Associate Scientist - Biochemistry & Vascular Biology

Mallikarjun Badadani Ph.D.

Vascular Biology Center

**** ***** ****** ****

Augusta University

Augusta GA-30912

Contact No.: 706-***-****

Email Id.: ********@*****.***

Objective:

To understand the science and applying to ameliorate human sufferings (diseases)

Long term goals:

To be placed in respectable position to serve in science and health field

Education:

Ph.D (Biochemistry / analytical biochemistry) – August 2002 to May 2007.

Department of Neurochemistry, National Institute of Mental Health And Neuro Sciences. (NIMHANS),

Bangalore, India.

Project: Biochemical studies on Handigodu disease: A familial syndrome of Epi (Meta) physeal dysplasia.

M.Sc. (Biochemistry/ Chemistry) (1997-1999) Karnataka University, Dharwad, India.

(Biological macromolecules, Intermediary metabolism, Biochemical genetics and molecular biology, Microbial biochemistry, Organic chemistry, Physical chemistry)

B.Sc. (Chemistry, Botany, Zoology) (1994-1997) Karnataka University, Dharwad, India.

Work and Research Experience:

Assistant research scientist: ( May 2023 to present) : Vascular Biology Center,

Dept of Medicine Medical College of Georgia, Augusta University, Augusta.

Project : Heterogeneity of B lymphocyte in Peripheral artery disease(PAD) and effect of infiltration on endothelial cells and myocytes

Visiting Researcher (September 2020- June 2021) : Dept of Neurology, Washington University school of Medicine at Saint Louis.

Project: Engineering intrabodies therapy in Alzeimer Disease.

Senior Research Fellow (2019 August-2020 August) : New York Medical college Valhalla, Dept of Pharmacology

Project: Immunobiology of Porphyromonas gingivallis and Akkermansia muciniphila on Bone Marrow Macrophages.

Postdoctoral research fellow ( 2017 February -2019 July ) : Stony Brook University – SUNY New York Dept of Neurobiology and behavior.

Project: Nucleocytoplasmic Transport Defects and Stress Response in Nuclear Export Mediation Factor (NEMF) Mutant Mice and Cell Culture.

Visiting Scholar (2014 JAN- 2014 March.): University of Pennsylvania, Philadelphia. School of Dental Medicine, Dept of Periodontics

PROJECT: Impact of diabetes and inflammation on wound healing and apoptosis.

Postdoctoral Research Fellow (July2008 – March 2011), University of California, Irvine. Dept of Pediatrics.

Project: Molecular mechanism of Inclusion body myopathy, Paget Disease of bone and Frontotemporal dementia, Amyotropic Lateral Sclerosis.

Postdoctoral Research Fellow (Oct 2007-June 2008), University of Texas Health Center, Tyler

Project: Molecular mechanism of apoptosis in cancer cells.

Research assistant in ICMR project from 2000 to 2002:

Department of Neurochemistry, National Institute of Mental Health and Neuro Sciences (NIMHANS), Bangalore, India.

Project: Biochemical assays and identification of biomarkers in Bone disease

Academic Support Specialist (May 2014-aug 2014) : Biotecnika Information Pvt Ltd Bangalore. India.

Project : Preparation of notes, teaching materials and presentation through videoconference

Technical skills:

Analytical, HPLC and Biomarkers: Method development and validation, Peptides and Amino acid analysis from proteins, peptides, serum and urine specimens. Fluorescence detector and UV detector. C18 column chromatography. ( J.CHROM B and ANNALS OF CLIN BIOCHEM)

Purification, lyophilisation, analysis

Spectrometry: UV-VIS Spectrophotometry, Spectrofluorimetry and Chemiluminescence.

Cell Culture, Drug treatments, cell based assays, immunoassays: Maintenance of Various cancer cell lines, Myoblasts, U20S, MEFs, Transfection with SiRNA’s, Plasmids, Transient and permanent cell lines using selection markers, Treatment with drugs, Cell survival assays, Protein extraction, XTT assay, apoptotic assays, bioassays, enzyme assays, immunoassays.

