Zareena Begum

Antitumor activity of curcumin is mediated through the induction of apoptosis in AK-5 tumor cells.

Curcumin, the yellow pigment of turmeric (Curcuma longa), used commonly as a spice, has been shown to possess anticarcinogenic activity. Curcumin inhibited AK‐5 tumor growth and induced apoptosis in AK‐5 cells. Curcumin induced apoptosis is mediated through the activation of caspase‐3, which is specifically inhibited by the tetrapeptide Ac‐DEVD‐CHO. In addition, curcumin induced tumor cell death is caused through the generation of reactive oxygen intermediates which is inhibited by N‐acetyl‐l‐cysteine. Our studies suggest that the apoptotic process induced by curcumin is the mechanism mediating AK‐5 tumor cell death.

Janibacter hoylei sp. nov., Bacillus isronensis sp. nov. and Bacillus aryabhattai sp. nov., isolated from cryotubes used for collecting air from the upper atmosphere

Three novel bacterial strains, PVAS-1(T), B3W22(T) and B8W22(T), were isolated from cryotubes used to collect air samples at altitudes of between 27 and 41 km. Based on phenotypic characteristics, chemotaxonomic features, DNA-DNA hybridization with the nearest phylogenetic neighbours and phylogenetic analysis based on partial 16S rRNA gene sequences (PVAS-1(T), 1196 nt; B3W22(T), 1541 nt; B8W22(T), 1533 nt), the three strains were identified as representing novel species, and the names proposed are Janibacter hoylei sp. nov. (type strain PVAS-1(T) =MTCC 8307(T) =DSM 21601(T) =CCUG 56714(T)), Bacillus isronensis sp. nov. (type strain B3W22(T) =MTCC 7902(T) =JCM 13838(T)) and Bacillus aryabhattai sp. nov. (type strain B8W22(T) =MTCC 7755(T) =JCM 13839(T)).

Bacterial diversity of soil in the vicinity of Pindari glacier, Himalayan mountain ranges, India, using culturable bacteria and soil 16S rRNA gene clones.

Three 16S rRNA gene clone libraries (P1L, P4L and P8L) were constructed using three soil samples (P1S, P4S and P8S) collected near Pindari glacier, Himalayas. The three libraries yielded a total of 703 clones. Actinobacteria, Firmicutes and Proteobacteria were common to the three libraries. In addition to the above P1L and P8L shared the phyla Acidobacteria, Bacteroidetes, Gemmatimonadetes and Planctomycetes. Phyla Chlamydiae, Chlorobi, Chloroflexi, Dictyoglomi, Fibrobacteres, Nitrospirae, Verrucomicrobia, candidate division SPAM and candidate TM7s TM7a phylum were present only in P1L. Rarefaction analysis indicated that the bacterial diversity in P4S and P8S soil samples was representative of the sample. Principal component analysis (PCA) revealed that P1S and P8S were different from P4S soil sample. PCA also indicated that arsenic content, pH, Cr and altitude influence the observed differences in the percentage of specific OTUs in the three 16S rRNA gene clone libraries. The observed bacterial diversity was similar to that observed for other Himalayan and non-polar cold habitats. A total of 40 strains of bacteria were isolated from the above three soil samples and based on the morphology 20 bacterial strains were selected for further characterization. The 20 bacteria belonged to 12 different genera. All the isolates were psychro-, halo- and alkalitolerant. Amylase and urease activities were detected in majority of the strains but lipase and protease activities were not detected. Long chain, saturated, unsaturated and branched fatty acids were predominant in the psychrotolerant bacteria.

