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Genes involved in signaling are highly regulated at the level of transcription. Several factors have been known to play role in transcriptional modification of genes. Among these DNA sequence variations present in the regulatory region and aberrant methylation of CpG Iceland in promoter region are the most important factors modifying transcriptional regulation of genes. DNA sequence variation interferes with assembling of regulatory protein TF (transcription factor) on the cis elements TFBS (transcription factor binding site). Presence of variations in regulatory region may alter the level of gene product via interaction of TF to TFBS (transcriptional modification). Promoter hypermethylation causes gene silencing and responsible for transcriptional dysregulation of gene. JAK-1, STAT-3, IL-6, MAPK and AR genes participate in signaling pathway and are tightly regulated. Overexpression of IL-6 and activated STAT3 may contribute to the development of prostate cancer and possibly other human cancers. Indeed, constitutively activated STAT3 have been found in a growing number of human tumors. In the present work, we have predicted 34 regulatory polymorphisms that lies in TFBS of 5 (JAK-1, STAT-3, IL-6, MAPK and AR) signaling genes and compare the methylation of CpG Iceland in promoter region of above motioned genes. On the basis of these predictions, it has been hypothesized that transcriptional modification of gene resulting from the DNA sequence variations in regulatory region or promoter hypermethylation increases the susceptibility to diseases such as cancer by alteration in the level of signaling genes product. Presence of DNA sequence variations may also influence the response to a particular drug.

With the growing commercial availability of dyes, their range and scope of application is also expanding. Consequently, a large amount of unused dyes are released in the industrial effluents. Ninety percent of textile dyes entering modern activated sludge sewage treatment plants pass through unchanged. There is an intense environmental concern about the fate of these unbound dyes. These discharged dyes form toxic products and their strong color causes turbidity which even at very low concentrations has a huge impact on the aquatic environment. These synthetic reactive dyes bond covalently with fabric and contain chromophoric groups like anthraquinone, azo, triarylmethane etc. along with reactive groups viz., vinyl sulphone, chlorotriazine, trichloropyrimidine etc. (Sumathi & Manju, 2000; Keharia & Madamvar, 2003). Disperse dyes and acid dyes have low solubility in water. They are mainly used in the dyeing of polyesters and find minor use in dyeing cellulose acetates and polyamides. Azo dyes constitute the largest group of colorants used in industry. These dyes can be precipitated or adsorbed only in small amounts, while under anaerobic conditions they are cleaved by microorganisms forming potentially carcinogenic aromatic amines (Chung & Cerniglia, 1992). Several studies on decolorization of textile dyes used in industries have been conducted. Some non-textile dyes commonly present in industrial effluents have been studied. Dye removal from wastewaters with traditional physicochemical processes, such as coagulation, adsorption and oxidation with ozone is expensive, can generate large volumes of sludge and usually requires the addition of environmentally hazardous chemical additives (Chen, 2006). On the other hand, most of the synthetic dyes are xenobiotic compounds which are poorly removed by the use of conventional biological aerobic treatments (Marco et al., 2007a, b). Although, biodegradation appears to be a promising technology, unfortunately the analysis of contaminated soil and water has shown persistence of toxic pollutants even in the presence of microorganisms (Robinson et al., 2001; Keharia and Madamvar, 2003). Decolorization of dye wastewater is an area where innovative treatment technologies need to be investigated. The focus in recent times has shifted towards enzyme based treatment of colored wastewater/industrial textile effluents. The peroxidase and polyphenol oxidases

Salt fractionated proteins from Trichosanthes diocia was used to study decolorization of Reactive Orange 15 under multifarious experimental conditions like pH, temperature, time interval, enzyme concentration and in the presence of redox mediator 1-hydroxybenzotriazole (HOBT)/ vanillin. T. diocia peroxidase showed extensive decolorization of Reactive Orange15 in the presence of 1-hydroxybenzotriazole (HOBT) / vanillin. The dye was decolorized effectively with HOBT and so further studies were performed in the presence of HOBT. At an enzyme concentration of 0.45 EUmL-1 the peroxidase was able to remove the reactive dye up to a maximum of 94.6% with 1.0 mM 1-hydroxybenzotriazole. Maximum decolorization was achieved at a temperature of 40°C, pH 5.0 and with an incubation period of 90 min. Thus, the data indicates that T. diocia peroxidase could be a potential source for developing an inexpensive and efficient method for the treatment of recalcitrant Reactive Orange15 dyes that are potentially toxic. --- Department of Biochemistry (DST-FIST & UGC-SAP Supported), Dr. Ram Manohar Lohia Avadh University, Faizabad-224001, U.P. (India) --- Please Cite This Article As:  Farrukh Jamal, Tabish Qidwai and Prabhash K Pandey. 2010. Soluble Fraction of Trichosanthes diocia Peroxidase in Decolorization of Reactive Orange 15. J. Ecobiotechnol. 2(5):36-41. Â

