PROFORMA – I PROFORMA FOR SUBMISSION OF PROJECT...
Transcript of PROFORMA – I PROFORMA FOR SUBMISSION OF PROJECT...
PROFORMA – I
PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH
AND
DEVELOPMENT, PROGRAMME SUPPORT
(To be filled by the applicant)
PART I: GENERAL INFORMATION
1. Name of the Institute/University/Organization submitting the Project Proposal:
Tumkur University, Tumkur
2. State: Karnataka
3. Status of the Institute: State govt funded/UGC recognized
(Please see Annexure-I)
4. Name and designation of the Executive Authority of the Institute/University
forwarding the
application: Registrar, Tumkur University Tumkur
5. Project Title: Molecular cloning and expression of anticoagulant and antiplatelet
protease (s) from cucumber sap extract: Understanding the probable molecular
mechanism and therapeutic application in thrombotic disorders.
6. Category of the Project (Please tick): R&D
7. Specific Area (Please see Annexure - II): Life science
8. Duration: 03 Years
9. Total Cost (Rs.): 97.128 lakhs
10. Is the project Single Institutional or Multiple-Institutional (S/M)? : Single
institutional
11. If the project is multi-institutional, please furnish the following:
Name of Project Coordinator: Affiliation:
Address:
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12. Scope of application indicating anticipated product and processes
The most common cause of death world wide is thrombotic disorders such as, myocardial
infarctions, acute arterial thrombosis, atrial fibrillation (clot in heart), unstable angina,
deep vein thrombosis, pulmonary embolism. Several anti-coagulants, anti-platelets agents
are being used to treat thrombotic disorders besides their health threatening side effects.
Consequently, identifying the novel protein(s) exhibiting strong anti-coagulant anti-
platelet properties along with fibrin clot hydrolyzing activity (fibrinolytic activity) from
the plants or fruits helps in the better management of thrombotic disorders. Therefore,
The development of new antithrombotic agents has been inspired by clinical needs and
by progress in biotechnology that have made it potential to fabricate drugs that target
specific steps in thrombogenesis. It is well known that, Heparin has pharmacokinetic and
biophysical limitations that are overcome by new anticoagulants. Of these, low-
molecularweight heparin and direct inhibitors of thrombin have been evaluated clinically.
Coumarins require careful laboratory monitoring because of concerns about safety.
Orally active direct inhibitors of thrombin and factor Xa may replace coumarins. Aspirin
is of limited efficacy because it inhibits only one pathway of platelet activation.
Therefore, identifying the effective and safer anticoagulant and antiplatelet agent helps in
the better management of thrombotic disorders. Moreover, target specific (Factor Xa and
thrombin inhibitors) proteins/enzymes with strong anticoagulant and antiplatelet
properties along with fibrin clot hydrolyzing activities would be the better therapeutic
molecules to treat thrombotic disorders. Thus, purification pharmacological
characterization of protease that exhibit anticoagulant, antiplatelet and fibrinolytic
protease from cucumber sap extract becomes gateway to the researchers/scientist to
further explore the importance of unexplored/ignored medicinal plants. Our preliminary
studies indicated that proteolytic activity of cucumber sap extract found to cause anti-
coagulation, inhibition of agonist ADP and collagen induced platelet aggregation of
human platelet rich plasma and washed human platelets. Hence, this study ultimately
contributes for both academicians and researchers who are working on thrombotic
disorders. Anti-coagulants and anti-platelets agents and fibrin clot hydrolyzing enzymes
have enormous importance and they contribute for better management of thrombotic
disorders. Above all, they may serve as prototypes for designing anti-thrombotic drugs
and they can be used as therapeutic agents to treat hypercoagubility of blood leads to fatal
heart attack and strokes that leads to high rate of death.
13. Project Summary (Not to exceed one page. Please use separate sheet).
The main aim of the project is purification, cloning, expression and pharmacological
characterization of proteases (s) that exhibit anticoagulant, antiplatlelet and fibrinolytic
activities. In addition, establish the possible molecular mechanism of anticoagulant and
antiplatelet activities of purified protease (s). According to our preliminary studies,
cucumber sap extract showed two isoforms of proteases in casein zymogram, it
selectively hydrolyzed human fibrinogen and fibrin clot. Further more, cucumber sap
extract showed strong anticoagulant activity in plasma recalcification time and inhibited
agonist collagen and epinephrine induced platelet aggregation of human platelet rich
plasma in a dose dependent manner. Anticoagulants and antiplatelet agents have an
immense therapeutic value because the most common cause of death world wide is
thrombotic disorders that include myocardial infarctions, acute arterial thrombosis, atrial
fibrillation (clot in heart), unstable angina, deep vein thrombosis and pulmonary
embolism. Anti-coagulant and anti-platelet therapy is the effective therapy for the
prevention and treatment of thrombotic disorders. The currently established
anticoagulants and anti-platelet agents are having numerous limitations with life
threatening side effects. Thus currently available anti-coagulant and anti-platelet therapy
fails to recommend protection against thrombotic disorders. Hence, identifying the novel
anti-coagulant and anti-platelet agents from the natural sources with least side effects is
the challenging task to the researchers. In addition, proteases exhibit anticoagulant,
antiplatelet and fibrin clot hydrolyzing activities helps in the better management of
thrombotic disorders.
PART II: PARTICULARS OF INVESTIGATORS
(One or more co-investigators are preferred in every project. Inclusion of co-
investigator(s) is
mandatory for investigators retiring before completion of the project)
Principal Investigator:
14. Name : Dr. Devaraja. S
Date of Birth : 24-04-1976 Sex (M/F): M
Designation : Assistant Professor
Department : Biochemistry
Institute/University : Tumkur University
Address: Dr. Devaraja. S, Assistant Professor, Department of Studies and
Research in Biochemistry, Tumkur University, University Constituent College
Campus, B.H, Road, Tumkur, PIN: 572103, Telephone: 919945765028, 0816-
2254546 Fax: 0816-2270719 E-mail: [email protected]
Number of research projects being handled at present: 02
Co-Investigator
15. Name : Dr.Kemparaju.K
Date of Birth : 20-07- 1964 Sex (M/F): M
Designation : Associate Professor
Department : DOS in Biochemistry,
Institute/University : University of Mysore
Address : DOS in Biochemistry, University of Mysore, Manasagangothry,
Mysore,
Karnataka, India. PIN: 570 006
Telephone: 91-9945996543
E-mail: [email protected]
Number of Research projects being handled at present: 02
Co-Investigator
16. Name : Dr. K.S. Girish
Date of Birth : 22-06-1976 Sex (M/F): M
Designation : Assistant Professor
Department : DOS in Biochemistry
Institute/University : University of Mysore
Address: DOS in Biochemistry, University of Mysore, Manasagangothry,
Mysore,
Karnataka,
PIN :570006.
Telephone : 9972268633
E-mail : [email protected]
Number of Research projects being handled at present: 02
Co-Investigator
16. Name : Dr. Sudhrshan. B,G
Date of Birth : 18-04-1975 Sex(M/F) : M
Designation : Senior Medical Officer and Assistant Professor
Department : Biomedical and instrumentation
Institute/University : R.V. Engineering College
Address : R.V. Engineering College, Mysore road, Banglore, Karnataka,
PIN : 5600059
Telephone: 08067178100
E-mail: [email protected]
Number of Research projects being handled at present: 02
Co-Investigator
16. Name : T.G.Thippeswamy
Date of Birth : 28.06.1971 Sex (M/F): M
Designation : Assistant Professor
Department : DOS in Biochemistry
Institute/University : Tumkur University,
Address : DOS in Biochemistry, Tumkur University, Constituent college
campus,
B.H.Road, Tumkur, Karnataka, India.
PIN: 572103
Telephone: 0816-2254546 Fax: 0816-2270719
E-mail: [email protected]
Number of Research projects being handled at present: 02
Co-Investigator
16. Name : Bhagyalakshmi.M
Date of Birth :16-08-1981 Sex (M/F): F
Designation : Assistant Professor
Department : DOS in Biochemistry
Institute/University : Tumkur University,
Address : DOS in Biochemistry, Tumkur University, Constituent college
campus,
B.H.Road, Tumkur, Karnataka, India.
PIN: 572103
Telephone: 0816-2254546 Fax: 0816-2270719
E-mail: [email protected]
Number of Research projects being handled at present: 02
PART III : TECHNICAL DETAILS OF PROJECT
(Under the following heads on separate sheets)
16. Introduction (not to exceed 2 pages or 1000 words)
In healthy people, homeostatic balance exists between procoagulant (clotting) forces and
anticoagulant and fibrinolytic forces. Numerous genetic, acquired and environmental
factors can incline the balance in favor of coagulation, leading to the pathologic
formation of thrombi in veins (deep venous thrombosis), arteries (myocardial infarction,
ischemic stroke), or cardiac chambers. Thrombi can obstruct blood flow at the site of
formation or detach and embolize to block a distant blood vessel (eg, pulmonary
embolism, embolic stroke). As such,the diagnosis and management of thrombotic
disorders are increasingly falling down within the realm of the practice of hematology.
There are about 10 million cases annually reported world wide exceeding the number of
deaths from cancer and other diseases. It is estimated that annual mortality rate of 60,000
in United States and more than 50,000 in India. Thus thrombotic disorders represents a
major health problem world wide including India. Anti-platelet and anti-coagulant agents
have been increasingly becomes an important tool for preventing cardiovascular events.
While aspirin is the most widely used and best studied anti-platelet agent, the use of
clopidogrel (Plavix) and combination dipyridamole-aspirin (Aggrenox) have increased
substantially in recent years, and a 2007 study introduced a new agent, prasugrel (Effient).
Despite the lack of efficacy and safety heavy marketing to physicians and direct-to-
consumer advertising have made Plavix the second best-selling drug in the world. The
main side effect of Plavix is at low dose it increases the risk of unwanted clot formation
and at high dose the coagulation mechanisms in the blood are shut down too severely and
there is increased risk of internal and external bleeding episodes. Thus currently available
anti-coagulant and anti-platelet therapy fails to recommend protection against thrombotic
disorders. Hence, the researchers are extremely enthused about the usage of proteins
those exhibit anti-coagulant, anti-platelet and fibrin clot hydrolyzing proteins/proteases
from naturally occurring medicinal plants and they may complement the existing anti-
platelet and anti-coagulant therapy. Hence, these proteases may be act as powerful
weapons in the better management of thrombotic disorders.
There are several natural unexplored medicinal plants and fruits whose pharmacological
properties are not well established. Cucumber is one such fruit from the creeper Cucumis
sativa L and belongs to Cucurbitaceae family, which is native to Indian subcontinent.
Cucumber has been found to possess water content of about 96% while it is Scarce in
calorific value. However, it has been reported to contain wide varieties of nutritional
components in trace amount such as lipids, proteins, carbohydrates as macronutrients and
potassium, calcium, magnesium, iron, vitamins (Vit-C, Vit-B complex) as micronutrients.
It has got broad spectrum of health beneficial applications & found to protect from heat,
inflammation, of skin, acidity, gastritis, ulcer, arthritis, gout, high and low BP and
elimination of toxic substances from the blood. It has been associated with healing
properties in relation to diseases of the kidney, urinary bladder, liver & pancreas. Sap is a
juicy circulating fluid of cucumber & its biological importance has not been studied so
far. Our preliminary studies established the proteolytic activity of cucumber sap found to
exhibit fibrin (ogen) lytic acitivty, anti-coagulant and anti-platelet property in platelet rich
plasma. Therefore, this project has been under taken to dissect the therapeutic role and
molecular mechanism of protease (s) those exhibit antiplatelet, anticoagulant and fibrin
clot hydrolyzing properties followed by their cloning and expression.
16.1 Origin of the proposal
Blood coagulation cascade is the co-ordinated activation of plasma serine proteases and
their co-factors. The end product is the protease thrombin, which cleaves fibrinogen to
generate a fibrin clot. In addition, platelets which circulating in the blood plays an
important role in the hemostasis or formation of fibrin clot (Furie et al., 2005, 2008). If
the number of platelets are too low in circulating blood that leads to excessive bleeding
and too high leads to the formation of blood clots or thrombosis respectively and it may
obstruct blood vessels and results in stroke, myocardial infarction, pulmonary embolism
or the blockage of blood vessels (Savage et al., 2001: William et al., 2010). In addition,
the abnormality in haemostatic system leads to excessive bleeding or thrombosis. The
human body has a complex mechanism that causes blood to clot if a wound occurs.
