Synthesis and in vitro evaluations of 6-(hetero)-aryl-imidazo[1,2-b]pyridazine-3-sulfonamide's as an inhibitor of TNF-α production.

Tumor necrosis factor-α is an important pro-inflammatory cytokine having a key role in hosts defensive process of immune systems and its over expression led to a diverse range of inflammatory diseases such as Rheumatoid arthritis, Cronh's disease, psoriasis, etc. This paper describes our medicinal chemistry efforts on imidazo[1,2-b]pyridazine scaffold: design, synthesis and biological evaluation. By the introducing sulfonamide functionality at 3 positions and substituting 6 positions with (hetero)-aryl groups', a small library of compounds was prepared. All synthesized compounds were screened for lipopolysaccharide (LPS) mediated TNF-α production inhibitory activity. Biological data revealed that the majority of the compounds of this series showed moderate to potent TNF-α production inhibitory activity. Compound 5u and 5v are the most potent compounds from the series with activity of IC50 = 0.5 µM and 0.3 µM respectively. A short SAR demonstrates that 3-sulfonyl-4-arylpiperidine-4-carbonitrile moiety on imidazo[1,2-b]pyridazine showed better activity compared to the 3-(4-aryllpiperazin-1-yl) sulfonyl) in hPBMC assay. The molecular modeling studies revealed that the potent TNF-α production inhibitory activity 5v due to the extra stability of complex because of an extra pi-pi (π-π) stacking, hydrogen-bonding interactions.

Synthesis and biological evaluation of imidazo[1,2-[Formula: see text]]pyridazines as inhibitors of TNF-[Formula: see text] production.

Tumor necrosis factor-alpha (TNF-[Formula: see text] is an important pro-inflammatory cytokine responsible for a diverse range of inflammatory diseases including rheumatoid arthritis. In the present manuscript, our medicinal chemistry efforts on the design, synthesis and TNF-[Formula: see text] evaluation of a series of 3, 6-disubstituted imidazo[1,2-b]pyridazine is described. The best compounds were 3-pyridyl and (4-(methylsulfonyl)phenyl) analogs 8q and 8w, showing inhibition of TNF-[Formula: see text] production with IC[Formula: see text]values of 0.9 and 0.4 [Formula: see text]M, respectively. The identified leads have potential for further development for treatment of inflammatory diseases.

Methylsulfonyl benzothiazoles (MSBT) derivatives: Search for new potential antimicrobial and anticancer agents.

A series of novel 4 and 5-substituted methylsulfonyl benzothiazole (MSBT) compounds having amide, alkoxy, sulfonamide, nitro and amine functionality were synthesized from sequential reactions on 5-ethoxy-2-(methylsulfonyl)benzo[d]thiazole such as nitration, reduction, sulfonation, dealkylation, etc. All synthesized compounds were screened against antimicrobial and selected screened for anticancer activity. Antimicrobial activities studies reveled that among all compounds screened, out of MSBT-07, MSBT-11, MSBT-12, MSBT-14, MSBT-19, and MSBT-27 were found to have promising antimicrobial activity at MIC range of 4-50µg/ml against selected bacterial as well as fungal species. Compounds having good antimicrobial activity were screened for cervical cancer (HeLA cell lines). Of these MSBT-07 and MSBT-12 significantly reduced the cell growth. Consequently their calculated GI50 values were found to be 0.1 or <0.1µM.

Aquipuribacter nitratireducens sp. nov., isolated from a soil sample of a mud volcano.