Our interest in development of hyaluronidase inhibitors as male antifertility agents led to identification of Terminalia chebula (T. chebula) plant with hyaluronidase (HAase) inhibitory activity of human spermatozoa ( approximately 93% inhibition) and rat caudal epididymal spermatozoa ( approximately 86% inhibition) in vitro at 30 mg/ml. We further demonstrated inhibition of hyaluronidase activity of testis and epididymal spermatozoa in vivo coincident with antispermatogenic activity and contraceptive efficacy of TC extract administered at 50 and 100mg/kg/day orally for 60 days in male albino rats. The significant decrease in motility, count and increase in morphological abnormalities of epididymal spermatozoa and severe reduction in fertility (-100%) of male rats treated with T. chebula fruit extract at 100mg/kg dose could be attributed to either direct effect on testis or direct or indirect interference with sperm maturation in epididymis, and/or inhibition of testicular and epididymal sperm hyaluronidase enzyme in vivo probably caused by flavonoids like tannins present in T. chebula.

The present study reports data on post-translational modifications in the glycosylation status during epididymal passage and significance in fertility of a 33 kDa glycoprotein of rhesus monkey (Macaca mulatta), designated as MEF3 (monkey epididymal fluid protein 3). MEF3 exhibited strong affinity for N-linked α-d-mannose groups and O-linkedN-Ac-galactosamine linkages in epididymal fluids and exhibited moderate affinity forN-Ac-glucosaminylated (wheat germ agglutinin), fucosylated (Tetragonolotus purpurea), andN-Ac-galactosamine (peanut agglutinin) residues on more mature corpus and caudal spermatozoa in a maturation-dependent manner on Western blots probed with specific biotinylated lectins. Polyclonal antiserum raised against affinity-purified MEF3 from caudal epididymal fluid (CEF) cross-reacted specifically with CEF and caudal sperm membrane of macaque and with Triton X-100 extract of ejaculated human spermatozoa, suggesting the existence of antigenically related components in both species. The tangled agglutination caused by anti-33 kDa serum of human spermatozoa, along with localization of MEF3 on entire sperm surface of epididymal and testicular sperm of monkey and human spermatozoa, suggest the significance of MEF3 in sperm function. The 100% inhibition of fertility of immunized female rabbits with this proteinin vivoand inhibition of human sperm penetration in zona-free hamster eggsin vitrosuggests the functional significance of MEF3 in fertility. Together, these results clearly indicate that MEF3 has potential significance as a target for antibodies that inhibit sperm function and fertility.

The present study investigated regional modifications of glycosylation status, sperm association and functional significance of N- and O-linked glycoproteins in epididymal luminal fluid of the rhesus monkey (Macaca mulatta). The predominant glycoproteins of the epididymal luminal fluid that increase in the extent of glycosylation or unmasking of exposed epitopes in a region-specific, maturation-dependent manner, included those of 150, 116, 68, 64, 58 (N- and O-linked) and 170 kDa (O-linked). The higher expression of 40 (N-linked), 38 (N- and O-linked) and 60, 56 and 33 kDa (O-linked) glycoproteins in the proximal caput epididymal fluid was followed by alteration or reorganization of 60, 38 and 33 kDa (O-linked) glycoproteins in the distal segments of the epididymis. The association of epididymal fluid glycoproteins with maturing spermatozoa was identified by generating polyclonal antiserum against monkey caudal sperm membrane in female albino rabbits. The antiserum crossreacted strongly with 58 and 33 kDa epididymal fluid glycoproteins of monkeys and also reacted with 116, 68, 58, 56 and 33 kDa glycoproteins from Triton X-100 extracts of human spermatozoa, indicating the presence of antigenically related components in both species. The functional significance of epididymal fluid glycoproteins in sperm functions was investigated by raising antiserum against a heavily glycosylated 58 kDa glycoprotein (MEF1) of caudal epididymal fluid, which crossreacted with the Triton X-100 extracts of epididymal spermatozoa of monkey and ejaculated human spermatozoa on immunoblots. In an in vitro micro-sperm agglutination assay, anti-MEF1 serum agglutinated both rat caudal epididymal spermatozoa and human spermatozoa. MEF1 seemed to be involved in fertilization as demonstrated by inhibition of fertility (100%) in female albino rabbits and rats immunized with this protein. A sperm-agglutinating 58 kDa glycoprotein of rhesus monkey epididymis with functional significance in fertility was identified, thus indicating that it is a potential candidate for contraceptive vaccine development.