Qualification: 
Ph.D
sanjaypal@am.amrita.edu

Research Interests

We are trying to understand the biology of the extracellular matrix (ECM) in health and disease relevant to India with special reference to malnutrition. Focus is on cell adhesion protein fibronectin and its interaction with different cell surface proteins from both host (MMP-2, heparin, collagen) and guest [fibronectin/collagen binding proteins (FBP /CBP) from microbes]. The broad themes are:

  • Matrix binding proteins from probiotics and viruses:
    We are screening different potential probiotic microbes from popular and affordable fermented food and beverages, which bind to denatured collagen (gelatine) and fibronectin. The goal is to design better probiotics, peptides drugs which will compete against the host -pathogen interaction at cell surface and also to design bacteriophages which will compete/kill the enteric pathogens (Sanitation Biotechnology).

  • Characterization of Fibronectin isoforms and proteolytic fragments affecting cell behaviour (cell morphology, migration, proliferation)
     
  • Factors affecting gelatinase (MMP-2) mediated fragmentation of fibronectin
     

dr-sanjay-pal-amrita-research-fibronectin-interaction-proteins-school-biotechnology-faculty

Qualification

  • School: Ramakrishna Mission, Narendrapur
  • BSc & MSc: Visva Bharati, Santiniketan
  • MTech & PhD: IIT Kharagpur
     

Keywords: Plant Tissue Culture, Matrix Biology, Malnutrition, Sanitation, Bacteriophages, Wastewater Microbiome, Biocontrol

Projects

  1. Development of bacteriophages and lytic agents to target and kill pathogens and odour-producing bacteria in fecal waste / Funded by DBT-BIRAC-BMGF (Govt of India - Bill & Melinda Gates Foundation)
  2. An innovative green technology for treating municipal and industrial wastewater entering rivers and streams. IC-IMPACTS, Canada & DBT, Govt of India.
  3. Characterization of the fibronectin isoforms and its fragments. Funded by Amrita Vishwa Vidyapeetham.
  4. Matrix Binding microbes and bacteriophages to counter infection. Funded by Amrita Vishwa Vidyapeetham.

Awards and Scholarships

  1. Recipient of Bill and Melinda GATES Foundation – Department of Biotechnology, GOI/BIRAC Grand Challenge Award, March 2014
  2. Post-doctoral Fellowship, University of Texas Health Science Center at San Antonio, Texas, USA with Prof. Bjorn Steffensen (2006-2010)  and Prof. Goutam Ghosh Choudhury (2010).
  3. Post-doctoral Fellowship, University of Guelph, Canada (2005) with Prof Praveen K. Saxena.
  4. Research Fellowship at Borstel Research Centre, GERMANY with Professor Dr. Ulrich Zähringer (2001).
  5. Council of Scientific & Industrial Research (CSIR) -Research Associate (2003).
  6. State Level Eligibility Test (SLET) conducted by West Bengal College Service Commission for Lectureship. (1996).
  7. CSIR – National Eligibility Test (NET) for Research Fellowship and Eligibility for Lectureship in Indian Universities (1996).
  8. Fellowship from IIT, Kharagpur (1997-98) and CSIR (1998-2004), Govt. of India for graduate (MTech & PhD) studies.
  9. Qualified Graduate Aptitude Test in Engineering (GATE) (1995). Percentile: 97.77 (All India rank: 58).
  10. Govt. of India Merit Scholarship (1983-1989)
     

Teaching

  1. BIO317/Research Methodology/B.Sc. Biotechnology and B.Sc. Microbiology/2 Credits/Semester 5
  2. BIO317/Project/B.Sc. Biotechnology and B.Sc. Microbiology/10 Credits/Semester 6
  3. BIO401/Molecular Biology/M.Sc. Biotechnology and M.Sc. Microbiology/3 Credits/Semester 1
  4. BIO408/Recombinant DNA Technology/M.Sc. Biotechnology and M.Sc. Microbiology/3  Credits/Semester 2
  5. BIO481/Recombinant DNA Technology Lab/M.Sc. Biotechnology and M.Sc. Microbiology/2  Credits/Semester 2
  6. BIO517/ Plant Biotechnology/ M.Sc. Biotechnology /3 Credits/ Semester 2
  7. BIO588/Cell & Molecular Biology Lab/M.Sc. Biotechnology and M.Sc. Microbiology/2  Credits/Semester 3
  8. BIO599/Project/M.Sc. Biotechnology and M.Sc. Microbiology/10 Credits/Semester 4
     

Publications

Publication Type: Journal Article

Year of Publication Title

2018

A. Vijayakumar, Ajith Madhavan, Chinchu Bose, Nanjan Pandurangan, Sindhu Shetty K., Archana Palillamvedu, Megha Prasad, Dr. Sanjay Pal, and Dr. Bipin G. Nair, “Potent Chitin Synthase Inhibitors from Plants”, Current Bioactive Compounds, vol. 14, 2018.