A novel Gram-stain-positive, coccoid, non-motile bacterium, designated strain AMV4T, was isolated from a soil sample collected from a mud volcano located in the Andaman Islands, India. The colony was pale orange. Strain AMV4T was positive for oxidase, aesculinase, lysine decarboxylase and ornithine decarboxylase activities and negative for amylase, catalase, cellulase, protease, urease and lipase activities. 16S rRNA gene sequence analysis indicated that strain AMV4T was a member of the order Actinomycetales and was closely related to Aquipuribacter hungaricus with a sequence similarity of 97.13% (pairwise alignment). Phylogenetic analyses showed that strain AMV4T clustered with Aquipuribacter hungaricus and was distantly related to the other genera of the family Intrasporangiaceae. DNA-DNA hybridization between strains AMV4T and Aquipuribacter hungaricus IV-75T showed a relatedness of 28%. The predominant cellular fatty acids were iso-C15 : 0 (6.9%), anteiso-C15 : 0 (25.3%), C16 : 0 (12.9%), anteiso-C16 : 0 (5.6%), C18 : 1ω9c (19.8%) and C18 : 3ω6,9,12c (9.1%). The diagnostic diamino acid in the cell-wall peptidoglycan of strain AMV4T was meso-diaminopimelic acid. Strain AMV4T contained MK-10(H4) as the predominant respiratory quinone. The polar lipids consisted of phosphatidylglycerol, one unidentified glycolipid, two unidentified phospholipids and five unidentified lipids. The DNA G+C content of strain AMV4T was 74.3 mol%. Based on data from this taxonomic study using a polyphasic approach, it is proposed that strain AMV4T represents a novel species of the genus Aquipuribacter, with the suggested name Aquipuribacter nitratireducens sp. nov. The type strain is AMV4T ( = CCUG 58430T = DSM 22863T = NBRC 107137T).

Paenisporosarcina indica sp. nov., a psychrophilic bacterium from a glacier, and reclassification of Sporosarcina antarctica Yu et al., 2008 as Paenisporosarcina antarctica comb. nov. and emended description of the genus Paenisporosarcina.

A Gram-stain-positive, aerobic, spore-forming, rod-shaped bacterium, PN2(T), was isolated from a soil sample collected near the Pindari glacier. It contained anteiso-C15 : 0, iso-C15 : 0 and C16 : 1ω7c alcohol as the predominant fatty acids, MK-7 as the major menaquinone and A4α type (l-Lys-d-Glu) peptidoglycan. Based on these characteristics, strain PN2(T) was assigned to the genus Paenisporosarcina. Phylogenetic analysis based on 16S rRNA gene sequence placed strain PN2(T) within the genus Paenisporosarcina and showed a sequence similarity of 98.5-99.0 % with members of this genus. Paenisporosarcina macmurdoensis CMS 21w(T), Paenisporosarcina quisquiliarum SK 55(T) and Sporosarcina antarctica N-05(T) were identified as the most closely related species with 16S rRNA gene sequence similarities of 98.6 %, 99.0 % and 98.4 %, respectively. The values for DNA-DNA relatedness between strain PN2(T) and P. macmurdoensis, P. quisquiliarum and S. antarctica were below the 70 % threshold value (32.0 %, 42.0 % and 38.0 % respectively). In addition, strain PN2(T) exhibited a number of phenotypic differences from P. macmurdoensis, P. quisquiliarum and S. antarctica. Based on the cumulative differences, strain PN2(T) was identified as representing a novel species and the name Paenisporosarcina indica sp. nov. was proposed. The type strain of Paenisporosarcina indica sp. nov. is PN2(T) (LMG 23933(T) = JCM 15114(T)). Furthermore, based on the morphological and chemotaxonomic characteristics, the species Sporosarcina antarctica was reclassified as a species of the genus Paenisporosarcina and renamed Paenisporosarcina antarctica comb. nov. In addition, an emended description of the genus Paenisporosarcina is presented.

Exiguobacterium alkaliphilum sp. nov. isolated from alkaline wastewater drained sludge of a beverage factory.

