Hydrogeochemical characterization and evaluation of subsurface water quality in the Proterozoic Cuddapah Basin, Andhra Pradesh, India.

In the current survey, different hydrogeochemical processes governing the geochemistry of aquifers, the usefulness of groundwater for regular consumption, and agricultural purposes were evaluated around the Tummalapalle area. One hundred forty-four borehole locations were chosen to characterize the major physicochemical components of the aquifer water. The analysis results of pH inferred that the groundwater is nominally acidic to basic, and pH ranged from 6.6 to 8.4. The average concentrations of TDS, Ca2+, Mg2+, total hardness (TH), HCO3-, and total alkalinity (TA) are within the allowable limits of potable water quality as prescribed by the Bureau of Indian Standards (BIS) and WHO. However, the average concentrations of Na+, K+, Cl-, and SO42- were all below the permissible limit. All samples were analyzed with the help of Piper and Chadha charts to determine the dominant hydrogeochemical components of groundwater. The dominance of cations in groundwater in this region is in the sequence of Ca2+ > Na+ > Mg2+ > K+, followed by anions HCO3- > Cl- > SO42-. The Gibbs plot analysis suggested the predominance of rock aquifer interaction as the major hydrogeochemical process governing groundwater geochemistry in this region. The water quality index (WQI) of all groundwater samples in the Tummalapalle region was estimated, with 55% of the samples being potable grade. The different irrigation indices were analyzed for the groundwater samples to estimate their desirability for agriculture. The maximum number of water samples was found to be well-suited for cultivation.

The 2017 International Joint Working Group White Paper by INDUSEM, the Emergency Medicine Association and the Academic College of Emergency Experts on Establishing Standardized Regulations, Operational Mechanisms, and Accreditation Pathways for Education and Care Provided by the Prehospital Emergency Medical Service Systems in India.

The government of India has done remarkable work on commissioning a government funded prehospital emergency ambulance service in India. This has both public health implications and an economic impact on the nation. With the establishment of these services, there is an acute need for standardization of education and quality assurance regarding prehospital care provided. The International Joint Working Group has been actively involved in designing guidelines and establishing a comprehensive framework for ensuring high-quality education and clinical standards of care for prehospital services in India. This paper provides an independent expert opinion and a proposed framework for general operations and administration of a standardized, national prehospital emergency medical systems program. Program implementation, operational details, and regulations will require close collaboration between key stakeholders, including local, regional, and national governmental agencies of India.

The cholinesterases: analysis by pharmacogenomics in man.

We have undertaken a study on variations in cholinesterase (ChE) genes in relation to cardiovascular (CV) function and the metabolic syndrome. Peripheral and central nervous system control of cardiovascular (CV) function mediated through cholinergic pathways is critical in homeostatic maintenance of blood pressure and responsiveness to stress. For acetylcholinesterase (AChE; EC 3.1.1.7) our focus is to identify single nucleotide polymorphisms (SNPs) in the gene that are linked to cardiovascular function. For butyrylcholinesterase (BChE; EC 3.1.1.8) we examined whether BChE activity correlated with parameters of the metabolic syndrome and cardiovascular function. ChE can be found in whole blood enabling a characterization of biochemical phenotype in addition to correlating genotype with phenotypic physiologic responses. Analysis of enzymatic activity was determined spectrophotometrically in blood samples from twin and other subject registries. Correlation analysis revealed significant relationships between enzyme activity and certain CV endpoints. Linkage analysis with data from a dizygotic (DZ) twin set showed a suggestive linkage at the BChE locus, and statistical analysis revealed a high correlation between BChE activity and variables associated with cardiovascular risk and the metabolic syndrome. Pattern of within-pair twin correlations by zygosity and the ACE model-fitting findings suggest the major source of this variation (65%) is attributable to an additive genetic component. To date 19 SNPs have been identified by the re-sequencing of AChE including four nonsynonymous coding SNPs (cSNPs).

A quantitative study of seminiferous tubular cells in the developing Murrah buffalo testis.

We report here a systematic quantitative study of the seminiferous tubular cells of Murrah buffaloes. The most advanced germ cell types in the different age groups (months) were A(0) spermatogonia (SG) (1 and 3), early pachytene (6 and 9), late pachytene (12), secondary spermatocytes (15 and 18), elongating spermatids (21 and 24), elongated spermatids attached to Sertoli cells (30), elongated spermatids detached from Sertoli cells (36) and spermatozoa (42 and 48). Central primitive Sertoli cells (CPSC) and basal primitive Sertoli cells (BPSC) were present in the sex cord of one-month-old calves, while Sertoli cells (SC) were first seen in nine-month-old calves. The number of gonocytes were maximal at six months but they were not seen after this time. Prespermatogonia (PSG) and SG were at a maximum at nine months of age but PSG were not seen after 36 months. The number of SG decreased significantly after nine months up to 36 months of age. Although spermatocytes and spermatids appeared in earlier developmental stages, a rapid increase in their number was recorded after 36 months. The number of SC was maximal in 18-month-old animals. BPSC predominated in the sex cord of animals aged one to six months, SG at 9-12 months of age, primary spermatocytes from 15-30 months and spermatids from 36 to 72 months and in older animals. We concluded that a decrease in the number of SG in buffalo calves after nine months of age might be responsible for a delay in sexual maturity. Moreover, the small number of spermatocytes and spermatids present before 36 months of age may be associated with the low yield of different germ cell divisions and with the cellular degeneration. A rapid increase in the number of spermatocytes and spermatids after 36 months resulted in sexual maturity between 42 and 48 months.

