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1.
Indian J Biochem Biophys ; 2011 Feb; 48(1): 29-34
Article in English | IMSEAR | ID: sea-135297

ABSTRACT

Bacterial organophosphate hydrolases (OPH) have been shown to hydrolyze structurally diverse group of organophosphate (OP) compounds and nerve agents. Due to broad substrate range and unusual catalytic properties, the OPH has successfully been used to develop eco-friendly strategies for detection and decontamination of OP compounds. However, their usage has failed to gain necessary acceptance, due to short half-life of the enzyme and loss of activity during process development. In the present study, we report a simple procedure for immobilization of OPH on biocompatible gelatin pads. The covalent coupling of OPH using glutaraldehyde spacer has been found to dramatically improve the enzyme stability. There is no apparent loss of OPH activity in OPH-gelatin pads stored at room temperature for more than six months. As revealed by a number of kinetic parameters, the catalytic properties of immobilized enzyme are found to be comparable to the free enzyme. Further, the OPH‑gelatin pads effectively eliminate OP insecticide methyl parathion and nerve agent sarin.


Subject(s)
Enzyme Stability , Enzymes, Immobilized/chemistry , Escherichia coli/enzymology , Escherichia coli/genetics , Gelatin/chemistry , Hydrolysis , Insecticides/poisoning , Methyl Parathion/chemistry , Organophosphorus Compounds/chemistry , Phosphoric Monoester Hydrolases/genetics , Phosphoric Monoester Hydrolases/isolation & purification , Phosphoric Monoester Hydrolases/metabolism , Sarin/chemistry , Substrate Specificity
3.
Article in English | WPRIM | ID: wpr-115058

ABSTRACT

PURPOSE: The purpose of this study was to identify differences in signal transduction gene expression between normal and diabetic keratocytes stimulated with interleukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha). METHODS: Normal and diabetic keratocytes were primarily cultured and treated with 20 ng/ml IL-1alpha and TNF-alpha for 6 h. cDNA was hybridized to an oligonucleotide microarray. Genes identified by the microarray were further evaluated by real-time PCR. RESULTS: Diabetic keratocytes over-expressed components of the MAPK and Notch pathways, and under-expressed components of the insulin, calcium, and TGF-beta pathways. Cytokine treated diabetic keratocytes differentially expressed components of the TGF-beta and MAPK pathways. After IL-1alpha and TNF-alpha treatment, nine genes were under-expressed, falling in the insulin, TGF-beta, and Toll-like receptor pathways. Real-time PCR showed a significant decrease in the IL-6 and TGF-beta2 genes and a significant increase in the Ppm1a gene. CONCLUSIONS: There were some differences in gene expression between normal and diabetic keratocytes related to signal transduction pathways, such as the insulin, MAPK, calcium, and TGF-beta pathways. In addition, IL-1alpha and TNF-alpha stimulating the insulin, TGF-beta, and Toll-like receptor signaling pathways may have different effects in diabetic keratocytes.


Subject(s)
Animals , Apoptosis , Cells, Cultured , Cornea/drug effects , DNA/genetics , Diabetes Mellitus, Experimental/genetics , Gene Expression Profiling , Insulin/genetics , Interleukin-1alpha/pharmacology , Mitogen-Activated Protein Kinase Kinases/genetics , Nuclear Proteins/genetics , Oligonucleotide Array Sequence Analysis/methods , Phosphoric Monoester Hydrolases/genetics , Polymerase Chain Reaction , Prolactin/genetics , Rats , Rats, Long-Evans , Receptors, Notch/genetics , Signal Transduction/drug effects , Transforming Growth Factor beta/genetics , Tumor Necrosis Factor-alpha/pharmacology , Ubiquitin-Protein Ligases/genetics
4.
J Genet ; 2002 Aug; 81(2): 65-71
Article in English | IMSEAR | ID: sea-114253

ABSTRACT

Knowledge of candidate gene polymorphisms in a population is useful for a variety of gene-disease association studies, particularly for some complex traits. A number of candidate genes, a majority of them from the monoaminergic pathway in the brain, have been very popular in association studies with schizophrenia, a neuropsychiatric disorder. In this study diallelic/multiallelic polymorphisms in some dopaminergic, serotonergic and membrane-phospholipid-related genes have been evaluated in a control population recruited from North India. Association, if any, of these allelic variants with schizophrenia has been tested using a case-control approach. The case data have been taken from our published family-based association studies in schizophrenia. Of the eight genes tested in this study, association with schizophrenia was observed for only two gene polymorphisms, one in the promoter region of the serotonin 2A receptor gene and the other in the tryptophan hydroxylase gene. One new allele for the dopamine transporter gene (with eight repeats, 570-bp size), not reported in any population so far, has been identified in one individual in our sample. The data generated in this study, besides providing a normative background for various disease association studies, are a significant contribution to the population-specific genome database, a currently growing requirement.


Subject(s)
Adult , Case-Control Studies , Catechol O-Methyltransferase/genetics , Dopamine/metabolism , Female , Gene Frequency , Humans , India , Male , Phospholipases A/genetics , Phosphoric Monoester Hydrolases/genetics , Polymorphism, Genetic , Receptors, Serotonin/genetics , Schizophrenia/genetics , Tryptophan Hydroxylase/genetics
5.
Article in English | WPRIM | ID: wpr-36359

ABSTRACT

To characterize the TGF-beta1 response of monocytic leukemia cells, we analyzed the effects of TGF-beta1 on cell proliferation, differentiation, and apoptosis of human monoblastic U937 cells. Treatment of cells with TGF-beta1 in the absence of growth factors significantly enhanced cell viability. Flow cytometric analysis of DNA content and CD14 expression revealed that TGF-beta1 does not affect cell proliferation and differentiation. Consistent with these results was the finding that no transcriptional induction of Cdk inhibitors such as p21Waf1, p15Ink4b, and p27Kip1 was detected following TGF-beta1 treatment. Interestingly, however, pretreatment of TGF-beta1 significantly inhibited Fas-, DNA damage-, and growth factor deprivation-induced apoptosis. This antiapoptotic effect was totally abrogated by anti-TGF-beta1 antibody. Quantitative RT-PCR analysis demonstrated a dose- and time-dependent transcriptional up-regulation of Bcl-X(L), suggesting its implication in the TGF-1-mediated antiapoptotic pathway. We also observed elevated expression of c-Fos and PTEN/MMAC1. But, no detectable change was recognized in expression of c-Jun, Fas, Fadd, Fap-1, Bcl-2, and Bax. Taken together, our study shows that TGF-beta1 enhancement of cellular viability is associated with its antiapoptotic effect, which may result from the transcriptional up-regulation of Bcl-X(L).


Subject(s)
Lipopolysaccharide Receptors/metabolism , fas Receptor/metabolism , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Division/drug effects , Cell Survival/drug effects , DNA/analysis , DNA Damage , Gene Expression Regulation, Neoplastic/genetics , Genes, Tumor Suppressor/genetics , Humans , Leukemia, Myeloid/genetics , Neoplasm Proteins/metabolism , Phosphoric Monoester Hydrolases/genetics , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/biosynthesis , RNA, Messenger/metabolism , Receptors, Antigen, T-Cell/genetics , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Transforming Growth Factor beta/pharmacology , U937 Cells , Up-Regulation
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