Translational, xenograft and In-vivo studies: Maintenance of colony for disease models, Animal handling, breeding, genotyping and toxicology experiments on animals, various tissue harvesting to study the effect of drugs on signal transduction and metabolic pathways (Mice).

Cell Biology, Immunohistochemistry, Confocal Microscopy and Fluorescence Microscopy: Paraffin and Cryosectioning of tissues, Confocal imaging, Inverted fluorescence microscopy, immunofluorescence staining and analysis, TUNEL assay, apoptotic assays, autophagy, mitochondria, nuclear architecture, Endoplasmic reticulum stress, stress granules, p-bodies etc.

Molecular biology techniques: Transformation and cloning of SiRNA in E.Coli DH5a (FADD, PTEN, DR4-DR5 etc), plasmid purification, Transfection into various cell lines, RNA isolation from tissues, PCR, genotyping, Real time q-PCR.

Protein Biochemistry: Recombinant Protein expression, protein extraction from cell lines and tissues, immunoprecipitation, SDS-PAGE Electrophoresis, Western immunoblotting and dialysis etc.

Neuroscience: Harvesting brain, spinal cord, muscle and bone tissues from mouse, analysis with stainings, Extraction of proteins, isolation of m-RNA, DNA.

Clinical studies: Collection and processing of overnight fasting serum samples, 24 hrs urine sample, serum profile, urinary profile, semi autoanalysers, SOP

Bioinformatics: Windows-98/2000/Xp, Windows Office: Word, XL, Power points, BLAST analysis, Swiss Prot analysis.

In vivo Micro CT Imaging and diagnosis: Imaging, Anatomy of bone and morphometry, Diagnosis and analysis, Inveon micro CT on mice.

Behaviour studies : Novel objective recognition test, Rotarod testing in mice, grip strength.

Flow cytometry: Enrichechemnt of be cell population from PBMC and checking the purity with CD19, CD20, CD14, CD16, CD235a and CD43

Awards and Fellowships:

1.National Eligibility Test (NET-2000): Examination qualified From University Grants Commission-Council of Scientific and Industrial Research (UGC-CSIR) and National Eligibility Test (NET), Govt. of India.

2.Senior Research Fellowship (2004-06): Awarded from Indian Council of Medical Research (ICMR). Govt. of India.

Academic recognition:

1.Biographical profile has been selected for Marquis Who’s Who in the World 2012 (29th edition)

2.Biographical profile has been selected for The Stanford Who’s Who Black Book 2012 edition

3.Who's Who in Science and Engineering 2016-2017 (12th Edition).

4.Two of articles published are among Top 10 and Top 20 Articles Published in the Same Domain by Biomed library: -

Disease Markers journal (Top 10)

International journal of Clinical and Experimental Medicine. (Top 10)

Prep Biochem Biotechnol; 2010;40(1):83-7 (Top 20)

Membership:

American Society for Matrix Biology (ASMB)

Society for Glycobiology

American society for Human Genetics

Reviewer for Journals:

Brain Sciences

Clinical Biochemistry

Connective Tissue research

Biochimica Biophysica Acta- Molecular Basis of Disease

American Journal of Medical Genetics Part-A

Publications:

1.Mallikarjun Badadani, K. Taranath Shetty and S.S Agarwal (2021) A Spondylo Epi (meta) Physeal Dysplasia:Hypocalcitonemia in Handigodu Disease: Highlights on Medicine and Medical Research Vol 13, by Book Publisher International Chapter1 Print ISBN: 978-93-91215-39-2, eBook ISBN: 978-93-91215-47-7

2.Nalbandian A, Nguyen C, Katheria V, Llewellyn KJ, Badadani M, et al. (2013) Exercise Training Reverses Skeletal Muscle Atrophy in an Experimental Model of VCP Disease. PLoS ONE 8(10): e76187. doi:10.1371/journal.pone.0076187. (IF: 4.411) (Citations : 13)