Physical mapping of autonomic/sympathetic candidate genetic loci for hypertension in the human genome: a somatic cell radiation hybrid library approach.

Allelic variation at multiple genetic loci may contribute to hypertension. Since autonomic/sympathetic dysfunction may play an early, pathogenic, heritable role in hypertension, we evaluated candidate loci likely to contribute to such dysfunction, including catecholamine biosynthetic enzymes, catecholamine transporters, neuropeptides, and adrenergic receptors. Since chromosomal locations and physical map positions of many of these loci had not yet been identified, we used the GeneBridge4 human/hamster radiation (somatic cell) hybrid library panel (resolution approximately 1 to approximately 1.5 Mb), along with specifically designed oligonucleotide primers and PCR (200-400 bp products) to position these loci in the human genome. Primers were designed from sequences outside the coding regions (3'-flanking or intronic segments) to avoid cross-species (hamster) amplification. Chromosomal positions were assigned in cR (centi-Ray) units ( approximately 270 Kbp/cR(3000) for GeneBridge 4). A total of 13 loci were newly assigned chromosomal positions; of particular interest was a cluster of adrenergic candidate loci on chromosome 5q (including ADRB2, ADRA1A, DRD1, GPRK6, and NPY6R), a region harbouring linkage peaks for blood pressure. Such physical map positions will enable more precise selection of polymorphic microsatellite and single nucleotide polymorphism markers at these loci, to aid in linkage and association studies of autonomic/sympathetic dysfunction in human hypertension.

Distribution analysis of nonsynonymous polymorphisms within the G-protein-coupled receptor gene family.

The G-protein-coupled receptor (GPCR) superfamily is one of the largest classes of proteins in mammalian genomes. GPCRs mediate diverse physiological functions and are the targets of >50% of all clinical drugs. The sequencing of the human genome and large-scale polymorphism discovery efforts have established an abundant source of single nucleotide polymorphisms (SNPs), particularly those that result in a change in the encoded amino acids (cSNPs), many are of which in GPCRs. Although the majority of these cSNPs are assumed not to be disease-causing (nDCs), experimental data on their functional impact are lacking. Here, we have computationally analyzed the distribution of 454 cSNPs within the GPCR gene family and have found that disease-causing cSNPs (DCs) are overrepresented, whereas nDCs are underrepresented or neutral in transmembrane and extracellular loop domains, respectively. This finding reflects the relative importance of these domains to GPCR function and implies different biological characteristics for the two sets of human polymorphisms.

Key role for constitutive cyclooxygenase-2 of MDCK cells in basal signaling and response to released ATP.

Madin-Darby canine kidney (MDCK) cells release ATP upon mechanical or biochemical activation, initiating P2Y receptor signaling that regulates basal levels of multiple second messengers, including cAMP (J Biol Chem 275: 11735--11739, 2000). Data shown here document inhibition of cAMP formation by Gd(3+) and niflumic acid, channel inhibitors that block ATP release. cAMP production is stimulated via Ca(2+)-dependent activation of cytosolic phospholipase A(2), release of arachidonic acid (AA), and cyclooxygenase (COX)-dependent production of prostaglandins, which activate prostanoid receptors coupled to G(s) and adenylyl cyclase. In the current investigation, we assessed the expression and functional role of the two known isoforms of COX, COX-1 and COX-2. Treatment of cells with either a COX-1-selective inhibitor, SC-560, or COX-2-selective inhibitors, SC-58125 or NS-398, inhibited basal and UTP-stimulated cAMP levels. COX inhibitors also decreased forskolin-stimulated cAMP formation, implying this response is in part attributable to an action of AA metabolites. These findings imply an important role for the inducible form of COX, COX-2, under basal conditions. Indeed, COX-2 expression was readily detectable by immunoblot, and treatments that induce or reduce COX-2 expression in other cells (interleukin-1beta, tumor necrosis factor-alpha, phorbol ester, or dexamethasone) had minimal or no effect on the levels of COX-2 immunoreactivity. RT-PCR using isoform-specific primers detected COX-2 mRNA. We conclude that COX-2 is constitutively expressed in MDCK-D(1) cells and participates in basal and P2Y(2)-mediated signaling, implying a key role for COX-2 in regulation of epithelial cell function.

Use of an in silico approach to define the gene structure of eukaryotic adenylyl cyclases.

