UNIVERSITY AND INSTITUTE OF ADVANCED RESEARCHiiar.res.in/form/annual_report2013.pdf · 11. Dr....

UNIVERSITY AND INSTITUTEOF ADVANCED RESEARCH

Annual Report

www.iar.ac.inwww.iiar.res.in

2013

M I S S I O NTo conduct fundamental/applied/analytical research in the frontier areas of science with the ultimate objective of contributing to high quality basic research and human resource development and to find application of the research results for the benefit of mankind

Fig.1. Axonal connections in Drosophila retina and brain complex.

Fig.2. Photoreceptor cells of Drosophila retina.

C O V E R P A G E

C O N T E N T S

Annual Report

2013

1

Foreword 2

1. Introduction 3

1.1 DBT Program Support 3

1.2 Faculty 4

1.3 Research Staff 6

1.4 Research Infrastructure 7

1.5 Lab Equipment 7

2. Institutional Committees 8

2.1 Governing Body 8

2.2 Research Advisory Committee 8

3. Research & Development 10

3.1 Human Health & Disease 10

3.2 Genetics & Developmental Biology 19

3.3 Cell Biology 21

3.4 Plant Developmental Biology 23

3.5 Bioinformatics & Structural Biology 25

3.6 Environmental Science 31

4. Publications 34

4.1 Presentations 34

5. Awards 37

6. Human Resource Development 38

7. Financials 40

7.1 Income and Expenses 40

7.2 Research Grants 41

FOREWORD

Provost, UIAR

University And Institute of Advanced Research is one of its kind in Gujarat,

where various choice and credit based courses are offered providing a

unique opportunity for students for learning in a research intensive

environment. It also offers students the flexibility to choose courses of their

choice.

The University has well equipped laboratories, library and other support

facilities for the students. The first centre of excellence “School of Biological

Sciences & Biotechnology” is carrying out research in frontier areas of

Biology to contribute towards the advancement of human knowledge and alleviating human

suffering. University has large number of students coming from different states of country for research

and Ph.D. course at UIAR campus. Faculties and students at UIAR are actively involved in high quality

research in different areas of Science.

The faculties with their research team have published more than 10 research articles in highly reputed

peer reviewed international journals and presented more than 25 papers in national and international

conference / symposia. The University has initiated training programs for school students and teachers

conducted by various departments. The programs were highly appreciated by the school, college

teachers and students.

stDr. Kailash Paliwal, the first Provost of the University passed away on May 31 2013. He had set up the

Department of Environmental Science and Climate change. We sincerely acknowledge his

contribution for development of the University in the early years.

I would like to congratulate Prof. Rajani Nadgauda for having been appointed as Professor Emeritus at

the University. I would also like to wish Dr. Sunil Kumar Singh and Dr. Rajesh Singh who have relocated

on promotion to other institutions.

This report is a summarized document of ongoing research activities at UIAR during the year 2013. It

highlights the update of the research activities, publications of various departments, educational

activities and finances of the University.

These achievements would not have been possible without the continuous support of the University

President, Prof. N R Puri, the guidance of the Board of Governors, Board of Management, Academic

Council and active participation of the faculty and staff of the university. We would also like to thank the

trustees of The Puri Foundation for Education in India for the generous financial support. I convey my

gratitude to all of them.

In the coming years we intend to increase our teaching and research collaboration at national and

international level. I wish UIAR will progress rapidly in coming years and will be University of choice for

higher education and research.

Shri. Ashwani Puri,Provost, UIAR

Annual Report

2013

2

1. INTRODUCTION:

The University and Institute of Advanced Research was established under the “Gujarat Private University

Amendment Act 2011. The University has initiated various academic programs to provide a unique

opportunity for students to learn in a research intensive environment. The University is promoted by The

Puri Foundation for Education in India, a registered charitable trust.

The First Centre of Excellence, School of Biological Sciences and Biotechnology, part of Indian Institute

of Advanced Research (IIAR), carries out cutting edge complementary research in various areas of

biology. It has departments of Human health and disease, Cell biology, Plant molecular biology,

Developmental biology, Bioinformatics and structural biology and Environmental Science. The school

also carries out doctoral and postdoctoral research.

The trust is registered with the Department of Science and Technology as a SIRO.

The University is also developing a faculty of Humanities and Social Sciences. The University will be

offering various choice and credit based undergraduate, postgraduate and doctoral programs.

1.1 DBT Program Support:

The institute currently receives Program Support grant from DBT. Five projects are being carried

out under this grant. Two new departments- Department of Genetics and Developmental Biology

and Department of Immunology has been established under this program. Establishment of a

central facility of scientific equipment utilizing this grant has helped in boosting the research

output. These facilities are also being extended to other Universities/ Institutes on a no charge

basis. The grant has additionally helped the Principle Investigators fetching new research funding

using the infrastructure strengthened by the same. The last assessment of the research work was rd

carried out by the Research Advisory Committee on 3 February, 2013. The work was considered

satisfactory.

Annual Report

2013

3

There�are�currently�10�faculty�members at�the�institute.st

Dr. Kailash Paliwal, respected director of the institute passed away on May 31 , 2013 after prolonged

illness.

Dr. Rajesh Singh and Dr. Sunil Singh left the institute on promotion during this period.

Dr. Desh Deepak Singh is presently at Central University of Punjab, Bathinda on lien from UIAR.

The details of the faculty and their areas of research interests are placed below.

1.2 Faculty

Annual Report

2013

S. No.

Scientist's Name, Qualification & DesignationDepartment Name

Full Time/ Part Time, Year of Joining, Experience

Research Interests

1. Dr. Kailash Paliwal, PhDDirector & ProfessorEnvironmental Sciences

Full Time 28/05/2010 30 yrs

Climate change plant biology. Study of effect of land use change in communities composition and invasiveness. Biodiversity conservation and ecosystem services.

2. Dr. Dheeraj Naik, PhDAssistant Professor,Environmental Sciences

Full Time05/06/20127 yrs

Molecular and physiological mechanisms underlying adaptation and acclimation of plants to their stressful environment, which are accelerated due to changing climate conditions. Affect of drought and edaphic stresses on forest and grassland ecosystems.

3. Dr. Rajani Nadgauda, PhDProfessor Emeritus,Plant Cell and MolecularBiology

Part Time 08/02/2006 28 yrs

In vitro plant cell culture and isolation of active principals from cell cultures / Hairy root cultures. Conservation of plants though micropropagation. Understanding the somatic embryogenesis process technology development and dissemination.

4. Dr. Sunil Singh, PhDAssistant ProfessorPlant Cell and MolecularBiology


Understanding molecular genetic that control programmed cell death, vascular tissues differentiation and cell wall formation in plants. Plant developmental biology and plantfunctional genomics.

5. Dr. Rajesh Singh, PhDAssistant ProfessorCell Biology


To understand the cell death and inflammatory pathways activated during physiological and pathological conditions. To study the mitochondrial targeted mitochondrial targeted miRNA, their physiological relevance in terms of cell death and survival. Regulation of mitochondrial dynamics and physiology by ubiquitination system.

4

Annual Report

2013

6. Dr. Chandramani Pathak, PhD Assistant ProfessorCell Biology

Full Time 21-10-2009 5 yrs

Understanding the molecular mechanism and cross talk between apoptosis and inflammatory signaling in cancer. Nano-particle based drug delivery in human cancer cells and evaluating their apoptotic potential for therapeutic intervention.

7. Dr. Rajendra TK, PhD Associate ProfessorGenetics and Development Biology


Global regulation of gene expression, molecular understanding of nuclear organization in relation to regulation of gene expression, biology of non-coding RNAs, protein and RNP assembly dynamics during germ-line development and differentiation, understanding developmental and molecular mechanisms of Spinal Muscular atrophy, Spinal and Bulbar Muscular Atrophy.

8. Dr. Anand Tiwari, PhDAssistant Professor Genetics and Development Biology


Role of molecular chaperon and ubiquitin ligases during eye development of Drosophila melanogaster

and the progression of Alzheimer disease

using Drosophila melanogaster as a model organism.

Effect of medicinal plants on development

of Drosophila melanogaster .

9. Dr. Ashima Bhardwaj, PhDAssistant ProfessorHuman Health and Disease


Deciphering the molecular mechanisms that govern the multidrug resistance phenotype of the isolates of Vibrio sp. and Shigella sp. Study of mobile genetic elements like SXT elements, integrons and plasmids that lead to fast acquisition and dissemination of the genes responsible for the observed drug resistance.

10. Dr. Reena Rajput, PhDAssistant Professor, Human Health and Disease

Full Time 1/7/2010 6 years

Study of Toll-like Receptors mediated immune outcomes in various disease models and during vaccine reactogenicity. Study of parallels between immune and the nervous system to reprogram immune cells to neurons.

11. Dr. Desh Deepak Singh, PhD,Associate Professor Bioinformatics and Structural Biology


Adhesins and surface proteins from Leishmania involved in host-pathogen interactions. Study of the glycome expression and its interactions in biological systems for mediating cellular interactions. Genome / proteome profiling, annotations and development of related tools and databases.

5

Annual Report

2013

12. Dr. Anju Pappachan, PhD Assistant Professor, Bioinformatics and Structural Biology


Characterization of proteins belonging to the purine salvage pathway of L. donovani Study of kinases involved in carbon metabolism, proteins belonging to trypanothione metabolism and methyl glyoxal pathways of L. donovani.

13. Dr. Neeraj Jain, PhDAssistant Professor,Plant Cell and Molecular Biology

Full Time1/12/20057 yrs

Plant functional genomics, DNA fingerprinting, marker development and in vitro regeneration studies with practical applications in plant conservation, biofortification, phytoremediation and production of elite planting material of economic and ecological importance like medicinal plants and biofuel.

1.3 Research Staff:

There are 49 research staff currently assisting the faculty in their research work. Research

staff includes Post Doctoral Fellows, PhD Scholars, SRF, JRF, and Project Assistants. The

details of the current research staff are placed below.

Project Investigator 10

Research Associate 4

SRF 9

JRF 30

Project Assistant 4

Technical Assistant 2

Total 59

6

1.4 Research Infrastructure:

The institute has the following facilities at the School of Biological Sciences and Biotechnology.

i) Plant cell and Molecular biology lab with green house, plant tissue culture facilities, growth chamber, laminar flows.

ii) Bioinformatics and Structural Biology lab with complement of servers, computers, cluster computer system and wet lab.

iii) Human Health and Disease laboratory with laminar flow, biosafety cabinets and allied microbial culture facilities.

iv) Cell biology lab with class II cell culture facility.

v) Genetics and Development Biology lab with Drosophilae model system.

vi) An Environment and Ecology laboratory with Eddy Covariance Instrument to measure climate change parameters.

vii) Cold room.

1.5 Lab Equipments:

The school of Biological Sciences and Biotechnology is equipped with state of the art facilities with

support from the Department of Biotechnology, Department of Science and Technology, ICMR,

Govt. of India and the trust- The Puri Foundation for Education in India.

List of key equipments is placed below.

Besides the above the departments have independent research specific facilities including small and

medium equipment to their specific requirements.

1

AKTA protein purification

system

2 Autoclave

3 Automated Cell Counter

4 Binocular Microscope

5 Biosafety Cabinets

6 Biosafety hoods

7 cDNA Synthesis Kit

8 Centrifuge Kubota

9 Circulatory water bath

10 CO2 incubators

11 Confocal Microscope

12 Cooling Incubator

13 Deep Freezers

14 Electronic orbital shaker

15 Fume Hoods

16 GC

17 Gel Doc Systems

18 HPLC

19 Incubator Shaker

20 Laminar Flow

21 Microscopes

22 Multi Mode Micro Plate

23 PCRs

24 Real Time PCR

25 Rota Vapour

26 Sorval centrifuge

27 Shaker

28 Spectrophotometers

29 Ultra centrifuge

30 Vacuum concentrator

31 Water Purification System

32 Water Treatment Plant

S. S. No. No. Name Name

Annual Report

2013

7

2. INSTITUTIONAL COMMITTEES

2.1 Governing Body:

The second governing body meeting of the university was held on 3rd December, 2013.

The following members constitute the Governing Body.

1 Mr. NR Puri President/Chancellor Chairman

2 Mr. Upendra Puri Trustee by rotation

3. Dr. G. C. Mishra Chairman of RAC, IIAR

4. Dr. R.C. Maheshwari

Chairman of University Development Committee

5. Dr. Rajendra TK Dean, Ex-Officio Member Secretary

6. Dr. B.Rao Executive Dean, London South Bank University

7. Mr. Ashwani Puri CEO, IIAR

8. Smt. Kirtidaben Bhrambhatt,

Joint

Sec,

Education

Deptt.