Bacterial biodiversity from Roopkund Glacier, Himalayan mountain ranges, India

The bacterial diversity of two soil samples collected from the periphery of the Roopkund glacial lake and one soil sample from the surface of the Roopkund Glacier in the Himalayan ranges was determined by constructing three 16S rRNA gene clone libraries. The three clone libraries yielded a total of 798 clones belonging to 25 classes.Actinobacteria was the most predominant class (>10% of the clones) in the three libraries. In the library from the glacial soil, class Betaproteobacteria (24.2%) was the most predominant. The rarefaction analysis indicated coverage of 43.4 and 41.2% in the samples collected from the periphery of the lake thus indicating a limited bacterial diversity covered; at the same time, the coverage of 98.4% in the glacier sample indicated most of the diversity was covered. Further, the bacterial diversity in the Roopkund glacier soil was low, but was comparable with the bacterial diversity of a few other glaciers. The results of principal component analysis based on the 16S rRNA gene clone library data, percentages of OTUs and biogeochemical data revealed that the lake soil samples were different from the glacier soil sample and the biogeochemical properties affected the diversity of microbial communities in the soil samples.

Diversity of Yeasts from Puddles in the Vicinity of Midre Lovénbreen Glacier, Arctic and Bioprospecting for Enzymes and Fatty Acids

A total of 132 yeast strains were characterised from 4 sediment samples collected from small puddles in the vicinity of Midre Lovénbreen glacier, Arctic. Based on the D1/D2 domain sequence similarity, the isolates could be categorised into 6 groups. The nearest phylogenetic neighbour of groups I to VI were identified as Cryptococcus gastricus, Cryptococcus terricolus, Rhodotorula muscorum, Mrakia psychrophila, Mrakia gelida and Rhodotorula glacialis, respectively. Strains representative of the six groups were psychrophilic and salt tolerant but varied in their ability to produce cold-active extracellular enzymes such as lipase, protease, pectinase, cellulase and amylase. C18:1 (w9C) and C18:2 (w9,12C) were the only two fatty acids common to all the yeasts and branched and (or) unsaturated fatty acids increased in yeasts growing at 8°C compared to 22°C, probably as an adaptation to low temperature. The present study establishes that psychrophilic yeasts are predominant in Arctic and could be used as work horses to produce cold-active enzymes and poly unsaturated fatty acids which have been implicated in low temperature adaptation and also for their use in biotechnology.

Paenibacillus glacialis sp. nov., isolated from the Kafni glacier of the Himalayas, India

A novel strain of the genus Paenibacillus, KFC91T, was isolated from the Kafni glacier of the Himalayas. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KFC91T clustered with Paenibacillus antarcticus LMG 22078T (98.9%) and Paenibacillus macquariensis LMG 6935T (98.7%). The cell wall peptidoglycan contained meso-diaminopimelic acid as the diamino acid, anteiso-C15:0 as the predominant fatty acid and MK-7 as the major isoprenoid quinone. Based on the phylogenetic analysis, chemotaxonomic characteristics and other phenotypic traits, strain KFC91T was assigned to the genus Paenibacillus. DNA-DNA hybridization experiments of KFC91T with P. antarcticus and P. macquariensis showed reassociation values of 39 and 52%, respectively. Thus, it is proposed that strain KFC91T should be assigned the status of a novel species of the genus Paenibacillus and the name proposed is Paenibacillus glacialis sp. nov., with KFC91T (=NCCB 100252T =DSM 22343T) as the type strain.

Comparison of bacterial diversity in proglacial soil from Kafni Glacier, Himalayan Mountain ranges, India, with the bacterial diversity of other glaciers in the world

Two 16S rRNA gene clone libraries (KF and KS) were constructed using two soil samples (K7s and K8s) collected near Kafni Glacier, Himalayas. The two libraries yielded a total of 648 clones. Phyla Actinobacteria, Bacteroidetes, Chloroflexi Firmicutes, Proteobacteria, Spirochaetae, Tenericutes and Verrucomicrobia were common to the two libraries. Phyla Acidobacteria, Chlamydiae and Nitrospirae were present only in KF library, whereas Lentisphaerae and TM7 were detected only in KS. In the two libraries, clones belonging to phyla Bacteroidetes and Proteobacteria were the most predominant. Principal component analysis (PCA) revealed that KF and KS were different and arsenic content influenced the differences in the percentage of OTUs. PCA indicated that high water content in the K8s sample results in high total bacterial count. PCA also indicated that bacterial diversity of KF and KS was similar to soils from the Pindari Glacier, Himalayas; Samoylov Island, Siberia; Schrimacher Oasis, Antarctica and Siberian tundra. The eleven bacterial strains isolated from the above two soil samples were phylogenetically related to six different genera. All the isolates were psychro-, halo- and alkalitolerant. Amylase, lipase and urease activities were detected in the majority of the strains. Long chain, saturated, unsaturated and branched fatty acids were predominant in the psychrotolerant bacteria.