2018

C. Porayath, M. K. Suresh, Dr. Raja Biswas, Dr. Bipin G. Nair, Dr. Nandita Mishra, and Dr. Sanjay Pal, “Autolysin mediated adherence of Staphylococcus aureus with Fibronectin, Gelatin and Heparin.”, International Journal of Biological Macromolecules, vol. 110, pp. 179-184, 2018.[Abstract]


Major autolysin (Atl) of Staphylococcus aureusis a cell surface associated peptidoglycan hydrolase with amidase and glucosaminidase domains. Atl enzymes (amidase and glucosaminidase) are known to participate in biofilm formation and also can bind with host matrices. Earlier studies demonstrated the binding of Atlwithfibronectin, thrombospondin 1, vitronectin and heat shock cognate protein Hsc70. Here, we have shown, Atl mediates attachment of S.aureus to heparin and gelatine as well. The atl mutant strain demonstrated around 2.5 fold decreased adherence with fibronectin, gelatin and heparin coated microtiter plates. The microscopic studies confirmed the reduced binding of atl mutant with them compared to its parental wild type and complemented mutant strains. Amidase and glucosaminidase were expressed as N-terminal histidine tagged proteins from Escherichia coli, purified and refolded. We found refolded amidase bind with fibronectin, gelatin and heparin; whereas refolded glucosaminidase binds with only fibronectin and heparin but not gelatin. These results reemphasize Atl as one of the crucial proteins from Staphylococcus that facilitate their binding with multiple host cellular components during colonization and infection.

More »»

2018

Vidhya Prakash, H. Sreetha, K. H. Poornima, K. N. Lakshmimol, R. Regma, H. Fathima, T. V. Vishnu, S. Venu, Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Antagonistic Effects of Bacteriocins from Lactobacillus Spp. against Enteric Pathogens”, Pollution Research Paper, vol. 37, pp. 128-134, 2018.[Abstract]


Bacteriocins from lactic acid bacteria (LAB) are natural antimicrobial peptides or proteins with potential applications in food preservation and healthcare. The study aimed at screening and characterization of bacteriocins from the secretome of five Lactobacillus strains. We were able to screen the antagonistic potential of two strains, L. fermentum and L. casei 335 against the multidrug resistant pathogens, Escherichia coli MDR (multidrug resistant) and Salmonella typhi isolated from sewage. Different dialysate fractions of the Lactobacillus strains were screened by soft agar diffusion and microtitre inhibition assay. Dialysate fractions from ammonium sulfate precipitate, at 65% (LF 65) and at 90% (LF 90) from L. fermentum were tested. LF60 (40μg/mL) was found to be inhibitory to the growth of E. coli MDR strain. Minimum Inhibitory Concentrations (MIC) against biofilm formation were found to be 40μg/mL for both E. coli MDR and Salmonella typhi, though inhibition was found at sub MIC of 20 μg/mL. Biofilm dispersal inhibition of E. coli MDR and S. typhi demonstrated a percentage inhibition rate of 50%. For initial characterization of the proteome of LAB strains, Tricine SDS-PAGE was performed which yielded a protein band of molecular size of 11 kDa from LF 60 fractions. Gel overlay assay showed an inhibitory effect on growth of E. coli MDR, indicating the presence of bacteriocin activity. Caenorhabditis elegans was employed as a nematode model to study the in vivo protection against E. coli MDR. Significant enhanced survival (91%) of the nematode was observed on treatment with LF 60. The efficacy of these partially purified preparations in inhibiting enteric pathogens showcase a prospective strategy of application of these antimicrobials in food and other environmental applications where cost of purification is of concern.

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2014

A. Bera, Das, F., Ghosh-Choudhury, N., Li, X., Gorin, Y., Kasinath, B. S., Abboud, H. E., Choudhury, G. Ghosh, and Dr. Sanjay Pal, “A positive feedback loop involving Erk5 and Akt turns on mesangial cell proliferation in response to PDGF”, American Journal of Physiology-Cell Physiology, vol. 306, pp. C1089–C1100, 2014.[Abstract]


Platelet-derived growth factor BB and its receptor (PDGFRβ) play a pivotal role in the development of renal glomerular mesangial cells. Their roles in increased mesangial cell proliferation during mesangioproliferative glomerulonephritis have long been noted, but the operating logic of signaling mechanisms regulating these changes remains poorly understood. We examined the role of a recently identified MAPK, Erk5, in this process. PDGF increased the activating phosphorylation of Erk5 and tyrosine phosphorylation of proteins in a time-dependent manner. A pharmacologic inhibitor of Erk5, XMD8-92, abrogated PDGF-induced DNA synthesis and mesangial cell proliferation. Similarly, expression of dominant negative Erk5 or siRNAs against Erk5 blocked PDGF-stimulated DNA synthesis and proliferation. Inhibition of Erk5 attenuated expression of cyclin D1 mRNA and protein, resulting in suppression of CDK4-mediated phosphorylation of the tumor suppressor protein pRb. Expression of cyclin D1 or CDK4 prevented the dominant negative Erk5- or siErk5-mediated inhibition of DNA synthesis and mesangial cell proliferation induced by PDGF. We have previously shown that phosphatidylinositol 3-kinase (PI3-kinase) contributes to PDGF-induced proliferation of mesangial cells. Inhibition of PI3-kinase blocked PDGF-induced phosphorylation of Erk5. Since PI3-kinase acts through Akt, we determined the role of Erk5 on Akt phosphorylation. XMD8-92, dominant negative Erk5, and siErk5 inhibited phosphorylation of Akt by PDGF. Interestingly, we found inhibition of PDGF-induced Erk5 phosphorylation by a pharmacological inhibitor of Akt kinase and kinase dead Akt in mesangial cells. Thus our data unfold the presence of a positive feedback microcircuit between Erk5 and Akt downstream of PI3-kinase nodal point for PDGF-induced mesangial cell proliferation.

