Screening for Indian isolates of egg-parasitic fungi for use in biological control of fascioliasis and amphistomiasis in ruminant livestock

Wild isolates of the egg-parasitic fungi Paecilomyces lilacinus and Verticillium chlamydosporium, obtained from the organic environment of Durg, Chhattisgarh, India, were subjected to screening for in vitro growth using different media types, range of incubation temperature and pH, and their predatory activity to the eggs of Fasciola gigantica and Gigantocotyle explanatum. Maximum growth of P. lilacinus was obtained in corn-meal agar compared to any other media types. The preferred medium for growth of V. chlamydosporium was corn-meal agar, followed by potato-dextrose agar. After initial growth for 16 h of incubation, no growth was observed in water agar for both the fungi. Six different temperatures--4 degrees C, 10 degrees C, 18 degrees C, 26 degrees C, 34 degrees C and 40 degrees C--were used to observe growth profiles of the fungi in corn-meal agar medium. While no and very little growth of P. lilacinus and V. chlamydosporium was observed at 4 degrees C and 10 degrees C, respectively, growth profiles of both the fungi were optimal at 26-40 degrees C. A range of pH (pH 4-8) supported growth of both P. lilacinus and V. chlamydosporium. Full-grown plates of the fungi baited with viable eggs of F. gigantica and G. explanatum revealed that V. chlamydosporium was more vigorous in its egg-parasitic ability compared to P. lilacinus. Distortion of the eggs started on day 2-3 of egg baiting in culture plates of V. chlamydosporium, with complete distortion by day 7. On the contrary, P. lilacinus exhibited very limited egg-parasitic ability and some of the baited eggs even showed development of miracidia.     

Gag-processing defect of human immunodeficiency virus type 1 integrase E246 and G247 mutants is caused by activation of an overlapping 5' splice site

We have previously described several human immunodeficiency virus type 1 (HIV-1) mutants that are characterized by an excessive-RNA-splicing phenotype and reduced virus particle production. In one of these mutants (NLD2up), the sequence of 5′ splice site D2 was changed to a consensus splice donor site. This splice site overlaps the HIV-1 integrase reading frame, and thus, the NLD2up mutant also bears a G-to-W change at amino acid 247 of the integrase. A previously described E-to-K mutant at position 246 of the C-terminal domain of the integrase, which resulted in a G-to-A mutation at the +3 position of overlapping splice donor D2 (NLD2A3), was also shown to affect virus particle production and Gag protein processing. By using second-site mutations to revert the excessive-splicing phenotype, we show that the effects on Gag protein processing and virus particle production of both the NLD2up and NLD2A3 mutants are caused by excessive viral RNA splicing due to the activation of the overlapping 5′ splice site and not to the changes in the integrase protein. Both integrase protein mutations, however, are lethal for virus infectivity. These studies suggest that changes in the usage of overlapping splice sites may be a possible alternative explanation for a defective virus phenotype resulting from changes in protein-coding sequences or in the nucleotide sequence during codon optimization.     

Concise synthesis of the pentasaccharide O-antigen of Escherichia coli O83:K24:H31 present in the Colinfant vaccine

A block synthetic approach is presented for the synthesis of the pentasaccharide repeating unit of the O-antigen of E. coli O83:K24:H31 strain, present in the “Colifant” vaccine. The target pentasaccharide has been synthesized by coupling a disaccharide with a trisaccharide in excellent yield. Yields are quite satisfactory in all intermediate steps. A concise synthesis of the pentasaccharide repeating unit of the O-antigen of E. coli O83:K24:H31 strain, present in the COLINFANT vaccine is presented. The target pentasaccharide has been synthesized following a block synthetic strategy by coupling a disaccharide with a trisaccharide in excellent yield.     

Simultaneous determination of metoprolol succinate and amlodipine besylate in human plasma by liquid chromatography-tandem mass spectrometry method and its application in bioequivalence study

A simple, sensitive and specific liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of metoprolol succinate (MPS) and amlodipine besylate (AM) using hydrochlorothiazide (HCTZ) as IS in human plasma. Both the drugs were extracted by simple liquid-liquid extraction with chloroform. The chromatographic separation was performed on a reversed-phase peerless basic C18 column with a mobile phase of methanol-water containing 0.5% formic acid (8:2, v/v). The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The method was validated over the concentration range of 1-100ng/ml for MPS and 1-15ng/ml AM in human plasma. The MRM transition of m/z 268.10-103.10, m/z 409.10-334.20 and m/z 296.00-205.10 were used to measure MPS, AM and HCTZ (IS), respectively. This method was successfully applied to the pharmacokinetic study of fixed dose combination (FDC) of MPS and AM formulation product after an oral administration to Indian healthy human volunteers.     

Modeling tyrosinase activity. Effect of ligand topology on aromatic ring hydroxylation: an overview

Synthetic modeling of tyrosinase (o-phenol ring hydroxylation) has emerged as a novel class of successful biomimetic studies. It is a well-established fact that the reaction of dioxygen with copper(I) complexes of m-xylyl-based ligands generate putative copper-oxygen intermediate species such as side-on peroxo {CuII2(mu-O2)}2+ [in some cases bis-oxo {CuIII2(mu-O)2}2+ in equilibrium with isomeric side-on peroxo], due to oxygen activation. Electrophilic attack of such species brings about monooxygenase activity by incorporating one of the oxygens to m-xylyl ring of the ligand and the other oxygen is reduced to hydroxide ion. The goal of this review is to provide a concise overview of the present day knowledge in this field of research to emphasize the important role the designed ligands play in eliciting the desired tyrosinase-like chemistry.     