3.Mallikarjun Badadani, Autophagy mechanism, regulation, function and disorders. Review article ISRN Cell Biology, vol 2012. Article ID 927064, doi:10.5402/2012/927064, 2012,1-11 (IF 1.000) (Citations : 17)

4.Mallikarjun Badadani, S.V.SureshBabu, K.Taranath.Shetty, S.S.Agarwal. Presence of Aggrecan fragments in the serum precipitated by DMB dye: Application in the study of Handigodu disease. Journal of Biochemical Technology, 2012, 3(4),438-441 (I.F: 0.962).

5.Nalbandian A, Llewellyn KJ, Kitazawa M, Yin HZ, Badadani M, et al. (2012) The Homozygote VCPR155H/R155H Mouse Model Exhibits Accelerated Human VCP-Associated Disease Pathology. PLoS ONE 7(9): e46308. doi:10.1371/journal.pone.0046308 (IF 4.411) (Citations : 25)

6.A. Nalbandian, K. Lavelline, Mallikarjun Badadani et al A progressive translational mouse model of Human VCP disease: The VCP R155H /+ Mouse. Muscle and Nerve, November 2012, DOI: 10.1002/mus.23522 (IF 2.367) (Citations : 36).

7.A. Nalbandian, S. Donkervoort, E. Dec, M. Badadani, et al. The multiple faces of valosin-containing protein-associated diseases: inclusion body myopathy with Paget disease of bone, frontotemporal dementia and amyotropic lateral sclerosis (Review). Journal of Molecular Neuroscience.2011, 45, 522-531. (IF: 2.922) (Citations : 95)

8.Mallikarjun Badadani, A. Nalbandian, Giles D. Watts, Jouni Vesa, Hailing Su, E Dec, Masashi Kitazawa, Douglas C. Wallace, Vincent Caiozzo and Virginia E. Kimonis et al. VCP associated inclusion body myopathy and paget disease of bone knock-in mouse model exhibits tissue pathology typical of human disease. PLoS ONE 5(10): e13183. doi:10.1371/journal.pone.0013183 (October 2010). (IF: 4.411) (Citations : 86)

9.Mallikarjun Badadani, K. Taranath Shetty, S. S. Agarwal. Hypocalcitonimia in Handigodu Disease: A spondylo epi (meta) physeal disorder. Int.J.Clin.Exp.Med. 2010, 3(2), 115-121. (IF: 1.875) (Citations : 4)

10.Mallikarjun Badadani, K. Taranath Shetty, S. V. Sureshbabu., S. S. Agarwal. Peptide bound hypohydroxyprolinuria in Handigodu Disease: A familial syndrome of spondylo epi (meta) physeal dysplasia. Disease Markers. 2009, 27, 7-12. (IF:1.723) (Citations : 4)

11.Mallikarjun Badadani and K. Taranath Shetty. Optimum Pyrolytic Conditions for furan derivative formation from poly aminosugars. Preparative biochem and biotechnology.2009, 40, 1,83-87 (IF:0.603)

12.Mallikarjun Badadani, S.V.Suresh Babu, K. Taranath Shetty, S. S. Agarwal. Metabolic status of Magnesium and Ceruloplasmin in Handigodu Joint Disease: a variety of spondylo epi (meta) physeal dysplasia. Clin Chim Acta. 2008 Sep; 395(1-2):170-171. (IF: 2.388) (Citations : 10)

13.Mallikarjun Badadani, S.V.Suresh Babu and K. Taranath Shetty. Optimum conditions of autoclaving for hydrolysis of proteins and urinary peptides of prolyl and hydroxyprolyl residues and HPLC Analysis. Journal of Chromatography B, 2007, 847, 2, 267-274. (IF: 2.971) (Citations: 12)