Nominee

Govt.

of

Gujarat

2.2 Research Advisory Committee

The following members constitute the Research Advisory Committee.

Chairman

G. C. Mishra, Former Director, NCCS & Scientist of Eminence, NCCS Complex, Ganeshkhind,

Pune-411 007.

Members:

• Dr. K. Muniyappa, Professor, Dept. of Biochemistry, Indian Institute of Science,

Bangalore-560012.

• Dr. K. Gurumurthi, Former Director, Institute of Forest Genetics & Tree Breeding

(IFGTB), Coimbatore- 641001.

• Dr. J. K. Roy, Cytogenetics Laboratory, Dept. of Zoology, Banaras Hindu, University,

Varanasi – 221005.

• Dr. B. B. Chattoo, Dept. of Microbiology & Biotechnology Centre, Faculty of Science, M.S.

University, Baroda – 390 002.

• Dr. Rajani Nadgauda, Dean, Indian Institute of Advanced Research, School of Biological

Sciences and Biotechnology, Village Koba, Gandhinagar-382 007.

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2013

8

• Mr. Ashwani Puri, CEO, Indian Institute of Advanced Research, School of Biological


• Dr. Kailash Paliwal, Director, Indian Institute of Advanced Research, School of Biological


• Dr. Jitendra P. Khurana, Tata Innovation Fellow (DBT), Professor & Coordinator (UGC-

SAP), Dept. of Plant & Molecular Biology, University of Delhi, South Campus, New Delhi –

110 021.

• Dr. K. Dharmalingam, DBT Distinguished Biotechnology Research Professor, School of

Biotechnology, Madurai Kamaraj University, Palkalainagar, Madurai- 625 021.

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2013

9

3. RESEARCH AND DEVELOPMENT

3.1 Human Health and Diseases:

Principal Investigators:

Dr. Ashima Bhardwaj (Associate Professor,

Group Head)

Research Fellows:

Mr. Priyabrata Mohanty (ICMR- SRF)

Ms. Neha Rajpara (ICMR- SRF)

Mr. Braj Mohan (CSIR- SRF)

Mr. K. Vinoth Kumar (GSBTM-JRF)

Ms. Aneri Shah (DBT programme

support-JRF)

Trainees:

Ms. Maitry Shah (Trainee)

Ms. Dhvani Patel (Trainee)

Ms. Nishtha Bhatt (Trainee)

Mr. Vivek Pathak (Trainee)

3.1.1 Description of research work

Project 1: Molecular characterization of factors

governing antibiotic resistance in the Indian

isolates of Vibrio spp.

As presented in the last year's report, detailed

analysis was carried out for unraveling the

mechanisms of antibiotic resistance in 119 clinical

isolates of Vibrio cholerae obtained from National

Institute of Cholera and Enteric Diseases (NICED),

Kolkata, India, 2009 (Kind courtesy Dr. T.

Ramamurthy). Among these, one of the clinical

isolate IDH02365 showed mixed growth on TCBS

and LB agar. Further study was carried out for

identification of the constituent strains by various

experiments described below.

When antibiogram assay was performed for clinical

isolate IDH02365, it showed double zones with

antibiotic discs like ampicillin, chloramphenicol,

ciprofloxacin, gentamycin, kanamycin, norfloxacin

and tetracycline. When this bacterial culture was

grown on TCBS agar plates, bluish green as well as

yellow colonies appeared indicating the presence

of two different bacteria in this clinical isolate.

Different type of growth was obtained on other

selective media such as HEA and XLD (Table

3.1.1.1). Both the bacteria were subjected to a

variety of biochemical tests and the results were fed

i n to AB IS on l i ne (Advanced Bac te r i a l

Identification Software, http://www.tgw1916.net /

bacteria_logare.html). Results showed its 90%

similarity to Providencia rettgeri. Therefore, the

clinical isolate actually consisted of Vibrio cholerae

and Providencia spp. 16s rRNA sequencing

revealed that the smal l co lony actual ly

corresponded to P. vermicola.

Table 3.1.1.1: Growth of Mixed culture, Vibrio cholerae (big colony) and Providencia spp. (small colony) on various media

Strains TCBS LB Agar HEA XLD

Mixed IDH02365 Yellow

colonies

Big and Small Merged Dark green Dark red

V. cholerae Yellow

colonies

Big sized mucoid

yellowish

No Growth No Growth

Providencia spp.

Bluish green

colonies

Small off-white Dark

Green

Dark red

Annual Report

2013

10

The genomic DNA was isolated from mixed, big

and small colonies. V. cholerae outer membrane

protein-specific OmpW primers were used for

identification of V. cholerae. An amplified band of

expected size (586 bp) was obtained only from

mixed and big-sized colonies and no amplification

was obtained from small-sized colonies (Figure

3.1.1.2). This result confirmed that big colonies

were V. cholerae.

Figure 3.1.1.2: Confirmation of V. cholerae bacteria through Outer membrane

proteins (OmpW)-specific primer. In this result big colonies, mixed cultures and

IDH01526 showed expected band size (586 bp) but small colonies showed no

amplicon. The IDH01526 was used as +ve V. cholerae strain.

RAPD experiments were performed with 1281

and 1283 random primers. BAB 812 and BAB

813 P. vermicola isolates were obtained from

Gujarat State Biotechology Mission, (GSBTM)

Gandhiagar, Gujarat, India. The MTCC 8089 and

MTCC 8929 P. rettgeri isolates were obtained from

MTCC

(Microbia l Type Cul ture Col lect ion ) an

international repository at IMTECH Chandigarh,

India. These isolates were used as positive controls

for P. vermicola and P. rettgeri respectively. The

IDH01304 clinical isolate of V. cholerae was used

as positive control for V. cholerae from mixed

infection. Results of RAPD showed different

patterns with both these primers for DNA from

mixed cultures, P. vermicola and V. cholerae,

hence confirming again that they were two distinct

Further work is in progress to understand the

organisms (Figure 3.1.1.3).

Figure 3.1.1.3: RAPD profile of mixed, big (V. cholerae) and small (P. vermicola)

colonies with their positive and negative controls. In these gels, mixed, small and

big colonies showed different amplification patterns.

molecular mechanisms governing the drug

resistance phenotype for this V. cholerae and P.

vermicola.

Project 2: Cloning and characterization of efflux

pumps responsible for multiple drug resistance in

clinical isolates of various Vibrio species

Two MATE-type efflux pumps (H- and D-

type) were cloned in the arabinose-inducible pBAD

E. coli expression vectors (Figure 3.1.1.4).

Optimisation studies with different inducer

concentrations and at different time points of

induction were carried out to determine the

conditions that yielded maximum expression of the

recombinant proteins (Figure 3.1.1.5). Further, to

assess the functionality of the recombinant efflux

pumps, a host strain was selected with deletion in

one of the outer membrane protein TolC. The

recombinant proteins are being characterized in

terms of their capacity to confer resistance towards

various antibiotics using MIC studies and drug

transport assays using this host.

Figure 3.1.1.4: (A). Coomassie blue stained SDS-PAGE showing expression from

pBAD VFD along with Lac-His positive control. (B). Western blot to detect

expression of recombinant protein. 0.2% L-arabinose was used for 4 hour induction.

Goat polyclonal Anti-V5 Primary antibody (Pierce) at 1:5000 dilution was used. Anti-

Goat HRP conjugated Secondary antibody (Jackson Immunoresearch) was used

for detection.

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11

Figure 3.1.1.5: Inference of optimal inducer concentration for recombinant protein

expression. Goat polyclonal Anti-V5 Primary antibody at 1:5000 dilution was used.

Anti-Goat HRP conjugated Secondary antibody was used for detection. Marker:

Spectra multicolor marker (Fermentas).

Project 3: Study of multidrug resistance in clinical

isolates of Shigella spp.

The work was carried out with 95 clinical isolates

of Shigella spp. which were procured from NICED

(courtesy Dr. T. Ramamurthy), Kolkata, India in

the form of stabs. The bacteria from stabs were

regenerated and confirmed on selective media

such as HEA, XLD, MacConkey and LB agar.

Analysis with these 95 isolates (years 2001-2010)

indicated the predominance of Shigella flexneri

and Shigella sonnei in these isolates. Antibiotic

susceptibility patterns were analysed for 91 of

these clinical isolates. Results revealed that all the

isolates were multidrug resistant and resistance to

drugs l ike trimethoprim, co- tr imoxazole,

streptomycin, nalidixic acid was very common

(Figure 3.1.1.6).

Figure 3.1.1.6: Antibiotic susceptibility profile of 91 clinical isolates of Shigella spp.

Amp, ampicillin; Azm, azithromycin; Cfx, ceftriaxone; Chl, chloramphenicol; Cip,

ciprofloxacin; Gen, gentamicin; Cxm, cefuroxime; Nal, nalidixic acid; Nor,

norfloxacin; Kan, kanamycin; Str, streptomycin; Ofx, ofloxacin; Tet, tetracycline; Tri,

trimethoprim; Cot, co-trimoxazole.

Further analysis of genomic and plasmid DNA from

66 shigella isolates showed the presence of multiple

plasmids which could be the possible reason for the

drug resistance phenotype. To detect the presence

of class 1 integrons, PCR experiments were carried

out using primers specific for 5' conserved region

(L2/L3), 3' conserved region (qacEΔ1/sul1B) and

variable region (inF/inB). Out of 66 isolates, 29

isolates were found to be positive for 5' conserved

region (L2/L3) (Figure 3.1.1.7). PCR analysis for 3'

conserved region revealed that out of 29 isolates

only one isolate gave expected 0.8 kb amplicon.

Figure 3.1.1.7: Agarose gel (1%) analysis of PCR products of 5' conserved region of

class 1 integron with L2, L3 primers (A) and 3' conserved region of class 1 integron

with qacEA1, sul1-B primers (B).

Further work is in progress to understand the other

possible drug resistance mechanisms like

mutations in topoisomerases and/or involvement

of mobile genetic elements in drug resistance

mechanisms.

Project 4: Unraveling the mechanisms underlying

quinolone resistance in multidrug resistant clinical

isolates of Vibrio and Shigella species from India

This project was sanctioned for financial support by

GSBTM, DST, Govt. of Gujarat, starting from 1st

April, 2013. Preliminary screening for this project

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was initiated last year. Quinolone resistant V.

fluvialis and Shigella isolates were screened for

quinolone-resistance-determining genes like

qnrVC, qnrB, qnrS, aac (6') Ib-cr, qepA and

oqxAB using PCR. The qnrVC gene was found in

three V. fluvialis isolates (BD146, L10734 and

L9978) and aac (6') Ib-cr was found in BD146

only. Sequencing of the QRDR regions of the four

topoisomerase genes (gyrA, gyrB, parC and parE)

revealed that two isolates (BD146 and L15318)

carried mutation in GyrA protein (Ser83→ Ile). As

clear from the above data, BD146 carried three

important quinolone resistance determinants

qnrVC, aac (6') Ib-cr and topoisomerase

mutations. Therefore, BD146 appeared to be an

interesting strain for study of quinolone resistance

and further detailed analysis was pursued with this

strain. The nucleotide sequences of the above

mentioned genes from V. fluvialis BD146 strain

were submitted to GenBank. In Shigella isolates,

S.flexneri M11560 was found to have qnrS.

The qnrVC gene from V. fluvialis BD146 was

cloned in pET28a expression vector to functionally

characterize the gene. The authenticity of the

recombinants was confirmed by restriction

enzyme analysis and PCR assays. Expression

studies were carried out in BL21(DE3) cells

harboring the recombinant plasmid. The protein

of band size ~27 kDa in SDS-PAGE was found to

be over expressed by the recombinants on IPTG

induction. The Coomassie blue stained protein

band at ~27 kDa position was excised from the gel

and analysed by BrukerUltraflex III MALDI

instrument. Trypsin digestion and peptide mass

fingerprinting followed by Protein identification

using Mascot software confirmed the protein to be

QnrVC5, a pentapeptide repeat protein. The

minimum inhibitory concentration assays (MIC)

were standardized to establish the functionality of

the recombinant QnrVCprotein (Table 3.1.1.8).

Table 3.1.1.8: Minimum Inhibitory Concentration (MIC) for qnrVC-pET28a

recombinant

Antibiotic qnrVC-pET28a in

E.coli BL21

(λDE3)

pET28a in E.coliBL21

(λDE3)

Elevation

in MIC

Nalidixic

acid

5

µg/ml

0.625

µg/ml 8 fold

Norfloxacin

0.125 µg/ml

0.0078 µg/ml 16 fold

Ciprofloxacin 0.0156 µg/ml < 0.00195 µg/ml >8 fold

More than 8 fold elevation in MIC was observed for

the quinolones tested in the study. The MIC results

clearly indicated that the recombinant protein was

expressed and functional. Therefore, it conferred

the elevation in MIC (resistance) to the host cells for

quinolones.