Under normal circumstances this is a desirable response that enables the body to heal
itself, but under certain clinical conditions, called “blood clot disorders or thrombotic
disorders", this same mechanism can cause an unwanted clot or "thrombus" that can be
life threatening. Thrombosis the formation of a blood clot within a blood vessel or cavity
of the heart is a maladaptive process of vascular occlusion and remains a primary cause
of cardiovascular morbidity and mortality. The primary etiology of myocardial
infarctions, and approximately 80% of strokes, is acute arterial thrombosis. In addition,
atrial fibrillation (clot in heart), myocardial infarction (heart attack), unstable angina,
deep vein thrombosis, pulmonary embolism and replacements of heart valves also leads
to the formation of clot with in the heart and hence represents the most common cause of
death world wide (Webster et L., 2005:Canhão et al., 2005:Hatzinikolaou-Kotsakou
et al., 2005: Lars Maegdefessel et al., 2010).
16.2 (a) Rationale of the study supported by cited literature (b) Hypothesis (c) Key
questions.
Thrombotic disorders increase the risk of formation of fibrin clot with in a blood vessel
or cavity of the heart and it is a leading cause of death and disability world wide. About
20 to 30% of patients hospitalized for an acute medical illness is due to arterial
thrombosis, atrial fibrillation, myocardial infarction, unstable angina, deep vein
thrombosis, pulmonary embolism. Anti-coagulants those inhibits the clot formation by
blocking the action of clotting factors/coagulation factors and anti-platelets agents those
blocks the formation of blood clot by preventing the clumping of platelets are extensively
used in treatment of thrombotic disorders (Leng et al.,1996: Anand et al.,
2010). Thus Anti-coagulant and anti-platelet therapy is the effective therapy for the
prevention and treatment of thrombotic disorders. While, the currently established
anticoagulants such as, warfarin, dabigatran, unfractionated heparin (UFH), enoxaparin,
fondaparinux, bivalirudin (thrombin inhibitors), low molecular weight heparin and anti-
platelet agents such as, aspirin, thienopyridines, dipyridamole, dlopidogrel, dpoprostenol,
abciximab, eptifibatide and tirofiban (glycoprotein IIb/IIIa inhibitors) are having
numerous limitations with several side effects, including lack of reversibility, a sheer
dose response, food and multiple drug-drug interactions, narrow therapeutic index and
cause bleeding (Rosendaal et al.,1993: CAPRIE Steering Committee 1996: van et al.,
2002: Hurlen et al., 2002: Antman et al., 2004). Therefore, identifying the novel anti-
coagulant and anti-platelet agents from the natural sources with least side effects is the
challenging task to the researchers. In addition, the other hypothesis is that targeting
proteases upstream from the common pathway provides a reduction in thrombin
generation slow down the formation of fibrin clot or thrombosis. Investigating the target
specific (factor Xa and thrombin) anti-coagulant, anti-platelet and fibrin clot hydrolyzing
proteins from the naturally occurring plants and fruits with least side effects help in the
better management of thrombotic disorders.
16.5 Current status of research and development in the subject (both international
and
national status)
Plants sustain our daily life, as they are the important source of food, agricultural
crops, and medicines. The value of medicinal plants to human livelihood is essentially
infinite. Medicinal plants are the most exclusive sources of life saving drugs for the
majority of the world’s population. The world health organization has estimated that
more than 80% of the world’s population in developing countries depends primarily
on herbal medicines for basic health care needs. Plant products also play an important
role in the health care systems of the remaining 20% of the population, mainly
residing in developed countries. A large proportion of drugs used in modern medicine
are either directly isolated from plants or synthetically modified from a lead
compound of natural origin. Medicinal plants and their extracts are often used as an
alternative to specifically targeted drugs in the treatment and prevention of many
diseases.
Finding healing powers in plants is an ancient idea. People on all continents have long
applied poultices and imbibed infusions of hundreds of indigenous plants dating back
to prehistory. Plants have the richest source of several products which are immense
use to man. Noteworthy among them and next to those of food value are substances of
medicinal importance. An estimate suggests that about 25 million plant species known
for their medicinal uses. Phytochemical tests have been performed in about 5000 and
nearly 1100 species are extensively exploited in 80% of ayurvedic, 46% of unani and
33% of allopathic medicines.
The world market of pharmaceutical products operates a business turnover of
US$ 320,000,000/year from which US$ 20,000,000 originates from vegetal active
substances. The current world market of phytomedicines operates a business turnover
of US$ 40 billion/year from which US$ 2 billion originates from India.
In more recent history, the use of plants as medicines has involved the isolation of
active compounds, beginning with the isolation of morphine from opium in the early
19th century (Kinghorn, 2001; Samuelsson, 2004). Drug discovery from medicinal
plants led to the isolation of early drugs such as cocaine, codeine, digitoxin, and
quinine, in addition to morphine, of which some are still in use (Newman et al., 2000;
Butler, 2004; Samuelsson, 2004). Isolation and characterization of pharmacologically
active compounds from medicinal plants continue today. More recently, drug
discovery techniques have been applied to the standardization of herbal medicines, to
elucidate analytical marker compounds. The search for new molecules, nowadays, has
taken a slightly different route where the science of ethnobotany and
ethnopharmacology are being used as guide to lead the chemist towards different
sources and classes of compounds. Current research in molecular medicine includes
the drug discovery from medicinal plants involves a multifaceted approach combining
botanical, phytochemical, biological, and molecular techniques. Medicinal plant drug
discovery continues to provide new and important leads against various
pharmacological targets including cancer, inflammation, wound healing, HIV/AIDS,
Alzheimer’s, malaria, pain, cardiovascular disorders including thrombolytic disorders.
Several natural product drug(s) of plant origin have either recently been introduced to
the world market.
16.6 The relevance and expected outcome of the proposed study
1. This project may provide preliminary data about the efficacy and safety use of
proteolytic enzymes that exhibit anticoagulant, antiplatelet properties along with
fibrinolytic activities from cucumber sap extract.
2. Cloning and expression of this protease will be evaluated for antithrombotic
activities along with its mechanism of action
3. Expressed protease will be tested for its non- toxicity
4. The success in cloning and expression of proteases with anticoagulant, antiplatelet
and fibrinolytic activities could definitely helps in the development of safer and
effective antithrombotic drugs with least side effects.
5. The results obtained in this project will be published in the peer reviewed
international journals.
16.7 Preliminary work done so far
According to our preliminary studies the cucumber sap extract found to show varied
proteins bands from the molecular weight range from 200 to 16 kDa (Fig 1). In addition,
the cucumber sap extract found to cause anti-coagulation as it increased normal clotting
time 250 sec to 430 sec (Fig 2). Cucumber sap extracts selectively hydrolyzed Aα and Bβ
chains of fibrinogen with out affecting γ chain over the incubation of 8h, suggesting its
specificity on Aα and Bβ chains of fibrinogen (Fig 3). Interestingly, cucumber extracts
specifically degraded α-polymer and α-chain with out affecting γ- γ dimmer and β-chain
of fibrin clot (Fig 4). Further more, the extract inhibited the collagen and epinephrine
induced aggregation of platelet rich human plasma dose dependently (Fig.5 and 6).
RESULTS
Fig.1.SDS-PAGE pattern of cucumber sap extract.
Cucumber sap extract (100 µg) under non-reduced condition (1), Cucumber sap extract
(100 µg ) under reduced condition (2).Lane M: Molecular mass markers (M) in kDa from
top to bottom: myosin-H-chain (200), phosphorylase b (97.8), BSA (68), ovalbumin (43)
carbonic anhydrase (29), β-lactalbumin (18.4) and lysozyme (14.3).Fig.1b.Activity
staining on casein zymogram
Fig. 2. Effect of cucumber sap extarct on re-calcification time of human plasma.
Cucumber sap extract ranging from 10 to 80 µg was pre-incubated with 300 µl of human
plasma in the presence of 30µl tris-Hcl buffer(10 mM; pH 7.4) for 1 min at 37˚C, 30 µl of
CaCl2 (0.25 M) was added to the pre-incubated mixture. The time for clot formation was
recorded
Fig. 3. Fibrinogenolytic activity of cucumber sap extract: Fibrinogen incubated for 8
h at 37o and run in 10% SDS-PAGE under reduced condition. Fibrinogen (50 µg) alone
(1), fibrinogen (50 µg), + 4µg cucumber sap extract (2), fibrinogen (50 µg), + 8µg
cucumber sap extract (3), fibrinogen (50 µg) + 12µg cucumber sap extract (4), fibrinogen
(50 µg) + 16µg cucumber sap extract(5),fibrinogen (50 µg) + 24µg cucumber sap extract
(6), fibrinogen (50 µg) + 24µg cucumber sap extract (7). M represents the molecular
weight marker in kDa from top to bottom: myosin-H-chain (200), phosphorylase b (97.8),
BSA (68), ovalbumin (43) carbonic anhydrase (29), β-lactalbumin (18.4) and lysozyme
(14.3).
Fig.4. Fibrinolytic activity cucumber sap extract: Washed plasma clot was incubated
with cucumber sap extract for 8 h and then separated on 7.5 % SDS-PAGE under reduced
condition. Plasma clot alone (1), plasma clot + 8 µg cucumber extract (2), 12 µg (3), 16
µg (4) 20 µg (5), 24µg (60), 30µg (7) M represents the molecular weight markers in kDa
from top to bottom: myosin H chain (200), phosphorylase b (97.8), BSA (68), ovalbumin
(45), carbonic anhydrase (31), and trypsin inhibitor (21.5).
Fig. 5. Effect of cucumber sap extract on collagen induced aggregation of platelet rich human
plasma.
Fig. 5. Effect of cucumber sap extract on epinephrine induced aggregation of platelet rich human plasma.
17. Specific objectives (should be written in bulleted form, a short paragraph
indicating the
methods to be followed for achieving the objective and verifiable indicators of
progress shouldfollow each specific objective)
• Isolation, biochemical and pharmacological characterization of anti-
coagulant, anti- platelet and fibrin clot hydrolyzing protease (s) from
cucumber sap extract
• Establish the site of action of purified protease(s) on blood coagulation
cascade using specific clotting factor /deficient clinical
plasma/reconstituted plasma samples
• Cloning and expression of protease (s) that possess anticoagulant,
antiplatelet and fibrinolytic activities
• Investigate the kinetic effects of protease(s) in the various agonist induced
platelet function of human platelet rich plasma and washed human
platelets
• Establish the probable molecular mechanism involved in the anti-coagulant
and anti-platelet properties of protease (s).
18. Work Plan: should not exceed 3-4 pages (the section can be divided according to
the specific aims and under each specific aim, the following should be stated clearly
as sub headings)
18.1 Work plan (methodology/experimental design to accomplish the stated aim
Preparation of sap extract from cucumber.
Cucumbers will be purchased from the Market; both the ends will cut to obtain the sap
material. The ends will be squeezed and the juicy material will be collected. The sap
material will be dissolved further in water and centrifuge at 2000 rpm and the supernatant
will be stored at -20OC until use.
Protein estimation:
The protein estimation will be done according to the method of Lowry et al., (1951) using
the bovine serum albumin (BSA) as standard.
Coagulant activity:
The plasma re-calcification time will be determined according to the method of Quick et
al. (1935). Briefly, the various concentration of cucumber sap extract will be pre-
incubated with 0.2 ml of citrated human plasma in the presence of 10 mM Tris-HCl (20
ml) buffer pH 7.4 for 1 min at 37o C. Twenty microlitres of 0.25 M CaCl2 was added to
the pre-incubated mixture and clotting time will be recorded.
Preparation of platelet rich plasma (PRP) and platelet poor plasma (PPP).
The method of Ardlie and Han (1974) will be employed. Nine volumes of human blood
from healthy donors (who are non-smokers and non-medicated at least for the previous
15 days) in to one volume of acid citrate dextrose (93 mM sodium citrate, 7 mM citric
acid and 140 mM glucose pH 6.5) followed by centrifugation at 90 g for 10 min at room
temperature. The supernatant is called platelet rich plasma (PRP). The remaining blood
will be centrifuged at 500 g for 15 min and the supernatant obtained is the platelet poor
plasma (PPP). The platelet concentration of PRP will be adjusted to 3.1 X 108
platelets/ml with PPP. The PRP maintained at 37o C will be used with in 2 hr. All the
above preparations will be carried out using plastic wares or siliconized glassware’s.
Preparation of washed platelets
The washed platelets will be prepared according the method described by Cazenave et al.
Briefly, platelet rich plasma (PRP) will be obtained by mixing fresh blood sample with
acid citrate dextrose solution –ACD (85 mM sodium citrate; 71 mM citric acid; 111 mM
dextrose) in a 6:1volume ratio followed by centrifugation at 120 × g for 15 min. The
supernatant portion obtained is the PRP and is transferre to plastic tube. Then the PRP
will be centrifuged at 1700 × g for 15 min at 37°C, the supernatant obtaine is to be
discarded and the platelet pellet will suspended in Tyrode's albumin buffer (145 mM
NaCl, 5 mM KCl, 10 mM HEPES, 0.5 mM Na2HPO4, 1 mM MgCl2, 6 mM glucose,
0.3 % human serum albumin) pH 6.5 containing 10 U/mL heparin and 0.5 µM
prostacyclin. After 10 min of incubation at 37°C, the suspension is centrifuge at 1700 ×
g for 15 min, the platelet pellet obtained is again suspended in Tyrode’s albumin buffer
containing 0.5 µM prostacyclin and is again centrifuge at 1700 × g for 15 min. Finally the
platelet pellets obtaine are suspended in Tyrode’s albumin buffer containing 0.02 U/mL
apyrase pH 7.4 and platelets will be counted using a Neubauer chamber and adjuste to
the concentration required using Tyrode's albumin buffer pH 7.4.