14.Mallikarjun Badadani, S.V.Suresh Babu and K. Taranath Shetty. “Dinitrophenyl Derivatization of Imino Acids, Spectral Characteristics and HPLC Analysis: Application in Urinary Peptide Derived Hydroxyproline and Proline Assay” Annals of Clinical Biochemistry 2007, 44, 2, 164-172. (IF: 2.209) (Citations : 3)

15.Dana TG, Wu Y, Badadani M. PYK2 contributes to re-epithialization by promoting MMP expression. Am.J.Physiol-Cell Physiol, 2014. (accepted) (I.F. 3.78) (Citations : 5)

Book Chapters:-

16.VE Kimonis, E Dec, M. Badadani, A. Nalbandian, J. Vesa, V Caiozzo, D. Wallace, B. Martin, C. Smith, GD Watts. Clinical spectrum of VCP mopathy, Pagets disease and fronto-temporal dementia, experimental models and potential treatments (chapter#15). Muscle Ageing, Inclusion-Body Myositis and Myopathies, eds. Valerie Askanas and W. King Engel, Blackwell publishing. pp 219-229. Publishing on 06 Jan 2012. (Citations : 4)

17.CD Smith, M. Badadani, A Nalbandian, E. Dec, J. Vesa, S. Donkervoort, B. Martin, GD Watts, V. Caiozzo and V. Kimonis. “Valosin-Containing Protein (VCP) Disease and Familial Alzheimer’s Disease: Contrasts and Overlaps (Chapter #16)” in “The Clinical Spectrum of Alzheimer’s Disease -The Charge Toward Comprehensive Diagnostic and Therapeutic Strategies” Edited by: Suzanne De La Monte, Publisher: InTech, Subject:Clinical Neuroscience. pp 331-346. Publication date: September 2011.

(Total impact factor : 35.783, Total citations: 314)

In Preparation

18. Mallikarjun Badadani Roger Sher et al. Role of Nuclear Export mediation Factor(NEMF) in stress invitro.

19.Roger Sher, Mallikarjun Badadani et al Nucleocytoplasmic transport defects with Nuclear Export mediation Factor(NEMF) mutation.

20.Mallikarjun Badadani, TDP-43 pathogenesis in musculoskeletal disorders. Review article (In Preparation).

21. Mallikarjun Badadani, Genomics and proteomics approaches to understand bone and muscle disorders. Review article (In Preparation).

Abstracts/Posters/ Platform presentations:

1.American Society of Human Genetics 60th Annual Meeting in Washington DC, November 2-6, 2010. M. Badadani, A. Nalbandian, J. Vesa, G. Watts, D. Eric, V. Caiozzo, D. Wallace, V. Kimonis. A VCP R155H Knock-In Mouse Model exhibits progressive muscle, bone and brain pathology typical of human IBMPFD Syndrome.

2.The American Society of Human Genetics 60th Annual Meeting November 2-6, 2010 Washington, DC A. Nalbandian, E. Dec, S. Ghimbovschi, S. Aizik, J. Vesa, M. Badadani, G. Watts, C. Smith, B. Martin, E. Hoffman, V. Kimonis. Global Differential Gene Profiling of Patients with VCP-Associated Myopathy and Paget Disease of Bone.

3.American Society of Human Genetics 59th Annual Meeting in Honolulu, Hawaii, October 20- 24, 2009. M. Badadani, J. Vesa, H. Su, G.D. Watts, V.J. Caiozzo, V.E. Kimonos. A VCP R155H Knock-In Mouse Model mimics human IBMPFD (Inclusion Body Myopathy and Paget disease of bone and Frontotemporal Dementia) disease.

4.7th International Imaging Genetics Conference, University of California at Irvine, January 17-18, 2011. Mary Lan, Tugan Muftuler, Sandra Donkervoort, Eric Dec, Angèle Nalbandian, Mallikarjun Badadani, Charles Smith, Barbara Martin, and Virginia E. Kimonis A study of MRI changes in individuals with VCP associated inclusion body myopathy, Paget and frontotemporal dementia.

