Haplotype data for 17 Y-STR loci in the population of Himachal Pradesh, India.

We report here the data of Y chromosome haplotypes of 259 unrelated males from the population of Himachal Pradesh, India, using the Yfiler® multiplex kit. A total of 188 haplotypes were detected, out of which 148 were unique. Three samples showed bi-allelic pattern on locus DYS448. The observed genetic diversity and discrimination capacity were 0.996 and 0.73, respectively. In order to compare the genetic distance of the studied population with the published populations, multidimensional scaling (MDS) plot was constructed. The reported data is expected to be valuable for both forensic and population genetics.

Stem cell/cellular interventions in human spinal cord injury: Is it time to move from guidelines to regulations and legislations? Literature review and Spinal Cord Society position statement.

PURPOSE: In preclinical studies, many stem cell/cellular interventions demonstrated robust regeneration and/or repair in case of SCI and were considered a promising therapeutic candidate. However, data from clinical studies are not robust. Despite lack of substantial evidence for the efficacy of these interventions in spinal cord injury (SCI), many clinics around the world offer them as "therapy." These "clinics" claim efficacy through patient testimonials and self-advertisement without any scientific evidence to validate their claims. Thus, SCS established a panel of experts to review published preclinical studies, clinical studies and current global guidelines/regulations on usage of cellular transplants and make recommendations for their clinical use. METHODS: The literature review and draft position statement was compiled and circulated among the panel and relevant suggestions incorporated to reach consensus. This was discussed and finalized in an open forum during the SCS Annual Meeting, ISSICON. RESULTS: Preclinical evidence suggests safety and clinical potency of cellular interventions after SCI. However, evidence from clinical studies consisted of mostly case reports or uncontrolled case series/studies. Data from animal studies cannot be generalized to human SCI with regard to toxicity prediction after auto/allograft transplantation. CONCLUSIONS: Currently, cellular/stem cell transplantation for human SCI is experimental and needs to be tested through a valid clinical trial program. It is not ethical to provide unproven transplantation as therapy with commercial implications. To stop the malpractice of marketing such "unproven therapies" to a vulnerable population, it is crucial that all countries unite to form common, well-defined regulations/legislation on their use in SCI. These slides can be retrieved from Electronic Supplementary Material.

A genomic exploration of 15 autosomal STR loci for establishment of a DNA profile database of the population of Himachal Pradesh.

In order to create an autosomal STR loci population database for Himachal Pradesh, 259 blood samples were taken from people residing in various regions of the state and AmpFlSTR® Identifiler® Plus PCR amplification kit was used for evaluation of 15 autosomal STR markers. A total of 149 alleles were investigated in this study with a mean allele number of 9.933 per locus. The locus D2S1338 was most informative in our data, as it had the highest discrimination power (PD-0.967) and the highest polymorphic information content (PIC-0.86). The matching probability and typical paternity index for all the studied loci were observed as 2.9x10-18 and 4.7x105, respectively. Discrimination power (CPD) and exclusion power (CPE) for all the studied loci were observed as 1 and 0.999998.

The CCTTT pentanucleotide microsatellite in iNOS promoter influences the clinical outcome in P. falciparum infection.

To assess the hypothesis that nitric oxide (NO) is critical in the pathogenesis of cerebral malaria, we analyzed those single nucleotide polymorphisms (SNPs) and microsatellite (MS) of the promoter region of inducible nitric oxide synthase (iNOS) gene which are known to enhance the NO production in vivo. A total of 428 (204 severe, 224 mild) adult patients living in the eastern part of India were analyzed. The single nucleotide substitutions -954G-->C was found to be very rare, and -1173C-->T was absent in this population. But interestingly, longer forms of MS were found to be significantly associated with severe malaria (OR = 2.89, 95% CI = 1.955-4.295, P < 0.0001), and the linear regression analysis revealed that the risk of severe malaria significantly increases as the summed repeat number in an individual increase (OR = 1.16, P = 0.0013). Further, the median plasma level of nitrate/nitrite (NOx) was observed to be high in mild patients compared to severe patients, and the level of parasitemia was significantly low among mild patients than severe ones. These findings suggest that the CCTTT repeats in iNOS may play a key role in the pathogenesis of severe malaria.

Comparative efficacy of five different rep-PCR methods to discriminate Escherichia coli populations in aquatic environments.