Platelet aggregation
The turbidometric method of Born (1962) will be followed for using a Chronolog dual
channel aggregometer connected to an omniscrible dual pen recorder to record the light
transmission as a function of time. Aliquotes of PRP (0.45 ml)/human platelet rich
plasma will be pre-incubated with different concentrations of various concentration of
sap extract for 3 min in a cylindrical glass cuvette under constant stirring. The
aggregation will be initiated by the addition of agonist such as collagen, ADP, thrombin
and epinephrine followed for 8 min. As platelets aggregate in response to an added
agonist, light transmission increases progressively producing an aggregation trace on the
recorder paper. The aggregation trace is the plot of light transmission between platelet
rich plasma (PRP) and platelet poor plasma (PPP) base line, which represent 0 % and
100 % aggregation respectively.
cDNA library construction
The cucumber sap extract cDNA library will be constructed accoding to the method of
Chaim et al. Briefly, cucumber sap extract mRNAs will be purified using the FastTrack
2.0 Mrna isolation kit (Invitrogen). The cDNAs will be then synthesized using the Super
Script Plasmid System with Gateway Technology for cDNA Synthesis and Cloning
(Invitrogen), cloned into NotI and SalI pre-cut pSPORT1 vector and transformed into
Escherichia coli DH5α cells. Transformants will be selected on LB (Luria– Bertani) agar
plates containing 100 µg/ml ampicillin.
cDNA library screening
Randomly chosen colonies (approx. 100 clones) will be inoculated into LB broth
containing 100 µg/ml ampicillin and grow overnight at 37◦C (with aeration). From this,
recombinant plasmids were purified using QIAprep Spin Miniprep Kit (Qiagen). The
cloned cDNAs will be sequenced on both strands using ABI PRISM® BigDye
Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems). Reactions will
be analysed using an ABI 377 automatic sequencer (Applied Biosystems). The promoter
regions will be used for priming the sequencing reactions. The cDNA sequences will be
analysed, and the putative protein products from these sequences.
Expression and purification of recombinant protease
Selected transfermants will be pregrown at 30 ℃ in 0.5 L baffled shake flasks
containing 0.15 L rich medium (20 g/L tryptone, 13.4 g/L yeast nitrogen base, 4 × 10-4
g/L biotin, 1 % glycerol, 0.1 mol/L potassium phosphate buffer, pH 6.0). The cells will
be grown for 24 h with an additional 1 % glycerol after 12 h. The culture will be
centrifuged and the pellet containing the cells will be resuspended in rich medium with
methanol (20 g/L tryptone, 13.4 g/L yeast nitrogen base, 4 × 10-4 g/L biotin, 1 %
methanol, 0.1 mol/L potassium phosphate buffer, pH 6.0). After induction for 72 h, the
culture will be collected and adjusted pH to 5.0 and then centrifuged to remove cells and
precipitates. Exchanging buffer to 20 mmol/L methylpiperazin-HCl, pH 5.0, by using
Sephedex G 25, the supernatant will be applied to Q-Sepharose Fast Flow (Pharmacia
Biotech AB, Sweden) and the target protein will be eluted at the gradient of 0-0.5 mol/L
NaCl in 20 mmol/L methylpiperazin-HCl, pH 5.0 in 20 column volume. The activity
peak will be pooled and then applied to Phenyl HP (Amersham Pharmacia Biotech AB,
Sweden) in a start buffer of 1 mol/L ammonium sulfate in 20 mmol/L Tris-HCl, pH 7.2.
The active fractions collected were concentrated by ultralfiltration (Amicon, USA) up to
5 times and 0.5 ml of them was applied onto Superdex 200 column. The active material
obtained will be loaded to ConA Hitrap (Pharmacia Biotech AB, Sweden), eluted by 0.5
mol/L NaCl and 0.5 mol/L methyl- a -D-glycopyanoside in 20 mmol/L Tris-HCl, pH 7.4
Proteolytic activity:
Proteolytic activity will be determined according to the method of Satake et al., (1963)
using 2 % casein in 0.2 M Tris-HCl buffer, pH 8.5. Various concentration of purified
protein/s from ion exchange chromatography will be incubated separately with 0.4 mL of
substrate in a total volume of 1 mL for 2 hr and 30 min at 37OC. Undigested casein will
be precipitated by adding 1.5 ml of 0.44 M trichloroacetic acid. The digested casein in
supernatant (1 ml)will be determined using Folin-Ciocalteu’s reagent. The method is
initially standardized using increasing amount of venom samples in the assay mixture.
One unit of activity was defined as the amount of enzyme required to cause an increase in
O.D. by 0.01 at 660 nm/min.
Substrate gel assay (zymogram):
SDS-PAGE (10%) will be prepared according to the method of Laemmli (1970). and
polymerized at a final concentration of 0.2 % with casein/gelatin.purified protein (10 µg
each) will be prepared under non-reduced condition. Electrophoresis will be carried out
using Tris (25 mM), glycine (192 mM) and SDS (0.1%) for 3h at 90V at room
temperature. After electrophoresis, gels will be washed with 10mM sodium phosphate
buffer containing 2.5%of Triton X-100 with shaking for 1h to remove SDS and incubated
overnight at 37oC in Tris-HCl buffer (50 mM) pH 7.6 containing 10 mM CaCl2 and 150
mM NaCl. Gels are then stained with Coomassie brilliant blue R-250 as described by
Nagaraju et al., (2006).
Coagulant activity:
Recalcification time will be determined according to the method of Quick (1935). by
incubating different concentration of purified protease in a total volume of 0.2 ml of
citrated human plasma with of 10 Mm Tris Hcl buffer pH 7.4 for 1 min, and then clotting
time will be determined by adding 20µl of 0.25 M CaCl2.
Fibrinogenolytic activity:
Fibrinogenolytic activity will be determined according to the method of Ouyang and
Teng (1976). Different concentrations of purified protein/s (0-20 µg) from ion exchange
column chromatography incubated independently with 50 µg human fibrinogen in 40 µL
of 10 mM Tris-HCl buffer pH 7.4. After 8 h, reaction was terminated by adding 20 µL
denaturing buffer containing 1 M urea, 4% SDS and 4% β-mercaptoethanol. It was
analyzed by 10% SDS-PAGE.
Fibrinolytic activity
Method of Rajesh et al (2006) will be employed for fibrinolytic activity. EDTA (2
mg/ml) treated blood is centrifuge for 15 min at 500g to separate plasma. Plasma (100 µl)
will be mixed with equal volume of 10 mM CaCl2 for 15 min at 37O C to get soft fibrin
clot. The fibrin clot is washe thoroughly 5-6 times with phosphate buffered saline (PBS)
and suspended in a final volume of 40 µl 10 mM Tris-HCl buffer pH 7.4 containing
different concentrations of purified protein of cucumber. The reaction will be stopped by
adding 20 µl of sample buffer containing 4% SDS, 1 M urea and 4% β-mercaptoethanol.
It is then boile for 3 min and centrifuge to settle the debris of plasma clot. An aliquot of
20 µl supernatant will be analyzed in 7.5% SDS-PAGE for fibrin degradation studied.
Platelet aggregation
The turbidometric method of Born (1962) will be followed for using a Chronolog dual
channel aggregometer connected to an omniscrible dual pen recorder to record the light
transmission as a function of time. Aliquotes of PRP (0.45 ml)/human platelet rich
plasma will be pre-incubated with different concentrations of various concentration of
sap extract for 3 min in a cylindrical glass cuvette under constant stirring. The
aggregation will be initiated by the addition of agonist such as collagen, ADP, thrombin
and epinephrine followed for 8 min. As platelets aggregate in response to an added
agonist, light transmission increases progressively producing an aggregation trace on the
recorder paper. The aggregation trace is the plot of light transmission between platelet
rich plasma (PRP) and platelet poor plasma (PPP) base line, which represent 0 % and
100 % aggregation respectively.
Platelet adhesion assay
Platelet adhesion assay will be performed according the method described by Bellavite et
al. Briefly, 20 µg each of collagen type I, fibrinogen and fibronectin in 200 µl PBS will
be added independently to 96-well polystyrene microplates and kept for 16 h at 4oC. The
wells will be blocked by adding 200 µl of 1% BSA in PBS for 1 h at 37oC, and will be
washed three times with 200 µl PBS. This will be followed by the addition of 100 µl
aliquot of washed platelets stock (1 x 108 washed platelets suspension was pretreated with
10 µg of purified protein from cucumber sap or anti-integrin α2β1 mAb 6F1 or PBS
alone in a final volume of 1 ml Tyrode’s buffer for 10 min at 37oC). In case of purified
cucumber protease, the activity will be inhibited by EDTA at a final concentration of 5
mM before aliquoting. The reaction mixture was incubated at 37oC for 90 min and this
will be followed by washing three times with 200 µl PBS. The adherent platelets are then
treated with 150 µl lysis buffer (0.1 M citrate buffer pH 5.4 containing 5 mM p-
nitrophenyl phosphate and 0.1% Triton X-100) and incubated at 37oC for 90 min.
Addition of 2 N NaOH (100 µl) terminated the reaction by inactivating the platelet
membrane acid phosphatase activity and the optical density is measure at 405 nm.
Platelets adhesion is expressed as % adhesion taking the platelets suspension treated with
PBS as 100%.
In vitro platelet viability and in vivo platelet count
The method of Da Silva will be used to test the platelet viability. Briefly, the human
platelet rich plasma is incubate with increasing dose (0 – 8 µg) of purified protein of
cucumber sap in PBS for 30 min at 37°C in a volume of 100 µl containing 1×105 platelets.
Aliquots of 20 µl are mixe independently with 20 µl trypan blue (0.4%) and the platelet
morphology is observe with a microscope using a Neubauer chamber. The data expressed
as % viability, considering 100% viability in the absence of purified protein (control).
For in vivo platelet count, the method of Loria et al will be followed. Briefly, the purified
protein (µg/g body weight; defibrinogenating dose) will be injected intravenously in to
groups of mice (n=5) in 50 ml PBS, and then the animals will be allowed to bleed by
cardiac puncture at different time intervals of 1, 3 and 5 h after anaesthetized by diethyl
ether in to sodium citrate solution. The platelet counts/µl is determined by using
Neubauer chamber and a group of mice received the PBS alone served as control.
18.2 Connectivity of the participating institutions and investigators (in case of
multiinstitutional projects only)
Not applicable
18.3 Alternate strategies (if the proposed experimental design or method does not
work
what is the alternate strategy)
The principal investigator has the basic knowledge and background for carrying out this
project as he acquainted with the methodologies used for plasma coagulation and platelet
aggregation including protein purification techniques and has publications addressing
thrombosis and hemostasis. The co-principal investigators has enormous experience on
thrombosis and hemostasis related research work and got several publications on that area
and their assistance is essential to complete the project successfully. The team has
investigators with vast research knowledge on the objectives of the project. Therefore, we
are very confident in successful completion of the proposed work with a meaningful
outcome. Certainly, the proposed work will definitely bring out the meaning full
results/product.
Timelines: (Please provide quantifiable outputs) Period of study Achievable targets
First six months Second six months
Year 1
Chemicals and proposed
instruments will be purchased. All
the proposed parameters will be
standardized.
Aim 1: Isolation, biochemical and
pharmacological characterization of
anti-coagulant, anti-platelet and fibrin
clot hydrolyzing protease(s) from
cucumber sap
Year 2
Aim 2: Establish the site of action
of purified protease(s) on blood
coagulation cascade using specific
clotting factor deficient clinical
plasma samples and reconstituted
plasma
Aim 4: Investigate the kinetic
effects of protease(s) in the various
agonist induced platelet function of
human platelet rich plasma and
washed human platelets.
Aim 4. Cloning and expression of
protease(s) that possess anticoagulant,
antiplatelet and fibrinolytic activities
&
Manuscript will be prepared for the
publication.
Year 3
Aim 5: Establish the probable
molecular mechanism involved in
the anti-coagulant and anti-platelet
properties of purified protease (s).
Any unfinished work from previous
period will be done. The second/third
manuscript will be prepared for the
publication.
20. Name and address of 5 experts in the field
Sl no Name Designation Address
1 Dr. V.R. Devaraj
Associate Professor, Banglore University, Central college Campus,
Banglore, Karnataka, India.
2 Dr. Narender, Scientist E, Central Drug Research Institute,
Luknow, UP, India.
3 Dr. Raghu, P, Senior Scientist, National Institute of Hyderbad, AP,
India.