5.Oral presentation at 32nd Association of Clinical Biochemists of India (ACBI) conference from 18th to 21st December 2005 At Patna. Mallikarjun Badadani, K.Taranath Shetty, S.V.Suresh Babu, and S. S. Agarwal. Defective hydroxylation of prolyl residues: A disorder of collagen metabolism.

6.32nd Association of Clinical Biochemists of India (ACBI) conference from 18th to 21st December 2005 At Patna. Mallikarjun Badadani, K. Taranath Shetty, S.V.Suresh Babu, and S. S. Agarwal Handigodu disease: A Metabolic disorder of skeletal system.

7.31st Association of Clinical Biochemists of India (ACBI) Conference from 1st - 4th December 2004 at Belgaum. Mallikarjun Badadani, S.V.Sureshbabu, A.Pavankumar Shetty, S.S.Agarwal and K.Taranath Shetty. Method for urinary peptide derived amino acids with special reference to the assay of proline and hydroxy proline.

8.30th Association of Clinical Biochemists of India (ACBI) conference from 19th to 22nd January 2004 At Bangalore. Mallikarjun Badadani., S.V.Sureshbabu., Seetharamaiah Chittiprol., Devaraju, K. S., and K.Taranath Shetty. Assay Method for Hydroxyproline and Proline in urinary peptides.

9.Taranath Shetty. S.V.Sureshbabu, M. Badadani, S.S.Agarwal. Inborn (neuro) metabolic disorder in Indian Scenario. Proceedings of 10th Asian Pacific Congress of Clinical Biochemistry (APCCB) Perth, Australia.19th-23rd September 2004, S32.

Summary of Research Experience:

Heterogeneity of B lymphocyte in Peripheral artery disease(PAD) and effect of infiltration on endothelial cells and myocytes

ScRNA SEQ from PAD (Peripheral Artery Disease) muscle biopsies Shows increased level of B cells. Based on this HLI (hind limb ischemia) for mouse was done, and data shows increased level of CD19 positive staining from Gastrocnemius muscle (Ischemic GA) biopsies from HLI compared to nonischemic gastrocnemius muscle. These results signify the B cells have infiltrated the muscle and can cause harmful or beneficial endothelial cells.

To address whether B cells have cytotoxic effect by interacting directly or by secretions. To answer this question B cells have been isolated /enriched from PBMC of human donor with cd19 negative selection cocktail. Purification was confirmed by CD19 flow cytometry.

The enriched B cells were subjected to 12 hrs HSS ( hypoxia serum starvation) and 24 hrs HSS in plain RPMI and Condition medium (CM) was collected.12 HRS HSS found to be optimal experimental condition, The CM was used to experiment on EC (endothelial cells) viability after EC and CM were incubated together for 12 hrs HSS and was compared to control (plain RPMI) . The viability of EC was going down in B cell derived CM. This implies B cells secrete proteins and cytokines that are cytotoxic to EC. The B cells HSS is confirmed by staining with HIF1a and is located to nucleus

B cells were enriched from PBMC of PAD samples and stained for fluorescence imaging for microscopy. The percentage of cell populations ; cd19+ only(89%), cd20+ only(88%), cd20+cd19+ (87%), cd20-cd20- (10% ) in control specimens were significantly higher when compared to PAD specimens {cd19+ only(70%), cd20+ only(70%), cd20+cd19+ (69%)}, In PAD specimens population of cd19-cd20- (27% )is significantly higher when compared to control specimens {cd19-cd20- (10%)}

The similar results were observed by flow cytometry .The percentage of cell populations cd19+ only(75%), cd20+ only(62%), cd20+cd19+ (57%) in control specimens were significantly higher when compared to b cell population from PAD specimens{ cd19+ only(58%), cd20+ only(34%), cd20+cd19+ (24%)}, In PAD specimens populations of cd19-cd20- (50% )is significantly higher when compared to control specimens {cd19-cd20- (23% )}.

These data signify plasma cell or plasma blast population have been increasing among PAD specimens and needs to be understood the importance of these cell populations.