A facultatively anaerobic, alkaliphilic, Gram-stain-positive, rod-shaped bacterium, designated strain 12/1(T), isolated from alkaline wastewater drained sludge of a beverage industry facility located near New Delhi, India, was subjected to a polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain 12/1(T) belonged to the genus Exiguobacterium and was most closely related to Exiguobacterium aurantiacum DSM 6208(T) (99.46 %), E. aquaticum IMTB-3094(T) (99.18 %), E. mexicanum 8N(T) (99.06 %), E. profundum 10C(T) (98.17 %), E. aestuarii TF-16(T) (98.1 %) and E. marinum TF-80(T) (98.03 %). The DNA G+C content of strain 12/1(T) was 55.6 mol%, major respiratory isoprenoid quinone was MK-7, major polar lipids were phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine and the cell-wall peptidoglycan was of the A3α l-Lys-Gly type, characteristics consistent with its affiliation to the genus Exiguobacterium. Strain 12/1(T) showed levels of DNA-DNA hybridization of less than 70 % with the closely related species of the genus Exiguobacterium. Overall, the phenotypic, chemotaxonomic and phylogenetic data presented in this study suggest that strain 12/1(T) represents a novel species of the genus Exiguobacterium, for which the name Exiguobacterium alkaliphilum sp. nov. is proposed. The type strain is 12/1(T) ( = CCM 8459(T) = DSM 21148(T)).

Lacinutrix himadriensis sp. nov., a psychrophilic bacterium isolated from a marine sediment, and emended description of the genus Lacinutrix.

A novel gram-negative, rod-shaped, non-motile, psychrophilic bacterium, designated strain E4-9a(T), was isolated from a marine sediment sample collected at a depth of 276 m from Kongsfjorden, Svalbard, in the Arctic Ocean. The colony colour was golden yellow. Strain E4-9a(T) was positive for amylase activity at 5 °C. The predominant fatty acids were iso-C(15 : 1) G (21.8 %), anteiso-C(15 : 0) (19.1 %), anteiso-C(15 : 1) A (18.6 %), iso-C(15 : 0) (13.8 %) and iso-C(16 : 1) H (6.4 %). Strain E4-9a(T) contained MK-6 as the major respiratory quinone. The polar lipids consisted of phosphatidylethanolamine, three unidentified aminolipids (AL1, AL4 and AL5), an unidentified phospholipid and four unidentified lipids (L1, L4 to L6). Based on 16S rRNA gene sequence similarity, it was ascertained that the closest related species to E4-9a(T) were Lacinutrix copepodicola, L. algicola and L. mariniflava, with sequence similarity to the respective type strains of 98.5, 96.5 and 95.8 %. Phylogenetic analysis showed that strain E4-9a(T) clustered with the type strain of L. copepodicola and with those of L. algicola and L. mariniflava at distances of 1.5 and 4.8 % (98.5 and 95.2 % similarity), respectively. However, DNA-DNA hybridization with L. copepodicola DJ3(T) showed 59 % relatedness with respect to strain E4-9a(T). The DNA G+C content of strain E4-9a(T) was 29 mol%. Based on the results of DNA-DNA hybridization and phenotypic data, it appears that strain E4-9a(T) represents a novel species of the genus Lacinutrix, for which the name Lacinutrix himadriensis sp. nov. is proposed. The type strain is E4-9a(T) ( = CIP 110310(T)  = KCTC 23612(T)).

Winogradskyella psychrotolerans sp. nov., a marine bacterium of the family Flavobacteriaceae isolated from Arctic sediment.

A novel Gram-negative, rod-coccus shaped, non-motile, strain, RS-3(T), was isolated from a sediment sample collected from the marine transect of Kongsfjorden, Ny-Ålesund, Svalbard, Arctic. Colonies and broth cultures were yellowish in colour due to the presence of carotenoids. Strain RS-3(T) was positive for oxidase, aesculinase, caseinase, gelatinase and urease activities and negative for amylase, catalase, lipase, lysine decarboxylase, ornithine decarboxylase, DNase and ß-galactosidase activities. The predominant fatty acids were iso-C15 : 0 (18.0), anteiso-C15 : 0 (16.8), iso-C15 : 1 G (14.2), anteiso-C15 : 1 A (6.0) and iso-C15 : 0 3-OH (6.8). Strain RS-3(T) contained MK-6 (72.42 %) and MK-7 (27.58 %) as the major respiratory quinones and phosphatidylethanolamine, two unidentified aminolipids and two unidentified lipids make up the polar lipid composition. The DNA G+C content of strain RS-3(T) was 34.7±1.2 mol%. The 16S rRNA gene sequence analysis indicated that Winogradskyella pacifica and Winogradskyella thalassocola are the most closely related species with sequence similarities to the type strains of these species of 98.5 and 97.7 %, respectively. However, DNA-DNA hybridization with Winogradskyella pacifica KCTC 22997(T) and Winogradskyella thalassocola DSM 15363(T) showed a relatedness of 22 and 42.5 % with respect to strain RS-3(T). Based on the DNA-DNA hybridization values, phenotypic and chemotaxonomic characteristics and phylogenetic inference, strain RS-3(T) is proposed as a novel species of the genus Winogradskyella, for which the name Winogradskyella psychrotolerans sp. nov. is proposed. The type strain of Winogradskyella psychrotolerans sp. nov. is RS-3(T) ( = CIP 110154(T) = NBRC 106169(T)). An emended description of the genus Winogradskyella is provided.

