Arun Kumar Jain

Natural rubber latex allergy

Natural rubber latex (NRL) is a ubiquitous allergen as it is a component of > 40,000 products in everyday life. Latex allergy might be attributed to skin contact or inhalation of latex particles. Latex allergy is an IgE-mediated hypersensitivity to NRL, presenting a wide range of clinical symptoms such as angioedema, swelling, cough, asthma, and anaphylactic reactions. Until 1979, latex allergy appeared only as type IV delayed hypersensitivity; subsequently, the proportion of different allergy types drifted towards type IV contact allergy reactions. Several risk factors for sensitization to NRL are already known and well documented. Some authors have established a positive correlation between a history of multiple surgical interventions, atopy, spina bifida malformation, and latex allergy incidence. We suspect an increase in latex allergy incidence in association with increased atopy and sensitivity to environmental allergens in the industrial population. It is often postulated in literature that the groups of workers at risk for this allergy are essentially workers in the latex industry and healthcare professionals. In this population, direct internal and mucosal contact with NRL medical devices may be the route of sensitization as factors such as the number of procedures and use of NRL materials (catheters and tubes) were associated with increased risk of latex sensitization and allergy.

Apoptosis and Bcl-2 protein expression in human placenta over the course of normal pregnancy.

With 2 figures and 2 tables

Altered Expression of Succinic Dehydrogenase in Asthenozoospermia Infertile Male

There is a possibility that mitochondrial respiration defects may contribute to asthenozoospermia.

Thickness of glomerular and tubular basement membranes in preclinical and clinical stages of diabetic nephropathy

Aims: This study aimed to elucidate the early renal changes in diabetes mellitus (DM) with and without clinical symptoms related to renal damage. Methods: Renal biopsy was studied in 25 patients (14 with microalbuminuria and 11 with albuminuria) both by light and electron microscopies (LM and EM, respectively) for renal changes and morphometry was performed to study glomerular and tubular basement membranes (GBM and TBM, respectively) width using a Soft Imaging System GmBH (analysis 3). Results: A significant increase was noted in the mean GBM and TBM thickness in both the preclinical and clinical groups compared to the control group. The changes in the TBM were noted to be predominant in both preclinical and clinical patients. Conclusions: This study indicates the importance of morphometric evaluation of the GBM and TBM width in the elucidation of early renal damage in diabetic nephropathy, especially in the absence of LM changes. The significance of identification of early renal changes using morphometric techniques for better management of these patients requires further studies.

Role of Steroid Hormone and Growth Factor Receptors and Proto-Oncogenes in the Behavior of Human Mammary Epithelial Cancer Cells in vitro

The cultivation of cells from primary breast cancers is very unpredictable. The majority of breast-cancer-derived cell lines are of metastatic origin. To define the characteristics of tumor cells which govern their ability to grow in vitro as primary cultures as well as continuous or established culture cell lineages, human mammary epithelial cancer (HMEC) cells from 18 cases of unselected primary breast cancer were propagated in culture. Propagation of HMEC cells in vitro as monolayers in primary culture was successful in 10 out of 18 (55.5%) cases, which showed continous proliferation of tumor cells only up to 6-8 passages before they reached senescence. An investigation of the effects of phenotypic expression of estrogen receptors (ER), the progesterone receptors, c-erbB-2 oncoprotein and epidermal growth factor receptors (EGFR) on the capacity of HMEC cells to grow in vitro as monolayers showed that expression of ER and EGFR is required for controlling tumor proliferative activity in vitro. Expression of ER protein made the growth of HMEC cells more difficult, while expression of EGFR protein made their growth in vitro easier. Phenotypic characteristics of floating HMEC cells were found to be different from those grown on cover slips as adherent cultures, suggesting a selective growth of HMEC cells of a specific phenotype in culture. Cultured HMEC cells in subsequent passages showed a decrease in their proliferative capacity, alterations in phenotypic characteristics and development of morphologic features of terminal differentiation, resulting in senescence.

Discovery of T-cell Driven Subunit Vaccines from Zika Virus Genome: An Immunoinformatics Approach

The recent outbreaks of Zika virus and the absence of a specific therapy have necessitated to identify T-cell-stimulating antigenic peptides as potential subunit vaccine candidates. The translated ssRNA (+) genome of Zika virus was explored in EMBOSS antigenic and VaxiJen to predict 63 peptides as potential antigens. Three MHC-II binding peptide prediction tools, viz. NetMHCIIpan, PREDIVAC and immune epitope database (IEDB) were employed in consensus on 63 antigenic peptides to propose 14 T-helper cell epitopes. Similarly, analysis on 63 antigenic peptides through NetMHC, NetCTL and IEDB MHC-I binding peptide prediction tool led to identification of 14 CTL epitopes. Seven T-cell epitopes, C:44-66, M:135-149, NS2A:124-144, NS3:421-453, NS3:540-554, NS4B:90-134 and NS4B:171-188, are observed to share overlapping MHC-I and MHC-II binding motifs and hence, are being proposed to constitute minimum T-cell antigens to elicit protective T-cell immune response against Zika. Three of them, C:44-66, NS3:421-453 and NS3:540-554 are identified to be conserved across all the selected strains of Zika virus. Moreover, the 21 T-cell epitopes are non-self to humans and exhibited good coverage in variable populations of 14 geographical locations. Therefore, 21 T-cell epitopes are proposed as potential subunit vaccines against Zika virus.