More »»

2012

M. Mikhailova, Xu, X., Robichaud, T. K., Dr. Sanjay Pal, Fields, G. B., and Steffensen, B., “Identification of collagen binding domain residues that govern catalytic activities of matrix metalloproteinase-2 (MMP-2)”, Matrix Biology, vol. 31, pp. 380–388, 2012.[Abstract]


An innovative approach to enhance the selectivity of matrix metalloproteinase (MMP) inhibitors comprises targeting these inhibitors to catalytically required substrate binding sites (exosites) that are located outside the catalytic cleft. In MMP-2, positioning of collagen substrate molecules occurs via a unique fibronectin-like domain (CBD) that contains three distinct modular collagen binding sites. To characterize the contributions of these exosites to gelatinolysis by MMP-2, seven MMP-2 variants were generated with single, or concurrent double and triple alanine substitutions in the three fibronectin type II modules of the CBD. Circular dichroism spectroscopy verified that recombinant MMP-2 wild-type (WT) and variants had the same fold. Moreover, the MMP-2 WT and variants had the same activity on a short FRET peptide substrate that is hydrolyzed independently of CBD binding. Among single-point variants, substitution in the module 3 binding site had greatest impact on the affinity of MMP-2 for gelatin. Simultaneous substitutions in two or three CBD modules further reduced gelatin binding. The rates of gelatinolysis of MMP-2 variants were reduced by 20–40% following single-point substitutions, by 60–75% after double-point modifications, and by > 90% for triple-point variants. Intriguingly, the three CBD modules contributed differentially to cleavage of dissociated α-1(I) and α-2(I) collagen chains. Importantly, kinetic analyses (kcat/Km) revealed that catalysis of a triple-helical FRET peptide substrate by MMP-2 relied primarily on the module 3 binding site. Thus, we have identified three collagen binding site residues that are essential for gelatinolysis and constitute promising targets for selective inhibition of MMP-2.

More »»

2011

B. Steffensen, Chen, Z., Dr. Sanjay Pal, Mikhailova, M., Su, J., Wang, Y., and Xu, X., “Fragmentation of fibronectin by inherent autolytic and matrix metalloproteinase activities”, Matrix Biology, vol. 30, pp. 34–42, 2011.[Abstract]


Fibronectin (FN) purified by gelatin affinity chromatography is unstable and undergoes fragmentation. The cleavage has been ascribed to inherent autolytic protease activities as well as co-purified matrix metalloproteinases (MMP). Understanding the mechanism by which the proteolysis of FN occurs is important, because the FN fragments have biological activities that differ from those of intact FN. Having excluded contributions of other plasma-derived proteases, the present experiments demonstrated that cleavage of FN by MMP-2 to distinct fragments occurred in synergy with inherent FN activities. Limited heat treatment of FN at 56 °C for 30 min inactivated the inherent protease activities sharply reducing autolysis of FN in a manner similar to that seen in the presence of serine proteinase inhibitors. Heat treatment did not alter cell attachment to FN, but significantly increased the susceptibility of FN to enzymatic cleavage by MMP-2. The carboxyl-terminal hemopexin-like domain (PEX) of MMP-2 was shown to possess critical exodomain properties required for the interactions of MMP-2 with FN, and FN was cleaved at a significantly reduced rate by an MMP-2 variant with deletion of PEX. Verifying the specificity of interactions, isolated PEX competed FN cleavage by MMP-2 in a concentration-dependent manner. These results have further elucidated the synergistic contributions of inherent autolytic serine protease-like activities and MMP-2 to fragmentation of FN and provide the rationale and basis for modified preparation and handling of FN used in biological research. More »»

2011

X. Xu, Mikhailova, M., Chen, Z., Dr. Sanjay Pal, Robichaud, T. K., Lafer, E. M., Baber, S., and Steffensen, B., “Peptide from the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (TIMP-2) inhibits membrane activation of matrix metalloproteinase-2 (MMP-2)”, Matrix Biology, vol. 30, pp. 404–412, 2011.[Abstract]


Cellular activation of latent matrix metalloproteinase-2 (proMMP-2) requires formation of a cell membrane-associated activation complex that involves specific binding between the hemopexin domain of proMMP-2 (PEX) and the C-terminal domain of tissue inhibitor of matrix metalloproteinases-2 (C-TIMP-2). In this study, we tested the feasibility of inhibiting activation of proMMP-2 by exogenous inhibitors, which block the binding between PEX and TIMP-2. The recombinant C-TIMP-2 and synthetic peptides from C-TIMP-2 were used as inhibitors for proMMP-2 activation. Recombinant C-TIMP-2 bound specifically to both the catalytically inactive MMP-2(E404A) and the C-terminal domain of MMP-2 (PEX) in a concentration dependent manner with apparent K(d) of 3.9×10(-7)M and 1.7×10(-7)M, respectively. Moreover, C-TIMP-2 competed the binding between MMP-2(E404A) and full-length TIMP-2. Finally, activity assays showed that addition of C-TIMP-2 to HT-1080 fibrosarcoma cells inhibited proMMP-2 activation in a concentration-dependent manner. We then designed a synthetic peptide, P175L, consisting of 20 residues from the PEX-binding tail region of C-TIMP-2. P175L bound PEX and inhibited cell membrane-mediated activation of proMMP-2 in a concentration dependent manner. Deletion of the last 9 tail residues of C-TIMP-2 in P175L abrogated the inhibitory activities of the peptide showing that these residues were essential for function. Overall, these experiments have demonstrated that proMMP-2 activation can be inhibited by exogenous inhibitors which points to a potential strategy for MMP-2 specific inhibition.

More »»

2010

X. ZHU, Xu, X., Chen, Z., Mikhailova, M., Dr. Sanjay Pal, and Steffensen, B., “Fibronectin and Collagen Glycation Alter MMP Expression in Periodontal Fibroblasts”, J Dent Res, vol. 89, no. A, p. #1458, 2010.