Selective involvement of caspase-3 in ceramide induced apoptosis in AK-5 tumor cells

Ceramide, a product of sphingomyelin metabolism, is a novel lipid second messenger that mediates diverse cellular functions. The present study demonstrates the activation of caspase‐3/CPP‐32β, during apoptosis induced by cell permeable exogenous ceramides, in AK‐5 tumor, a spontaneously regressing rat histiocytoma. The apoptotic events were suppressed by the caspase‐3 specific tetrapeptide inhibitor DEVD‐CHO but not by the caspase‐1 inhibitor YVAD‐CMK. In cells overexpressing Bcl‐2, a significant decrease in cell death was observed after exogenous addition of ceramides. Furthermore the processing of caspase‐3 to its active form upon apoptotic stimulus, and the subsequent cleavage of the substrate PARP, suggested a central role for caspase‐3 in the ceramide mediated apoptosis in AK‐5 tumor cells.

Exiguobacterium soli sp. nov., a psychrophilic bacterium from the McMurdo Dry Valleys, Antarctica

A novel psychrophilic bacterium, designated strain DVS 3Y(T), was isolated from a moraine sample from the McMurdo Dry Valleys, Antarctica. Phenotypic and chemotaxonomic characteristics and data from a phylogenetic analysis based on 16S rRNA gene sequences indicated that strain DVS 3Y(T) was related to the genus Exiguobacterium. Strain DVS 3Y(T) exhibited >97 % gene sequence similarity with respect to Exiguobacterium acetylicum DSM 20416(T) (97.4 %), Exiguobacterium oxidotolerans DSM 17272(T) (97.4 %), Exiguobacterium indicum IAM 15368(T) (97.6 %), Exiguobacterium undae DSM 14481(T) (98.2 %), Exiguobacterium sibiricum DSM 17290(T) (98.6 %) and Exiguobacterium antarcticum DSM 14480(T) (99.6 %). However, the levels of DNA-DNA relatedness between strain DVS 3Y(T) and E. acetylicum DSM 20416(T), E. oxidotolerans JCM 12280(T), E. indicum IAM 15368(T), E. undae DSM 14481(T), E. sibiricum DSM 17290(T) and E. antarcticum DSM 14480(T) were less than 70 % (52, 25, 16, 33, 16 and 38 %, respectively). Strain DVS 3Y(T) also differed from these six closely related species in a number of phenotypic traits. Phenotypic, chemotaxonomic and phylogenetic data suggest that strain DVS 3Y(T) represents a novel species of the genus Exiguobacterium, for which the name Exiguobacterium soli sp. nov. is proposed. The type strain is DVS 3Y(T) (=MTCC 4816(T)=JCM 14376(T)).

Lack of heat shock response triggers programmed cell death in a rat histiocytic cell line

Stress response is a universal phenomenon. However, a rat histiocytic cell line, BC‐8, showed no heat shock response and failed to synthesize heat shock protein 70 (hsp70) upon heat shock at 42°C for 30 min. BC‐8 is a clone of AK‐5, a rat macrophage tumor line that is adapted to grow in culture and has the same chromosome number and tumorigenic potential as AK‐5. An increase in either the incubation temperature or time or both to BC‐8 cells leads to loss of cell viability. In addition, heat shock conditions activated apoptotic cell death in these cells as observed by cell fragmentation, formation of nuclear comets, apoptotic bodies, DNA fragmentation and activation of ICE‐like cysteine proteases. Results presented here demonstrate that BC‐8 cells cannot mount a typical heat shock response unlike all other eukaryotic cells and that in the absence of induction of hsps upon stress, these cells undergo apoptosis at 42°C.