TiD: Standalone software for mining putative drug targets from bacterial proteome.

TiD is a standalone application, which relies on basic assumption that a protein must be essential for pathogens survival and non-homologous with host to qualify as putative target. With an input bacterial proteome, TiD removes paralogous proteins, picks essential ones, and excludes proteins homologous with host organisms. The targets illustrate non-homology with at least 40 out of 84 gut microbes, considered safe for human. TiD classifies proposed targets as known, novel and virulent. Users can perform pathway analysis, choke point analysis, interactome analysis, subcellular localization and functional annotations through web servers cross-referenced with the application. Drug targets identified by TiD for Listeria monocytogenes, Bacillus anthracis and Pseudomonas aeruginosa have revealed significant overlaps with previous studies. TiD takes <2h to scan putative targets from a bacterial proteome with ~5000 proteins; hence, we propose it as a useful tool for rational drug design. TiD is available at http://bmicnip.in/TiD/.

Probing binding mechanism of interleukin-6 and olokizumab: in silico design of potential lead antibodies for autoimmune and inflammatory diseases

Abstract Computer-aided antibody engineering has been successful in the design of new biologics for disease diagnosis and therapeutic interventions. Interleukin-6 (IL-6), a well-recognized drug target for various autoimmune and inflammatory diseases such as rheumatoid arthritis, multiple sclerosis, and psoriasis, was investigated in silico to design potential lead antibodies. Here, crystal structure of IL-6 along with monoclonal antibody olokizumab was explored to predict antigen–antibody (Ag − Ab)-interacting residues using DiscoTope, Paratome, and PyMOL. Tyr56, Tyr103 in heavy chain and Gly30, Ile31 in light chain of olokizumab were mutated with residues Ser, Thr, Tyr, Trp, and Phe. A set of 899 mutant macromolecules were designed, and binding affinity of these macromolecules to IL-6 was evaluated through Ag − Ab docking (ZDOCK, ClusPro, and Rosetta server), binding free-energy calculations using Molecular Mechanics/Poisson Boltzman Surface Area (MM/PBSA) method, and interaction energy estimation. In comparison to olokizumab, eight newly designed theoretical antibodies demonstrated better result in all assessments. Therefore, these newly designed macromolecules were proposed as potential lead antibodies to serve as a therapeutics option for IL-6-mediated diseases.

Dual therapy of rosiglitazone/pioglitazone with glimepiride on diabetic nephropathy in experimentally induced type 2 diabetes rats

Diabetic nephropathy is a major cause of end-stage renal disease (ESRD) in the general population. It is estimated that diabetic nephropathy will eventually develop in about 40% of all patients with diabetes; therefore, prevention is critical for delaying the development and progression of diabetic kidney disease. Despite extensive efforts, medical advances are still not successful enough to prevent the progression of the disease. In the present study, we focused on the comparison of combination therapies and whether they offered additional renoprotection. Type 2 diabetes mellitus was induced by intraperitoneally administering streptozotocin (90 mg/kg) in neonatal rats and then these rats were treated with rosiglitazone (1.0 mg/kg) in combination with glimepiride (0.5 mg/kg) or with pioglitazone (2.5 mg/kg) in combination with glimepiride (0.5 mg/kg). Diabetic nephropathy markers were evaluated by biochemical and ELISA kits and renal structural changes were examined by light microscopy and transmission electron microscopy. Results show that the combination of pioglitazone with glimepiride is more effective in amelioration of diabetic nephropathy than rosiglitazone with glimepiride drug therapy due to glycemic control, suppressing albumin excretion rate, total protein excretion rate and augmented TNF-a signaling during the development of streptozotocin induced type 2 diabetic nephropathy.

181 Discovery of potential inhibitors of BMX non-receptor tyrosine kinase through e-pharmacophore based virtual screening.

Cellular functioning depends on proper coordination between proteins, and much of this communication is mediated by so-called peptide-protein interactions in which a short linear motif is bound by a corresponding peptide-binding domain. These interactions provide major challenges both to experiment as well as modeling due to their often weak, transient character, but our ability to study them is critical for improved understanding, and precise modulation, of cellular regulation. A major focus of our research lies in the development and application of protocols for the accurate modeling of peptide-protein interactions. Rosetta FlexPepDock takes as input a receptor structure and an approximate starting structure of the peptide, and refines this structure to near-native resolution using a Monte-Carlo sampling approach coupled with minimization that starts with an attraction–dominated energy function and gradually increases repulsion to generate well-packed, clash-free models (Raveh et al., 2010, 2011; London et al., 2011a). These models can then be used to rank different peptide sequences for their ability to bind to a given receptor (London et al., 2011b). In my talk I will describe recent new developments and applications of peptide-protein modeling, including initial and advanced steps towards the full ab initio modeling of peptide-protein interactions in which no prior information about the peptide-binding site is available. Most exciting, for peptides with characterized binding motifs, we show that a motif-based search for fragments in solved structures and their subsequent docking allows us to identify an acceptable model of interaction, which can then be further refined using e.g. FlexPepDock. We anticipate that this novel approach and implementation will significantly extend the number of peptide-mediated interactions that can be accurately modeled, and thus characterized and manipulated.