Iodobacter arcticus sp. nov., a psychrotolerant bacterium isolated from meltwater stream sediment of an Arctic glacier.

Two novel violet-pigmented, Gram-negative, rod-shaped, non-motile bacteria, designated strains M4-16(T) and M4-9, were isolated from sediment from an Arctic glacier. The predominant fatty acids of both strains were C16 : 1ω7c and/or C16 : 1ω6c (summed feature 3), C16 : 0, C14 : 0 and C18 : 1ω7c and/or C18 : 1ω6c (summed feature 8) and both strains contained ubiquinone-8 as the respiratory quinone. The polar lipids consisted of phosphatidylethanolamine, two unidentified phospholipids and one unidentified aminolipid. 16S rRNA gene sequence analysis indicated that strains M4-16(T) and M4-9 were members of the genus Iodobacter and closely related to Iodobacter fluviatilis ATCC 33051(T) with pairwise sequence similarity of 98.9 %. The DNA-DNA relatedness between strains M4-9 and M4-16(T) was 92.5 %, while strains M4-9 and M4-16(T) had DNA-DNA relatedness values of 21.5 and 18.2 %, respectively, with Iodobacter fluviatilis JCM 9044(T). The RAPD-PCR banding patterns of strains M4-9 and M4-16(T) were similar but differed from that of Iodobacter fluviatilis JCM 9044(T). Based on data from the current polyphasic study, strains M4-16(T) and M4-9 represent a novel species of the genus Iodobacter, for which the name Iodobacter arcticus sp. nov. is proposed. The type strain of Iodobacter arcticus is M4-16(T) ( = CIP 1103011(T) = MTCC 11351(T)).

Detection of pregnancy and fertility status in big cats using an enzyme immunoassay based on 5α-pregnan-3α-ol-20-one.

Development of non-invasive steroid hormone assays using fecal samples is crucial for detection of pregnancy and monitoring of fertility status in big cats and thus facilitates conservation and management of wild animals. Due to changes in metabolism and excretory pattern, animals excrete different steroid metabolites in feces and urine. The present study is an attempt to develop a common enzyme immunoassay for 5α-pregnan-3α-ol-20-one one of the predominant progestogen metabolites in the feces samples of big cats. The developed ELISA showed a high sensitivity and low cross reactivity to other hormones compared to commercially available RIA kits based on progesterone antibody. It could be used in a wide range of animals for monitoring fertility status and pregnancy detection by measuring fecal steroid metabolites.

Draft genome sequence of Cecembia lonarensis strain LW9T, isolated from Lonar Lake, a haloalkaline lake in India.

The draft genome sequence (4.84 Mb) of Cecembia lonarensis strain LW9(T), isolated from a water sample (4.5-m depth) from Lonar Lake, a meteorite-created haloalkaline lake in India, is reported. The enzymes produced by these microorganisms need to be stable under alkaline conditions prevailing in its habitat. Such enzymes would be of immense importance for enzymatic processes operating at high pH.

Genome sequence of the alkaliphilic bacterium Nitritalea halalkaliphila type strain LW7, isolated from Lonar Lake, India.