Acknowledgements:

The laboratory is supported by the grants from the

Department of Biotechnology (DBT), Ministry of

Science and Technology, Government of India (No.

BT/PR/11634/INF/22/104/2008), Indian Council

of Medical Research, New Delhi, India (No.

A M R / 4 9 / 1 1 - E C D I ) a n d G u j a r a t S t a t e

Biotechnology Mission, Department of Science

and Technology, Government of Gujarat (No.

GSBTM/MD/PROJECTS/SSA/1535/2013-14).

We gratefully acknowledge Dr. T. Ramamurthy, Dr.

S. K. Niyogi and Dr. Amit Ghosh, National Institute

of Cholera and Enteric Diseases, Kolkata, for

providing us the Vibrio and Shigella strains and for

their constant support and encouragement.

Thanks are due to Prof. V. K. Chaudhary,

University of Delhi South Campus, for his

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continued support for DNA sequencing.

3.1.2 Principal Investigator:

Dr. Reena A. Rajput (Assistant Professor)

Research Fellows:

Mr. Sagar Gaikwad (DST-JRF)

Ms. Dipeeka Mandaliya (DBT programme

support-JRF)

Ms. Kshama Jain (CSIR-JRF)

Mr. Manthan Patel (GSBTM-JRF)

Mr. Divyesh Patel (DBT-JRF)

Ms. Farha Memon (DBT-JRF)

Mr. Omkar Naik (DBT-JRF,

Left in July 2013)

Mr. Ranjeet (Lab. Attendant)

Trainees:

Ms. Krumali Chokshi (Trainee)

Ms. Nilam Gori (Trainee)

Ms. Ekta Patel (Trainee)

3.1.2.1. Description of research work

Despite of extensive research, neurodegenerative

disorders account for a major toll to human health

and cannot be fully treated using conventional

approaches. Many neurodegenerative diseases

occur as a result of neuroinflammatory processes.

Central nervous system (CNS) has little capacity

for self-repair after the loss of cellular elements

hence protection and regeneration of neurons is

palliative. In that context, we are studying TLR4

mediated neuroinflammation and possible rescue

mechanism. Further we are extending our study to

regenerate neurons from cancer stem cells and

immune cells.

Pattern recognition receptors (PRRs) are involved

in the recognition of pathogen-associated

molecular patterns (PAMPs) and danger-

associated molecular patterns (DAMPs). These

PRRs, on the innate immune cells, are the

molecules responsible for mounting inflammatory

responses leading to a novel avenue of

investigation aimed at developing a new

generation of immunotherapy. However,

persistent inflammation induced, as a result of PRR

activation, may pose deleterious consequences to

the host as well which has not been examined in

details. We are studying the molecular mechanism

of TLR5 mediated cholera vaccine reactogenicity

and also trying to delineate the active players

involved in dectin activation during fungal

infection. The major focus of the group is to explore

novel strategies that will reverse the extent and

severity of PRR activation and activation of

appropriate T-cell subset that would dictate better

immune outcomes.

Project 1: Neuroprotective effects of Toll like

receptor 4 (TLR-4) antagonists and/ or signaling

inhibitors on LPS induced neuronal insults.

Microglia cells are the resident macrophages of the

nervous system with pivotal role in innate immune

regulation and neuronal homeostasis. Prolonged

activation of microglia can cause the chronic

neuroinflammation and promote the neuronal

injury due to increase in the production of

neurotoxic pro- inf lammatory mediators .

Neuroinflammation is an important defence

mechanism against infectious agents and neuronal

injuries in the central nervous system. Chronic neu-

roinflammation may result in the neuronal damage

observed in many neurodegenerative disorders,

such as Alzheimer's, Parkinson's and Huntington's

diseases. Several evidences suggest that toll-like

r e c e p t o r 4 ( T L R 4 ) p l a y k e y r o l e i n

neuroinflammation by microglial activation and

cytokines production, a major hallmark of

neurodegeneration. Here, our study focuses on

achieving neuroprotection by targeting TLR4

mediated neuronal injury.

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In the present study, we investigated the effects of

RS-LPS, a TLR4 antagonist, and MAPK inhibitors

on LPS induced inflammatory responses in BV2

microglial cells. RS-LPS and MAPK inhibitors

decreased lipopolysaccharide (LPS)-induced

TLR4 expression. We also found that RS-LPS and

MAPK inhibitors reduced the expression of NF-κB

target genes, such as those for COX-2, iNOS, TNF-

α and IL-1β in LPS-stimulated BV2 microglial cells

(Figure 3.1.2.1). RS-LPS inhibited LPS-induced

phosphorylation of p65/RelA, resulting in

inhibition of the nuclear translocation and

transacting activity of NF- κB in BV2 microglial

cells. Interestingly, RS-LPS blocked LPS-induced

p38 MAPK, JNK phosphorylation but not ERK 1/ 2

and inhibition of JNK/p38 MAPKs abrogated LPS-

induced p65/NF-kB activation (Figure 3.1.2.2).

Figure 3.1.2.1 RS-LPS abrogated LPS induced neuroinflammation in BV-2

microglial cell

Figure 3.1.2.2. Rs-LPS blocks LPS induced MAPK and NF-κB pathways

Collectively, our results suggest that TLR4 plays

k e y r o l e i n m i c r o g l i a a c t i v a t i o n ,

neuroinflammation and neurotoxicity. RS-LPS a

TLR4 antagonis t prevents LPS induced

neuroinflammation. Moreover, RS-LPS prevent

microglial cell death by inhibiting the JNK/ p38

MAPK and NF-κB signaling pathways. We are

further trying to gain insight into the TLR4

induced phagocytic loss of neurons and its possible

rescue with TLR antagonist and/or MAPK

inhibitors. We also plan to explore whether

microglia mediated neurotoxicity can be abrogated

using TLR4 antagonism and MAPK inhibitors in

microglia-neuron coculture system. The in vivo

study needs to be carried out to confirm the

potential of RS-LPS and MAPK inhibitors in

neuroprotection.

Project 2: Role of Berberine in inhibition of cancer

cell stemness and possible potential in transforming

cancer Stem Cells to neurons.

Neuroblastoma is an aggressive childhood cancer

and current therapies remain unsuccessful as most

of neuroblastoma originates from cancer stem cells

(cSCs), which establish a tumor through

continuous self-renewal and proliferation. The

successful elimination of cSCs and generation of

new functional neurons can serve as an effective

strategy to achieve complete remission for

neuroblastoma. Reports from the 1980s have

shown that neuroblastoma cell lines can often be

induced to terminally differentiate on exposure to

retinoid compounds. In this context we are

exploiting a natural product, Berberine, for its role

in inhibiting cancer cell stemness and inducing

neurodifferentation.

The potential of berberine in inducing cellular

differentiation was first observed in the form of

gradually increased neurite outgrowth and

branching in berberine treated Neuro 2a cells.

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Berberine induced Neuro 2a differentiation was

further supported by increased expression of

Map2, β-III Tubulin and NCAM (neural cell

adhesion molecule). Increase in the expression of

differentiation marker Hsp -70, which is a

chaperone that supports maturation of neuronal

cells, was observed in berberine treated cells. The

treatment also leads to induction of neural cell

adhesion molecule (NCAM) expression and

reduction in its polysialylation, thus elucidating

its anti migratory potential, which is also

supported by down regulation of MMP-2 and 9

activity (Figure 3.1.2.3).

Figure 3.1.2.3.: Berberine induces differentiation of Neuro 2a cells

Berberine significantly decreased the expression of

cyclin D, cyclin E cdk2, and cdk4 (cyclin

dependent kinases). The expression of two

important CKIs (cdk inhibitors) p21 and p27 was

found to be up regulated by berberine in a dose

dependent manner. Similarly p53 expression was

also increased while the decrease in expression of

Top 2 (topoisomnerase 2) was observed. To

investigate the role of berberine in apoptosis, we

studied the expression of pro-apoptotic and anti-

apoptotic genes at transcriptional level .

Expression of proapoptotic genes, bak and bax

increased while the expression of anti apoptotic

genes, bcl2 and bcl xl was decreased in a dose

dependent manner. Further the data was

supported by the results of flow cytometry where,

the percentage of cells arrested at G0/G1 phase

increased to 63.95% cells compared to the

untreated group where 19.55 % of cells were in

G0/G1 phase. (Fig 3.1.2.4).

Figure 3.1.2.4.: Berberine induces G0/G1 cell cycle arrest and

apoptosis in Neuro 2a cells

To check the role of berberine in reducing stemness

in cancer cells, expression of different stem cell

markers was studied at transcriptional level. Results

indicate decrease in the expression of N-myc,

which is otherwise found to be over expressed in

cancer cells. Berberine reduced sphere formation

with significant suppression of CD133(+) nestin(+)

cell population and also suppressed expression of

the stemness-regulating transcription factors sox2,

Notch-2, and β-catenin in Neuro 2a cells.

PI3/AKT and MAPK pathways are known to play

pivotal role in cell survival, proliferation and

migration. Berberine treatment significantly drops

the expression of phosphorylated form of PI3

Kinase (pPI3K) and AKT (pAKT) in a dose

dependent manner. Similarly, expression of

phosphorylated ERK and its upstream regulators

Ras1 and Raf1 was also found to be decreased

following Berberine treatment. These results

suggest that berberine prevents activation of

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PI3/AKT and MAPK pathways suggesting

apoptosis induction and reduction in the cancer

cell stemness (Figure 3.1.2.5).

Figure 3.1.2.5.: Fig. Berberine reduces stemness in Neuro2a cells.

Collectively, our results suggest that berberine

significantly suppressed two fundamental

characteristics of cancer stem cells: self renewal

and tumorigenicity in neuro2a cells. We further

plan to isolate cCS population from Neuro 2A cells.

The isolated cells would then be used for

generating neurons and the generated neurons

would then be characterized phenotypically and

functionally.

Project 3: To study cholera toxin and flagellin

induced TLR5 mediated immune response in

HT29 human intestinal epithelial cell lines.

We are trying to delineate the mechanism behind

cholera vaccine reactogenicity which has

previously been attributed to TLR5 mediated MAP

Kinase and NFκB activation. These findings

provide further insight into the interaction of

reactogenic V. cholerae vaccine candidates with

host cells. Understanding the mechanisms

involved in the interaction between V. cholerae

and host cells should facilitate the development of

improved attenuated V. cholerae vaccines. We

plan to check the interaction between GM1

receptor and TLR5 since both of these are

activated by components of Cholera toxin. The

findings would also potentiate cholera toxin

mediated immunomodulation and adjuvancy and

can be exploited to other models of inflammation.

Expression pattern of TLR5 on mouse macrophage

cell line (RAW 264.7) was checked upon activation

with Flagellin (100ng/ml) and CTB (100ng/ml and

500ng/ml). The expression study of TLR5 was

carried out by semi-quantitative Reverse

Transcriptase - Polymerase Chain Reaction (RT-

PCR) and immunofluorescence (Figure 3.1.2.6).

Figure 3.1.2.6.: Cholera toxin modulates Flagellin induced TLR5 expression

We have tried to decipher the immuno-deviating

mechanism of cholera toxin at various doses. It was

interesting to observe that the utilization of TLR5

by Cholera toxin is dependent on the dose of toxin.

At lower doses CTB ligates TLR5 activating pro-

inflammatory cytokines whereas at higher doses

the involvement of TLR5 was not prominent and it

led to decrease in pro-inflammatory cytokines

(Figure 3.1.2.7.).

Figure 3.1.2.7.: CTB and Flagellin induced cytokines in HT29 cells.

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The role of CTB in differential T-cell subset

activation is under active investigation. We want to

extend our study further towards looking at TLR5

and GM-1 interaction and the possible role of CTB

in reducing the reactogenicity. Once we establish

the role of GM1 as co-receptor we further want to

look at different T cell subsets generated when

CTB is used along with Flagellin during TLR5

activation. Future experiments will also focus on

inducing regulatory or tolerogenic T-cell

population that would further be isolated and used

for re-challenge experiments.

Project 4 : Diversifying the dectins and

inflammasomes in Aspergillus fumigatus specific

immune response.

Aspergillus fumigatus can cause not only invasive

infection but also triggers allergic aspergillosis. The

pattern associated molecular patterns for most of

the fungi are β- glucans that are recognized by

many of the pattern recognition receptors

inc lud ing dec t in s . The s tud ie s a im a t

understanding host immune response against A.

fumigatus with respect to its recognition by dectins

and fu r the r downs t ream ac t i va t ion o f

inflammasome. The manipulation of these signals

may allow the host to overcome infection

mediated by appropriate activation of the T cell

subsets.