2010

Dr. Sanjay Pal, Chen, Z., Xu, X., Mikhailova, M., and Steffensen, B., “Co-purified gelatinases alter the stability and biological activities of human plasma fibronectin preparations”, Journal of periodontal research, vol. 45, pp. 292–295, 2010.[Abstract]


Background and Objective: Fibronectin (FN) is an important cell adhesion molecule that is used widely to characterize cell behavior. Preparations of FN purified from human plasma by gelatin-Sepharose affinity chromatography typically also contain gelatin-binding gelatinases that may cleave FN, reduce its stability and alter its biological activities. Available methods for separating gelatinases from FN are resource demanding. Therefore, our objective was to devise a time- and cost-efficient protocol for purification of gelatinase-free FN. Material and Methods: Experiments tested the elution profiles for FN and gelatinases from gelatin-Sepharose using a concentration range (1-7%) of dimethyl sulfoxide (DMSO) and 4 m urea as eluants. Subsequently, we explored the sequential application of those eluants for differential elution of gelatinases and FN using a single affinity column. Finally, experiments characterized the stability of purified FN with or without contaminating gelatinases, as well as the effects of FN degradation on cell attachment and migration. Results: Assay optimization demonstrated that pre-elution with 3% DMSO efficiently eliminated gelatinases but not FN from gelatin-Sepharose, whereas subsequent elution with 4 m urea released FN. Sequential elutions with DMSO and urea produced gelatinase-free FN, which was more stable than FN eluted by urea only. Fibronectin degradation did not affect human gingival fibroblast attachment, but increased cell migration significantly. Conclusion: The present experiments devised a time- and cost-efficient protocol for eliminating gelatinases during purification of human plasma FN. Gelatinase-free FN preparations had greater stability, which may be essential for experiments because FN fragments have altered biological activities compared with intact FN. © 2009 The Authors. Journal compilation © 2009 Blackwell Munksgaard. More »»

2009

X. Xu, Mikhailova, M., Ilangovan, U., Chen, Z., Yu, A., Dr. Sanjay Pal, Hinck, A. P., and Steffensen, B., “Nuclear magnetic resonance mapping and functional confirmation of the collagen binding sites of matrix metalloproteinase-2”, Biochemistry, vol. 48, pp. 5822–5831, 2009.[Abstract]


Interactions of matrix metalloproteinase-2 (MMP-2) with native and denatured forms of several types of collagen are mediated by the collagen binding domain (CBD). CBD positions substrates relative to the catalytic site and is essential for their cleavage. Our previous studies identified a CBD binding site on the α1(I) collagen chain. The corresponding synthetic collagen peptide P713 bound CBD with high affinity and was used in this study to identify specific collagen binding residues by NMR analysis of 15N-labeled CBD complexed with P713. Results obtained showed that P713 caused chemical shift perturbations of several surface-exposed CBD backbone amide resonances in a concentration-dependent manner. The 10 residues that underwent the largest chemical shift perturbations (R252 in module 1, R296, F297, Y302, E321, Y323, and Y329 in module 2, and R368, W374, and Y381 in module 3) were investigated by site-specific substitution with alanine. The structural integrity of the CBD variants was also analyzed by one-dimensional 1H NMR. Surface plasmon resonance and microwell protein binding assays of control and CBD variants showed that residues in all three CBD modules contributed to collagen binding. Single-residue substitutions altered the affinity for peptide P713, as well as native and denatured type I collagen, with the greatest effects observed for residues in modules 2 and 3. Additional alanine substitutions involving residues in two or three modules simultaneously further reduced the level of binding of CBD to native and denatured type I collagen and demonstrated that all three modules contribute to substrate binding. These results have localized and confirmed the key collagen binding site residues in the three fibronectin type II-like modules of MMP-2.

More »»

2008

S. BABER, B, S., Z, C., M, M., Dr. Sanjay Pal, and Xu, X., “Competitive Inhibition of proMMP-2 Activation”, J Dent Res , vol. 87, no. A, p. 1012, 2008.[Abstract]


Objectives: Activation of proMMP-2 by membrane type matrix metalloproteinase-1 (MT1-MMP) requires formation of an activation complex involving the hemopexin-like domain of proMMP-2 (PEX), the carboxyl-terminal domain of TIMP-2 (C-TIMP-2) and MT1-MMP. The goal of these experiments was to demonstrate that excess of extraneous recombinant (r) PEX or rC-TIMP-2 may interrupt the activation complex and therefore inhibit MMP-2 activation. Methods: rC-TIMP-2 and rPEX were expressed in E. coli and purified to homogeneity by Ni3+ affinity chromatography as determined by SDS-PAGE. Human fibrosarcoma (HT1080) and rat osteosarcoma (ROS) cell lines were co-cultured with a concentration range of either purified rC-TIMP-2 or rPEX for 24 hours. The level of activation of proMMP-2 secreted into the culture medium was analyzed by gelatin zymography for relative amounts of latent 66 kDa and activated 59-kDa forms of MMP-2. Gel images were digitized and intensities of the two forms of MMP-2 were quantified. Results: With concanavlin-A induction, the ratio of active:latent MMP-2 was 78.6 in HT1080 and 2.5 in ROS culture media. When treated with 7 μM rC-TIMP-2, these ratios decreased to 2.8 in HT1080 and 0.5 in ROS conditioned media, reflecting 96% and 80% inhibition of proMMP-2 activation for HT1080 and ROS cells, respectively. Addition of 2.2 μM rPEX decreased the ratio to 1.5 and 0.22 for HT1080 and ROS, respectively, corresponding to 98% and 91% inhibition of proMMP-2 activation. The inhibition of proMMP-2 activation by rC-TIMP-2 and rPEX was concentration dependent. Conclusion: These results showed that rC-TIMP-2 and rPEX can inhibit activation of proMMP-2 in cancer cells by a mechanism that most likely involves competitive interruption of the interactions between PEX and TIMP-2. This approach constitutes a potential future strategy for MMP-2 inhibition in periodontal disease and oral cancer. (Supported by DE 017139 (COSTAR), DE 018135 and DE 017139). More »»