An alkaliphilic bacterium, Nitritalea halalkaliphila LW7, which belongs to the family Cyclobacteriacae in the phylum Bacteroidetes, was isolated from Lonar Lake in Maharastra, India. Here we announce the draft genome sequence of the type strain LW7, which contains 3,633,701 bp with a G+C content of 48.58%.

Draft genome sequence of Bacillus isronensis strain B3W22, isolated from the upper atmosphere.

We report the 4.0-Mb genome sequence of Bacillus isronensis strain B3W22 isolated from air collected at an altitude ranging from 27 to 30 km above the city of Hyderabad, in India. This genome sequence will contribute to the objective of determining the microbial diversity of the upper atmosphere.

Oceanisphaera arctica sp. nov., isolated from Arctic marine sediment, and emended description of the genus Oceanisphaera.

A novel Gram-staining-negative, coccoid, non-motile bacterium, designated strain V1-41(T), was isolated from a sample of marine sediment collected, at a depth of 200 m, from Kongsfjorden (an inlet on the west coast of Spitsbergen, an island that forms part of the Svalbard archipelago in the Arctic Ocean). The strain formed cream-brown colonies on marine agar. Cells of the novel strain were positive in tests for catalase, oxidase, lysine decarboxylase and ornithine decarboxylase activities but negative for gelatinase and lipase activities. They hydrolysed aesculin, starch and urea, but not casein or DNA. Most of the cellular fatty acids were medium-chain and saturated (37.1%) or long-chain and unsaturated (27.8%), with C(12:0) (37.1%), C(18:1)ω7c, and summed features 2 (19.3%) and 3 (24.1%) predominating. The major respiratory quinone was Q-8. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, two unidentified aminophospholipids, four unidentified phospholipids and one other unidentified lipid. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the novel strain's closest known relatives were Oceanisphaera litoralis DSM 15406(T) (98.5% sequence similarity) and Oceanisphaera donghaensis BL1(T) (98.3%). In DNA-DNA hybridizations, however, the levels of relatedness between strain V1-41(T) and O. litoralis DSM 15406(T) and between the novel strain and O. donghaensis DSM 17589(T) were found to be only 19% and 29%, respectively. Based on these low levels of similarity at the DNA-DNA level and the phenotypic and chemotaxonomic differences from O. litoralis DSM 15406(T) and O. donghaensis DSM 17589(T), strain V1-41(T) represents a novel species of the genus Oceanisphaera for which the name Oceanisphaera arctica sp. nov. is proposed. The type strain is V1-41(T) ( = CCUG 58690(T) = KCTC 23013(T) = NBRC 106171(T)).

Lutibaculum baratangense gen. nov., sp. nov., a proteobacterium isolated from a mud volcano.

A novel Gram-negative, oval to rod-shaped, motile bacterium, strain AMV1(T), was isolated from a soil sample collected from a mud volcano of Baratang Island, Andamans, India. The predominant fatty acids were C(16:0) (5.7%), C(18:1)ω7c (78.6%) and C(19:0) cyclo ω8c (6.3%). Strain AMV1(T) contained ubiquinone 10 (Q-10) as the major respiratory quinone and minor quantities of ubiquinone 9 (Q-9). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three unidentified lipids, one unidentified phospholipid and one unidentified aminolipid. 16S rRNA gene sequence analysis indicated that strain AMV1(T) was related most closely to the type strains of Tepidamorphus gemmatus, Bauldia consociata, Afifella pfennigii and Amorphus coralli, four members of the order Rhizobiales (class Alphaproteobacteria), with pairwise sequence similarities of 95.0, 94.5, 94.4 and 94.0%, respectively; it shared <94% 16S rRNA gene sequence similarity with all the other members of the order Rhizobiales. Phylogenetic analyses indicated that strain AMV1(T) clustered with Tepidamorphus gemmatus and with species of the genera Amorphus, Rhodobium and Afifella. Phenotypic and phylogenetic characteristics thus suggest that strain AMV1(T) is a representative of a novel species of a new genus, for which the name Lutibaculum baratangense gen. nov., sp. nov. is proposed. The type strain of Lutibaculum baratangense is AMV1(T) ( = KCTC 22669(T) = NBRC 105799(T) = CCUG 58046(T)).