Nucleocytoplasmic Transport Defects and Stress Response in Nuclear Export Mediation Factor (NEMF) Mutant Mice and Cell Culture.

NEMF is 122kd protein thought to be functioning as exporter from nucleus to cytoplasm. To understand the nucleo-cytoplasm transport mediated by NEMF, R86S mutant mouse created by Jackson lab that shows paddle foot syndrome similar to ALS (Amyotropic Lateral Sclerosis). We used both spinal cord from mutant mouse and Mouse embryonic fibroblasts(MEFs) and studied nucleocytolasmic location of RanGAP, TDP-43, phospho-TDP43, Lamin A/C, Nucleoporins (Nups), and several stress markers like eIF3s, DCP1, DCP2, TIA1, HSF1, Ribosomal proteins S6 and L3. We observed 1.Loss of femoral motor nerve and ubiquitination of spinal cord motor neuron. 2.Denervation of Neuromuscular junction 3. Mislocalization of RanGAP1 and collapse of Ran gradient.4. Ran and RanGAP are more cytoplasmic in MEFs. 5. Nup’s are more nuclear. 6.Importin-B pathway gets affected but not exportin pathway.

We studied NUPs in NEMF mutant MEFs and observed following changes 1.Mislocalization of TDP43 RANBP1, RANBP2, NUP98, SFPQ & RCC1 2. Aberrant changes in the Nuclear Envelope formation with NUP62, NUP50, GLE1 & RANGAP1.

To understand the transport of RBP in NEMF mutant mice we created stress condition in MEFs by inducing with 250 um Na- Arsenite and looked translocation of different proteins, their stress granular and p-body formation. We observed the following results 1. loss of cytoplasmic NEMF 2.No changes in TDP43 3.Less stress granular formation by TIA1,HSF1and eIF3a 4. Less P-body formation by DCP2 and eIF2b. 5. No changes in ribosomal proteins RPS6 and RPL3 6. Increased accumulation of Trascription Factor EB (TFEB).These results implies NEMF is required for nucleocytoplasmic shuttling of RBPs and lack of response mechanism to stress in ALS. The study needs to be addressed in other cell lines like iPSC neurons,U2OS HEK293 and MEFs with NEMF mutant plasmid transfection.

A Knock-In VCP R155H Mouse Model mimics Inclusion Body Myopathy Paget disease, fronto temporal dementia and amyotropic lateral sclerosis replicating human IBMPFD Syndrome.

Dominant mutations in the valosin containing protein (VCP) gene cause inclusion body myopathy associated with Paget’s disease of bone and frontotemporal dementia (IBMPFD), which is characterized by progressive muscle weakness, dysfunction in bone remodeling, and premature frontotemporal dementia. (IBMPFD) is caused by mutations in the Valosin Containing Protein (VCP) gene on chromosome 9p12-13.

VCP is a ubiquitously and highly expressed in cells and is involved in several cellular processes including post-mitotic nuclear envelope reformation and Golgi reassembly, cell cycle progression, endoplasmic reticulum-associated degradation of defective proteins and DNA damage repair. VCP plays an essential role in the endoplasmic reticulum associated degradation of proteins by binding to ubiquitinated substrates and transferring them to the 26S proteasomes of the ubiquitin proteasome system. The majority of mutations are located in the ubiquitin-binding domain of VCP, R155H being the most common disease mutation.

To elucidate the pathological mechanisms underlying the disease progression of IBMPFD, a knock-in mouse model with the R155H mutation and analyzed the progression of disease up to 15 months of age.