Attenuation of Zn-Induced Acute Pancreatitis in Wistar Rat Fed on Cu- and Mg- Enriched Modified Poultry Egg^{\Psi }

Zinc (Zn) consumption has increased in many populations either due to the increased intake of Zn-fortified foods as in the USA or in agricultural food stuffs as in some Indian states during the last decade. Its excessive intake has been reported to induce acute pancreatitis (AP) in many studies due to increase in oxidative stress that was further reported to cause Cu and Mg deficiencies. This led us to design a modified poultry egg (ME^Ψ) enriched with Cu and Mg along with other antioxidants, and its efficacy on Zn-induced AP was studied in male Wistar rats. In one set, the rats were fed on equacaloric semi-synthetic basal diet containing 20 mg Zn/kg diet (control, group I), and Zn-induced AP-I diet and AP-II diet containing 40 and 80 mg Zn/kg diet (groups II and III) for 180 days, respectively. In another set, the rats were initially fed on Zn-induced AP-I and AP-II diets for 90 days and then shifted to -mixed Zn-induced AP-I and AP-II diets in groups IIME and IIIME for another 90 days. At the end of the experiment, data displayed increased serum and urinary Zn, Cu, and Mg levels in groups II and III rats, which were reduced and approached closer to control group I after feeding in groups IIME and IIIME rats. Transmission electron microscopic studies of acinar cells revealed progressive dilation, vesicularization, and degeneration of endoplasmic reticulum (ER), and decrease in zymogen granules (ZG) in groups II and III rats in contrast to their curvilinear or concentric long parallel running cisternal profile of ER in control group I. The treatment of helped in the restoration of the ER profile and ZG number, approaching closer to the control group I. The degree of recovery was dependent upon the degree of toxicity caused by the amount of Zn given in the diet. The results of this study suggest that -mixed diet can protect the acinar cells from the deleterious effects of Zn by decreasing the oxidative stress. Indian patent application no. 2264/Del/2005.     

Assessment of Minerals in Obesity-related Diseases in the Chandigarh (India) Population

Excessive Zn but normal Cu and Mg in the staple food consumed by the people of Chandigarh (Union territory and capital of Punjab and Haryana States of India) has been considered to be the major risk factor for the prevalence of obesity (33.15%) and obesity-related diseases in this region. Therefore, in the present investigations, in obesity-related diseases, the status of these minerals was estimated in their tissues, including hair, nails, and blood serum and urine, and compared with those of normal subjects. They were grouped as: normal subjects in control Group A, middle-aged diabetics in Group D_M, older diabetics in Group D_O, and diabetics with osteoarthritis in Group D+ OA, osteoarthritis in Group OA and rheumatoid arthritis in Group RA, respectively. The results evaluated in the order as: hair Zn, group D+OA>D_M>OA>A (control)>RA>D_O (p < 0.001); hair Cu, group A (control)>D_M>OA>D+OA>D_O>RA (p < 0.001); hair Mg, group A (control)>D_M>OA>D+OA>RA>D_O (p < 0.001, 0.01); hair Mn, group A (control)>RA>OA>D-OA>D_M>D_O (p < 0.001); nail Zn, group D_M>D+OA>OA>A (control)>RA>D_O (p < 0.001, 0.05); nail Cu, group A (control)>OA>D_M>D+OA>RA>D_O (p < 0.001); nail Mg, group A (control)>OA>D_M>D_O>D+OA >RA (p < 0.001); nail Mn, group A (control) >RA>OA>D+OA>D_M>D_O (p < 0.01); urine Zn, group D_O>D_M>D+OA>A (control)>RA>OA (p < 0.01); urine Cu, group RA>D+OA>D_O>OA> D_M>A (control) (p<0.001); urine Mg, group RA>OA>D+OA>D_O>D_M>A (control; p < 0.001); urine Mn, group D_O>D_M>OA>D+OA>RA>A (control; p < 0.001), respectively. The analysis of the mineral status in serum of diabetics further showed their highly significant rise from lower mean age subgroup to higher mean age subgroup than their control counter parts (p < 0.001, 0.01, and 0.05) with coincident deficiencies of Cu, Mg, and Mn in their tissues. This study would be helpful considering the status of minerals in these obesity-related diseases depending on the choice of the food consumed to improve the quality of life and prognosis for the diseases.     

Optimized culture conditions for bacteriocin production by Pediococcus acidilactici LAB 5 and its characterization

A strain of Pediococcus acidilactici LAB 5 was isolated from vacuum-packed fermented meat product, in order to obtain a novel bacteriocin from food-grade organisms. Optimized culture conditions for bacteriocin production in different media (viz., MRS, TGE, TGE + buffer, TGE + Tween 80, and TGE + Tween 80 + buffer) and at different temperatures and pH conditions were reported. TGE + Tween 80 + buffer medium was found to be most effective for bacteriocin production (about 2400 AU/ml) by this strain, when incubated at 37 degrees C for 24 h. Bacteriocin, partially purified by adsorption-desorption method showed molecular mass of 10.3 kDa and produced prominent inhibition zone in activity gel. It showed significant storage stability both at high as well as in low temperatures for up to 6 months and retained its activity in a number of organic solvents, except in 2-mercaptoethanol. The treatment with amylase and lysozyme did not change its activity, but it lost its activity on proteinase K treatment. Antibacterial efficacy of bacteriocin was proved against some food spoilage and human pathogenic bacteria like Enterococcus, Leuconostoc, Listeria, Staphylococcus and Streptococcus.