Development of efficient techniques to discriminate the sources of E. coli in aquatic environments is essential to improve the surveillance of fecal pollution indicators, to develop strategies to identify the sources of fecal contamination, and to implement appropriate management practices to minimize gastrointestinal disease transmission. In this study the robustness of five different rep-PCR methods, such as REP-PCR, ERIC-PCR, ERIC2-PCR, BOX-PCR and (GTG)(5)-PCR were evaluated to discriminate 271 E. coli strains isolated from two watersheds (Lakelse Lake and Okanagan Lake) located in British Columbia, Canada. Cluster analysis of (GTG)(5)-PCR, BOX-PCR, REP-PCR, ERIC-PCR and ERIC2-PCR profiles of 271 E. coli revealed 43 clusters, 35 clusters, 28 clusters, 23 clusters and 14 clusters, respectively. The discriminant analysis of rep-PCR genomic fingerprints of 271 E. coli isolates yielded an average rate of correct classification (watershed-specific) of 86.8%, 82.3%, 78.4%, 72.6% and 55.8% for (GTG)(5)-PCR, BOX-PCR, REP-PCR, ERIC-PCR and ERIC2-PCR, respectively. Based on the results of cluster analysis and discriminant function analysis, (GTG)(5)-PCR was found to be the most robust molecular tool for differentiation of E. coli populations in aquatic environments.

Targeted disruption of the transition protein 2 gene affects sperm chromatin structure and reduces fertility in mice.

During mammalian spermiogenesis, major restructuring of chromatin takes place. In the mouse, the histones are replaced by the transition proteins, TP1 and TP2, which are in turn replaced by the protamines, P1 and P2. To investigate the role of TP2, we generated mice with a targeted deletion of its gene, Tnp2. Spermatogenesis in Tnp2 null mice was almost normal, with testis weights and epididymal sperm counts being unaffected. The only abnormality in testicular histology was a slight increase of sperm retention in stage IX to XI tubules. Epididymal sperm from Tnp2-null mice showed an increase in abnormal tail, but not head, morphology. The mice were fertile but produced small litters. In step 12 to 16 spermatid nuclei from Tnp2-null mice, there was normal displacement of histones, a compensatory translationally regulated increase in TP1 levels, and elevated levels of precursor and partially processed forms of P2. Electron microscopy revealed abnormal focal condensations of chromatin in step 11 to 13 spermatids and progressive chromatin condensation in later spermatids, but condensation was still incomplete in epididymal sperm. Compared to that of the wild type, the sperm chromatin of these mutants was more accessible to intercalating dyes and more susceptible to acid denaturation, which is believed to indicate DNA strand breaks. We conclude that TP2 is not a critical factor for shaping of the sperm nucleus, histone displacement, initiation of chromatin condensation, binding of protamines to DNA, or fertility but that it is necessary for maintaining the normal processing of P2 and, consequently, the completion of chromatin condensation.

Purification and characterization of a novel caffeine oxidase from Alcaligenes species.

Alcaligenes species CF8 isolated from surface water of a lake produced a novel serine type metallo-caffeine oxidase. The optimal medium for caffeine oxidase production by this strain was (w/v) NaNO(3), 0.4%; KH(2)PO(4), 0.15%; Na(2)HPO(4), 0.05%; FeCl(3).6H(2)O, 0.0005%; CaCl(2).2H(2)O, 0.001%; MgSO(4).7H(2)O, 0.02%; glucose, 0.2%; caffeine, 0.05%, pH 7.5. The enzyme was purified to 63-fold by using ammonium sulfate precipitation, dialysis, ion exchange (diethylaminoethyl-cellulose) and gel filtration (Sephadex G-100) chromatographic techniques. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the purified caffeine oxidase was monomeric with a molecular mass of 65 kDa. The purified caffeine oxidase with a half-life of 20 min at 50 degrees C had maximal activity at pH 7.5 and 35 degrees C. The purified caffeine oxidase had strict substrate specificity towards caffeine (K(m) 8.94 microM and V(max) 47.62 U mg protein(-1)) and was not able to oxidize xanthine and hypoxanthine. The enzyme activity was not inhibited by para-chloromercuribenzoic acid, iodoacetamide, n-methylmaleimide, salicylic acid and sodium arsenite indicating the enzyme did not belong to xanthine oxidase family. The enzyme was not affected by Ca(+2), Mg(+2) and Na(+), but was completely inhibited by Co(+2), Cu(+2) and Mn(+2) at 1mM level. The novel caffeine oxidase isolated here from Alcaligenes species CF8 may be useful in biotechnological processes including waste treatment and biosensor development.

Optimization of culture conditions and properties of immobilized sulfide oxidase from Arthrobacter species.

Arthrobacter species strain FR-3, isolated from sediments of a swamp, produced a novel serine-type sulfide oxidase. The production of sulfide oxidase was maximal at pH 7.5 and 30 degrees C. Among various carbon and nitrogen sources tested, glucose and yeast extract were found to be the most effective substrates for the secretion of sulfide oxidase. The sulfide oxidase was purified to homogeneity and the molecular weight of the purified enzyme was 43 kDa when estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified sulfide oxidase can be effectively immobilized in DEAE (diethylaminoethyl)-cellulose matrix with a yield of 66%. The purified free and immobilized enzyme had optimum activity at pH 7.5 and 6.0, respectively. Immobilization increases the stability of the enzyme with respect to temperature. The half-life of the immobilized enzyme was 30 min at 45 degrees C, longer than that of the free enzyme (10 min). The purified free sulfide oxidase activity was completely inhibited by 1 mM Co2+ and Zn2+ and sulfhydryl group reagents (para-chloromercuribenzoic acid and iodoacetic acid). Catalytic activity was not affected by 1 mM Ca2+, Mg2+, Na+ and metal-chelating agent (EDTA).