The limited information available regarding the gene structure of adenylyl cyclases (AC), which catalyze the synthesis of cAMP, suggests a complex arrangement with many exons and large introns such that molecular techniques to define these gene structures are time- and labor-intensive. We report here the use of a computer-based approach involving the assembly of fragmented sequence data generated by the Human Genome Project and nucleic acid analysis software to decipher the gene structure of human and murine AC 6 and other human AC isoforms (ACs 3, 7, and 8). The results, which document 21 exons in human and murine AC 6, human AC 3, 18 exons in AC 8, and 24 exons in AC 7, show substantial conservation of exon organization in the AC family and in particular regions of the AC protein. Application of such in silico methods should prove useful to characterize genes for other ACs and protein families and data provided here should facilitate studies of polymorphisms in AC genes.

Genetic variations and polymorphisms of G protein-coupled receptors: functional and therapeutic implications.

G protein-coupled receptors (GPCRs) represent a major class of proteins in the genome of many species, including humans. In addition to the mapping of a number of human disorders to regions of the genome containing GPCRs, a growing body of literature has documented frequently occurring variations (i.e. polymorphisms) in GPCR loci. In this article, we use a domain-based approach to systematically examine examples of genetic variation in the coding and noncoding regions of GPCR loci. Data to date indicate that residues in GPCRs are involved in ligand binding and coupling to G proteins and that regulation can be altered by polymorphisms. Studies of GPCR polymorphisms have also uncovered the functional importance of residues not previously implicated from other approaches that are involved in the function of GPCRs. We predict that studies of GPCR polymorphisms will have a significant impact on medicine and pharmacology, in particular, by providing new means to subclassify patients in terms of both diagnosis and treatment.

High polymorphism at the human melanocortin 1 receptor locus.

Variation in human skin/hair pigmentation is due to varied amounts of eumelanin (brown/black melanins) and phaeomelanin (red/yellow melanins) produced by the melanocytes. The melanocortin 1 receptor (MC1R) is a regulator of eu- and phaeomelanin production in the melanocytes, and MC1R mutations causing coat color changes are known in many mammals. We have sequenced the MC1R gene in 121 individuals sampled from world populations with an emphasis on Asian populations. We found variation at five nonsynonymous sites (resulting in the variants Arg67Gln, Asp84Glu, Val92Met, Arg151Cys, and Arg163Gln), but at only one synonymous site (A942G). Interestingly, the human consensus protein sequence is observed in all 25 African individuals studied, but at lower frequencies in the other populations examined, especially in East and Southeast Asians. The Arg163Gln variant is absent in the Africans studied, almost absent in Europeans, and at a low frequency (7%) in Indians, but is at an exceptionally high frequency (70%) in East and Southeast Asians. The MC1R gene in common and pygmy chimpanzees, gorilla, orangutan, and baboon was sequenced to study the evolution of MC1R. The ancestral human MC1R sequence is identical to the human consensus protein sequence, while MC1R varies considerably among higher primates. A comparison of the rates of substitution in genes in the melanocortin receptor family indicates that MC1R has evolved the fastest. In addition, the nucleotide diversity at the MC1R locus is shown to be several times higher than the average nucleotide diversity in human populations, possibly due to diversifying selection.

Antifungal activity and kinetics of inhibition by essential oil isolated from leaves of Aegle marmelos.

The antifungal activity of essential oil isolated from the leaves of bael (Aegle marmelos (L.) Correa ex Roxb., Rutaceae) has been evaluated using spore germination assay. The oil exhibited variable efficacy against different fungal isolates and 100% inhibition of spore germination of all the fungi tested was observed at 500 ppm. However, the most resistant fungus, Fusarium udum was inhibited 80% at 400 ppm. Kinetic studies showed concentration as well as time dependent complex inhibition of spore germination by the essential oil.

Formation and activities of xylanhydrolysing enzymes of Humicola grisea var thermoidea.

A thermophilic fungus, Humicola grisea var thermoidea, produced in liquid culture two endoxylanases (1,4-ß-D-xylan-xylanohydrolase, EC 3.2.1.8) with M r of 95 (Xyl I) and 13 (Xyl II) kDa. PAGE of the crude culture filtrate and of each fraction obtained by gel filtration produced three and one band, respectively. Cross-reaction of the culture filtrate and each fraction with polyclonal antibodies prepared against Xyl II produced two and one precipitin bands, respectively. Hydrolysis of wheat straw and rice husk xylan was maximal using a combination of Xyl I and Xyl II. The products formed after hydrolysis, xylo-oligosaccharides and traces of xylose, indicated an endotype enzyme action and the co-operative activities of the xylanases.

A rapid method for detecting fungi-toxic substances.

A simple and rapid technique is reported for the preliminary screening of fungi-toxic extracts/samples by direct spotting onto silica gel plates and subsequent over-spraying with a fungal spore suspension. After incubation fungi-toxicity is indicated by a growth inhibition zone, the area of which is related to the concentration of the sample.