Marked change in Dectin 1 and 2 expression was

observed upon stimulation with β-glucans

(0.5μg/ml).While the expression pattern of TLR2

was not significantly increased with β-glucans as

with LPS ruling out the possibility of TLR2

involvement.

Figure 3.1. 2.8.: β- glucan Modulates dectin expression and

inflammatory response

The study shows that β-Glucan induces

proinflammatory cytokines like TNF-α and IL-1β in

a dose dependent manner and inhibits anti-

inflammatory cytokines IL-10 (Figure 3.1.2.8).

Figure 3.1.2.9: β- glucan Modulates dectin expression and inflammatory response

To assess the role of NFκB dependence during

inflammasome activation, inhibitors targeting the

signaling pathways were analyzed. Cells were pre-

treated with inhibitors and their effect on dectin

expression and phosphorylation was observed.

Dectin expression as well as the key signalling

molecules are modulated differentially when the

pathways are inhibited selectively (Figure 3.1.2.9).

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Figure 3.1.2.10: Modulation of dectins and signaling cascades during interference with various signaling and oxidative pathways inhibition

To summarize, β-glucan regulates dectin

expression as well as the key signalling molecules.

Selective inhibition of signalling cascade results in

differential modulation of dectin expression and

phosphorylation yielding a possibility of their

potential role regulating further inflammation

which is yet to be explored. The concept would be

extended to orchestrate the immune response by

differential T cell activation. We plan to fine-tune

the Th17/Treg population to achieve better

protection in models of asthama and allergy.

3.2 Genetics & Developmental Biology

Principal Investigator:

Dr. Anand Kumar Tiwari, Assistant Professor

Research Fellow:

Mr. Ajay Kumar (DST, JRF)

Ms. Pearl Christian (DBT PS, JRF)

Mr. Bhavin Uttekar (GSBTM, JRF)

Mr. Ranjeet (Lab attendant)

Research Interest:

1. Study of the role of molecular chaperone during

eye development in Drosophila melanogaster.

2. Study of the role of molecular chaperone and

ubiquitin ligases in the progression of Alzheimer's

disease using Drosophila melanogaster as a model

organism.

3. Identification of medicinal property of selected

Indian plants (Catharanthus roseus L. & Triticum

aestivum L.) and cyniobacteria (Arthrospira

platensis) using Drosophila melanogaster as an

alternate animal model.

A summary of the progress made under above

mentioned projects are as follows:

Project 1: Study of the role of molecular chaperone

dur ing eye deve lopment in Drosophi la

melanogaster.

Progress made: The following results were

obtained from the present study:

1) The cell death shown by Hsp70/Hsc70

m u t a n t s w a s s i g n i f i c a n t l y re s c u e d b y

overexpression of Drosophila inhibitor of apoptosis

(DIAP1) and baculovirus anti-apoptotic gene UAS-

p35.

In our previous report we have shown that

mutation in Hsp70/Hsc70 results in degenerated

eye phenotype, thus in order to study whether the

degenera ted eye pheno type shown by

Hsp70/Hsc70 mutants was due caspase

dependent cell death, genetic interaction study was

performed between Drosophila inhibitor of

apoptosis protein 1 (DIAP1) and baculovirus anti-

apoptotic gene UAS-p35. A, significant rescue was

observed in eye size of UAS-Hsp70DN-GMR-

GAL4/DIAP1 (Figure 3.2.1.E & F) and UAS-

Hsc70DN-GMR-GAL4/UAS-p35 (Figure 3.2.1.G

& H), suggested that mutation in Hsp70/Hsc70

induces caspase dependent cell death. The eye

phenotype was not completely rescued because

over-expression of DIAP1 & p35 cannot restore the

normal Hsp function in eye cells.

Figure 3.2.1.: Genetic interaction between Hsp70/Hsc70 mutants and DIAP1 & baculovirus inhibitor UAS-p35. (A-B) Oregon R+ (A) and GMR-GAL4 (B) control flies showing normal ommatidial arrangement in eye. (C-D) UAS-HSP70DN-GMR-GAL4 (C) & UAS-HSC70DN-GMR-GAL4 (D) flies showing reduced eye and degenerated eye phenotype. (E-H) UAS-HSP70DN-GMR-GAL4 and UAS-HSC70DN-GMR-GAL4 flies showing in DIAP1 (E & F) and UAS-p35 (G & H) background showing a significant rescue in eye size and degenerated eye phenotype.

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1) Mutation in Hsp70/Hsc70 results in

activation of JNK signalling in Drosophila eye cells

Since, JNK signalling is a key signalling pathway,

participate in cellular stress and apoptosis, genetic

interaction study was performed to study the

status of JNK signaling in Hsp70/Hsc70 mutant

eyes. For this, pucE69 (a negative regulator of JNK

signalling), UAS-eiger (Activator of JNK signaling)

& UAS-bskDN (Dominant negative mutation of

Drosophila JNK signalling) candidate genes of

JNK signalling was chosen and genetic study was

performed. When JNK activity was inhabited in

eye in UAS-Hsp70DN-GMR-GAL4/UAS-bskDN

flies a significant rescue in eye size was observed

(Figure 3.2.2.H) while pupal lethality was

observed when JNK was overexpressed in UAS-

Hsp70DN-GMR-GAL4/UAS-eiger flies (Figure

3.2.2.I). These results, suggest that mutation in

Hsp70/Hsc70 results in activation of JNK

signalling in Drosophila eye.

Figure 3.2.2.: Genetic interaction study of Hsp70/Hsc70 mutants with candidate genes of JNK signalling pathway. (A-B) Oregon R+ (A) and GMR-GAL4 (B) control flies showing normal ommatidial arrangement in eye. (C) UAS-Hsp70DN-GMR-GAL4 flies showing reduced eye and degenerated eye phenotype. (D-F) pucE69/GMR-GAL4 (D), UAS-bskDN/GMR-GAL4 (E) and UAS-eiger-GMR-GAL4 (F) control flies. (G-I) Eye phenotype of UAS-Hsp70DN-GMR-GAL4 flies in pucE69 (G), UAS-bskDN (H) & UAS-eiger (I) background, showing rescue in reduced eye size phenotype, when JNK was down regulated using UAS-bskDN a severe pupal lethal phenotype when JNK was overexpressed (I).

Project 2: Study of the role of molecular chaperone

and ubiquitin ligases in the progression of

Alzheimer's disease using Drosophila melanogaster

as a model organism.

Progress made: The following results were

obtained from the present study

1. Inhibition of Appl gene/overexpression of

Aβ42 results in induction of cell death in fly brain:

Since, neuronal cell death is a key feature in

neurodegenerative diseases thus, to observe

whether the inhibition of Appl gene results in

induction of cell death, acridine orange staining

was performed in 40 days old adult brain from

Oregon R+ (Control), UAS-ApplRNAi>GMR-

GAL4 and GMRAβ42K52; GMRAβ42K53 flies. A

significant increase in dead cell population was

observed in neuropil region of Alzheimeric fly brain

(Figure 3.2.3.B & C) as compared to the control

flies (Oregon R+) (Figure 3.2.3.A). This result

suggested that cell death in Alzheimeric fly brain is

an additive consequence in alzheimeric conditions

in flies

Figure 3.2.3.: Acridine orange (AO) staining the 40 days old adult fly brain: (A-C) AO staining in 40 days old adult fly brain showing developemtal apoptosis in Oregon R+ (A) and excessive cell death in GMR-GAL4>UAS-ApplRNAi (B) & GMRAβ42K52; GMRAβ42K53 brain.

Since, death of neurons is a key feature of

neurodegenerative disease, thus acridine orange

(AO) staining was performed in 30 & 40 days old

adult fly brain to observe the neuronal death. A

significant induction in dead cell population was

observed in adult brain as compared to the control

group.

Alzheimeric flies show plaque formation in the

adult fly brain:

Thioflavin S (TS) staining was performed in 50

days old adult brain of control and AD mutant flies.

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I t was observed that GMR-GAL4>UAS-

ApplRNAi flies show plaque formation in the

Kenyon cell region of the brain (Figure 3.2.4B)

while it was absent in the control brain (Figure

3.2.4.A).

Figure 3.2.4.: Thioflavin S (TS) staining in 50 days old adult fly brain. (A) TS staining in control files with no plaque in the brain. (B) TS staining in GMR-GAL4>UAS-ApplRNAi fly brain showing plaque in the Kenyon cell region of the brain.

Project 3: Identification of medicinal property of

selected Indian plants (Catharanthus roseus L. &

Tri t icum aest ivum L.) us ing Drosophi la

melanogaster as an alternate animal model.

Acknowledgements:

We are thankful to Prof. J. K. Roy, Dr. S.

Srikrishna, Banaras Hindu University, Varanasi,

Prof. K. Vijay Raghavan & Dr. Roshan F. Begum,

NCBS, Bangalore for fly stocks. The work was

suppor ted by funds from Gujarat State

Biotechnology Mission (GSBTM), DST, DBT,

Government of India & Puri Foundation for

Education in India.

3.3. Department of Cell Biology


Dr. Chandramani Pathak (Assistant Professor)

Research Students:

Mr. Kishu Ranjan- (ICMR-SRF)

Mr. Bhargav N. Waghela (JRF – DBT-NNT

Project)

Ms Anupama Sharma- (CSIR-SRF)

Ms Suhashini Dhumale- (JRF - DST SERB

Project)

Ms. Kavita Shirsath(JRF – DBT-NNT Project)

Ms. Rinky Prakara-(JRF - DBT RGYI Project)

Ms. Shubita Tripathi - (JRF - DBT RGYI Project)

Our major focus is to understand the cell death and

inflammatory signaling pathways activated during

physiological and pathological conditions.

Programmed cell death (PCD) is a physiological

process that is responsible for removal of unwanted

cells during embryonic development, tissue

homeostasis; immune cell maturation and

e l iminat ion of pathogen in fec ted ce l l s .

Dysregulation of PCD leads to many diseases like

neurodegeneration, cancer, inflammatory and

metablic disorders.

Apoptosis is one of the most important mechanism

of cell death. The cells of multi-cellular organisms

have the inherent capacity to undergo death by a

highly organized manner mechanism known as

programmed cell death or apoptosis. This highly

regulated cellular process is utilized during

embryonic development or upon tissue injury or

disturbance of tissue homeostasis. Thus, apoptosis

is a key regulatory mechanism for regulating

various physiological events including eliminating

the unwanted cells and defense against infections

and maintaining the homeostasis of normal

tissues. Alteration in apoptosis not only contributes

to the promotion of malignancy but can also

enhance drug resistance in response to anti-cancer

therapies. Therefore, regulation of apoptosis

during pathological conditions is important for

therapeutic intervention.

Apoptosis and inflammation are two closely related

processess o f mul t i -ce l lu lar organism.

Inflammation is one of the spontaneous cellular

responses which may have dual effects. Under

some conditions it may activate host immune

defenses, while under others in rest, it may induce

opposite effects. Acute inflammation is the innate

immune response that leads to adaptive immunity;

but when it becomes chronic it increases the risk to

develop several diseases including cancer,

cardiovascular diseases, diabetes and neurological

disorder. Several lines of the evidences support the

assumption that inflammation plays an important

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role in the progression of malignancy by providing

tumor micro environment. Thus, elevation of

inflammatory mediators and inhibition of

apoptosis contributes cell proliferation and

survival in cancer. Therefore, regulation of

apoptosis and inflammatory mediators might be

important for critical targets in both prevention

and therapy. Towards this aim we are focusing our

research on regulation of apoptosis and

inflammation in cancer.

The major objectives of our research group are:

1. Find out a new molecular target and explore

cellular mechanism which can control

activation of inflammatory mediators and cell

proliferation in cancer.

2. Induction of apoptosis in cancer cells by

modulating the apoptotic signaling.

3. Apoptotic potential of nanoparticulate anti-

cancerous molecules.

3.3.1 Crosstalk of FADD and cFLIP in regulation of

death receptor mediated apoptosis

Fas-associated death domain protein (FADD) is an

adaptor protein molecule which play a crucial role

in transducing the apoptotic signals. FADD

provides docking site for hemophilic interaction,

oligomerization and autocatalytic processing for

activation or regulation of downstream apoptotic

signaling. The Death Domain (DD) of FADD

interacts with DD of the death receptors and Death

Effector Domain (DED) allows recruitment of

DEDs containing carrying proteins like pro-

caspase-8/ 10, which in turn initiates the formation

of a death inducing signaling complex (DISC) to

further progression of death receptor signaling for

apoptosis. The death receptor mediated apoptosis

is effectively regulated by anti-apoptotic protein

cFLIP, which is structurally similar to procaspase-8

and -10 but lacks cysteine residue for autocatalytic

activity. Thus, FADD and cFLIP both are

important component for cell death and survival.