2008

J. Murillo, Wang, Y., Xu, X., Klebe, R. J., Chen, Z., Zardeneta, G., Dr. Sanjay Pal, Mikhailova, M., and Steffensen, B., “Advanced glycation of type I collagen and fibronectin modifies periodontal cell behavior”, Journal of periodontology, vol. 79, pp. 2190–2199, 2008.[Abstract]


Background: Advanced glycation end products (AGEs) have been linked to pathogenic mechanisms of diabetes mellitus. However, little is known about the contribution of protein glycation to periodontal disease in patients with diabetes. Therefore, this study investigated whether glycation of type I collagen (COLI) and fibronectin (FN) modified the behavior of human gingival fibroblasts (hGFs) and periodontal ligament fibroblasts (hPDLs). Methods: Procedures for rapid in vitro glycation of COLI and FN used methylglyoxal (MG). Formation of AGEs was analyzed by changes in protein migration using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with antibodies specific for MG-glycated proteins. Experiments then characterized the effects of glycated FN and COLI on the behavior of hGFs and hPDLs. Results: MG glycated COLI and FN in <6 hours. Confirming the specificity of the reactions, antibodies specific for MG-induced AGEs reacted with glycated FN and COLI but not with control proteins. In cell culture experiments, glycated FN was significantly less efficient in supporting the attachment of hGFs and hPDLs (P <0.05). Moreover, the morphologic parameters, including length, area, perimeter, and shape factor, were altered (P <0.001) for cells on both glycated proteins. Finally, cell migration was reduced on glycated FN and COLI (P <0.001). Conclusions: MG treatment efficiently glycated COLI and FN, providing a new tool to study the effects of diabetes on periodontal disease. The substantial effects of glycated COLI and FN on hGF and hPDL behavior indicated that protein glycation contributed to the pathogenesis and altered periodontal wound healing observed in patients with diabetes. More »»

2008

C. M. Stanley, Wang, Y., Dr. Sanjay Pal, Klebe, R. J., Harkless, L. B., Xu, X., Chen, Z., and Steffensen, B., “Fibronectin fragmentation is a feature of periodontal disease sites and diabetic foot and leg wounds and modifies cell behavior”, Journal of periodontology, vol. 79, pp. 861–875, 2008.[Abstract]


Background: Fibronectin (FN) undergoes fragmentation in periodontal disease sites and in poorly healing diabetic wounds. The biologic effects of FN fragments on wound healing remain unresolved. This study characterized the pattern of FN fragmentation and its effects on cellular behavior compared to intact FN. Methods: Polyclonal antibodies were raised against FN and three defined recombinant segments of FN and used to analyze gingival crevicular fluid from periodontal disease sites in systemically healthy subjects and in subjects with diabetes, as well as chronic leg and foot wound exudates from subjects with diabetes. Subsequently, the behavior of human gingival fibroblasts (hGFs) and HT1080 reference cells were analyzed by measuring cell attachment, migration, and chemotaxis in the presence of intact FN or recombinant FN fragments. Results: FN fragmentation was evident in fluids from periodontal disease sites and diabetic leg and foot wounds. However, no fragmentation pattern distinguished systemically healthy subjects from subjects with diabetes. hGFs and HT1080 cells required significantly higher concentrations of FN fragments to achieve attachment comparable to intact FN. Cells cultured on FN fragments also were morphologically different from cells cultured on full-length FN. Migration was reduced for hGFs cultured on FN fragments relative to full-length FN. In contrast, FN fragments increased HT1080 fibrosarcoma cell migration over intact FN. Conclusions: FN fragmentation is a prominent feature of periodontal and chronic leg and foot wounds in diabetes. Furthermore, cell culture assays confirmed the hypothesis that exposure to defined FN fragments significantly alters cell behavior. More »»

2007

M. R, B, S., Z, C., A, Y., Dr. Sanjay Pal, E, K., and X, X., “Cloning and Expression of Treponema Denticola Fibronectin-binding Protein (Fbp)”, J Dent Res , vol. 86, no. A, p. 2873, 2007.[Abstract]


Objectives: The bacterium T. denticola is a periodontal pathogen. The ability of T. denticola to bind fibronectin (FN) may be critical to its pathogenicity. Several T. denticola FN binding proteins (Fbps) have been identified previously by Western Blotting, but have not been characterized in detail. A gene (fbp) with significant homology to known Fbps genes from other bacteria was identified from the T. denticola genome sequence. To investigate this potential virulence factor, we cloned the fbp gene and initiated characterization of the encoded protein. Methods: The fbp gene was amplified from T. denticola strain 35405 genomic DNA by high fidelity PCR after digestion with HindIII, cloned into pRSETA with an N-terminal His tag, and verified by DNA sequencing. Fbp was then expressed in E. coli BL21(DE3) and purified by metal affinity chromatography. Interactions of Fbp with FN were analyzed in microwell-based protein-protein binding assays. Results: The amplified fbp gene from T. denticola ATCC 35405 genomic DNA had a size of 1.4 Kb. The fbp reading frame encoded a 471 amino acid Fbp, precisely matching the GenBank database sequence (accession number AE017226.1). Recombinant Fbp expressed in soluble form with a mass of 54 kDa after induction with IPTG. Fbp was purified to a purity >90% as assessed by scanning of images of SDS-PAGE gels. Initial protein binding assays demonstrated specific interactions between the T. denticola Fbp and FN. Studies are now verifying the role of Fbp in binding T. denticola to FN. Conclusion: These experiments cloned a gene (fbp) from T. denticola which encoded a FN binding protein (Fbp). Understanding the specific contribution of Fbp to T. denticola interactions with FN may provide important clues to the virulence mechanisms of this microorganism. Supported by NIDCR grants DE12818, DE14236, and DE016312 as well as DE14318 for CO STAR. More »»