4 Prof, Sreeramulu, Department of Biochemistry, Gulbarga
University, Gulabarga, Karnataka, India.
5 Shylaja Dharmesh, Senior Scientist, Central Food Technological
Research Institute, Mysore, Karnataka, India.
PART IV: BUDGET PARTICULARS
Budget (In Rupees)
A. Non-Recurring (e.g. equipments, accessories, etc.)
Sl
No
Equipment required
1st year 2
nd year 3
rd year Total
1
2
3
4
5
6
7
8
9
10
11
12
FPLC (SCL10 Shimadzu)
Flurimetry (F5301PCShimadzu)
Lyophilizer (CRIST ALPHA)
Microscope with camera
Western blot unit
Electrophoresis unit
Ph meter
Incubator
weighing balance
Hot air oven
Computer, printer and Xerox
machine
Yes
Yes
Yes
----------------
Yes
Yes
--------------
-------------
--------------
-------------
Yes
-----------
--------------
--------------
---------------
Yes
--------------
------------
Yes
-----------
-----------
Yes
-------------
Yes
-------------
-------------
-------------
--------------
--------------
--------------
Yes
Yes
Yes
-----------
------------
------------
Rs. 18,00000
Rs, 10,00000
Rs, 4,00000
Rs, 700000
Rs, 50,000
Rs, 50,000
Rs, 20,000
Rs, 50,000
Rs, 100000
Rs, 50,000
Rs, 200000
Rs,300000
13
14
15
16
17
18
Laminar air flow
Centrifuge with variable rotors
PCR
Gel doc
Gel dryer
Transiluminator
Electrophorator
Yes
--------------
-------------
-------------
-------------
-------------
------------
Yes
Yes
Yes
Yes
yes
-------------
-------------
------------
-------------
-------------
------------
Rs,200000
Rs, 400000
Rs,300000
Rs, 200000
Rs, 100000
Rs, 200000
Total Rs. 61,20,000
Budget A Rs. 61,20,000
Overhead Costs (@20% of
project cost)
---------------- --------------- -------------- Rs, 12,24,000
Sub-Total (A) = Rs, 61, 20,000
B. Recurring
B.1 Manpower (See guidelines at Annexure-III)
Sl no Position
Consolidated
Emolument
Year 1
(Lakhs)
Year 2
(Lakhs)
Year 3
(Lakhs)
Total
(Lakhs)
1 JRF 2 14,000+HRA 3.696 3.696 3.696 11.088
Total -------------
---
------------- 3.696 3.696 3.696 11.088
Sub-Total (B.1) = 11.088 lakhs
B.2 Consumables
Sl
no
Items Year 1
(Lakhs)
Year 2
(Lakhs)
Year 3
(Lakhs)
Total
(Lakhs)
01 Chemicals &
Glassware/plastic
wares
3 3 3 9
Sub-Total (B.2) =9 lakhs
B.3 Travel
Travel Type Year 1
(Lakhs)
Year 2
(Lakhs)
Year 3
(Lakhs)
Total
(Lakhs)
Total 0.2 0.2 0.2 0.6
Sub –Total (B.3) = 0.6 lakhs
B.4 Contingency
Travel
Type
Year 1
(Lakhs)
Year 2
(Lakhs)
Year 3
(Lakhs)
Total
(Lakhs)
Total 1.0 1.0 1.0 3.00
Sub-total (B.4)= 3 lakhs
B.5 Overhead (If applicable)
Item Year 1
(Lakhs)
Year 2
(Lakhs)
Year 3
(Lakhs)
Total
(Lakhs)
4.08 4.08 4.08 12.24
Sub-total of B.5= 12.24lakhs
(B.1+B.2+B.3+B.4+B.5)
(11.088 lakh+9.0 lakh+0.6lakh+ 3.0 lakh+ 12.24lakh)= 35.928 lakhs
Sub-total of B= 35.928 lakhs
Grand Total (A + B)
(61.20 lakh+ 35.928) = 97.128 lakhs
Grand total=97.128 lakhs
Note: Please give justification for each head and sub-head separately mentioned in
the above table.
Financial Year: April - March
In case of multi-institutional project, the budget estimate to be given separately for each
institution.
JUSTIFICATION:
The principle investigator himself works at bench, in addition proposed staff is
essential to share the responsibilities to achieve the proposed project in time.
Consumable proposed are essential, chemicals such as reagent kits, factor deficient
plasma, human fibrinogen, thrombin, collagen, ADP, epinephrine, pepstatin,
leupeptin, column materials etc and other glassware’s.
Traveling grant is essential for attending symposia/ seminar/works shops/ training
programmes/consulting rare references in nearest universities/ research institutions.
Contingency is essential to meet expenses such as postal charges, typing of reports
and research articles, acquisition of books and documents of relevance, photo and
copying of reports and reprints etc.
Although, the preliminary work was standardized with the help of co-principle
investigators lab in Mysore University. Mysore, Karnataka, India. Currently working
Tumkur University is newly established and Department of PG studies and research
in Biochemistry lack all the basic instruments. Hence, all equipments are very much
essential for carrying out this type of work.
PART V: EXISTING FACILITIES
Resources and additional information
1. Laboratory: Available
a. Manpower: 2 JRFs will be recruited
b. Equipments
2. Other resources such as clinical material, animal house facility, glass house.
Experimental garden, pilot plant facility etc.
Basic facilities are available in the host institute (Tumkur University) Animal studies
will be carried out with the help of co-investigators from University of Mysore.
Sl No Equipments available
1 pH meter
2 Spectrophotometer
3 Centrifuge
4 Electronic balance
5 Colorimeter
6 Water bath
7 Incubator
8 Hot air oven
9 Electrophoresis unit
10 ELISA reader
11 Western blot
PART VI: DECLARATION/CERTIFICATION
It is certified that a) The research work proposed in the scheme/project does not in any way duplicate
the work already done or being carried out elsewhere on the subject. b) The same project proposal has not been submitted to any other agency for financial
support. c) The emoluments for the manpower proposed are those admissible to persons of
corresponding status employed in the institute/university or as per the Ministry of Science & Technologyguidelines (Annexure-III)
d) Necessary provision for the scheme/project will be made in the Institute/University/State budget in anticipation of the sanction of the scheme/project.
e) if the project involves the utilisation of genetically engineered organisms, we agree to submitan application through our Institutional Biosafety Committee. We also declare that while conducting experiments, the Biosafety Guidelines of the Department of Biotechnology would be followed in toto.
f) If the project involves field trials/experiments/exchange of specimens, etc. we will ensure that ethical clearances would be taken from concerned ethical Committees/Competent authorities and the same would be conveyed to the Department of Biotechnology before implementing the project.
g) it is agreed that any research outcome or intellectual property right(s) on the invention(s) arising out of the project shall be taken in accordance with the instructions issued with the approval of the Ministry of Finance, Department of Expenditure, as contained in Annexure-V.
h) We agree to accept the terms and conditions as enclosed in Annexure-IV. The same is signed and enclosed.
i) The institute/university agrees that the equipment, other basic facilities and such other administrative facilities as per terms and conditions of the grant will be extended to Investigator throughout the duration of the project.
j) The Institute assumes to undertake the financial and other management responsibilities of the project.
Signature of Principal Investigator: Signature of Executive
Authority
Date: Institute/University with
seal
Date:
Signature of Co-Investigator Signature of Co-
Investigator
Date: Date:
Signature of Co-Investigator Signature of Co-
Investigator
Date: Date:
Signature of Co-Investigator
Date :
PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF
INVESTIGATORS
Provide the following information for the key personnel in the order listed on PART II.
Follow this format for each person. DO NOT EXCEED THREE PAGES
Name : Dr. Devaraja. S
Designation : Assistant Professor
Department/Institute/University: Tumkur University
Date of Birth : 24-04-1976 Sex (M/F): M, SC/ST: SC
Education (Post-Graduation onwards & Professional Career)
A. Position and Honors
Position and Employment (Starting with the most recent employment)
Sl
No
Institution
Place
Position From (Date) To
(date)
Degree Year University/Institute Field of Specialization
M. Sc., 2003 University of Mysore Biochemistry
Ph. D., 2010 University of Mysore Biochemistry
Post doc 2010 Cleveland Clinic,
Ohio State University, USA
Molecular cardiology
1 University of
Mysore
Department of
Biochemistry
Junior Research
Fellow
July 2005-august
2006
2 University of
Mysore
Department of
Biochemistry
Senior Research
Fellow
August 2006-January
2009
3 Learners Research
Institute ,
Cleveland clinic,
Ohio, USA
Department of
Molecular
Cardiology
Post doctoral
Fellow
14th May2010 to 2nd
July 2010
4 Tumkur University Department of
Biochemistry
Assistant
Professor
5th July2010 to till
date
Honors/Awards
Title of Award Year
1. Best teacher award from Dayananda Sagar College of 2003- 2004 Life Sciences, Bangalore 2. UPG Fellowship (Mysore University) 2005-2006 3. University Grand Commission Fellowship (UGC, Govt of India) 2006-2009 4. Post doctoral Fellowship, Learners Research Institute, 2010 Cleveland Clinic, Cleveland, Ohio, USA.
Professional Experience and Training relevant to the Project
The principal investigator has h vast research knowledge on the objectives of the
project which is accounted by their peer reviewed publications. Therefore, we are very
confident in successful completion of the proposed work with a meaningful outcome.
More over, Dr. K. Kemparaju and Dr. K.S. Girish, University of Mysore has lot of
expertise in the field of hemostatsis and platelet biology research. They have
standardized several protocols related to hemostasis. In addition, they have isolated
enzymatic toxins and elucidated their involvement in the hemostasis (coagulation
cascade). The data generated by the investigators are well appreciated by scientific
community as suggested by the citations for their peer reviewed publications in
international reputed journals. In addition, other co-investigators namely, Dr,
Sudharshan, Senior medical Officer and Thippeswamy and Bagyalakshmi also
committed to involve in the successful completion of this piece of work.
B. Publications (Numbers only): 08
Books : Nil, . Research Papers, Reports:16 , General articles :Nil
Patents : Nil, .Others (Please specify) : Nil
Selected peer-reviewed publications (Ten best publications in chronological order)
1. Devaraja S, Girish K.S, Gowtham Y.M.J, Kemparaju K. The Hag-protease-II is a
fibrin(ogen)ase from Hippasa agelenoides spider venom gland extract:
Purification, characterization and its role in hemostasis.. Toxicon. 2011.
2. Devaraja. S., Nagaraju. S., Mahadeswaraswamy. Y.H., Girish. K.S., Kemparaju.
K., A low molecular weight serine protease: purification and characterization
from Hippasa agelenoides spider venom gland extract. Toxicon 2008 52(1): 130-
138.
3. Nagaraju. S., Devaraja. S., Kemparaju. K., Purification and properties of
hyaluronidase from Hippasa partita (funnel web spider) venom. Toxicon 2007
50(3):383-393.
4. S.Devaraja, K. S. Girish, V. R. Devaraj and K. Kemparaju. Factor Xa-like and
fibrinogenolytic activities of a serine protease from Hippasa agelenoides spider
venom gland extract. J of Thromb Thrombolysis, 2010 Jan; 29(1): 119-126.
5. R.Sharma, Y.H. Mahadeswaraswamy, K. Harish Kumar, S. Devaraja, K.
Kemparaju, B.S. Vishwanath and K.S. Girish. Effect of Anticoagulants on the
Plasma Hyaluronidase activities. Journal of Clinical Laboratory Analysis 2008,
22, 1–5.
6. Mahadeswaraswamy YH, Devaraja S, Kumar MS, Goutham YNJ and Kemparaju
K. Inhibition of local effects of Indian Daboia/Vipera russellii venom by the
methanolic extract of grape seeds (Vitis vinifera L.). Indian Journal of
Biochemistry and Biophysics, 2009, 46, 154-160.
7. Hemshekhar M, Sunitha, Sebastin Santhosh M, Devaraja S, Kemparaju K, Girish
K.S. An overview on genus garcinia: phytochemical and therapeutical aspects.
Phytochemistry review 2011.
8. Y. H. Mahadeswaraswamy, B. Manjula, S. Devaraja, K.S. Girish and K.
Kemparaju: Daboia Russelli Venom Hyaluronidase: Purification, Charcterization
and inhibiton by β-3-(3-hydroxy – 4 – oxopyridyl) α-amino- propionic acid.
Current Topics in medicinal chemistry 2011.
List maximum of five recent publications relevant to the proposed area of work.
1. Devaraja S, Girish K.S, Gowtham Y.M.J, Kemparaju K. The Hag-protease-II is a
fibrin(ogen)ase from Hippasa agelenoides spider venom gland extract:
Purification, characterization and its role in hemostasis.. Toxicon. 2011.