Dysregulated expression of FADD and cFLIP is

associated with inhibition of apoptosis and other

signaling for cell death which leads to progression

of malignancy (Figure.3.3.1). Moreover, elevated

intracellular level of cFLIP competitively excludes

the binding of procaspase-8 to the death effector

domain of FADD at the DISC, thus blocking the

activation of death receptor signaling of apoptosis.

In the present study we have explored the crosstalk

of FADD and cFLIP in regulation of death receptor

mediated apoptosis (Figure.3.3.2). The aim of

present study was investigating the cross talk

between cFLIP and FADD for regulation of cell

death and survival. Our findings reveal that low

endogenous level of FADD doesn't provide a

sufficient binding platform to caspase-8/10 at the

DISC. Interestingly, overexpression of FADD in

HEK293T and MCF-7 cells down regulates

express ion of cFLIP.Moreover, se lec t ive

knockdown of cFLIP using siRNA promotes cell

death. Further, knockdown of cFLIP during FADD

over expressed conditions showed rapid loss of

mitochondrial integrity with simultaneous release

of cytochrome c and activation caspase cascade for

apoptosis along with further inactivation of PARP

through its cleavage. Collectively, our data suggest

that aberrant expression of cFLIP and FADD

promote cell survival and inhibit apoptotic cell

death. Taken together, these results suggest that

gaining insights into the regulatory mechanisms of

FADD and cFLIP in death receptor mediated cell

death might open up a way towards a novel

approach for therapeutic intervention of cancer.

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Figure.3.3.1.: Role of FADD in cell proliferation

Figure 3.3.2.: Involvement of FADD and cFLIP in regulation of death receptor mediated apoptosis

3.3.2 Apoptotic potential of nanoparticulate anti-

cancerous molecules.

Recent advances in Nanotechnology have paved

way for therapeutic potential using different

strategies and pharmacological manipulation.

More importantly, recent reports highlight that

nanoparticulate drugs may contribute as a novel

target for drug delivery for treatment of cancer.

Nanoparticle based therapy might be able to

improve the therapeutic index of anti-cancer

drugs for treatment of cancer. We also focusing

our research on nano-particle based drug delivery

and evaluating their apoptotic potential in human

cancer cells using dendrimers as a delivery tools.

Dendrimers are nano-sized, radially symmetric,

well-defined, homogeneous and monodisperse

consisting of tree-like branched structure. These

highly branched three-dimensional structures

provide a high degree of surface functionality and

versatility for improved drug delivery . Dendrimes

have been reported to have functional diversity but

its inherent toxicity limits its application for

therapeutic intervetion. As the free functional

groups on the surface contribute to its toxicity, the

surface engineering of dendrimers can lead to its

improved properties, especially in the context of

biomedical applications.

3.4. Plant Developmental Biology


Dr. Sunil Kumar Singh ( Assistant professor)

Group Members:

Suresh Varsani (DBT-JRF)

Manoj Kumar Patel ( DBT-JRF)

Manisha Farsodia (Inspire DST-JRF)

Misheal Zimik ( M.Sc. Project Student)

Sweety Shah (M.Sc. Project Student)

Shamal Desai (Lab Assistant)

3.4.1 Understanding the role of genes involved in

programmed cell death in plants:

PCD is genetically controlled physiological process

that plays an important role during plant

development and also during stress responses. We

aim to identify genetic regulators that are involved

in developmental as well as stress induced cell

death. We are using Arabidopsis thaliana and

Solanum lycopersicum as model plants to

understand the role of candidate metacaspases. In

addition to these, we are also characterizing the

role of a Ubiquitin-Conjugating Enzyme and a

MYB transcription factor that are coexpressed with

one of the candidate metacaspase in Arabidopsis

thaliana.

Arabidopsis thaliana Metacaspases:

Our main interest in Arabidopsis metacaspases is to

understand their roles in PCD. Since metacaspases

are distantly related to animal caspases, they can

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be in teres t ing candidates that regula te

developmental or stress induced cell death in

plants. We are using molecular genomics based

approaches to characterize the role of selected

Arabidopsis metacaspases (AtMCs).

For silencing of Arabidopsis AtMC3, artificial

microRNAs (amiRNA) expressing homozygous T2

lines were generated. In order to know the

silencing of AtMC3 gene transcript in amiRNA

expressing lines, Quantitative RT-PCR analysis on

mRNA isolated from 6 days old seedlings and

flower tissues of 35 days old mature plants of

AtMC3 amiRNA expressing T2 lines was

performed. qPCR results were normalized to the

expression of reference genes, UBQ5 and APT1.

Results showed that AtMC3 gene transcript was

strongly suppressed in AtMC3 amiRNA lines as

compared to Col-0. Selected AtMC3 amiRNA

expressing T2 line was grown on MS plates and in

pots for phenotypic analysis.

AtMC3 overexpressing homozygous T2 lines were

generated for overexpression of AtMC3.

Quantitative RT-PCR analysis on 6 days old

seedlings and flowers of 35 days old mature plants

of AtMC3 expressing T2 lines was performed to

confirm the level of AtMC3 overexpression. Three

lines which showed very high level of AtMC3

overexpression were selected for detail phenotypic

analysis.

To quantify mRNA expression of Arabidopsis

Metacaspases, 6 days old seedlings and different

vegetative and reproductive tissues isolated from

35 day old pot grown plants were used. Expression

results were normalized to the expression of

selected reference genes, UBQ5 and APT1.

Expression values were calculated using the ∆∆Ct

method. Results showed that AtMC1 and AtMC4

were expressed constitutively in different

vegetative and reproductive tissues of wild type

plants. Expression of AtMC2 was stronger in

rosette leaves. While, AtMC9 expression was

stronger in flower buds, open flowers and siliques.

AtMC3 was expressed strongly in rosette leaves,

seedlings, flowers and siliques.

We have also generated several AtMC1

p r o m : : G U S , A t M C 2 p r o m : : G U S , a n d

AtMC3prom::GUS lines to analyse detail cell and

tissues specific expression pattern. Promoter

expression analysis showed that AtMC1 was

expressed in the cotyledons, leaves, hypocotyl of 6

days old seedlings. Strong AtMC2 expression was

observed in the cotyledons, leaves, hypocotyl and

vascular tissue of primary root. AtMC2 expression

was also observed in differentiating stomata cells of

cotyledons and leaves. AtMC3 was expressed very

specifically in vasculature of cotyledons, leaves,

hypocotyl and roots.

(ii) Solanum lycopersicum Metacaspases:

Tomato genome encodes eight metacaspases, six

of them are type I and rest two are type II

Metacaspases, which is unlike Arabidopsis which

encode three type I and six type II metacaspases.

For silencing of tomato Metacaspases, artificial

microRNAs (amiRNA) specific to SolycMC4 and

SolycMC9 were designed and cloned in vector

pMDC32. We have also made overexpresser

constructs for both SolycMC4 and SolycMC9 in

tomato. We are transforming these genes

constructs in tomato to understand the effects of

silencing and over expression.

(iii) Understanding the role of Ubiquitin-

Conjugating Enzyme/ Ubiquitin Ligase (AtUBCx)

and MYBx transcriptional factor in Arabidopsis.

We are also characterizing a Ubiquitin-Conjugating

Enzyme and a AtMYBx transcriptional factor which

are shown to be co-expressed with one

Metacaspase in reproductive and vascular tissues

of Arabidopsis.

To characterize the function of AtUBCx, promoter

and gene specific primers were designed and

cloned into vectors pMDC32 and pMDC107/

pMDC163, respectively, for gene overexpression

and promoter expression analysis. Artificial

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microRNA (amiRNA) for silencing of this gene was

also designed. Designed oligos was used to

amplifiy amiRNA through overlapping PCR using

miRNA precursor miR319a and recombined in to

express ion vector pMDC32.These gene

constructs, i.e, over-expressor, amiRNA and two

promoter constructs, were transformed in

Agrobacterium tumafecians and subsequently

transformed in Arabidopsis Col-0. Gene

overexpresser and UBCxprom::GFP and

UBCxprom::GUS transgenic lines selected on

hygromycin and T1 and T2 lines were generated.

Expression analysis using semi-quantitative PCR

method shows that this gene was expressed in

most of the tissues and organs analysed so far with

sl ightly higher expression in leaves and

reproductive organs. Prom::GFP expression of

young seedlings was also analysed using confocal

microscope. GFP expression was observed mainly

in vascular tissues of primary root and in the

trichome of the leaves. On the basis of both

expression studies, we show that UBCx is

expressed in most of the tissues and organs of the

plants with slightly higher expression in the

vascular tissues of root, leaf, leaf trichomes and

reproductive organs.

Global expression data shows that selected MYBx

transcriptional factor was expressed in the

vasculature of Arabidopsis plant. To characterize

the function of this gene, its promoter and coding

sequences were PCR amplified and cloned in to

Promoter expression and gene over expression

vectors and transformed into Agrobacterium

tumefaciens strain. This Agrobacterium cells were

subsequently used to genetically transformed Col-

0 plants by floral dip method. The putatively

transformed seeds were harvested and screened

on Hygromycin and T1 lines were isolated. We are

fur ther screening these l ines to isolate

homozygous T2 promoter::GUS, promoter::GFP

and over expresser lines for detailed functional

analysis.

3.4.2 The role of Tetraamine Synthases during

plant development and stress responses in tomato:

Ro le o f te t raamines syn thases such as

thermospermine synthase (SolycTSPMS) and

spermine synthase (SolycSPMS), are not well

characterized specifically during reproductive

development of plants. Candidate tetraamine

biosynthetic genes were indentified from tomato

genome and will be silenced and over expressed in

tomato to understand their roles during plant

development and stress responses.

Expression pattern of these genes in different

reproductive tissues of tomato plant were analysed

using quantitative RT-PCR. We examined that

SolycSPMS was consistently expressed throughout

all the flower developmental stages, while

SolycTSPMS was expression was relatively high in

flower but than the other developmental stages

which indicate that thermospermine has some

important role during early flower development.

During tomato fruit development SolycSPMS

expression was quite higher in immature and

young fruits and expression decreases gradulaly

down to very low in fully grown mature

fruit.SolycTSPMS expression was varying

throughout all the developmental stages of fruit.

To understand the role of these tetraamines during

reproductive development of tomato we have

initiated work to generate gene silencing and

overexpressor lines. SolycSPMS and SolycTSPMS

specific amiRNA and overexpressor constructs

were made. Once these lines are generated we will

explore the effect of gene silencing and gene

overexpression in tomato.

3.5 Bioinformatics & Structural Biology :


Dr. Desh Deepak Singh (Associate Professor,

Group head)

Dr. Anju Pappachan (Assistant Professor)

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Research Fellows :

Mr. Radhey Shyam Kaushal (CSIR -SRF)

Mr. Manoj Kumar, (DBT Project SRF)

Mr. Kunal Shah (PhD Student)

Mr. Dhaval Patel, (Technical Assistant)

Ms. Bhumi Patel, (DST Project JRF)

Ms. Jhanvi Jani, (Project Assistant)

Mr. Bhavesh Darji, (Lab Assistant)

3.5.1. Characterization of Leishmanial surface

proteins, lectins adhesion proteins from

Lactobacillus spp. and developing bioinformatics

tools .

Project 1 – Characterization of Surface proteins

from Leishmania donovani

C-type lectin Protein (CTL):

Leishmaniasis is one of the most significant

neglected tropical disease, with 350 million people

in 88 countries worldwide living at risk with few

therapeutic options. Surface virulence factors on

Leishmania parasite are important for the host-

pathogen interaction. Lectins are ubiquitous in

nature and, thus, the increasing number of

protozoan genera in which these carbohydrate-

binding proteins have been reported is not

s u rp r i s i ng . I n t he genus Le i shman ia ,

carbohydrate-binding proteins were first reported

to play a role in Leishmania – macrophage

interaction . LmjF04.0430 is a 4123 amino acid

(a. a) long hypothetical protein having a C-type

lectin domain in the region 443–585. Multiple

alignment with hits from FUGUE (1MSB:A &

1H8U:A) and model generated with 1H8U:A as

template show a good overlap of residues and

topology of C-type lectin domain as shown in

figure 3.5.1.1

Figure 3.5.1.1 : Multiple alignment and model of putative C-type lectin region of LmjF04.0430

Cloning , Protein expression and purification

strategies for CTL:

We have cloned this C-type lectin domain as

shown in figure 3.5.1.2 of LmjF04.0430 gene in

pET28a vector and sequencing result shows more

than 99% match with LmjF04.0430 (geneDB).