We created three knock-in mouse models pertaining to VCP 1. Heterozygous Neo +ve R155H Knock-in

2. Heterozygous Neo –ve R155H Knock-in 3. Homozygous R155H / R155H knock-in.

Muscle strength measurements in heterozygous mice demonstrated progressive muscle weakness in mutant mice starting before the age of 6 months. Histological analyses of mutant muscle performed by hematoxylin and eosin staining, and electron microscopy showed vacuolization of myofibrils, centrally located nuclei, and disorganized muscle fibers. Immuno histochemical analyses of the quadriceps muscle tissues from mutant mice showed accumulation of TDP-43- and ubiquitin-positive inclusion bodies in 4% of quadriceps myofibrils. LC3 staining of muscle sections showed increased number of autophagosomes, and increased apoptosis was demonstrated by elevated Caspase-3 activity (2.8-fold increase) and increased number of TUNEL-positive nuclei in mutant quadriceps. The tissue lysates from quadriceps and brain have been analyzed for VCP, ubiquitin and LC3 by western blot. The ubiquitin and LC3 have been expressed more in Knock-in mice than control except VCP. The expression of VCP have been confirmed by q-RT PCR analysis and no changes have been observed. The brain analysis from knock-in mouse shows similar pathology of humans like accumulation of extra nuclear tdp-43, VCP and Ubiquitin positive inclusions.

The ionized calcium binding adaptor protein 1 (IBA1) is thought to play a role in regulating the function of microglia and considered to be specific biomarker. Microglial primary function involves maintenance of normal tissue brain homeostasis and production of variety of neurotransmitters. Higher levels of IBA 1 expression was observed in frontal cortex of knock-in mouse. Hence homeostasis of brain and neurotransmission may be altered

X-ray micro tomography (micro CT) images show radiolucency of distal femur and proximal tibiae in knock-in mice and bone morphometry shows decreased trabecular pattern factor and increased cortical wall thickness. Bone histology and bone marrow derived macrophage (BMDM) culture in these mice revealed increased osteoclastogenesis observed by TRAP staining suggestive of Paget disease of bone. Hence the mouse model mimicks paget disease in addition to IBMPFD.

Sporadic PDB and familial PDB have Sequestosome 1 (SQSTM1) gene mutations, which encodes p62 and has an important role in autophagy. The SQSTM/P62 P394L mutant mice 64 revealed focal osteolytic lesions on hind limbs by micro CT pathology which resembles the lesions seen in our new Paget disease like mouse model.

The homozygous VCPR155H/R155H mice manifest prominent muscle, heart, brain and spinal cord pathology, including striking mitochondrial abnormalities, in addition to disrupted autophagy and ubiquitin pathologies. VCPR155H/R155H mouse spinal cords and discovered significant motor neuron degeneration and TDP-43-positive cytoplasmic accumulation, associated with ALS pathology. The mice are able to survive upto 21 days shows severity of disease. Upon analysis all these mice show similar characteristics to Amyotropic Lateral sclerosis (ALS) a neurodegenerative disorder.

Our analyses revealed that the generated R155H knock-in mice replicate human disease, and therefore can be used to clarify the mechanisms causing IBMPFD and ALS, as well as to develop novel therapies for this currently untreatable disease. In conclusion, the R155H knock-in generated mouse model replicates human disease, and therefore can be used to clarify the mechanisms causing IBMPFD. The knock-in mice can also be used in the development of novel therapies for this important disease.

Treatment

investigation was undertaken to examine the effects of uphill and downhill exercise training on muscle histopathology and the autophagy cascade in an experimental VCP mouse model carrying the R155H mutation. Progressive uphill exercise in VCPR155H/+ mice revealed significant improvement in muscle strength and performance by grip strength and rotarod analyses when compared to the sedentary mice. In contrast, mice exercised to run downhill did not show any significant improvement. Histologically, the uphill exercised VCPR155H/+ mice displayed an improvement in muscle atrophy, and decreased expression levels of ubiquitin, P62/SQSTM1, LC3I/II, and TDP-43 autophagy markers, suggesting an alleviation of disease-induced myopathy phenotypes. There was also an improvement in the Paget like phenotype.