Cecembia lonarensis gen. nov., sp. nov., a haloalkalitolerant bacterium of the family Cyclobacteriaceae, isolated from a haloalkaline lake and emended descriptions of the genera Indibacter, Nitritalea and Belliella.

A novel Gram-staining-negative, rod-shaped, non-motile bacterium, designated strain LW9(T), was isolated from a water sample collected from Lonar Lake of Buldhana district, Maharashtra, India. Colonies and broth cultures were reddish orange due to the presence of carotenoid pigments. Strain LW9(T) was positive for catalase, ornithine decarboxylase and lysine decarboxylase activities and negative for gelatinase, oxidase, urease and lipase activities. The predominant fatty acids were iso-C(15 : 0) (31.3 %), iso-C(16 : 0) (9.3 %), anteiso-C(15 : 0) (7.3 %), iso-C(16 : 1) H (6.1 %), summed feature 3 (comprising C(16 : 1)ω7c/C(16 : 1)ω6c; 5.9 %), iso-C(17 : 1)ω9c (5.4 %) and iso-C(17 : 0) 3-OH (5.0 %). Strain LW9(T) contained MK-7 as the major respiratory quinone. The polar lipids consisted of phosphatidylethanolamine, two unidentified aminolipids and seven unidentified lipids. The DNA G+C content of strain LW9(T) was 40.5 mol%. 16S rRNA gene sequence analysis indicated that the type strains of Indibacter alkaliphilus and Aquiflexum balticum, two members of the family Cyclobacteriaceae (phylum 'Bacteroidetes') were the most closely related strains with sequence similarities of 93.0 and 94.0 %, respectively. Other members of the family Cyclobacteriaceae showed sequence similarities <93.0 %. Based on these phenotypic characteristics and on phylogenetic inference, strain LW9(T) is proposed as the representative of novel species in a new genus, Cecembia lonarensis gen. nov., sp. nov. The type strain of the type species, Cecembia lonarensis, is LW9(T) (= CCUG 58316(T) = KCTC 22772(T)). Emended descriptions of the genera Indibacter, Nitritalea and Belliella are also proposed.

Cyclobacterium qasimii sp. nov., a psychrotolerant bacterium isolated from Arctic marine sediment.

A novel Gram-stain-negative, horseshoe-shaped, non-motile bacterium, designated strain M12-11B(T), was isolated from a marine sediment sample collected at a depth of 200 m from Kongsfjorden, Svalbard. The colony colour was orangish red due to the presence of carotenoids. Fatty acids were dominated by branched and unsaturated fatty acids (90.8 %), with a high abundance of iso-C(15 : 0) (14.9 %), anteiso-C(15 : 0) (11.4 %), iso-C(15 : 1) G (13.1 %), C(15 : 1)ω6c (5.4 %), C(17 : 1)ω6c (6.7 %), summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c; 9.3 %) and summed feature 9 (10-methyl C(16 : 0) and/or iso-C(17 : 1)ω9c; 5.9 %). Strain M12-11B(T) contained MK-7 as the major respiratory quinone. The polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, one unidentified aminolipid and three unidentified lipids. Based on 16S rRNA gene sequence similarities, the type strains of Cyclobacterium amurskyense, Cyclobacterium marinum and Cyclobacterium lianum were most closely related to M12-11B(T) with sequence similarities of 98.2, 96.8 and 93.3 %, respectively. Other members of the family Cyclobacteriaceae had sequence similarities of <92.0 %. However, DNA-DNA hybridization with Cyclobacterium amurskyense KCTC 12363(T) and Cyclobacterium marinum DSM 745(T) showed relatedness values of only 24.5 and 32.5 % with respect to strain M12-11B(T). Based on the results of DNA-DNA hybridization experiments and phenotypic and chemotaxonomic data, it appears that strain M12-11B(T) represents a novel species of the genus Cyclobacterium, for which the name Cyclobacterium qasimii sp. nov. is proposed; the type strain is M12-11B(T) (= KCTC 23011(T) = NBRC 106168(T)) and it has a DNA G+C content of 40.5 mol%.