Over expression was achieved in insoluble form in

Rossetta (DE3) strain with 1mM IPTG as inducing

agent and SDS-PAGE analysis shows a band

corresponding to 26kDa, the expected molecular

weight of the protein. Even after extensive

optimization through IPTG & temperature,

presence of protein was seen in insoluble inclusion

bodies only. Protein could be successfully

solubilized by on-column refolding using different

buffers as shown in figure 3.5.1.3 . Bioinformatics

analysis and modeling using I-TASSER program

confirms the loop-helix-sheet structure, which is

reported for c-type lectins.

PCR amplification & clone confirmation

Figure 3.5.1. 2 : PCR amplification & restriction digestion of C-type lectin clone

Expression & refolding studies:

Figure 3.5.1.3 : Expression & refolding of CTL protein

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Proteophosphoglycan 3 (PPG3):

Proteophosphoglycans (PPGs) are mucin-like

glycoproteins present on the surface and secreted

by both promastigote and amastigote stages of

Leishmania and play impor tant role in

transmission, invasion and intracellular survival of

the parasites (Ilg et al. 1999). PPGs were reported

to share amino acid sequence similarity (restricted

to a leucine rich repeat (LRR) motif) with members

of the parasite surface antigen -2 (PSA-2) family of

glycoproteins (Montgomery et al. 2000) .LRR

motifs have functions in protein-protein

interaction, signal transduction, as well as

pathogen recognition. Earlier bioinformatics

studies from our lab have identified LRR proteins

with putative adhesion like role (Singh & Singh,

2008). Thus, structural characterization of LRR

motif containing PPGs of Leishmania is

particularly important.

Cloning, protein expression and purification

strategies for PPG3:

N-terminal region of PPG3 (L. infantum) (530

residue long) showing maximum antigenic

properties was selected for cloning from L.

donovani genome for detailed structural

characterization. A 864 bp region of L. donovani

putative ppg3 was amplified, cloned in pGEX4T-2

vector and a 288 amino acid long partial protein

was expressed in Rosetta (DE3) in soluble form.

Protein was purified through GST binding affinity

column and identity of the expressed protein was

confirmed through MALDI-TOF analysis. In-silico

structural characterization of the expressed

putative PPG3 (partial) using PSIPRED and I-

TASSER was performed.

Partial N-terminal domain of L. donovani putative

PPG3 which was selected in the present study for

structural characterization was found to possess a

significant number of antigenic determinants and

could act as a potential prophylactic vaccine

against visceral leishmaniasis. In-silico analysis

using PSIPRED software predicted the presence of

16 β-sheets and 7α-helices in the partial PPG3

(putative) protein from L. donovani which was

expressed in E .coli as a GST-tagged recombinant

protein. The expressed partial PPG3 (putative)

protein showed 97% homology with L. infantum

PPG3 and conserved domain search revealed the

presence of PLN00113 domain which corresponds

to the Leucine rich repeat receptor like protein

kinase (provisional) from 54th to 177th residue.

(Fig 3.5.1.4)

Fig 3.5.1.4 : (a) Lane 1: 1kb ladder, lane 2: ppg3 amplicon. (b) Lane 1: 1 kb ladder, lane 2: Restriction digestion of positive recombinant clones showing vector and ppg3 insert. (c) Lane 1: Medium range protein marker, Lane 2: Purified L. donovani partial PPG3 (putative) protein with N-terminal GST tag.(d) MALDI-TOF analysis of the expressed and purified L. donovani PPG3 (putative) confirms its identity by showing 3 peptide match with L. infantum PPG3 and a highly significant MS/MS score of 222.

Project 2 - Isolation and purification of plant lectins

Beta Glucosidase Aggregating Factor (BGAF) from

Sorghum bicolor:

Beta Glucosidase plays a very important role in

plant defense by converting the sugar moieties into

secondary metabolites which go and fights with the

infection. These beta glucosidases are very

vulnerable to proteolytic degradation by proteases

released by infecting organisms. Beta Glucosidase

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Aggregative factors are a chimeric lectin. BGAF

contains N-terminal dirigent domain (disease

response gene) and C-terminal jacalin like lectin

domain. As name suggests they, aggregate beta

Glucosidase in plants and prevent them from

proteolytic degradation by proteases released by

infecting organisms. Lectin domain is involved

with binding with Beta Glucosidase while N-

terminal dirigent domain is responsible for the

dimerization of BGAF molecule, which is very

important step in aggregation of beta glucosidase.


strategies used for BGAF:

BGAF gene from Sorgum bicolor was synthesized

and was supplied in pUC57 vector containing the

R.E site of EcoRI and NotI. After initial digestion

the gene was ligated to pET30a and the positive

clones were confirmed through restriction

digestion, colony PCR, gel shifting and finally

through nucleotide sequencing to check proper

orientation and sequence similarity as shown in

figure 3.5.1.5 and figure 3.5.1.6 . Protein was

induced by using auto-induction media for O/N at O24 C. Induced supernatant containing the

recombinant protein was purified through Ni-NTA

column. All eluted fractions were analyzed through

15% SDS-PAGE as shown in figure 3.5.1.7 .

Purified protein was excised from SDS-PAGE and

the identity of the induced protein was confirmed

by MALDI-TOF analysis.

Figure 3.5.1.5 : Plasmid Shift of BGAF

Figure 3.5.1.6 : Clone confirmation by digestion

Figure 3.5.1.7 : Expression & Purification of BGAF

Project 3 – Adhesion proteins from Lactobacillus

spp.

Glyceraldehyde 3-phosphate dehydrogenase

(GAPDH) from Lactobacillus acidophilus:

GAPDH is intracellularly located enzyme, a

housekeeping enzyme essential for glycolysis,

which has also been identified on the outer surface

of several pathogens, including group A

streptococci Staphylococcus epidermis and

Staphylococcus aureus, also in pathogenic fungi

and parasites such as Candida albicans and

Schistosoma mansoni. Some pathogens possess a

GAPDH on their cell surface and they have been

identified as adhesins. GAPDH is also identified on

the outer surface of Lactobacillus crispatus and a

variety of other Lactobacillus strains. Cell surface

Lactobacillus plantarum LA 318 GAPDH is found

to adheres human colonic mucin. However, it is not

known why the GAPDH exists on the cell surface

without a conventional N-terminal signal peptide.

The secretion and anchoring mechanisms of

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GAPDH on the bacterial surface have not been

characterized. In case of Lactobacilli genus besides

its metabolic functions, GAPDH undergo

'moonlighting' when they are exposed on the

bacterial surface, developing additional functions.


strategies used for GAPDH – LBA0698:

L. acidophilus strain was procured from NDRI,

Karnal and the genomic DNA was isolated in

house using standard protocol and the strain was

confirmed as L. acidophilus La-14/NCFM through

16S rRNA sequencing. GAPDH gene from L.

acidophilus was amplified by PCR with set of

forward and reverse primer containing the

restriction sites site of Nde1 and BamH1. After

initial digestion the gene product was ligated to

pET28a and the positive clones were confirmed

through restriction digestion, colony PCR, gel

shifting and finally through nucleotide sequencing

to check proper orientation and sequence

similarity as shown in figure 3.5.1.8 and figure

3.5.1.9. Protein was induced by using 0.25 mM

IPTG O/N at 18°C. Induced supernatant

containing the recombinant protein was purified

through Ni-NTA column. All eluted fractions were

analyzed through 15% SDS-PAGE as shown in

figure 3.5.1.10 Purified protein was excised from

SDS-PAGE and the identity of the induced protein

was confirmed by MALDI-TOF analysis.

Figure 3.5.1.8: (a)PCR amplification of GAPDH gene Figure 3.5.1.9: (b) Restriction digestion to confirm putative clone.

Figure 3.5.1.10 : SDS-PAGE analysis of GAPDH protein.

Project 4 – Bioinformatics tools

(a) Maintenance of biological database and tools:

i) GluD: A program to find distance between

s u g a r i n g l y c o p r o t e i n s .

ii) Protanno: Automated HMM and Sequence

Homology Based Protein Annotat ion.

iii) ANN/SVM tool: Artificial intelligence based

tool for annotation of adhesins from tritryps.

iv) ALU finder: For annotation of Alu elements.

( b ) D a t a b a s e & r e s o u r c e s :

i) Adhesin database: A comprehensive

database and knowledge point for adhesin

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class of proteins.

ii) Plant pathology database: Plant-Patho

Database is a database of important Plants

and crops which contains expertly curated

biological information of plants and related

disease and pathogen along with certain

virulence factors and genes with their

molecular and biological aspects.

iii) Plant lectin database: A web based database

& classification of plant lectins based on

protein domains, fold, sugar specificity and

domain architecture along with its structural

& crystallization information.

3.5.2. Characterization of Leishmanial metabolic

pathway proteins

Principal Investigator

Dr. Anju Pappachan (Assistant Professor)

Project 1: Purification of recombinant Leishmania

donovani Xanthine phosphoribosyl transferase

(LdXPRT)

Xanthine phosphoribosyl transferase (XPRT):

Leishmania donovani is a protozoan parasite

causing visceral leishmaniasis. It lacks the

metabolic machinery for synthesis of purine

nucleotides by de novo pathway and hence

depends on its human host for salvage of

nucleotides. XPRT which recycles xanthine from

the host to XMP is unique to the parasite and lacks

a human counterpart. XPRT lacks mammalian

counterpart and is, therefore, a potential target for

antiparasitic therapy.

Expression, Purification & crystallization of XPRT:

XPRT gene from L. donovani was cloned in

pET15b vector & it was expressed in Rosetta(DE3)

cell lines. Expressed protein was purified by Nickel

affinity chromatography for crystallization and

enzyme assay. Crystallization was carried out by

micro-batch method using Hampton crystal

screens. Crystals were obtained after 20 days as

shown in figure 3.5.2.1.

Figure. 3.5.2.1: Crystals obtained in Hampton screen 1.0.2M CaCl2 dihydrate +0.1M Sodium acetate trihydrate pH 4.6 +20% v/v 2-propanol

Project 2: Purification of recombinant L. donovani

Hexokinase

Hexokinase:

The putative hexokinase taken up for the study is

involved in glycolytic metabolism. A hexokinase is

an enzyme that phosphorylates hexoses, forming

hexose phosphate. Hexokinase catalyses the

transfer of a phospho group from ATP to glucose.

Almost all glucose 6-phosphate so formed is

metabolized by the glycolytic pathway, allowing for

the synthesis of ATP, whereas remainder enters the

mannogen metabolism and pentose phosphate

pathway. Thus, any inhibition of hexokinase would

directly interfere with the formation of both ATP, β-

mannan and nucleic acid precursors. Hexokinase

has been well-characterized kinetically in T. cruzi &

T. brucei. However in Leishmania many

characteristics of this enzyme that allow designing

a rational strategy of selective inhibition are

unknown.

Cloning, Expression and Puri f icat ion of

Hexokinase: Leishmania donovani putative

hexokinase (471 amino acid long) sequence

(gi|398014713) was retrieved from NCBI

database. The 1416bp long gene of hexokinase

was cloned in two different vectors i.e. pET 28a &

pET 15b(Novagen). Expression trials were carried

out in Rosetta(DE3) as well as pLysS cell lines at

various IPTG concentrations, auto induction & at

different temperature. The protein was going in

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insoluble fraction as shown in figure: 3.5.2.2. &

3.5.2.3.

Figure 3.5.2.2 : PCR amplification of L. donovani HexokinaseFigure 3.5.2.3 : Expression of Hexokinase in Rosetta(DE3) by auto induction at 18ºC.

3.6. Department of Environmental Science


Dr. Dhiraj Naik-(Assistant Professor, Group

head)

Research Fellow:

Dr. Usha Joshi - (DBT project- Research

Associate)

Mrs. Gayatri Gadhvi (PhD Scholar)

Mrs. Megha Pandya (PhD Scholar)

Mrs. Utsavi Agarwal (PhD Scholar)

Mr. Manharsinh Solanki (DST Project JRF)

Ms. Twinkle Solanki (DST Project JRF)

Mr. Ritesh Prajapati (DST project assistant-II)

Mr. Hemant Prajapati (DBT Project JRF)

Ms. Divya Patel (DBT Project JRF)

Mr. Jigar Thakar (DBT Field Assistant)

Ms. Priyanka Patel (DBT Field Assistant)

Project 1: Potential for Carbon Sequestration in

Grassland and Afforested Ecosystem using

Molecular and Eddy Covariance Techniques

Project Summary:

The present study provides the first insight on the

diurnal, daytime, nightime, and daily rates of net

ecosystem CO2 exchange based on the

measurements of EC in arid ecosystems with

alkaline soils in Gujarat. The results found that the

diurnal courses of NEE in each month followed

clear sinusoidal patterns during growing season.