2. Devaraja. S., Nagaraju. S., Mahadeswaraswamy. Y.H., Girish. K.S., Kemparaju.
K., A low molecular weight serine protease: purification and characterization
from Hippasa agelenoides spider venom gland extract. Toxicon 2008 52(1): 130-
138.
3. S.Devaraja, K. S. Girish, V. R. Devaraj and K. Kemparaju. Factor Xa-like and
fibrinogenolytic activities of a serine protease from Hippasa agelenoides spider
venom gland extract. J of Thromb Thrombolysis, 2010 Jan; 29(1): 119-126.
4. R.Sharma, Y.H. Mahadeswaraswamy, K. Harish Kumar, S. Devaraja, K.
Kemparaju, B.S. Vishwanath and K.S. Girish. Effect of Anticoagulants on the
Plasma Hyaluronidase activities. Journal of Clinical Laboratory Analysis 2008,
22, 1–5.
C. Research Support, Ongoing Research Projects
Sl
No.
Title of Project Funding
Agency
Amount Date of sanction and
duration
1 Fund for infrastructure
development in science
and technology
Vision Group
on Science
and
Technology,
Bangalore
40 lakh March 2011-March 2013
2 Screening of Indian
medicinal plants for
anticoagulant and
UGC Minor
project
2 lakh March 2012- March 2014
antiplatelet properties
Completed Research Projects (State only major projects of last 3 years)
Sl No. Title of Project Funding Agency Amount Date of
Completion
Not applicable
Place : Signature of Investigator
Date :
Dr. K. Kemparaju Biodata
Name : DR. K. Kemparaju
Designation : Associate Professor of Biochemistry
Date of Birth : 20 th July 1964
Organization : Department of Studies in Biochemistry
University of Mysore, Manasagangotri,
Mysore – 570 006, India.
Permanent address : As above
Educational Qualification:
Ph.D. Degree : Biochemistry (1996) University of Mysore
M.Sc. Degree : Biochemistry (1987) University of Mysore
B.Sc. Degree : CBZ (1985) University of Mysore
Teaching and research experience
2009 Due for : Professor of Biochemistry (yet to declare)
2001 to 2009 : Associate Professor of Biochemistry,
2002 to 2003 : Post doctoral fellow,
Albert Einstein College of Medicine,
Bronx, New York, USA.
1996-2001 : Senior Lecturer in Biochemistry, Univ. of Mysore.
1988-1996 : Lecturer in Biochemistry, Univ. of Mysore.
1987–1988 : Ph.D. student, Dept. of Biochemistry, Indian
Institute of Science, Bangalore.
Supervisor of PhD students
Awarded Submitted Working
8 1 5
Publications:
1. Kumar MS, Devaraj VR, Vishwanath BS, Kemparaju K. Anti-coagulant activity of a
metalloprotease: further characterization from the Indian cobra (Naja naja) venom. J
Thromb Thrombolysis. 2010 Apr;29(3):340-8.
2. Devaraja S, Girish KS, Devaraj VR, Kemparaju K. Factor Xa-like and
fibrin(ogen)olytic activities of a serine protease from Hippasa agelenoides spider
venom gland extract. J Thromb Thrombolysis. 2010 Jan; 29(1):119-26.
3. Ushanandini S, Nagaraju S, Nayaka SC, Kumar KH, Kemparaju K, Girish KS. The
anti-ophidian properties of Anacardium occidentale bark extract. Immunopharmacol
Immunotoxicol. 2009; 31(4):607-15.
4. Shashidharamurthy R, Mahadeswaraswamy YH, Ragupathi L, Vishwanath BS,
Kemparaju K. Systemic pathological effects induced by cobra (Naja naja) venom
from geographically distinct origins of Indian peninsula. Exp Toxicol Pathol. 2009
Sep 2.
5. Girish KS, Kemparaju K, Nagaraju S, Vishwanath BS. Hyaluronidase inhibitors: a
biological and therapeutic perspective. Curr Med Chem. 2009; 16(18):2261-88.
Review.
6. Mahadeswaraswamy YH, Devaraja S, Kumar MS, Goutham YN, Kemparaju. K.
Inhibition of local effects of Indian Daboia/Vipera russelli venom by the methanolic
extract of grape (Vitis vinifera L.) seeds. Indian J Biochem Biophys. 2009 Apr;
46(2):154-60.
7. Chandrashekara KT, Nagaraju S, Nandini SU; Basavaiah, Kemparaju K.
Neutralization of local and systemic toxicity of Daboia russelii venom by Morus alba
plant leaf extract. Phytother Res. 2009 Aug; 23(8):1082-7.
8. Sharma R, Mahadeswaraswamy YH, Harish Kumar K, Devaraja S, Kemparaju K,
Vishwanath BS, Girish KS. Effect of anticoagulants on the plasma hyaluronidase
activities. J Clin Lab Anal. 2009; 23(1):29-33.
9. Devaraja S, Nagaraju S, Mahadeswaraswamy YH, Girish KS, Kemparaju K. A low
molecular weight serine protease: Purification and characterization from Hippasa
agelenoides (funnel web) spider venom gland extract. Toxicon. 2008 Jul; 52(1):130-8.
10. Mahadeswaraswamy YH, Nagaraju S, Girish KS, Kemparaju K. Local tissue
destruction and procoagulation properties of Echis carinatus venom: inhibition by
Vitis vinifera seed methanol extract. Phytother Res. 2008 Jul; 22(7):963-9.
11. Nagaraju S, Girish KS, Fox JW and Kemparaju K. ‘Partitagin’ a hemorrhagic
metalloprotease from Hippasa partite spider venom: Role in tissue necrosis Biochimie
89, 1322-1331, 2007.
12. Nagaraju S, Devaraja S, Kemparaju K. Purification and properties of hyaluronidase
from Hippasa partita (funnel web spider) venom gland extract.Toxicon. 2007 Sep 1;
50(3):383-93. Epub 2007 Apr 24.
13. Girish KS, Kemparaju K. The magic glue hyaluronan and its eraser hyaluronidase: a
biological overview. Life Sci. 2007 May 1; 80(21):1921-43. Review.
14. Shashidharamurthy R, Kemparaju K. Region-specific neutralization of Indian cobra
(Naja naja) venom by polyclonal antibody raised against the eastern regional venom:
A comparative study of the venoms from three different geographical distributions. Int
Immunopharmacol. 2007 Jan; 7(1):61-9.
15. Girish KS, Machiah KD, Ushanandini S, Harish Kumar K, Nagaraju S, Govindappa
M, Vedavathi M, Kemparaju K. Antimicrobial properties of a non-toxic glycoprotein
(WSG) from Withania somnifera (Ashwagandha). J Basic Microbiol. 2006; 46(5):365-
74.
16. Nagaraju S, Mahadeswaraswamy YH, Girish KS, Kemparaju K. Venom from spiders
of the genus Hippasa: biochemical and pharmacological studies. Comp. Biochem
Physiol C Toxicol Pharmacol. 2006 Sep; 144(1):1-9.
17.Ushanandini S, Nagaraju S, Harish Kumar K, Vedavathi M, Machiah DK, Kemparaju
K, Vishwanath BS, Gowda TV, Girish KS. The anti-snake venom properties of
Tamarindus indica (leguminosae) seed extract. Phytother Res. 2006 Oct; 20(10):851-
8.
18. Shashidharamurthy R, Kemparaju K. A neurotoxic phospholipase A2 variant:
isolation and characterization from eastern regional Indian cobra (Naja naja) venom.
Toxicon. 2006 Jun 1; 47(7):727-33.
19. Girish KS, Kemparaju K. Inhibition of Naja naja venom hyaluronidase: role in the
management of poisonous bite. Life Sci. 2006 Feb 23; 78(13):1433-40.
20. Kemparaju K, Girish KS. Snake venom hyaluronidase: a therapeutic target. Cell
Biochem Funct. 2006 Jan-Feb; 24(1):7-12. Review.
21. Girish KS, Kemparaju K. Inhibition of Naja naja venom hyaluronidase by
plant-derived bioactive components and polysaccharides. Biochemistry (Mosc). 2005
Aug; 70(8):948-52.
22. Girish KS, Kemparaju K. A low molecular weight isoform of hyaluronidase:
purification from Indian cobra (Naja naja) venom and partial characterization.
Biochemistry (Mosc). 2005 Jun; 70(6):708-12.
23. Rajesh R, Raghavendra Gowda CD, Nataraju A, Dhananjaya BL, Kemparaju K,
Vishwanath BS. Procoagulant activity of Calotropis gigantea latex associated with
fibrin(ogen)olytic activity. Toxicon. 2005 Jul; 46(1):84-92.
Book chapter
1. K. Kemparaju, K.S. Girish and S. Nagaraju; Hyaluronidases, a Neglected Class of
Glycosidases from Snake Venom, Beyond a Spreading Factor. (Reptile Venoms and
Toxins by S. Meckessy, IRL Press, USA.) 235 to 256, 2009.
Invited talks delivered in national and international symposia
1. K. Kemparaju; Geographical variation of Naja naja venom: an important factor in
making therapeutic antivenom. 2005; Dept. of Biochemistry and Biotechnology,
Annamalai University, Annamalai nagar, Tamilnadu, India.
2. K. Kemparaju; Hyaluronidase : a potential target in the management of
snakebite,2007; School of Biotechnology, Chemical and Biomedical Engineering,
VIT University, Vellore, Tamilnadu, India.
3. K. Kemparaju; The metzincin Partitagin is a unique Beta fibrinogenase from
Hippasa
partita spider venom that inhibits platelet aggregation. 2008; Dept. of
Biochemistry
and Biotechnology, Annamalai University, Annamalai nagar, Tamilnadu, India.
4. K. Kemparaju; NN-PF3 is a fibrin(ogen)olytic enzyme from Naja naja venom.
2009, Dept. Biochemistry and Biotechnology, Annamalai University, Annamalai
nagar, Tamilnadu, India.
5. K.Kemparaju; Local effects of snakebite: possible therapeutic targets for the
better management. 2008, Little flower Hospital and Research Centre ,
Angamaly, Kochin , kerala India.
6. K.Kemparaju; Neutralization Vipera russellii venom and its purified
hemorrhagin by gallic acid, 2009, Medical Biotechnology on Clinical research, J,
N. Tata Auditorium, IISc Campus, Bangalore.
7. K.Kemparaju; The non toxic fibrin(ogen)olytic enzyme from Naja naja venom:
inhibition of collagen induced human platelet aggregation by affecting alpha-2,
beta-1 and GP-VI. 2010, State level Refresher Course for pre-University Teachers,
Suttur, Mysore, India.
Papers/Posters presented at national and international symposia:
1. Kumar M.S., Vishwanath B.S., Kemparaju K. NN-PF3, an anticoagulant and
antiplatelet metalloprotease from the Indian cobra (Naja naja) venom. International
anatomical sciences and cell biology conference. May 2009, Department of anatomy,
National university of Singpore, Singpore.
2. Mahadeswaraswamy YH and Kemparaju K. Screening of Indian medicinal plants for
anti-venom property against Indian Daboia/ Vipera russellii and Echis carinatus
venoms. Dr. Norman E. Borlaug commemoration national conference on “Plant
diversity and plant health”. March 2010, Department of Studies in Botany,
University of Mysore, Mysore. India.
3. Mahadeswraswamy Y.H, Girish K.S and Kemparaju, K. Antivenom potential of
Gallic acid against the Indian Daboia/Vipera russellii venom and its purified
hemorrhagic complex. State level conference on Recent advances in Environmental
Science and Engineering. October, 2009. Department of Biotechnology and
Engineering, Shridevi Institute of Engineering and Technology, Tumkur, India.
4. Devaraja S and Kemparaju K. A low molecular weight serine protease the Hag-
protease from Hippasaagelenoides spider venom gland extract: Role in
fibrin(ogen)olysis andplatelet function. International conference on `cardiovascular
diseasessecondary to the metabolic disorders: Mechanism and therapy`.December,
2009. Department of Studies in Biochemistry, University of Mysore, Mysore. India.
1. Mahadeswaraswamy YH and Kemparaju K. Gallic acid: a potent inhibitor of Indian
Daboia russellii venom and its purified hemorrhagic complex induced local toxicity.
National symposium on Bioactive molecules: from discovery to industry. April 2009.
Department of Studies in Biochemistry, University of Mysore, Mysore. India.
2. Mahadeswaraswamy YH and Kemparaju K. Local tissue destruction property of
Indian Daboia /Vipera russellii venom and its purified hemorrhagic complex:
Inhibition by Gallic acid. National conference on Recent trends in animal
physiology. October, 2009, Department of Studies in Zoology, University of Mysore,
Mysore. India.
3. Devaraja S and Kemparaju K. Local and systemic toxicity of Hag-protease a low
molecular weight serine protease from Hippasa agelenoides spider venom gland
extract. National symposium on Recent Trends in Animal Physiology. October 2009.