2006

Dr. Nandita Mishra, Dr. Sanjay Pal, K, M. T., and S, D., “Production and characterization of arabinogalactan protein (AGP) from a hairy root line of Catharanthus roseus (L.) G. Don”, Indian Journal of Biotechnology, vol. 5, pp. 211-216, 2006.[Abstract]


Arabinogalactan protein (AGP), a class of cell wall proteoglycan, was isolated from the hairy root cultures of a newly developed hairy root line IIT-BT/D1 of Catharanthus roseus (L.) G. Don. AGP was found to be present both in the roots (0.3 mg/g fresh weight) and in the spent media (47 mg/L). The compositional analysis revealed the predominance of arabinose and galactose sugar, a characteristic feature of AGP. More »»

2003

Dr. Sanjay Pal, Das, S., and Dey, S., “Peroxidase and arabinogalactan protein as by-products during somatic embryo cultivation in air-lift bioreactor”, Process Biochemistry, vol. 38, pp. 1471–1477, 2003.[Abstract]


The spent medium after somatic embryo production has been analysed with respect to various enzymes, arabinogalactan protein and sandal oil. Apart from various hydrolases, a high level of peroxidase activity (32 200 U/ltre medium, with specific activity of 1.3417 U/μg protein) has been obtained. The peroxidase with an optimum activity at 50 °C was reasonably stable at 70 °C. The enzyme was active in the pH range 5–11, with an optimum at pH 6. The enzyme showed a Km of 10.91 mM and Vmax of 14.88 μM/min. Arabinogalactan protein (26–35 mg/l) has also been recovered from the extracellular medium. Sandal oil could not be detected by thin layer chromatography in the spent biomass or somatic embryos or the cell free medium, probably indicating its very low concentration. More »»

1999

S. Das, Das, S., Dr. Sanjay Pal, Mujib, A., Sahoo, S. S., Ponde, N. R., S Gupta, D., and Dey, S., “A novel process for rapid mass propagation of the aromatic plant Santalum album in liquid media and bioreactor”, Acta Horticulturae,(Proc. WOCMAP II 1999) Volume, vol. 3, pp. 281–288, 1999.[Abstract]


A rapid mass propagation method for Santalum album through in vitro cultivation in liquid media and airlift bioreactor has been developed. Embryogenesis, maturation, and germination were performed in the same bioreactor by the removal of exhausted medium and the input of fresh medium without any subculturing. In comparison to the propagation in agar solidified medium, this process of propagation increases the efficiency of production reduces the time required, and minimizes abnormalities. In the bioreactor, about 3000 somatic seedlings were obtained (59.3% were abnormal) in 6 weeks time, per liter of medium, against about 800 (88.9% abnormality) in 12 weeks in the case of solid medium. About 70% of the somatic seedlings grew to healthy plantlets and tolerated hardening trials, something that was rarely possible for somatic seedlings raised on solid media. More »»

1998

D. S, S, D., A, M., Dr. Sanjay Pal, and S, D., “Optimization of sucrose and dissolved oxygen level for somatic embryo production of Santalum album in airlift bioreactor”, Prensa Aromatica, vol. 14, pp. 12-13, 1998.

Publication Type: Conference Paper

Year of Publication Title

2018

A. P. Veedu, R. N. Reshma, Dr. Bipin G. Nair, Dr. Sanjay Pal, and A. Madhavan, “Activity of probiotic strains against enteric pathogens”, in Innovative Strategies for Sustainable Water Management (ISSWM-2017) , Punjab,India. , 2018.

2018

D. Tharuvana, A. Sundaresh, V. J. Sreelakshmi, A. Das, Dr. Bipin G. Nair, Dr. Sanjay Pal, and Madhavan, A., “Sand and charcoal as matrices for immobilization of phages for wastewater treatment”, in Innovative Strategies for Sustainable Water Management (ISSWM-2017) , Punjab, India. , 2018.

2018

S. Subhash, A. B. Kuruvelil, P. V. Aswathi, K. Deepasree, C. D. Navyamol, Das N. P. V., P. Prasad, Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Screening of nematicidal activities of biocontrol fungi Aspergillus niger and Penicillium oxalicum using C. elegans as model”, in Innovative Strategies for Sustainable Water Management (ISSWM-2017) , Punjab, India. , 2018.

2018

M. Sreejith, A. P. Reghu, K. Anandakrishnan, P. J. Gopika, G. Kuriakose, M. J. Reshma, K. Vishnu, A. Madhavan, Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Screening potential antimicrobial compounds by resazurin dye based viability assay”, in Innovative Strategies for Sustainable Water Management (ISSWM-2017), Punjab, India, 2018.

2018

Pradeesh Babu, S. J. Poornendu, Amrita Salim, Madhavan, A., Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Resazurin based redox dye as an indicator for monitoring wastewater biological activity”, in Innovative Strategies for Sustainable Water Management (ISSWM-2017), Punjab, India, 2018.

2018

V. Amrutha., Prasad Megha, A. Lekshmija, R Anjana, S. Aleena, Dr. Bipin G. Nair, Madhavan, A., and Dr. Sanjay Pal, “Effect of compost derived lytic agents against enteric bacteria in sewage”, in InnovativeStrategies for Sustainable Water Management (ISSWM-2017), Punjab, India. , 2018.