Studies on manganese nodule leached residue 4. Physicochemical characterization and catalytic activity of acetic acid treated manganese nodule leached residue.

The catalytic activity of water-washed manganese nodule leached residue (WMNLR) samples improved by treating with acetic acid. The effects of acetic acid treatment on the physicochemical properties and catalytic activity of manganese nodule leached residue have been studied. The surface area, surface oxygen, surface hydroxyl groups, surface acidity, electron donating properties, etc., increase gradually with acid treatment up to 0.5 M and thereafter show a decreasing trend. The rate constant of H2O2 decomposition, catalytic activity of CO oxidation, and esterification of acetic acid also show a similar trend to that of surface properties.

Synthesis, characterization, and catalytic activity of phosphomolybdic acid supported on hydrous zirconia.

A series of ecofriendly solid acid catalyst was synthesized by phosphomolybdic acid impregnated on hydrous zirconia (ZPMA) by an incipient wetness impregnation technique. The support and the resulting catalysts were characterized by X-ray diffraction, FTIR, EPMA, BET surface area, acid sites, etc. These studies indicate that the phosphomolybdic acid keeps its Keggin-type structure unaltered up to 500 degrees C. Catalytic activities of the material were evaluated for the esterification of acetic acid reaction. The 12 wt% of phosphomolybdic acid supported on hydrous zirconia shows highest surface area, acid sites and gives about 86% conversion with 100% selectivity of n-butyl acetate.

Assessment of feeding response of tumor-bearing rats to hypothalamic injection and infusion of neuropeptide Y.

Tumor-bearing rats exhibited significant decreases in 1- to 4-h intake of rat chow following the intrahypothalamic injection of 2 micrograms neuropeptide Y (NPY). This refractory feeding response was present prior to the onset of anorexia and became more severe as anorexia worsened. The constant infusion of NPY (125 ng/h) into the perifornical hypothalamus of TB and control rats elicited increased feeding for only 2 days. Because chromatography revealed minipump NPY to be intact after 10 infusion days, downregulation of NPY receptors may have occurred. Daily injection of increasing doses of NPY stimulated ad lib feeding in non-TB rats, while having no effect on TB rats. Desensitization to NPY-induced feeding following daily injections of the peptide was suggested by the loss of feeding response to a dose (500 ng) of NPY that increased food intake prior to the daily NPY treatments. These results suggest that hypothalamic NPY feeding systems are refractory in TB rats, even before they exhibit anorexia. In addition, a rapid loss of the feeding response occurred in rats with constant infusion of NPY into hypothalamic tissue or with daily intrahypothalamic injections of the peptide, suggesting possible NPY receptor-mediated alterations. Therefore, control of obesity or anorexia through NPY feeding mechanisms may prove difficult due to rapid compensatory receptor changes.

Potentiation of bleomycin-induced chromosome aberrations by the radioprotector reduced glutathione.

In this study we investigated whether the radioprotector reduced glutathione (GSH) can reduce the frequency of chromosome aberrations induced by the radiomimetic antitumour drug bleomycin (BLM) in muntjac lymphocytes in vitro. Our results demonstrate that, instead of yielding any protection, the presence of GSH potentiates the clastogenic action of BLM. A significant enhancement in the frequency of rearrangements and deletions was observed and the number of aberrations per metaphase was also enhanced. We suggest that this potentiation may be due to GSH acting as a reducing agent in reactivating oxidised BLM.

High prevalence & heterogenicity of thalassaemias in Orissa.

High incidence of alpha thalassaemias (estimated from detection of Hb Bart's from cord blood), Hb constant spring (CS) and high A2 beta thalassaemia trait (estimated from normal adult subjects) were detected in 12.6, 15 and 8 per cent people of coastal Orissa (with less than 5% tribal population) respectively. Quantitation of Hb Bart's suggest that the alpha thalassaemia could be any of the genotypes such as, -alpha/alpha alpha, -alpha/-alpha, --/alpha alpha and alpha alpha/alpha alpha cs. Both heterozygotes and homozygotes for Hb CS were identified.

Double blind clinical trial on centperazine & DEC in bancroftian filariasis.

