivo, David G.Beer, Donald W Weaver and

Raj K Goyal.; Digestive Disease Week, San Diego, California, May 17-22, 2008

3. “Effect of a novel telomerase inhibitor, GRN163L on growth of Barrett’s esophageal

adenocarcinoma cells in vitro and in vivo.” Masood A Shammas, Aamer Qazi,

Ramesh B Batchu, Jason YY Wong, Manjula Y Rao, Christopher S Bryant, Sanjeev

Kumar, Madhu Prasad, Christopher P Saeffes, Immaculata De Vivo, David G.Beer,

Donald W Weaver and Raj K Goyal.; Digestive Disease Week, San Diego,

California, May 17-22, 2008

4. “Botulinum toxin A Inhibits Gastric Antrum Smooth Muscle by Acetycholine

Independent Muscle Hyperpolarization” Xue D.He, Bo Yang, Madhu Prasad,

Manjula Y. Rao, Hemant S.Thatte, Xiao Qi Zhang and Raj K. Goyal; Digestive

Disease Week, Los Angeles, California, May 20-25, 2006.

5. “Calmodulin bound and Calmodulin lacking forms of nNOS in the nerve terminals in

the gut: implications for nitrergic neurotransmission” Y. Manjula Rao,Yasuo

Watanabe and Raj K Goyal; Digestive Disease Week, Chicago, Illinois, May14-19,


6. “Role of PIN (protein inhibitor of nNOS) in the Regulation of nNOS in Nerve

Terminals” Y.Manjula Rao and Raj K Goyal; Digestive Disease Week, Chicago,

Illinois, May 14-19, 2005

7. “Transcriptional changes correlate with elevated protein expression of MeCP2 during

in vitro neuronal differentiation” Manjula Y.Rao and Janine M. LaSalle; 43rd

Annual Meeting of The American Society for Cell Biology, San Francisco,

California, December 13-17, 2003

8. “Quantification of free radicals in bioreactors employing the liquid-phase oxygen

supply strategy” Malabika Bairagi, Y Manjula Rao, A.K. Suresh and G.K.Suresh

Kumar. CHEMCON 2001

9. “Photolysis of Caged Dioxygen Drives Superoxide production by Xanthine Oxidase in

a model system for Radical Damage in Ischemia Reperfusion injury” M. Rao

and Kurt Warncke; 23rd International EPR Symposium at Department of Chemistry

and Biochemistry, University of Denver, CO, USA, July 2000.

10. “Oxidative Stress inducible xanthan gum is regulated by SoxRS Regulon in

Xanthomonas campestris ”, Y.Manjula Rao and G.K.Sureshkumar, International

Symposium on Transcription Assembly & Nucleic Acid-Protein Interactions,

Bangalore, India, June 7-9, 1999.

11. “Cellular Responses to Induced Free-Radicals in Xanthomonas campestris”,

Y.Manjula Rao and G.K.Sureshkumar, 39th Annual Conference of the Association of

Microbiologists of India, Mangalore, India, December 5-7, 1998.

12. “Studies on Method of Oxygen Supply Without Aeration to Bioreactors”,

G.Sriram, Y.Manjula Rao, G.K.Sureshkumar and A.K.Suresh, Conference of recent

Advances in Fermentation Technology II, San Diego, CA, USA, November 1997.

13. “Experimental Studies on Environmentally induced Mutations in and Generated Free-

Radicals on, the Plant Pathogenic Bacteria, Xanthomonas campestris”

Y.Manjula Rao and G.K.Sureshkumar, International Congress on Sustainable

Development of Environment and Wildlife, Ujjain, India, December 18-21, 1997.

14. “Effect of Pesticides, Endosulphan and Nuvan on Benthic Biota”

Y.Manjula Rao and U.Chaudhary, Conference of the National Academy of

Environmental Sciences, Indore, India, Dec. 1994.

Computational Skills

MS office, software to analyze electrophysiology data such as clampfit, cellular imaging systems such as Metamorph and statistical analysis software such as sigma plot etc. Molecular biology related software such as Accelrys, Vector NTI and BioEdit.

Reference: Research Associate Professor VA Puget Sound : Chang En Yu

Phone: (206) ***-****, e-mail

Associate Professor University of Washington : Nigel Bamford Phone: 425-***-**** e-mail

Associate Professor Chidren’s Research: Anne M Stevens: Phone: 206-***-****


Contact this candidate