Autophagy modifying drugs such as rapamycin at pharmacological doses have previously shown to alter the autophagic flux. Rapamycin treatment in 20 months old VCP R155H/+ mice showed improved muscle performance, quadriceps histological analysis, and improvement of the ubiquitin, and TDP-43 pathology and defective autophagy as indicated by decreased protein expression levels of autophagy markers LC3-I / II, P62/SQSTM1 and optineurin.

Engineering Intrabodies therapy in Alzeimers Disease.

We generated P301S -Tau HEK293t cells by lentiviral transduction.These cell lines were used for intrabody transfection. Intrabodies were created by fusing FvSc fragment and LIR domain of sequestosome (p62).These have been creted with different size of linkers of FvSc. Tgese intrabodies were tranfected to P301S -Tau HEK293t cells and the levels of tau have been quantified by western blot. The decrease in tau levels were observed with intrabodies fused with LIR domain compared to FvSc alone.The study needs to be applied to primary neurons.

Immunobiology of Porphyromonas gingivallis (PG) and Akkermansia muciniphila (AKK) on Bone Marrow Macrophages

I developed the method to quantify the PG and AAK bacteria in blood specimen using blood clot from mouse by employing Qpcr. I also isolated bone marrow macrophages from mouse and were incubated with PG and AKK bacteria and inflammatory markers were quantified like TNF-alfa, IL-10, . the levels of TNF-alfa and IL-10 were elevated in PG but were low in AKK. When PG is combined with AKK the levels of inflammatory markers were reduced compared to PG alone. This can be employed to reduce the inflammation in osteoarthritis etc.

Molecular Mechanism of Apoptosis in Cancer Cells:

The SiRNA s like DR4-DR5 SiRNA, Caspase 8-10 SiRNA, FADD SiRNA, PTEN SiRNA, and PUMA SiRNA etc have been transformed and cloned using DH5 alfa E.Coli bacteria. The isolated SiRNA’s have been transfected into various cancer cell lines like HCT116 colorectal, LNcap, HT29 COLON, jurkat, and HL 60 leukemia etc. The permanent and transient transfected cells have been made to study molecular mechanism of apoptosis in cancer cells. The drugs like EGCG, Resevetrol, LY29, akt inhibitor and small peptides have been treated to cell lines. The proteins have been isolated for western blotting. Apart from this the cell viability has been assayed using XTT salt. For example, the Resveratrol induces apoptosis revealed by inhibiting the expression of Bcl-2 and XIAP in human cancer cell lines. In the XTT assay the control cell lines have shown more values than drugs treated cell lines with SiRNA transfected. This implies drugs along with SiRNA induces apoptosis.

Animal models like KrasG12D transgenic mice and xenograft mouse models have been used for invivo studies. The drugs like EGCG, Resvetrol and small peptides have been given through intraperitonial and oral. The pancreas had been collected and used for immunohistochemistry. The tumor growth is suppressed in drug treated animal.

The use of SiRNA with combination of drugs may be beneficial in treating the cancers.

The xenografts have been created using SCID mice by injecting cancer cell lines. The drugs have been used treat these models to observe whether have any reducing effects on cancer. We observed that Reservetrol, EGCG, Curcumin have reducing effects on cancer.

Summary of Ph.D Thesis:

Thesis Title: Biochemical studies on Handigodu disease: A familial syndrome of Epi (Meta) physeal dysplasia:

Handigodu Disease (HD), a familial syndrome of Epi (Meta) physeal dysplasia, first reported during year 1975 from Handigodu village of Shimoga district, affects a particular socio-economically backward community habitated in Shimoga and Chickamagalore districts of Karnataka state, India. The clinical manifestation of the disease onset, characterized by pain in joints and spine, varies between individuals; symptoms appearing as early as the first decade of life and progressively leading to crippling condition unable to walk and confine to indoor. Earlier studies comprising clinical, eco-toxicological aspects spanning over three decades have failed to explain the etiology and or the biochemical/molecular basis of the disease. Nevertheless, based on the wide spectrum of the clinical presentations and the associated radiological findings on skeletal system has led to the classification of the patients as under three categories, viz., Type I (

Contact this candidate