Centperazine, an analogue of DEC, was subjected to a double blind controlled trial, to evaluate its efficacy as a newer antifilarial agent. Centperazine (300 mg/day) along with equivalent quantities of DEC and placebo were administered to different types of filariasis patients. DEC was found to be significantly effective in reducing peripheral microfilaraemia, in different weeks and months of follow-up, except at the end of 6th month, as compared to Centperazine. There was no significant difference between the placebo and Centperazine treated patients, in this respect, revealing that the drugs had no efficacy in eliminating peripheral microfilaraemia. Recurrence of acute attack within 6 months was nearly equal with both Centperazine and DEC, being 28.2 and 24 per cent respectively, whereas in the placebo group the recurrence rate was 48.9 per cent. Centperazine treated patients showed significantly less side effects (8.9%), as compared to DEC treated patients (34%). Giddiness, nausea and vomiting were the common adverse effects observed.

Studies on manganese-nodule leached residues; 1. Physicochemical characterization and its adsorption behavior toward Ni(2+) in aqueous system.

Physicochemical characterization of manganese-nodule leached residues was carried out by chemical analyses, XRD, TG-DTA, surface area measurement, and FTIR techniques. The material is very fine-grained (<75 microm), is cryptocrystalline to amorphous in nature, and contains mainly of delta-MnO(2), quartz (alpha-SiO(2)), and zeolite/feldspar minerals. Physically adsorbed sulfates in the leached residue are removed by repeated water washing and the washed sample shows an appreciable increase in surface area. This is indicated by the absence of 1387 and 1099 cm(-1) peaks in the IR spectrum of the washed sample. The adsorption behavior of the washed sample toward Ni(2+) was recorded as a function of time, pH, temperature, and concentrations of adsorbent and adsorbate.

Antifungal efficacy of bacteria isolated from marine sedentary organisms.

The antibiotic-producing ability of 57 bacteria isolated from 8 marine sedentary organisms, 6 sponges (Spirastrella sp., Phyllospongia sp., Ircinia sp., Aaptos sp., Azorica sp., Axinella sp.), 1 soft coral (Lobophytum sp.) and 1 alga (Sargassum sp.), was evaluated against 6 phytopathogenic fungi (Helminthosporium oryzae, Rhizoctonium solani, Pyricularia oryzae, Fusarium oxysporum, Aspergillus oryzae and A. fumigatus). Bacteria of the genus Bacillus (20%), Pseudomonas (33%) and Flavobacterium (40%) were predominant among the heterotrophic bacteria isolated from the marine sponges, soft coral and alga, respectively. Bioassay results revealed that 36 (63%) bacterial isolates displayed antifungal activity against at least one fungus, the alga (Sargassum sp.) being the source of highest number (80%) of producer strains. Twelve bacterial isolates inhibited all fungi. The MIC of the organic extracts of 12 bacteria ranged from 0.3 to 22.8 mg/L.

Serum immunoglobulins in sickle cell disease.

Immunoglobulins IgG, IgA and IgM were estimated in 20 cases of sickle cell disease in steady state, eleven cases of sickle cell disease in crisis and/or infection, 20 cases of sickle cell trait and 14 normal healthy controls. Immunoglobulin deficiency has not been observed in sickle cell disease. Significantly raised IgG but normal IgA and IgM found in sickle cell disease cases was probably due to chronic stimulation of reticuloendothelial cells secondary to chronic hemolysis. IgM levels were significantly higher in patients with splenic enlargement > 3 cm. There was no alteration in immunoglobulin levels in sickle cell trait.

Cholelithiasis in sickle cell disease in India.

Eighty cases (63 Males and 17 Females) of sickle cell disease were searched for gall stone by ultrasonography, 8 (10%) cases had gall stone. Gall stones were more common in males (11.1%) than females (5.8%). It was not found below 13 years of age. Three cases had single and five cases had multiple gall stones. Three of them had thickened gall bladder wall. Typical biliary colic was uncommon (1 case). There was no case of obstructive jaundice. HbF% level was significantly lower in cases with gall stone (12.31 +/- 3.95) than without gall stones (16.73 +/- 6.30). Episodes of aggravated anaemia and total serum bilirubin was significantly higher (1.27/pt and 4.12 +/- 1.34 mg%) in cases with gall stone than without gall stone (0.31/pt and 2.74 +/- 1.47 mg%). The above findings suggest association of greater degree of haemolysis with formation of gall stone in sickle cell disease cases.

Detection of a new focus of Brugia malayi infection in Orissa.

526 people were surveyed in a village called Chudamani, in Balasore district of Orissa, for detection of asymptomatic microfilaria (mf) carriers. Of these 36 (6.8 per cent) were cases found to harbour mf; 19 cases had Brugia malayi, 4 Wuchereria bancrofti and 5 cases had mixed infection. In 8 cases, species could not be ascertained. For the first time after 1955, a focus of B. malayi has been detected in Orissa.