2017

P. Nagarajan, K. S. Sruthy, V. P. Lal, V. P. Devan, A. Krishna, A. Lakshman, K M Vineetha, Dr. Bipin G. Nair, Dr. Sanjay Pal, and Madhavan, A., “Biological treatment of domestic wastewater by selected aquatic plants”, in IEEE International Conference on Technological Advancements in Power & Energy (TAP Energy 2017). , 2017.[Abstract]


Around 30 percent of the operating costs of conventional centralized wastewater treatment is required for energy to operate the machines for pump and aeration. Comparatively, constructed wetlands involving photosynthetic plants and other organisms have a lower energy requirement. The aim of the present work is to select aquatic plants which have high efficiency of removing infection in wastewater. Four plants were chosen from locally available plants growing in wastewater. They were put in raw wastewater from toilets for 24 h and were tested for their survival and growth measured by chlorophyll content. Brahmi (Bacopa monnieri) and Duckweed were found to be best compared to Azolla and Hydrilla. The efficiency of removal of enteric infections was done by culturing the treated wastewater on Eosin Methylene Blue (EMB) agar media, a selective and differential medium for enteric gram negative bacteria. The best performing plants were Brahmi and Hydrilla which reduced the load by 67-70%. But Hydrilla did not grow as well compared to Brahmi as evident from the chlorophyll content. Hence Brahmi and Duckweed can be considered as most effective for removal of infection in wastewater among the four tested plants and potentially may save energy and synthetic chemicals otherwise required for conventional treatment systems. Brahmi, famous for its high value medicinal compounds against neuronal problems, may be used for extraction of the compounds. The remaining biomass can be used as feedstock in anaerobic digester for methane/ethanol generation to be used as biofuel.

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2017

A. Madhavan, Sindhu Shetty K., G. Anjana, A. Das, A.S. Athira, H. Hari, K. S. Lekshmi, S. Babu, Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Pneumatophore inspired biomimetic approach to design an energy neutral aerator for application in composting”, in Technical Program Committee of the “2017 IEEE International ( Biennial) Conference on Technological Advancements in Power and Energy”, TAP Energy-2017. , 2017.

2017

A. Madhavan, Prasad Megha, S. Girish, Sindhu Shetty K., Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Caulobacter crescentus as a novel exoelectricigen in a dual chambered Microbial Fuel Cell (MFC)”, in Technical Program Committee of the “2017 IEEE International( Biennial) Conference on Technological Advancements in Power and Energy”, TAP Energy-2017. , 2017.

2016

A. Madhavan, Vishnu Nandakumar, Sindhu Shetty K., Dr. Bipin G. Nair, and Dr. Sanjay Pal, “Optimization of microbial fuel cell (MFC) operated with waste water as substrate”, in Electrical, Electronics, and Optimization Techniques (ICEEOT), International Conference on, Chennai, India, 2016.[Abstract]


In the present study bioelectricity generation from waste water was evaluated in a double chambered microbial fuel cell (MFC). Waste water and potassium permanganate solution (0.3%) was loaded in the anodic and the cathodic chamber respectively. The performance of MFC was studied with two different electrodes (aluminum and carbon as anodes and copper as cathode). Current and voltage measurements were carried out using Keithley source meter (2420) and multimeter respectively. The Voltammetric analysis was done to study the anodic oxidation rate. The resistance of the system was found to be low (0.466 Ω for the aluminum anode and 0.673 Ω carbon anode) which were deduced from the power density curves. The system showed a COD removal of 77% over a period of a week ofk operation.

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2013

A. L, K, A., S, A., MU, C., S, N., Dr. Sudarslal S., and Dr. Sanjay Pal, “Isolation and characterization of host binding proteins from Bacillus clausii using mass spectrometry”, in Proceedings of International Conference on Biotechnology for Innovative Applications (Amrita BioQuest 2013), 2013.

2013

R. M. Nair, Nambiar, S. S., TV, V., L, V. Prabhu, Menon, D., Gandhi, B. Balan, P, C., Dr. Nandita Mishra, and Dr. Sanjay Pal, “Characterization of Fibronectin Isoforms and Fragments”, in Proceedings of International Conference on Biotechnology for Innovative Applications (Amrita BioQuest 2013), 2013.

2013

Dr. Nandita Mishra, Suresh, A., S, A., P.V, A., .P, P., O.J, S., P, C., and Dr. Sanjay Pal, “Studies on Probiotic Strains from Fermented Foods & Beverages in Kerala”, in Proceedings of International Conference on Biotechnology for Innovative Applications (Amrita BioQuest 2013), 2013.

1997

D. S, S, D., Dr. Sanjay Pal, A, M., S, S., NR, P., S, D., and S, D., “A novel process for rapid mass propagation of the aromatic plant Sandal (Santalum album) in liquid media and bioreactor”, in World Congress on Medicinal and Aromatic Plants for Human Welfare, Nov.10-15, ICMAP-ISHS-SAIPA, Mendoza, Republica Argentina, 1997.

Publication Type: Conference Proceedings

Year of Publication Title

2015

C. P., P., B., N., M., V., A., C., N., G., V., S., S., A., M., B.G., N., N., M., and Dr. Sanjay Pal, “Transformation, expression and activity analysis of recombinant Staphylococcus autolysin in Bacillus”, The 39th All India Cell Biology Conference on “Cellular Organization and Dynamics", Thiruvananthapuram, Kerala, India, . Thiruvananthapuram, Kerala, India, 2015.

