Characterization and identification of Russula firmula and Russula postiana from Himalayan moist temperate forests of Kashmir

Two ectomycorrhizal species of genus Russula: Russula postiana and Russula firmula (Basidiomycota, Agaricales) have been characterized and identified from Kashmir Himalaya using morpho-anatomical and molecular methods targeting its rDNA. The target internal transcribe spacer (ITS)-rDNA of both species was amplified using polymerase chain reaction (PCR) with universal fungal primers (ITS1 and ITS4), which generated 700 bp fragments. After sequencing of amplified product, the initial blast analysis revealed and confirmed the identification of both species by comparing the sequences of these respective species present in GenBank. Further, in phylogenetic analysis both species distinctly clustered with their respective groups. Morphological characteristics like shape, size and colour of pileus, stipe and gills, basidiospore size of both the species was measured and compared with data given in literature.Keywords: Ectomycorrhizal, morpho-anatomical, sequencing, phylogeneticAfrican Journal of Biotechnology Vol. 12(23), pp. 3643-364

Genotypic variants of MYP2 Locus: Analysis for association with High Myopia

Identification of genes involved in the progression of myopia is largely hampered by challenges inherent in mapping genes due to high prevalence, genetic heterogeneity, and wide clinical spectrum of the condition. Genetic mapping studies have identified at least 24 chromosomal loci suspected of harboring genes for myopia progression, MYP1–MYP24 of which MYP2 is considered to be a strong candidate gene locus. Environmental and genetic factors together are attributed to explain the spectrum of geographical and population dependent variations in the incidence of high myopia. Incidently researchers have come up with controversial results with regard to the association of MYP2 locus despite a varied spectrum of polymorphic changes reported in the genes harboured by the locus. The controversy is largely attributed to population heterogeneity. The purity of genetic traits associated with Kashmiri population is likely to minimize the influence of mixed risk/resistance alleles to reliably establish their potential association. One of the three SNPs observed in codon 10 of TGFβ1 showed a significant difference between patients and control subjects (rs1982073: p genotype=0.003, p allele=0.001). There were no statistically significant differences between patients and control subjects for the other two SNPs, rs1800471 at codon 25 and a novel variant at codon 52. In TGIF1 three adjacent novel intronic variations (T>C/A; p=0.04: T>G; p=0.02: G>C; p=0.01) and one novel missense sequence variation G26A (p = <0.001) were observed that show possible association with high myopia. G26A also segregates with gender and degree of myopia (p = 0.05). DLGAP1 gene revealed a total of two polymorphic variations among which G507A (P=1) was novel and one reported polymorphic variation G517A with a significant (P=<0.001) occurrence in affected population. G517A show association with gender and degree of myopia (p=<0.0001). A previously reported variant T451C observed in EMILIN2 gene did not appear to associate with disease phenotype. MYOM1 showed five polymorphic variations; two in coding region (G333A; P=A; P=T & C>G; P= < 0.001) that potentially segregate with the disease phenotype. G333A shows a statistically significant association with gender (p = 0.01) and degree of myopia (p = 0.01) while G341C does not associate with any of the clinical parameters. Among intronic variations, G>T (rs55779127) and C>G (rs8096379) showed significant association with degree of myopia (p=<0.0001 & p=<0.001). The assessment of the ITASSER predicted protein structure showed change in energy for almost all mutants compared to wild type proteins. The results are indicative that the energy changes due to these polymorphic variations may have significant functional consequences

Mutations in OTOF, CLDN14 & SLC26A4 genes as major causes of hearing impairment in Dhadkai village, Jammu & Kashmir, India

Background & objectives: A high incidence of hearing impairment is reported from the village of Dhadkai in the State of Jammu and Kashmir, India. Prevalence of endogamy in this community suggested a common genetic basis for the disorder. A genetic study was undertaken to ascertain the basis for the high incidence of hearing impairment in this region. Methods: In a two-step approach to identify the causative mutation/s, a whole-genome-based linkage analysis of an extended family of 45 members was carried out, which included 23 affected and 22 unaffected members. Mutational analysis for the candidate deafness genes helped reveal causative mutations in the family. In addition, seven deafness-causing genes, Cx26, SLC26A4, CLDN14, TMPRSS3, TMC1, TMIE and USH1C, were analyzed in smaller families with hearing impairment. Results: In the 45-member extended family, the critical chromosomal region mapped to 2p24-p22.The c.2122C>T (p.R708X) mutation in OTOF in 2p24-p22was identified as being the causal change. Linkage to 2p24-p22 locus was not observed in a particular branch of this extended family. Analysis of seven known deafness-causing genes in this branch revealed a mutation, c.254T>A (p.V85D), in CLDN14. Among seven small families unrelated to the 45-member extended family, hearing loss was attributable to p.R708X in OTOF in three families and to p.V85D in CLDN14 in one family; a new mutation c.1668T>A (p.Y556X) SLC26A4 was identified in two families and the causative change could not be identified in one family. Interpretation & conclusions: This study suggested considerable genetic heterogeneity in the causation of hearing loss in Dhadkai. Recessive mutations were observed in at least three genes causing hearing loss: OTOF (p.R708X), SLC26A4 (p.Y556X) and CLDN14 (p.V85D). Mutation p.R708X appeared to be the major cause of hearing impairment in Dhadkai

Growth Inhibition by Bupivacaine Is Associated with Inactivation of Ribosomal Protein S6 Kinase 1

Bupivacaine is an amide type long acting local anesthetic used for epidural anesthesia and nerve blockade in patients. Use of bupivacaine is associated with severe cytotoxicity and apoptosis along with inhibition of cell growth and proliferation. Although inhibition of Erk, Akt, and AMPK seemingly appears to mediate some of the bupivacaine effects, potential downstream targets that mediate its effect remain unknown. S6 kinase 1 is a common downstream effector of several growth regulatory pathways involved in cell growth and proliferation known to be affected by bupivacaine. We have accordingly attempted to relate the growth inhibitory effects of bupivacaine with the status of S6K1 activity and we present evidence that decrease in cell growth and proliferation by bupivacaine is mediated through inactivation of S6 kinase 1 in a concentration and time dependent manner. We also show that ectopic expression of constitutively active S6 kinase 1 imparts substantial protection from bupivacaine induced cytotoxicity. Inactivation of S6K1 though associated with loss of putative mTOR mediated phosphorylation did not correspond with loss of similar phosphorylations in 4EBP1 indicating that S6K1 inhibition was not mediated through inactivation of mTORC1 signaling pathway or its down regulation