Department of Studies in Zoology, University of Mysore, Mysore. India
4. Mahadeswraswamy YH, Girish KS and Kemparaju K. Local toxicity of Indian
Daboia/Vipera russellii venom and its purified hemorrhagic complex: Contribution
for the better management. National conference on Medical Biotechnology and
Clinical research. October 2009. Department of Biotechnology, Sri. M.
Visvesvaraya Institute of Technology and Sri Krishnadevaraya Education Trust, JN
Tata Auditorium, Indian Institute of Science campus, Bangalore, India.
5. Yashonandana J Gowtham, Sadiq Saleh Ahmed, Naveen H M and K. Kemparaju.
The Indian King Cobra (Ophiophagus hannah) Venom: A preliminary study.
National symposium on Bioactive molecules: from discovery to industry. April 2009.
Department of Studies in Biochemistry, University of Mysore, Mysore. India.
6. Yashonandana J Gowtham and K. Kemparaju. The Indian King Cobra
(Ophiophagus hannah) Venom strongly interferes in Hemostasis. National
conference on Recent trends in animal physiology. October, 2009, Department of
Studies in Zoology, University of Mysore, Mysore. India.
7. Devaraja S and Kemparaju K. Hag-protease from Hippasa agelenoides spider
venom extract: Role in tissue necrosis and hemostasis. National symposium on
Bioactive molecules: from discovery to industry. April 2009. Department of Studies
in Biochemistry, University of Mysore, Mysore.
8. Rashmi K, Zahara Ashkavand, Devaraja S and K.Kemparaju. preliminary studies
on proteolytic activity of cucumber sap extract. Nationalsymposium on Bioactive
molecules: from discovery to industry. April2009. Department of Studies in
Biochemistry, University of Mysore, Mysore.
9. Mahadeswaraswamy YH and Kemparaju K. Local effects of snakebite: Possible
therapeutic targets for the better management. International symposium on Snakes,
venoms and snake bites. SNA-CON October 2008, Angamaly, Cochin.
10. Girish. K.S., Nagaraju, S., and Kemparaju, K. Hyaluronidase: a potential target in
the management of snakebite. 3rd National Symposium on Venoms and Toxins. Dept.
of Biochem. Univ. of Mysore (2004).
11. Nagaraju, S., Girish. K.S., and Kemparaju, K. Spider venom: comparative
characterization from three different species (Hippasa). 3rd National Symposium on
Venoms and Toxins. Dept. of Biochem. Univ. of Mysore (2004).
12. Kemparaju K. and Girish.K.S. A pharmacological examination of the Indian saw-
scaled viper venom. 28, Proc. National Symp. On Proteins in Biology, Mysore, India
(1999).
13. Jagadeesha, D.K., Shashidhara murthy, R and Kemparaju, K. Isolation and
characterization of a non-toxic fibrinogenase from Indian cobra (Naja naja) venom.
48, Proc. National Symp. On Proteins in Biology, Mysore, India (1999).
14. Shashidhara murthy, R., Jagadeesha, D.K. and Kemparaju, K. Isolation,
characterization and chemical modification of histidine and lysine residues of a
neurotoxic phospholipase A2 from Indian cobra (Naja naja) venom. 55, Proc.
National Symp. On Proteins in Biology, Mysore, India (1999).
15. Kemparaju, K. and Gowda, T.V. Comparative characterization of phospholipases
A2 from Indian saw-scaled viper (Echis carinatus) venom. 324, 64th Annual Meeting
of SBC, Lucknow, India (1995).
16. Kemparaju, K. and Gowda, T.V. Selective neutralization of enzymatic activity of a
toxic phospholipase A2 from Echis carinatus venom by polyclonal antibodies, P7-82,
XVIth IUBMB meeting, India (1994).
17. Kemparaju, K. and Gowda, T.V. Purification and characterization of a toxic
phospholipase A2 (EC-PLA2-IV) from Indian saw-scaled viper (Echis carinatus)
venom. PEFD 019, 197, 62nd Annual Meeting of SBC, Madurai, Inida (1993).
18. Kemparaju, K. and Gowda, T.V. Preliminary studies on Indian saw-scaled viper
(Echis carinuatus) venom. Abstract No. 365, 61st Annual Meeting of SBC,
Hyderabad, India (1992).
19. Kemparaju, K., Kasturi, S. and Gowda, T.V. Effect of trypsin digestion on the
enzymatic and edema inducing activities of Vipera russelli phospholipase A2 A4-9,
15, Second Asia-Pacific congress on Animal, plant and microbial Toxins, Varanasi,
India (1990).
Resource person in academic programmes
1.Immune system: an overview.
Science and Technology Rejuvenation program for Pre-University Teachers organized by
Government Pre-University College Board, (2010) .
2. Mechanisms of signal transduction.
Refresher Course for Zoology Teachers. Organized by DOS in Zoology at Academic
Staff College, Univ. of Mysore, Mysore (2002).
3. Immunological techniques.
Refresher Course for Zoology Teachers. Organized by DOS in Zoology at Academic
Staff College, Univ. of Mysore, Mysore (2000).
4. SDS-PAGE; Detection and Analysis of proteins.
Work Shop on “Techniques in protein purification” in DOS in Biochem., July 15th to 24th
1999.
5. Proteins: structure-function relationships.
In-service Training Course in Chemistry for Postgraduate Teachers of Kendriya
Vidyalaya Sanghatan, Organized by KVS, New Delhi conducted at Regional Institute of
Education, Mysore (1997).
6. Fundamentals of Biochemistry- special emphasis on enzymes and metabolism.
In-service Training Course in Chemistry for Postgraduate Teachers of Kendriya
Vidyalaya Sanghatan, Organized by KVS, New Delhi conducted at Regional Institute of
Education, Mysore (1996).
Workshop/symposia conducted
1. Joint Secretary for Work Shop on “Techniques in protein purification” in DOS in
Biochem., July 15th to 24th 1999.
2. Treasurer, National Symposium on “Proteins in Biology: Structure/Function,
Expression and Specificity of Action” at Dos in Biochem. Univ. of Mysore, Mysore,
November 15th to 16th 1999.
3. Joint Secretary, 3`rd National symposium on venoms and Toxins” at Dos in Biochem.
Univ. of Mysore, Mysore, Jan 23 rd and 24th 2004.
4. Joint Secretary, International Conference on Cardiovascular Diseases Secondary to the
Metabolic Disorder: Mechanisms and Therapy. Dept. of Biochemistry, University of
Mysore, Mysore, Dec. 17 – 19,
External Reviewer for the Journals
Toxicon, Comparative Biochemistry and Physiology, Gene, Basic and Clinical
Pharmacology and Toxicology, Molecular and Cellular Biochemistry, Indian Journal of
Biochemistry and Biophysics, Chinese Journal of Medicine, Indian Journal of Medical
Research (ICMR)
Evidence Based complementary Medicine, African Journal of Agricultural Sciences
Memberships
Society of Biological Chemists, India, People for Animals.
Dr. K. S. Girish biodata
Name : Dr. Girish. K.S
Birth Date : June 22, 1976
Home Address : Kesturu, Krishnarajanagara
Taluk, Mysore-Dist,
Karnataka-571602, India
Citizenship : India
Postal Address : Dr. Girish. K.S
Assistant Professor,
DOS in Biochemistry,
Manasagangothri,
University of Mysore,
Mysore, India-560 006.
Email : [email protected]
Current address : DOS in Biochemistry,
University of Mysore, MGM
Educational Qualification:
Ph.D. Degree : Biochemistry (2004) University of Mysore
M.Sc. Degree : Biochemistry (1998) University of Mysore
B.Sc. Degree : BBM (1996) University of Mysore
POST-DOC : Virology/Tendon biology/Cancer biology (2004-
2007)
University of Pittsburgh/University of Virginia,
USA
Appointments and positions
Dates
Attended
Name and Location of Organization Rank/Title
1999-2004
Manasagangothri,
Mysore University, Mysore, India
Post-Graduate Teaching
Assistant
2004-2005 Department of Surgery
University of Pittsburgh
Postdoctoral Associate
2005- 2007 Department of Orthopedics
University of Virginia
Research Associate
2008
DOS in Biochemistry
University of Mysore, India
Assistant Professo
Peer-Reviewed Publications:
1. Girish KS, Hogan M, James R, Balian G, Hurwitz S, Chhabra A. Growth
differentiation factor-5 regulation of extracellular matrix gene expression in murine
tendon fibroblasts. J. Tissue Engineering and Regenerative Med 2010 (Accepted).
Impact factor: 1.6
2. Devaraja S, Girish KS, Devaraj VR, Kemparaju K. Factor Xa-like and
fibrin(ogen)olytic activities of a serine protease from Hippasa agelenoides spider venom
gland extract. J Thromb Thrombolysis 29, 119-126, 2010. Impact factor: 1.8
3. Ushanandini S, Nagaraju S, Chandra Nayaka S, Harish Kumar K, Kemparaju K,
Girish KS. The Anti-ophidian Properties of Anacardium occidentale bark extract.
Immunopharmacology and Immunotoxicology 31, 607-615, 2009. Impact factor: 0.9
4. Girish KS, Kemparaju K, Nagaraju S and Vishwanath BS. Hyaluronidase
Inhibitors: A biological and Therapeutic Perspective. Current Medicinal Chemistry
16, 2261-2288, 2009. Impact factor: 4.9
5. Sharma R, Mahadeshwara swamy YH, Harish kumar K, Devaraja S, Vishwanath
BS, Kemparaju K and Girish KS. Effect of anticoagulants on plasma hyaluronidase
activities. Journal of Clinical Laboratory Analysis 23, 29-33, 2009. Impact factor: 1.2
6. Devaraja S, Nagaraju S, Mahadeswaraswamy YH, Girish KS, Kemparaju K. A
low molecular weight serine protease: Purification and characterization from Hippasa
agelenoides (funnel web) spider venom gland extract. Toxicon 52, 130-138, 2008.
Impact factor: 2.45
7. James R, Kesturu GS, Balian G, Chhabra AB. Tendon: Biology, Biomechanics,
Repair, Growth Factors, and Evolving Treatment Options. American Journal of Hand
Surgery 33, 102-112, 2008. Impact factor: 1.6
8. Mahadeshwara swamy YH, Nagaraju S, Girish KS and Kemparaju K.
Neutralization of Saw-scaled viper (Echis Carinatus) venom induced local effects and
some enzymes by Mimosa pudica root extract. Phytotherapy Research 22, 963-969,
2008. Impact factor: 1.3
9. Nagaraju S, Girish KS, Fox JW and Kemparaju K. ‘Partitagin’ a hemorrhagic
metalloprotease from Hippasa partite spider venom: Role in tissue necrosis Biochimie
89, 1322-1331, 2007. Impact factor: 3.1
10. Girish KS and Kemparaju K. The magic glue hyaluronan and its eraser
hyaluronidase: a biological overview. Life Sciences 80 1921-1943, 2007. Impact factor:
2.6
11. Girish KS, Deepa M, Ushanandini S, Harish Kumar K, Nagaraju S, Govindappa
M, Vedavathi M and Kemparaju K: Antimicrobial properties of a non-toxic
glycoprotein (WSG) from Withania somnifera (Ashwagandha). Journal of Basic
Microbiology 46 (5) 365-374, 2006. Impact factor: 0.8
12. Nagaraju S, Mahadeshwaraswamy YH, Girish KS and Kemparaju K:
Biochemical and pharmacological characterization of venom from the spiders H. partita,
H. aglenoides and H. mahabaleshwarensis tikadar and malhotra. Communicated to
Comparitive Biochemistry and Physiology (Part C) 144 (1), 1-9, 2006. Impact
factor: 2.4
13. Ushanandini S, Nagaraju S, Harish Kumar K, Vedavathi M, Deepa M, Kemparaju
K, Vishwanath BS, Gowda TV, Girish KS: The anti snake venom properties of
Tamarindus indica (Leguminosae) seed extract. Phytotherapy Research 20 (10) 851-
858, 2006. Impact factor: 1.3
14. Deepa K. Machiah, Girish KS and Gowda TV: A glycoprotein from a folk
medicinal plant, Withania somnifera, inhibits hyaluronidase activity of snake venoms.
Comparative Biochemistry and Physiology (Part C) 143, 158-161, 2006. Impact
factor: 2.4
15. Girish S Kesturu, Colleton BA, Liu Y, Heath L, Shakil OS, Rinaldo Jr CR,
Shankarappa R: Minimization of genetic distances by the Consensus, Ancestral and
Center-of-Tree (COT) sequences for HIV-1 variants within an infected individual and
the design of reagents to test immune reactivity. Virology 348 (2) 437-448, 2006.