Negative values of mean NEE were found at

daytime on sunny days, indicating a net carbon

uptake. In contrast, positive values of mean NEE

were observed on cloudy or rainy days and at

nighttime, which implied a net carbon source.

Furthermore, strong dependency of NEE on PAR

and the response of NEE to precipitation indicated

the ecosystems were still dominated by biotic

factors, similar to other ecosystems, and abiotic

CO2 absorption by soils may be trivial in terms of

magnitude and aptitude. Establishment of

Grassland ecosystem for measurement of eddy

covariance is in progress by carrying out gap filling

for the empty places.Biomet data collection and

analysis is in progress. Diurnal and seasonal

variation of data provided very valuable

information about impact of abiotic factors on the

grassland ecosystem. Preparation of measurement

of soil respiration using closed chamber is in

progress.Techniques for measurement of NEE

using closed chamber both opaque and

transparent, is in progress. This will allow the

comparative studies of NEE using open and closed

system.

Project Achievements:

The first objective of this study is to quantify the

growing season NEE of grassland ecosystems using

EC techniques. The second objective is to test the

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hypothesis that semi arid ecosystem acts as a

carbon sink at night during growing season. For

this purpose, we analyze half-hourly mean NEE

data at daytime and nightime. Finally, this study

i nve s t i ga t e s t he re sponse s o f NEE to

meteorological variables and soil moisture and

temperature, in order to interpret the magnitude of

maximum uptake ability of CO2 absorption by

alkaline soil. Site showed a typical pattern of net

carbon uptake at daytime and net carbon release

at nightime (Figure 3.6.2). Mean monthly −2 −1nightime NEE ranged from 1.65 μmol m s in

−2 −1May and June to 1.0 μmol m s in August

(Figure 3.6.3). At the site, daytime carbon uptake

rates during May–September months ranged from −2 −1 −2 −1−0.7 μmol m s in May to −7.2 μmol m s

in July. The differences among months were

obvious, and the peak carbon uptake rate was in

July. The mean nightime ecosystem respiration −2 −1ranged between 2.15 μmol m s in September

−2 −1 and 3.88 μmol m s in June (Figure 3.6.2). At

the site, the maximum mean nightime ecosystem

respiration occurred in June, different from the

months of maximum mean daytime NEE.

Figure 3.6.1: Diurnal courses of mean NEE during study months from May to September in year 2012 (upper panel) and in the year 2013 (lower panel) sites. Shaded areas represent ±1 standard deviation.

The mean diurnal NEE in each month followed a

clear sinusoidal dynamic during the growing

season (Figure 3.6.1). Mean diurnal variations of

NEE at each month showed a net carbon uptake

(negative NEE) at daytime and a net carbon

release (positive NEE) at nightime in both years.

EC-measured nightime NEE in low turbulence

conditions may be subject to systematic bias, and

the dependence of nightime NEE on friction

velocity (u*) could vary site by site. Relating

nighttime NEE and u* helps to identify the

uncertainty caused by low turbulence. At the site,

the

Figure 3.6.2: Mean monthly NEE at daytime and nightime in year 2013. Error bars represent ±1 SD.

nightime respiration was influenced by u*,

especially under very low turbulence conditions −1(u*<0.15 m s ). The dependence of nightime

NEE on u* may be partly explained by the

heterogeneous landscapes around the site and

wind direct ion dis tr ibut ion. At the s i te,

considerable wind flows were from the west where

oasis croplands were distributed and hence, EC

measurement may be impacted.

EC-measured nightime NEE in low turbulence

conditions may be subject to systematic bias, and

the dependence of nightime NEE on friction

velocity (u*) could vary site by site. Relating

nightime NEE and u* helps to identify the

uncertainty caused by low turbulence. At the site,

the

Figure 3.6.3: Dependence of normalized nightime NEE (defined as the ratio of NEE-min [NEE] to max [NEE]-min [NEE]) on friction velocity (left panel), and the wind rose diagram (right panel). Symbol with “x” represents half-hourly data during the study period, and open circle indicates bin-averages of 0.05 ms-1 width. Error bars refer to ±1 standard deviation.

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nightime respiration was influenced by u*,

especially under very low turbulence conditions −1(u*<0.15 m s ). The dependence of nightime

NEE on u* may be partly explained by the

heterogeneous landscapes around the site and

wind direct ion distr ibution. At the si te,

considerable wind flows were from the west where

oasis croplands were distributed and hence, EC

measurement may be impacted. Biomet data was

analyzed for the period of almost one year starting

from March 2013 to April 2014. Average of 5

minute was carried out in order to analyze the

diurnal variation in various parameters. Whereas

for observing the seasonal variation weekly means

of the variables were taken into account.

Project 1: Carbon Sequestration Potential of

Albizzia lebbeck in intercropping and co-

cultivation with economic crops and plant growth

promoting microorganism

Source of funding- DST, New Delhi

Project Summary:

• Biomass allocation studies have shown

that the native agroforestry tree species

such as Azadirachta indica, Albizzia

lebbeck and Ailanthus excelsa has higher

potential of carbon storage capacity as

compared to other species. Age- and size-

dependent responses on b iomass

allocation were observed. The older

plantation showed higher carbon storage

capacity as compared to younger one.

• The d iurna l measurement o f so i l

respiration under different vegetation sites

showed clear diurnal patterns. Highest

respiration rates were observed under

direct sunlight followed by shade and

lowest in drier area as compared to wet

areas of vegetations.

• Eddy covariance measurement of CO2 and

energy fluxes showed that the plantation

forest sites is very sensitive to changes in

environmental parameters, of which

precipitation pattern has the largest

influence on the next CO2 fluxes of the

vegetation. The preliminary data show the

tree vegetation is carbon sink during the dry

season.

• Ni t rogen fe r t i l i za t ion s tud ies and

nonstructural carbon allocations will be

carried out in coming season which will help

us in understanding the role of mobile

carbon reserves in tree carbon capacity.

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4. LIST OF PUBLICATIONS

The research findings were published in international peer reviewed indexed journals. The list of

publications is placed below.

a) Neha Rajpara, Kittappa Vinothkumar, Priyabrata Mohanty, Arun Kumar Singh, Rajesh Singh, Ritam

Sinha, Dhrubajyoti Nag, Hemanta Koley, Ashima Kushwaha Bhardwaj (2013). Synergistic Effect of

Various Virulence Factors Leading to High toxicity of Environmental V. cholerae Non-O1/ Non-O139

Isolates Lacking ctx Gene : Comparative Study with Clinical Strains. PLoS one. 8 (9): e76200.

b) Kittappa Vinothkumar, Ashima Kushwaha Bhardwaj, Thandavarayan Ramamurthy, Swapan Kumar

Niyog (2013). Triplex PCR assay for the rapid identification of three major Vibrio species, Vibrio

cholerae, Vibrio parahaemolyticus and Vibrio fluvialis. Diagnostic Microbiology and Infectious

Disease. 76:526-528.

c) Braj M.R.N.S. Kutar , Neha Rajpara , Hardik Upadhyay, Thandavarayan Ramamurthy, Ashima K.

Bhardwaj. (2013). Clinical Isolates of Vibrio cholerae O1 El Tor Ogawa of 2009 from Kolkata, India:

Preponderance of SXT Element and Presence of Haitian ctxB Variant. PLoS one. 8(2): e56477.

d) Ashima K. Bhardwaj, Kittappa Vinothkumar, and Neha Rajpara (2013). Bacterial Quorum Sensing

Inhibitors: Attractive Alternatives for Control of Infectious Pathogens Showing Multiple Drug

Resistance. Recent Patents on Anti-Infective Drug Discovery. 8:68-83.

e) Pant DC, Dave M, Tiwari AK. (2013). Wheatgrass (Triticum aestivum L.) supplementation promotes

longevity in Drosophila melanogaster. Annals of Plant Sciences, 49-54.

f) Kumar A, Dave M, Pant DC, Luxar R, Tiwari AK. (2013). Vinca rosea leaf extract supplementation

leads to developmental delay and several phenotypic anomalies in Drosophila melanogaster.

Toxicological & Environmental Chemistry, 95:635-645.

4.1 Presentations:

4.1.1. Invited Lectures:

• Ashima Bhardwaj Multidrug Resistance in Vibrios- Never Underestimate Bacteria. International

Conference on Advances in Biotechnology and Bioinformatics-2013. Pune . 26th November 2013.

• Anand Krishna Tiwari. Drosophila as a model organism for Genetics & Developmental Biology

studies at M.H. Degree College, Jaunpur, Uttar Pradesh.

• Chandramani Pathak and Kishu Ranjan. Crosstalk of FADD and cFLIP in regulation of death

receptor mediated apoptosis. International conference on recent trends in cancer prevention &

therapy, 19-20 Nov 2013, Central University Gujarat, PL-10:26

• Chandramani Pathak, Kishu Ranjan and Anupama Sharma. Regulation of HA14-1 mediated

oxidative stress, toxic response and autophagy by curcumin to augment apoptotic activity in human

embryonic kidney cells. National Conference on Emerging Trends and Challenges in Basic and

Translational Research in Biochemistry. February 4-5, BHU, Varanasi SL-5, P-10

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4.1.2. Oral Presentations:

• Kumar A, Tiwari AK. Molecular Chaperons (Hsp70/Hsc70) are required during eye development of

Drosophila melanogaster in “International Symposium on Conceptual advances in cellular

Homeostasis regulated by proteases and chaperons” held at ACTREC, Mumbai from 3-6 December

2013.

• Tiwari AK, Uttekar B. Overexpression/Downregulation of ubiquitin ligases & molecular chaperon

modulate Alzheimer's like phenotype in Drosophila melanogaster in “National Seminar on

Biochemistry: A Science beyond Compartmentalization” organized by M.G. Science Institute,

Ahmedabad from 20th-21st December 2013.

• Vijay Singh. ProtAnno: an open source online integrated tool for homology search. BTISNet

Bioinformatics National symposium, 2nd February 2013 at NBRI & CIMAP, Lucknow, UP.

• Radhey Shyam Kaushal. Kinetoplastid Membrane Protein-11 (KMP-11), a potential drug target

molecule from Leishmania species” 42nd National Seminar on Crystallography and International

Workshop on Application of X-ray Diffraction for Drug Discovery, 21-23 November, 2013, JNU,

New Delhi.

• Kunal R Shah. Isolation, purification and characterization of kunitz type serine protease inhibitor

with lectin like activity from Solanum tuberosum 42nd National Seminar on Crystallography and

International Workshop on Application of X-ray Diffraction for Drug Discovery, 21-23 November,

2013, JNU, New Delhi.

4.1.3. Poster Presentations:

• Vinothkumar K and Bhardwaj A K. Role of QnrVC protein in conferring quinolone resistance to the

multi-drug resistant clinical isolates of Vibrio fluvialis; Poster presentation in International

Conference on Plant Biotechnology, Molecular Medicine and Human Health, University of Delhi

South Campus, 18-20 October, 2013.

• Braj M. R. N. S. Kutar, Neha Rajpara, Thandavarayan Ramamurthy, Ashima K. Bhardwaj.

Unraveling the molecular mechanisms of multidrug resistance in Vibrio cholerae clinical isolates of

2009 from Kolkata, India. Poster presentation in International Conference on Plant Biotechnology,

Molecular Medicine and Human Health, University of Delhi South Campus, 18-20 October, 2013.

• Kishu Ranjan, Shubita Tripathi, Chandramani Patahk. A Novel function of Fas associated death

domain in non-inflammatory activation of IL1β and IL-18. International Conference on Host

Pathogen Interactions, July 12-15, NIAB Hyderabad, P-102.

• Anupama Sharma, Bhargav N. Waghela and Chandramani Pathak. Evaluation of in vitro

cytotoxicity and apoptotic cell death by surface modified dendrimers in human lung carcinoma

cells. International conference on Chemical Biology, CSIR-IICT, Hyderabad, P-99

• Kishu Ranjan, Rajesh Kumar Shah, Chandramani Pathak. Involvement of FADD in ubiquitin

mediated degradation of ant-apoptotic protein cFLIP. International symposium on conceptual

advances in cellular homeostasis regulated by proteases and chaperons, Dec 3-6, 2013, ACTREC,

Mumbai, PP-7:45

• Kishu Ranjan, Chandramani Pathak. Selective Knockdown of cFLIP Augments Death Receptor-

Annual Report

2013

35

mediated Apoptosis in cancer cells. The XXXVII AICB Conference and symposium on Cell

Dynamics and cell fate, 22-24 dec. 2013, NCBS, Banagalore, P001,AICB-56:63

♦ Kishu Ranjan and Chandramani Pathak. Knockdown of cFLIP enhances apoptosis cell death in

cancer. International conference on recent trends in cancer prevention & therapy, 19-20 Nov 2013,

Central University Gujarat, P-14:58

♦ Suhashini Dhumale and Chandramani Patahk. Inhibition of expression of RAGE and its ligand

HMGB-1 by Quercetin towards promotion of apoptosis in human breast adenocarcinoma cells.