2015

Salim Amrita, Ugesh, k.p., Chandni, P., Sreerangini, M. R., Bipin, N., Ajith, M., Dr. Sanjay Pal, and Sreetha, H., “Isolation and characterization of bacteriophage treatment of domestic wastewater”, Proceeding of the 56th International Annual Conference of The Association of Microbiologist of India . JNU, New Delhi, 2015.

2015

Vijayakumar Amrutha, Chinchu, B., Nanjan Pandurangan, Dr. Sanjay Pal, G. Bipin, N., Ajith, M., and Salim Amrita, “Potent chitin Synthase inhibitors from Plant sources”, Proceeding of National seminar on Recent advances in medicinal plant research. Bharat mata college , 2015.

2013

B. Steffensen, Mikhailova, M., Xu, X., Robichaud, T. K., Dr. Sanjay Pal, and Fields, G. S., “Molecular targets for selective inhibitors of MMP-2.”, J. Dent. Res., vol. 92. SAGE, p. 2861, 2013.

2013

Dr. Nandita Mishra, Dr. Sanjay Pal, Nair, R. R., Krishna, A., Ajith, A., V, A., KS, D., Nedungadi, D., and P, C., “Expression and refolding of recombinant staphylococcal amidases in E. coli”, Proceedings of International Conference on Biotechnology for Innovative Applications (Amrita BioQuest 2013). Elsevier Publications, p. 106, 2013.

2011

Dr. Sanjay Pal, “Characterization of the Probiotic attributes of a Gelatin binding Lactobacillus from local curd”, XXXV Cell biology conference, 16-18 Dec. NISER, Bhubaneswar, Orissa, 2011.

2010

M. M, X, X., Dr. Sanjay Pal, and B, S., “Collagen Binding Site Residues are Key to MMP-2 Enzymatic Activities”, 28th Annual Dental Science Symposium, UTHSCSA and Cancer Development and Progression Program Retreat, San Antonio. 2010.

2010

S. B, Dr. Sanjay Pal, Z, C., X, X., M, M., H, Y., and Y, W., “Proteolytic processing of fibronectin by MMP-2 produces fragments with distinct biological properties”, Gordon Research Conference on Proteases and Inhibitors. Il Ciocco, Italy, 2010.

2009

M. M, X, X., Z, C., Dr. Sanjay Pal, and B, S., “Identical amino acids on MMP-2 position different types of collagen for cleavage”, The 27th Annual Dental Science Symposium, UTHSCSA and The Cancer Development and Progression Program Retreat, San Antonio. 2009.

2009

X. X, M, M., U, I., Z, C., A, Y., Dr. Sanjay Pal, AP, H., and B, S., “Mapping and Functional Confirmation of the Collagen Binding Sites of MMP-2”, Gordon Research Conference on Matrix Metalloproteinases. Les Diablerets, Switzerland, 2009.

2008

X. X, S, B., Z, C., Dr. Sanjay Pal, M, M., and B, S., “Inhibition of proMMP-2 activation by carboxyl-terminal domain of TIMP-2 in vitro”, The American Society for Cell Biology Annual Meeting Special Poster Session. Washington, DC, 2008.

2008

Dr. Sanjay Pal, Z, C., X, X., M, M., Y, W., and B, S., “Cleavage of fibronectin by MMP-2 generates fragments with distinct biological activities”, The American Society for Cell Biology Annual Meeting Special Poster Session. Washington, DC, 2008.

2007

Dr. Sanjay Pal, Z, C., X, X., M, M., RJ, K., and B, S., “Enhanced strategy for separating human plasma fibronectin from MMPs by affinity chromatography”, The American Society for Cell Biology Annual Meeting Special Poster Session. Washington, DC, 2007.

2004

Dr. Sanjay Pal, S, D., A, M., and S, D., “Characterization Of Spent Media During Somatic Embryogenesis Of Sandalwood In Suspension Culture: Exploring Value Addition”, IUPAC International Conference on biodiversity and natural products: Chemistry and Medical Applications, Jan. 26-31. New Delhi, India, 2004.

2001

Dr. Sanjay Pal, S, D., and S, D., “Purification and characterization of arabinogalactan protein (AGP) during somatic embryogenesis in sandalwood (Santalum album L.)”, XXV All India Cell Biology Conference, Nov 1-3. IISc, Bangalore, India., 2001.

Publication Type: Patent

Year of Publication Title

2000

Dr. Sanjay Pal, Dey, S., and Das, S., “Herbal skin nourishing gel”, 2000.

Publication Type: Book Chapter

Year of Publication Title

1998

S. Das, Das, S., .Mujib, A., Dr. Sanjay Pal, and .Dey, S., “Influence of carbon and pH on rapid mass propagation of Santalum album through somatic embryogenesis: Biotechnology application in agroforestry”, in Sandal and Its Products, AICAR, Canberra, Australia, 1998, pp. 66-68.

1998

A. .Mujib, Das, S., Das, S., Dr. Sanjay Pal, and .Dey, S., “Biotechnological Routes of Mass Propagation of Santalum album”, in Role of biotechnology in medicinal and aromatic plants - I. A Khan (ed.) , vol. I, Ukaaz Publication, Hyderabad, India, 1998, pp. 83-94.

1998

S. Das, .Mujib, A., Das, S., Dr. Sanjay Pal, and .Dey, S., “Biotechnology of medicinal plants: Recent advances and potential”, in Role of biotechnology in medicinal and aromatic plants, vol. 2, Ukaaz Publication, Hyderabad, India, 1998, pp. 126-139.

Patents:

Year Title
2000 Herbal skin nourishing gel: Ref: 695/ Cal/ 2000, Publication dated, 19.12.2000