Impact factor: 3.2
16. Girish KS and Kemparaju K: The hyaluronidase enzyme a prime target for better
management of snakebite. Life sciences 78, 1433-1440, 2006. Impact factor: 2.6
17. Kemparaju K and Girish KS: Snake venom hyaluronidase: s therapeutic target.
Cell Biochemistry and Function 24 (1), 7-12, 2006. Impact factor: 1.5
18. Vedavathi M, Girish KS and Karunakumar M: A novel low molecular weight
alanine aminotransferase from fasted rat liver. Biochemistry (Moscow) 71 (Suple 1),
S105-S112, 2006. Impact factor: 1.2
19. Girish KS and Kemparaju K: Inhibition of Naja naja venom hyaluronidase by
plant derived bioactive components and polysaccharides. Biochemistry (Moscow) 70
(8), 948-952, 2005. Impact factor: 1.2
20. Girish KS and Kemparaju K: A low molecular weight isoform of hyaluronidase:
purification from Indian cobra (Naja naja) venom and partial characterization.
Biochemistry (Moscow) 70 (6), 708-712, 2005. Impact factor: 1.2
21. Vedavathi M, Girish KS and Karunakumar M: Isolation and characterization of
alanine aminotransfarase isoforms from starved male rat liver. Molecular and Cellular
Biochemistry 267, 13-23, 2004. Impact factor: 1.56
22. Girish KS, Mohan Kumari HP, Nagaraju S, Vishwanath BS and Kemparaju K.
Hyaluronidase and Protease activities from Indian snake venoms: Neutralization by
Mimosa pudica root extract. Fitotherapia, 75 (3-4), 378-380, 2004. Impact factor: 1.25
23. Girish KS, Shashidhara murthy R, Nagaraju S, Veerabasappa Gowda TV and
Kemparaju K. Isolation and characterization of hyaluronidase a “spreading factor” from
Indian cobra (Naja naja) venom. Biochimie, 86(3), 193-202, 2004. Impact factor: 3.1
24. Girish KS, Jagadeesh DK, Rajeev KB and Kemparaju K., Snake venom
hyaluronidase: An evidence for isoforms and extracellular matrix degradation.
Molecular and Cellular Biochemistry, 240, 105-110, 2002. Impact factor: 1.56
25. Jagadeesh DK, Shashidharamurthy R, Girish KS and Kemparaju K. A non-toxic
anticoagulant metalloprotease: purification and characterization from Indian cobra
(Naja naja naja) venom. Toxicon 40 (6), 667-675, 2002. Impact factor: 2.45
26. Shashidharamurthy R, Jagadeesh DK, Girish KS and Kemparaju K. Variations
in biochemical and pharamacological properties of Indian cobra (Naja naja naja) venom
due to geographical distribution. Molecular and Cellular Biochemistry, 229, 93-101,
2002. Impact factor: 1.56
Book Chapters:
Kemparaju K, Girish KS and Nagaraju S. Hyaluronidases, a neglected class of
glycosydases from snake venom: beyond a spreading factor in a book “ Hand Book of
Venoms and Toxins of Reptiles” (Cat # 9165), August 2009, CRC Press LLC.
Hemshekhar M, Kemparaju K, and Girish KS. An overview on remedial qualities of
Tamarindus indica seeds. Book Chapter in “Nuts and Seeds in Health and Disease”
Edited by Victor R Preedy, Academic Press-Elsevier, 2010.
National/International Symposia Presentation:
1. Girish KS, James R, Park A, Hogan MV, Balian G, Chhabra AB. Expression
of Matrix and Tendon Specific Cellular Markers by Rat Adipose Derived
Mesenchymal Stem Cells Treated with Growth Differentiation Factor-5. International
Conference on Anatomical and Cell Biology Sciences. National University of
Singapore, Singapore, May 26th to May 30th 2010.
2. Mahadeswraswamy YH, Girish KS and Kemparaju K. Local toxicity of
Indian Daboia/Vipera russellii venom and its purified hemorrhagic complex:
Contribution for the better management. National conference on Medical
Biotechnology and Clinical research. October 2009. Department of Biotechnology, Sri.
M. Visvesvaraya Institute of Technology and Sri Krishnadevaraya Education Trust, JN
Tata Auditorium, Indian Institute of Science campus, Bangalore, India. (Best poster
presentation award, second prize).
3. Hemshekhar, Sunitha K, Avinash S, Mouna N, Deepthi L, Soumya N, Girish
KS. Guggul proteins: A group of neglected bioactive molecules. National symposium
on “Bioactive molecules: from Discovery to Industry”. Organized by DOS in
Biochemistry, University of Mysore, Manasagangothri, April 2009, Mysore
4. Girish KS, James R, Balian G and Chhabra A. GDF-5 modulates the
synthesis and expression of extracellular matrix and cell adhesion related molecules of
rat Achilles tendon fibroblasts. 54th Orthopedic Research Society meeting, San
Francisco, CA, USA. Feb 2008
5. Hogan MV, Starnes T, Kesturu Girish, James R, Balian G, Chhabra A. Cell-
based Augmentation of Tendon Repair Using a Biodegradable Polymer Scaffold. 2008
AAOS Annual Meeting, San Francisco, CA. (Accepted Podium)
6. Girish KS, Roshan J, Balian G, Chhabra A: Treatment with GDF-5 induces
tendinogenic differentiation of adipose derived stromal cells. 53rd Orthopedic Research
Society meeting, San Deigo, CA, USA. March 2007
7. Hogan MV, Starnes T, Kesturu Girish, James R, Huang D, Balian G,
Chhabra A. GDF-5 Induced Tendon Repair and Regeneration. 2007 National Medical
Association Annual Convention and Scientific Assembly, Honolulu, HI (Podium)
8. Kwoon M, Girish KS, Meyers T and Diduch D: Intra-articular
chondroprotection against septic arthritis with the use of adenosine agonist (ATL-313).
53rd Orthopedic Research Society meeting, San Deigo, CA, USA. March 2007
9. Trevor S, Huang D, Girish KS, Balian G and Chhabra A: Immunolocalization
of GDF-5 in native and surgically repaired Rat Achilles Tendon. 53rd Orthopedic
Research Society meeting, San Deigo, CA, USA. March 2007
10. Trevor S, Huang D, Girish KS, James R, Balian G and Chhabra A:
Potentiation of tendon repair and regeneration. Annual meeting of orthopedic surgeons
(AAOS). San Deigo, CA, USA. March 2007
11. Meyers T, Girish KS, Kwoon M and Diduch D: Intravenous versus intra-
articular chonroprotection against septic arthritis with the use of an adenosine agonist
(ATL-313). 59th Orthopedic annual Virginia Orthopedic Society meeting, Norfolk,
Virginia, USA May 19th to May 21st 2006
12. James R, Huang D, Girish KS, McCulloch MD, O’Neal BL, Balian G,
Chhabra AB: GDF-5 improves collagen fiber organization and increases extracellular
matrix synthesis during tendon repair. 52nd Orthopedic Research Society meeting,
Chicago, IL, USA. March 19th to March 25th 2006
13. Girish Kesturu*, LianFu Wang, Charles R. Rinaldo, Jr., and Raj
Shankarappa: Quantitative Considerations in the Assessment of Hepatitis C Virus
Quasispecies. 3rd Annual Richard L. Simmons Lecture in Surgical Science and
Department of Surgery Research Day. University of Pittsburgh, Pittsburgh, USA. April
27th 2005
14. Girish KS and Kemparaju K. The hyaluronidase enzyme a prime target in
management of snakebite. 3rd National symposium on venoms and toxins. University of
Mysore, Mysore, India, Jan 23th to Jan 26th 2004
15. Nagaraju S, Girish KS and Kemparaju K. Comparative characterization of
three Indian funnel-web spider venoms. 3rd National symposium on venoms and toxins.
University of Mysore, Mysore, Mysore, India, Jan 23th to Jan 26th 2004
16. Dhananjaya BL, Girish KS and Cleatus DM Desouza. Partial characterization
of 5’ nucleotidase from Indian cobra (Naja naja) venom. 3rd National symposium on
venoms and toxins. University of Mysore, Mysore, Mysore, India, Jan 23th to Jan 26th
2004
17. Vedavathi M, Girish KS and Karunakumar M. Comparative characterization
of low and high molecular weight isoforms of alanine aminotransfarases from starved
rat liver. 3rd National symposium on venoms and toxins. University of Mysore, Mysore,
Mysore, India, Jan 23th to Jan 26th 2004
18. Kemparaju K and Girish KS. A pharmacological examination of the Indian
saw scaled viper venom. National Symposium on Proteins (Structure and Function).
Mysore, India, 1999
External Reviewer
Toxicology, Biochimie, Connective tissue research, Journal of Medicinal Chemistry,
Biomaterials, and Pesticide, Biochemistry and Physiology
Number of papers reviewed: 12
Dr.B.G.Sudharshan Biodata
PERSONAL PROFORMA
NATIONALITY : Indian D.O.B. : 18-04-1975 GENDER : MALE LANGUAGES KNOWN : English, Kannada, and Hindi ACADEMIC COURSE : MBBS, PhD (Bio-medical engineering)
ACADEMIC PERFORMANCES:
Course University Institution class Percentage
PhD Avinashlingam university , Coimbatore
CMRTU , R.V.C.E (MAY2007TO DEC 2009).
MBBS Bangalore University SRI SIDDARTHA MEDICAL COLLEGE TUMKUR
PASS
PHD THESIS TITLED:
“Electromagnetic and RF Interference on the Implanted Cardiac Pacemakers and Remedial Measures for their Effective Functioning”
PUBLICATIONS:
� Interference with Cardiac Pacemaker due to Cellular Phones, International Journal of
Computer Applications in Engineering, Technology and Science, issue: April 2009,
pp.289-291
� A Review on Electromagnetic Interference on Cardiac pacemakers, presented and
published at International conference on Emerging Technologies and Applications in
Engineering, Technology and Sciences, held on 13-14 January, 2008
� Electromagnetic Interference due to Mobile Phones on Artificial Cardiac Pacemaker
Patients, XXXII National Systems Conference, 03-11-2008, at Roorkee
� Heart rate variability ;A review paper to be presented in an international conference
in 2011 International Conference on Signal Processing, Communication,
Computing and Networking Technologies to be held on 21-22 july2011
� HRV under stress using two different methods of non-linear analysis in a national
conference held at Dr.AIT WIRELESS CONTROL AND Communication
technology on apr28-29.
� Understanding autonomic and dynamics under stress using non-linear analysis of
data page no page no152-155 National conference on emerging trends in bio-
medical signal processing 26feb -2011
WORK EXPERIENCE: 1. ONE YEAR ROTATORY INTERNSHIP IN DISTRICT HOSPITAL TUMKUR;
• Attending opd
• Emergencies
• Community health programmes
• Serving rural people
• Regular concerned department posting 2. RESIDENT DOCTOR DEPARTMENT OF NEUROLOGY MANIPAL
HOSPITAL;
• Attending neuro opd
• Treating patients in wards in absence of specialists
• Performing procedures like lumbar punctures etc .3. RESIDENT DOCTOR DEPARTMENT OF ICU BANGALORE HOSPITAL;
• Treating critically ill patients
• Treatment terminally ill patients
• Performing procedures like intubation etc 4. MEDICAL OFFICER MINISTRY OF LABOUR, GOVT OF INDIA;
• Regular opd
• Handling emergencies
• Had a mobile dispensing unit where in patients were treated at there working place
• Implementation of national programmes such as immunisation, vaccination 5. MEDICAL OFFICER R.V.C.E TILL TO DATE;
• Treating patients on opd and day care basis
• The strength of students ranges from 1200 in hostels and 4500 students on campus and staff members 450 and their families
• Maintaining hygiene and health of all these people
• Regular vaccination for students mainly hepatitis-a .hepatitis-b and chicken-pox vaccines
• Regular medical checkups for students and food handlers
• Attending emergencies during night time
• Shifting of patients to the nearby tertiary care centres and taking care of them till they are discharged
• Counseling for students
• Arranging health camp for students mainly eye check-up camps
• Oral hygiene camp annually POSITION HELD AT RVCE:
1. MEDICAL OFFICER IN RVCE Health centre and hostel Health centre.
2. ASSISTANT PROFESSOR IN Biomedical Instrumentation Signal Processing Department.
3. RESEARCH AND ACADEMICS; Guiding students of M-tech biomedical instrumentation and engineering for
research projects namely; � Heart rate variability on stress induced subjects by using non-linear
methods. � Heart rate variability in hypothyroid patients. � Introduction classes for bio-medical instrumentation students. � Handling theory classes such as Physiology for Engineers.
FUNDED PROJECTS:
1. As co-investigator, in a project titled insilico analysis of bone-tumors funded by DST.
2. Projects submitted for various funding agencies;
• Synthesis and characterization of plasmin and their thrombolytic applications submitted as CO-PI to ICMR.
• Development of Doppler based foetal ultrasound stethoscope submitted to DBT as CO-PI.
• Development of a healthcare monitoring device for early detection of cardio-vascular diseases. As principal investigator to DBT
3. An ergonomic study of working conditions in garment industry and their health complications and remedial measures. Submitted to ICSSR.