International conference on recent trends in cancer prevention & therapy, 19-20 Nov 2013, Central

University Gujarat, P-15:59

• Bhargav N. Waghela, Anupama Sharma and Chandramani Pathak. Curcumin conjugated with

PLGA augments cellular uptake and apoptosis in human colon carcinoma cells. 4th International

Conference on Stem Cells and Cancer (ICSCC-2013): Proliferation, Differentiation and Apoptosis,

19-22 October 2013, Mumbai. Abs No. P 99.

• Kishu Ranjan & Chandramani Pathak. Knockdown of cFLIP enhanced FADD and TNFα induced

apoptosis. 5th EMBO Meeting-2013 from 21-24th Sept 2013 at Amsterdam Netherland.

• Bhargav N. Waghela, Anupama Sharma, Suhashini Dhumale and Chandramani Pathak. Anti-

proliferative and apoptotic potential of Curcumin-PLGA conjugates in human colon carcinoma

cells. Internatl. Conference on Emerging Trends in Chemical Sciences, Central Univ. Gujarat, March

14-15, 2013, PP15:102

• Kishu Ranjan and Chandramani Patahk. The interaction of FADD and cFLIP proteins on regulation

of death receptor mediated apoptosis in cancer cells. International conference on molecular forms

and functions Jan 8-13, 2013,IISc Bangalore. SN 113, P162.

• Tiwari AK, Christian P, Kumar. A, Pant D. Supplementation of Wheatgrass & Spirulina increases

lifespan and health benefit in Drosophila melanogaster in “National Conference on New Frontiers

in Medicinal Plant Research”Organized at Sikkim University, Sikkim.

• Christian P, Tiwari AK. Investigation of the role of Arthrospira platensis as a therapeutic agent in

Parkinson's disease model in Drosophila melanogaster in “Annual Meeting of Indian Society of

Developmental Biologist” held at TIFR Mumbai from 01-04 December 2013.

• Uttekar B. & Tiwari AK. Drosophila, as an alternate animal model to study Human

neurodegenerative diseases in “National Seminar on Biochemistry: A Science beyond

Compartmentalization” organized by M.G. Science Institute, Ahmedabad from 20th-21st

December 2013.

• Varsani S, Zimik M, Shah S and Singh SK. 2013. Functional characterization of some phloem

expressed genes in Arabidopsis thaliana In: National Symposium on Plant Tissue Culture and

Biotechnology for Food and Nutritional Security, March 11-13, 2013, CFTRI, Mysore. p.58.

• Bhumi Patel. Xanthine phosphoribosyl transferase (XPRT) – A potential anti-leishmanial drug

target. International Conference on Biomolecules forms & functions, January 8-11, 2013, (IISc.

Bangalore).

• Dhaval Patel. Gene ontology mapping bioinformatics analysis & compilation of unique tritryp

Annual Report

2013

36

proteome. International Conference on Biomolecules forms & functions, January 8-11, 2013, (IISc.

Bangalore).

5. Awards:

The faculty and research staff received the following awards during the year.

• Best poster award was awarded to Kishu Ranjan and Chandramani Pathak. Knockdown of cFLIP

enhances apoptosis cell death in cancer. International conference on recent trends in cancer

prevention & therapy, 19-20 Nov 2013, Central University of Gujarat, P-14:58

• Best poster award was awarded to Bhargav N. Waghela, Anupama Sharma and Chandramani

Pathak. Curcumin conjugated with PLGA augments cellular uptake and apoptosis in human colon

carcinoma cells. 4th International Conference on Stem Cells and Cancer (ICSCC-2013):

Proliferation, Differentiation and Apoptosis, 19-22 October 2013, Mumbai. Abs No. P 99.

Annual Report

2013

37

6. HUMAN RESOURCE DEVELOPMENT:

6.1. PhD

The university PhD program has been initiated. Currently 16 students are registered with Pune University

& 4 students are registered with MS University, Baroda for PhD.

The following students have received independent funding to pursue PhD

External Funded Research Fellowships:

S.

No.

Name of Fellow Funding

Agency

Project Investigator Fellowship Position

1 Ms. Anupama Sharma

CSIR

Dr. Chandramani Pathak

160800

SRF

2 Mr. Arun Kumar

ICMR

Dr. Rajesh Singh

194400

SRF

3 Mr. Braj Mohan

CSIR

Dr. Ashima Bhardwaj

200800

SRF

4 Mr. Dhanendra Tomar

CSIR

Dr. Rajesh Singh

186300

SRF

5 Mr. Radhey Shyam Kaushal

CSIR Dr. Desh Deepak Singh

197200

SRF

6 Ms. Sripada Lakshmi

UGC Dr. Rajesh Singh

172800

JRF

7 Ms.Khyati Bhataliya

CSIR Dr. Rajesh Singh 165600

JRF

8. Mr. Kishu Ranjan

ICMR Dr. Chandramani Pathak

196400

SRF

9. Mr. Kritarth Singh

UGC Dr. Rajesh Singh 96000

JRF

10. Ms. Kshama Jain

CSIR Dr. Reena Rajput

177920

JRF

11. Ms. Manisha Farsodia

DST Dr. Sunil Kumar Singh

165600

JRF

12. Ms. Preeti Dabbas

CSIR Dr. Anand Krishna Tiwari

79531

JRF

Annual Report

2013

38

13. Mr. Priyabrat Mohanti

ICMR Dr. Ashima Bhardwaj

186300

SRF

15. Dr. Debashree Sengupta

DBT Dr. Dhiraj Naik

31355

RA

16. Dr. Harish Suthar

DBT Dr. Desh Deepak Singh

193200

RA

6.2. Trainees:

a) Ms. Nishtha Bhatt and Mr. Vivek Pathak (Veer Narmad South Gujarat University, Surat), Dhavani

Patel and Ms. Maitry Shah (Sardar Patel University, Anand) pursued their dissertation training with

Dr. Ashima Bhardwaj, Department of Human Health and Diseases.

b) Ms. Krumali Chokshi, Ms. Nilam Gori & Ms. Ekta Patel (Sardar Patel University, Anand) pursued

their dissertation training with Dr. Reena Rajput, Department of Human Health and Diseases.

c) Sweety Shah from Veer Narmad South Gujarat University, Surat pursued her dissertation training

with Dr. Sunil Kumar Singh, Plant Molecular Biology Department.

Annual Report

2013

39

7. FINANCIALS

7.1 Income and Expense

The Income and Expense for the financial year 2012-2013 is placed below.

INCOME (in

lakhs)

Grants

Donations

Project(s)

related

Foreign

Contribution

Testing

Others Total

Income in

Rs.in Lakhs

For R&D

Activities

196.00 328.00 0.00 0.00 0.00 524.00

For Non R & D

activities

0.00 0.00 0.00 0.00 9.00 9.00

Total income 196.00 328.00 0.00 0.00 9.00 533.00

EXPENDITURE

(in lakhs)

Capital Revenue exp

other than

salaries

Salari es

Others Total

Expenditure

Rs.in lakhs

For R & D

Activities

101.00 116.00 155.00 0 372.00

For Non R & D

activities

8.00 69.00 40.00 0 117.00

Total

Expenditure

109.00 185.00 195.00 0 489.00

Note 1 The support from the trust has been reflected in income head

under Grants and donations and not bifurcated under foreign

contributions.

Note 2 The research grants have been reflected under income head

project related.

Annual Report

2013

40

7.2 Research Grants

The Scientists have written independent research grants and have been awarded the following

projects. Some of these projects have been successfully completed during the year.

Annual Report

2013

S No.

Title and Scope of Project Sponsoring Agency

Project Leader Total Sanctioned Rs. In Lakhs

1 Establishment of distributed information sub center (DISC) at Indian Institute of Advanced Research Gandhinagar

DBT Govt. of India

Dr. D D Singh 51.46

2 Prospecting of genes for oil biosynthesis and storage in Jatropha curcus L.

DBT Govt. Of India

Dr. R Nadgauda 52.02

3 Identification and analysis of new and established virulence factors from Vibrio cholera, Leishmania major, Trypanosoma cruzi and Trypanosoma brucei through a bioinformatics study

Dept of Sc. and Tech. Govt. of India

Dr. D D Singh 27.19

4 Cloning and characterization of efflux pumps responsible for multiple drug resistance in clinical isolates of various Vibrio species


Dr. A Bhardwaj 19.78

5 Multiplication of elite planting material on a large scale of high value medicinal crops of western India using tissue culture and nursery techniques

CSIR complex Pusa , New Delhi


6 Isolation, purification and preliminary X ray structure analysis of lectins from some important medicinal plants


Dr. D D Singh 26.91

7 Program support for School of Biological Sciences and Biotechnology

DBT Govt. of India


8 Implementation of the Mitochondrial specific miRNA

DBT Govt. of India

Dr. Rajesh Singh 41.04

9 Structural characterization of glycoproteins from Leishmania, involved in host-pathogen interaction

DBT Govt. of India

Dr. D D Singh 42.79

10 To study the role of mitochondrial proteins in IKKe

breast cancer cell lines mediated NF b activation inκ

DBT Govt. of India


41

11 Study of apoptotic signaling pathways in mesenchymal stem cells during normal and differentiated states

DBT Govt. of India


12 Unraveling the cross talk between inflammation and apoptosis focusing to regulation of inflammatory mediators and apoptosis in cancer

DBT Govt. of India

Dr. C Pathak 25.36

13 The study of mitochondrial dynamics during S. aureus infection in human epithelial cells

ICMR Govt. of India


14 To study the role of ubiquitin proteosome system and molecular chaperon 70 in the

disease using Drosophila malanogaster as a model


Dr. A K Tiwari 6.98

15 GSBTM Financial assistance program

GSBTM Dr. Neeraj Jain 19.16

16 Nutrahelix NEFT Dr. K Paliwal 0.50 17 Targeting the mitochondria

dependent and independent apoptotic signaling by nano particulate drug conjugate to induce apoptosis in cancer cells

DBT Govt. of India

Dr. C Pathak 49.27

18 Study of miRNA in cell death Parkinson disease

DBT Govt. of India


19 Role of mitochondrial MAVS and NLRX1 interaction in regulation of TNF induced ROS and inflammation

INDO-RUSSIA, DST, Govt. of India


20 Potential for carbon sequestration in grassland afforested ecosystem using molecular and Eddy Covariance techniques

DBT Govt. of India

Dr. K Paliwal 72.36

21 Neuroprotective effects of Toll like receptor 4 antagonists and signaling inhibitors in LPS induced neuronal insults

DBT Govt. of India

Dr. Reena Rajput 25.20

22 Investigating the functional relationship of RAGE

SERB, Govt. of India

Dr. C Pathak 20.30

in the progression of Alzheimer’s

Annual Report

2013

42

23 Role of Trim proteins in regulation of autophagy pathway

DBT Govt. of India


24 Role of heat shock protein during eye development in Drosophila

Science and engineering Research Board, Govt. of India

Dr. A K Tiwari 16.45

25 Structural and functional characterization of unique pathway proteins from Leishmania donovani

Science and engineering Research Board, Govt. of India

Dr. A Pappachan 24.00

26 Cloning and structural studies of Kinases from Leishmania donovani

DBT Govt. of India


27 Characterizing the functional role of Metacaspases in Arabidopsis thaliana

DBT Govt. of India

Dr. S K Singh 53.74

28 Study of multidrug resistance in clinical isolates of Shigella Spp.

ICMR, Govt. of India

Dr. A Bhardwaj 26.94

29 Dectins and inflammasomes in Aspergillus fumigatus specific immune response

DBT Govt. of India

Dr. R Rajput 28.20

30 FADD protein in cancer cells ICMR, Govt. of India

Dr. C Pathak 7.86

31 Carbon sequestration of Albizzia lebbek and plant growth promoting microorganism

DST, Govt. of India

Dr. D Naik 34.23

32 Design synthesis and evaluation of novel autophagy inducers as potential anticancer agents

GSBTM Dr. Rajesh Singh 19.82

33 Reprogramming of Immune cell to Neuron

GSBTM Dr. Reena Rajput 19.99

34 Unraveling the mechanism underlying quinolone resistance in multi drug resistant clinical isolates of Vibrio and Shigella species from India

GSBTM Dr. A Bhardwaj 19.99

35 Studies on purine salvage pathway from L. donovani

DBT Govt. of India


36 TLR5 mediated T regulation in cholera

DBT Govt. of India

Dr. Reena Rajput 25.35

Total Research Grants 1,427.97

Annual Report

2013

43

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