Cell cycle synchronization of bison (Bos gaurus) fibroblasts derived from ear piece collected post-mortem.

Efficiency of the technique of somatic cell nuclear transfer critically depends on the cell cycle phase compatibility between the donor somatic cell nucleus and recipient cytoplasm. In this study, attempts were made to optimize conditions for cell cycle synchronization of bison ear fibroblasts at G0/G1 using different approaches such as using cells in confluency, after contact inhibition, serum starvation or treatment with dimethyl sulfoxide (DMSO) (0.5%, 1.0% and 2.0%), sodium butyrate (NaBu) (0.5, 1.0 and 2.0 mm), cytochalasin-B (CB) (7.5 µg/ml), cycloheximide (CHX) (7.5 µg/ml) and 6-dimethyl aminopurine (6-DMAP) (2.0 mm). A small piece of an ear of an adult female bison collected post-mortem 10 h after death was used for the preparation of fibroblast cells. The synchronization efficiency was determined by fluorescence-activated cell sorting. Higher proportion of G0/G1 phase was obtained when cells were subjected to serum starvation for 48 h (85.4%). Sodium butyrate had no effect on synchronization of cells at G0/G1 when the cells were treated for 24 and 48 h. Similarly, DMSO (0.5% and 1.0%) had also no effect on the proportion of cells at G0/G1 for 24 and 48 h. The synchronization ability of CB, CHX and 6-DMAP at G0/G1 phase was equally effective when cells were treated for 4 h (68.5%, 68.7% and 67.4%) and 24 h (67.8%, 66.1% and 67.5%). In conclusion, this study shows that cells subjected to serum starvation for 24-48 h or confluent monolayer, or cycling cells treated with 1.0% DMSO or 2.0 mm NaBu for 24 h showed best synchronization in G0/G1 phase of cell cycle.

Cesiribacter andamanensis gen. nov., sp. nov., isolated from a soil sample from a mud volcano.

A novel gram-staining-negative, rod-shaped, non-motile bacterium, strain AMV16(T), was isolated from a soil sample collected from a mud volcano located in the Andaman Islands, India. The cell suspension was pale orange. Cells of strain AMV16(T) were positive for catalase, oxidase, lipase, ornithine decarboxylase and lysine decarboxylase and negative for gelatinase and urease. The fatty acids present were anteiso-C(11 : 0) (5.4 %), anteiso-C(12 : 0) (4.1 %), C(12 : 0) (7.0 %), iso-C(15 : 0) (14.4 %), anteiso-C(15 : 0) (3.4 %), anteiso-C(16 : 0) (3.0 %), C(16 : 0) (2.6 %), anteiso-C(17 : 0) (3.7 %), iso-C(19 : 0) (9.7 %), C(13 : 1) (13.8 %), iso-C(15 : 1) G (15.9 %), iso-C(16 : 1) G (11.1 %) and summed feature 5 (anteiso-C(18 : 0) and/or C(18 : 2)ω6,9c; 5.9 %). Strain AMV16(T) contained MK-4 is [corrected] the major respiratory quinone and diphosphatidylglycerol and phosphatidylethanolamine made up the phospholipids. The G+C content of DNA of strain AMV16(T) was 50.9 mol%. blast sequence similarity searches based on the 16S rRNA gene sequence indicated that species of the genus Marivirga were the nearest phylogenetic neighbours, with pairwise sequence similarity ranging from 89.9 to 90.0 %. Phylogenetic analyses indicated that strain AMV16(T) clustered with the type strains of Marivirga tractuosa and Marivirga sericea at a phylogenetic distance of 14.6 % (85.4 % similarity), distinct from clades representing other genera of the family 'Flammeovirgaceae'. Based on the above-mentioned phenotypic and phylogenetic characteristics, strain AMV16(T) is proposed as a representative of a new genus and novel species, Cesiribacter andamanensis gen. nov., sp. nov. The type strain of Cesiribacter andamanensis is AMV16(T) ( = DSM 22818(T)  = CCUG 58431(T)).

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.