Thippeswamy.T.G. Biodata
1. Name : T.G.THIPPESWAMY 2. Date of Birth : 28.06.1971 3. Designation : Assistant Professor
4. Department : Department of Biochemistry 5. Residential Address : c/o Ravisha, Nalanda convent Road, Tumkur. 6. Phone Number: Office: Mobile: 9663417864 7. Email address : [email protected] 8. Personal Homepage:
9. Academic Qualification
Degree University Year of Passing Specialization (if any)
B.Sc Kuvempu 1995 Chemistry, Botany, Zoology
M.Sc Kuvempu 1997 Biochemistry
NET CSIR- UGC 2002 Biochemistry
10. Teaching / Industrial Experience
Teaching and research activities in Biochemistry
Designation Teaching/ Research
Experience
Duration Name of the organization
Technical Assistant
Fish nutrition and extension work
14.12.1998 - 14.01.2003
Central Marine Research Institute (CMFRI), Chennai (Tamil Nadu), Kochi (Kerala), India.
Senior Technical Assistant
Micronutrient Research
15.01.2003 - 08-04-2008
National Institute of Nutrition (NIN), Hyderabad, Andhrapradesh, India.
Technical Officer
Micronutrient Research
08.04.2008 - 24.06.2010
National Institute of Nutrition (NIN), Hyderabad, Andhrapradesh, India.
Assistant Professor
Biochemistry 25.06.2010 - Till date
University College of Science , Tumkur University,Tumkur, Karnataka.
1. Standardization and estimation of ascorbic acid in diets by High
Performance 2. Liquid Chromatography (HPLC) coupled with Electrochemical Detector
and UVdetector.
3. Proximate composition analysis in food 4. Standardization and estimation of 5-methyltetraydrofolate and ascorbic
acid using Dried Blood Spot technique from finger puncture blood samples by HPLC Method.
5. Purification and characterisation of protein
Publications:
1. Pullakhandam R, Nair, MK, Kasula S, Kilari S, Thippande TG: Ferric reductase
activity of low molecular weight human milk fraction is associated with enhanced
iron solubility and uptake in Caco-2 cells. Biochem Biophys Res Commun.2008
374(2):369-72. Impact Factor : 2.72
Papers National Conferences, Seminars, Symposia:
1. T.G.Thippeswaamy: Poster presentation title on “5- Methyl tetrahydofolate in
Dried Blood Spot as an Indicator of Folate status in Humans” at 36th annual
conference of Association of Clinical Biochemists of India (ACBICON 2009),
Kochi.
2. T.G.Thippeswaamy: Oral presentation title on “Stability of Ascorbic acid in
vegetables and fruits under different conditions of temperature and storage” at
Knowledge Utsav, Jain University, Global Campus on 28th August, 2010 at
Bangalore.
3. T.G.Thippeswamy: Poster Presentation entitled “Measurement of Iodine Value in
Vegetable oils: Stability of Shark liver oil” National Level one day conference on
“Recent Trends in Food Science &Nutrition Research ” organised by the Centre
for Nano Science Research in association with Dr.P.Sadananda Maiya Centre for
Food Science and Research, Bangalore and Jain University, Bangalore on15th
December, 2011.
4. T.G.Thippeswamy: Abstract entitled “Role of dihydrofolate reductase and folate
conjugase on folic acid absorption in gestational diabetes, arthritis and
pregnancy induced hypertension: Implication on neural tube defects” National
Level one day conference on “Recent Trends in Food Science &Nutrition
Research ” organised by the Centre for Nano Science Research in association with
Dr.P.Sadananda Maiya Centre for Food Science and Research, Bangalore and Jain
University, Bangalore on15th December, 2011.
5. T.G.Thippeswamy and D.Manjunatha: Abstract submitted entitled
“Nanobiosensors for Detection of Agricultural and food contaminants “at the
Conference on “Social Relevance of Nano Materials and Applications An
Interdisciplinary Approach” organised by the Centre for Nano Science Research
in association with Higher Education Department , Government of Karnataka,
Bangalore on31st December, 2011
6. D.Manjunatha and T.G.Thippeswamy: Abstract submitted entitled “Nanosensors: A
Technology for Biomedical Applications” at the conference on “Social Relevance of
Nano Materials and Applications an Interdisciplinary Approach” organised by the
Centre for Nano Science Research in association with Higher Education Department,
Government of Karnataka, Bangalore on31st December, 2011.
7. T.G.Thippeswamy, P.Raghu, K. Sreenivasalu and K.Madhavan Nair: Abstract
submitted entitled “Purification of iron binding low molecular weight human milk
factor” at the conference on “ Perceptive on health benefits of therapeutic
molecules” organized by Centre for Bioscience and Innovation, Tumkur University
and Karnataka state higher education council, Bangalore on 06-01-2012
8. Sowmyashree, Bhavana, T.G.Thippeswamy, M.Bhagyalaksmi and Dr.S.Devaraja:
Abstract submitted entitled “Anticoagulant properties of endocarp extract of
swietenia Macrophylla and Thespesia populnea sap” at the conference on
“Perceptive on health benefits of therapeutic molecules” organized by Centre for
Bioscience and Innovation, Tumkur University and Karnataka state higher education
council, Bangalore on 06-01-2012
Conferences, Seminars, Symposia, Workshops participated:
International events: 1. Attended International seminar on “Bangalore India Bio-2011” at Bangalore
International Exhibition Centre on May, 4-6,2011
National events: 1. Attended National Level Seminar on “Reforms in Higher Education and Public
Participation” organised by Karnataka State Higher Education Council,
Government of Karnataka, June 18th, 2011
2. Attended 5th National Conference of Ancient Sciences and Archaeology organised
by Post Graduate Department of Studies and Research in History & Archaeology,
Centre for public History and Archaeology , Tumkur University and Ancient
Sciences and Archaeological Society of India on 22- 23, July,2011.
3. Attended two days conference on “Biodiversity and Sustainable development” at
Siddaganga College of Arts, Science and Commerce on 20-21, August, 2011.
4. Attended National Level one day workshop on “Biotechnology for Health and
Environment in Future” organised by the Department of Biotechnology on
Saturday, 22 October, 2011 at Shridevi Institute of Engineering & Technology, Sira
Road, Tumkur- 572106.
5. Attended 28 days “Orientation Course” Batch No.102 conducted by UGC-
Academic Staff College ,University of Madras, Chennai- 600 005 from 02-09-2011
to 29-09-2011.
6. Participated in three days National conference on “VEDANTA: A HOLISTIC
APPROACH” at Tumkur University in association with Vedanta Bharathi,
K.R.Nagara,Mysore from 26th ,27th and 28th December,.2011.
7. Participated three days workshop on “Training on Research process and Writing a
scientific paper” sponsored by Indian Council of Medical Research(ICMR) , New
Delhi and Rajiv Gandhi University of Health Sciences(RDUHS) , Bangalore held
during 11th to 13th February 2012 at Sree Siddaganga College of Pharmacy,
B.H.Road, Tumkur-572102, Karnataka.
Regional events:
1. Attended State level one day workshop on “Recent scenario on biogas from food
wastes” organized by Department of Biotechnology , ShriShridevi institute of
engineering and technology ,10th march 2011,Tumkur-02
2. Attended 2 days work shop on “Handling of analytical instruments – HPLC,
GC, AAS, and UV-Spec & PCR” at Azyme Biosciences Private Limited,
Bangalore on 28, 29 November, 2010.
3. Attended “Inter university subject based conference on Bioscience and
Bioinformatics” on 23-24 May, 2011 at Tumkur University, Tumkur.
4. Attended State Level Conference on “Frontiers of Nanotechnology” organised
by Karnataka State Higher Education Council, Government of Karnataka, 31st
May, 2011
5. Attended “Inter university subject based conference on Chemistry and
Biochemistry” on 10-11June, 2011 at Central College, Bangalore University,
Bangalore.
6. Attended Seminar on “GM Crops –Future Perspectives” organised by Tumkur
University 16th July, 2011
7. Attended 3 days “Faculty development program” conducted at Jnana Sahydri,
Kuvempu University, Shankaragatta, Shimoga from 03-03-2011 to 05-03-2011 in
association with VGST and Colligate Education, Govt. of Karnataka.
8. Attended workshop on “Multi disciplinary project proposals” on 09 June, 2011 at
Tumkur University, Tumkur.
Events Organized (conferences, seminars, symposia, workshops, etc)
1. Work shop conducted on “Biochemical techniques” at Department
of PG Studies & Research in Biochemistry, Tumkur University, Tumkur.
2. Local organising committee member for one day national seminar
on “Recent Trends in Food Science &Nutrition Research ” organised by the
Centre for Nano Science Research in association with Dr.P.Sadananda Maiya
Centre for Food Science and Research, Bangalore and Jain University, Bangalore
on15th December, 2011.
3. Local organizing committee member for one day national conference on
“Perceptive on health benefits of therapeutic molecules” organized by Centre
for Bioscience and Innovation, Tumkur University and Karnataka state higher
education council, Bangalore on 06-01-2012
Membership of Professional Associations, academic bodies, etc.
Life member ship: Nutrition Society of India (NSI), National Institute of Nutrition, Hyderabad.
Bhagyalakshmi.M biodata
1. Name of the applicant : BHAGYALAKSHMI.M
2. Mailing address : Assistant Professor,
Department of Studies in Biochemistry
Tumkur University,Tumkur,Karnataka,
Mobile No: 9481343477
Email: [email protected]
3. Date of Birth : 16-08-1981
Academic Qualification:
Degree University Year of Passing Specialization
B.Sc Kuvempu University 2004 Biochemistry,
Microbiology, Botany
Teaching: 2006-2010- Worked as a Lecturer in the Department of Biochemistry, Kuvempu University. Projects Guided: More than 15 M.Sc students for their project work in various topics of Biochemistry. National Conferences, Seminars, Symposia:
1. Bhagyalakshmi. M and Chinmayee(2010):“Evaluation of tioxidantproperties of E. officinalis, T. chebula and T. bellirica berries”. Knowledge Utsav” National Conference. 28th august,2010. Conferences, Seminars, Symposia, Workshops participated: International events:
1. Bangalore INDIA BIO-2011, at Bangalore International Exhibition Centre(BIEC)
from 04 to 06 May 2011 organized by the Department of Information Technology,
Biotechnology and Science & Technology, Government of Karnataka, Vision Group
on Biotechnology.
National events:
1. Three days national conference on “VEDANTA; A holistic approach” organized
by Tumkur university and Vedanta Bharathi, on 26 to 28 dec 2011.
2. UGC sponsored national level conference on “Biodiversity and sustainable
development ”organized by Sree Siddaganga college on 20 to 21 Aug 2011
3. One day national seminar on “ Reforms in Higher Eduction and Public
participation” organized by Karnataka state higher education council ,on 18
June 2011
4. International symposium on” challenges in drug discovery program -2011
(ISCDDP)”, February 16, 17 in Mysore.
5. State level one day workshop on “Recent scenario on biogas from food
wastes” organized by Department of Biotechnology ,SriShridevi institute of
engineering and technology ,10th march 2011,Tumkur-02
6. “Knowledge Utsav” National Conference organized by Jain University and
Tumkur University, Global Campus, Kanakapura Taluk, Bangalore Rural,
Karnataka. On 28th august 2010.
7. National seminar on “Recent advances and challenges in Biochemistry and
Biotechnology” organized by the Department of Biochemistry, Jnansahyadri,
Kuvempu university, shankaraghatta, Shimoga. On 01, 02 April, 2009
M.Sc Kuvempu University 2006 Biochemistry
8. National seminar on “ Emerging areas in Biomedical Sciences” organized by
Department of Biochemistry, P.G Centre, Shivagangotri, Davangere, on 29 march
2008.
9. National seminar on “ Advances in Biochemistry, Biotechnology and
Nanotechnology Science for 21st Century” organized by Department of
Biochemistry, P.G Centre, Shivagangotri, Davangere, on Nov. 13, 14, 2006.
10. National seminar on “Trends and Technologies in the Biochemical sciences”
organized by Department of Biochemistry, P.G Centre, Shivagangotri,
Davangere, on 10, 11 Feb. 2006
Regional events:
1. One day workshop on formulation vision documentation of strategic road map of
Tumkur university on 14-7-2011
2. One day workshop on formulation of interdisciplinary projects organized by
Tumkur university on 09-07-2011
3. Second State level Science and Technology conference 2011, organized by
Karnataka state council for Science and Technology, Vision Group for Science
and Technology and Department of Science & technology, Govt. of Karnataka, on
26th to 28th may 2011.
4. One day workshop on current scenario on production of Biogas from food wastes
organized by Shridevi Institute of Engineering and Technology, Tumkur-02,on
10-003-2011.
5. Workshop on ‘‘Recent scenario in SCAR’’ (stem cells, cancer, AIDS and Raman
spectroscopy) organized by Department of Biotechnology, Shridevi institute of
engineering and technology, Tumkur-02 on 29-09-2010.