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Biomedical subjects

S Shivaji

Publications and source records attributed to S Shivaji.

At least 37 records · Page 2Linked to original sources

Psychrophilic Pseudomonas syringae requires trans-monounsaturated fatty acid for growth at higher temperature.

A psychrophilic bacterium, Pseudomonas syringae (Lz4W) from Antarctica, was used as a model system to establish a correlation, if any, between thermal adaptation, trans-fatty acid content and membrane fluidity. In addition, attempts were made to clone and sequence the cti gene of P. syringae (Lz4W) so as to establish its characteristics with respect to the cti of other Pseudomonas spp. and also to in vitro mutagenize the cti gene so as to generate a cti null mutant. The bacterium showed increased proportion of saturated and trans-monounsaturated fatty acids when grown at 28 degrees C compared to cells grown at 5 degrees C, and the membrane fluidity decreased with growth temperature. In the mutant, the trans-fatty acid was not synthesized, and the membrane fluidity also decreased with growth temperature, but the decrease was not to the extent that was observed in the wild-type cells. Thus, it would appear that synthesis of trans-fatty acid and modulation of membrane fluidity to levels comparable to the wild-type cells is essential for growth at higher temperatures since the mutant exhibits growth arrest at 28 degrees C. In fact, the cti null mutant-complemented strain of P. syringae (Lz4W-C30b) that was capable of synthesizing the trans-fatty acid was indeed capable of growth at 28 degrees C, thus confirming the above contention. The cti gene of P. syringae (Lz4W) that was cloned and sequenced exhibited high sequence identity with the cti of other Pseudomonas spp. and exhibited all the conserved features.

Bacteria↗

N-acetyl transferase 2 polymorphism and advanced stages of endometriosis in South Indian women.

Aylamine-N-acetyl transferase is a phase II detoxification enzyme encoded by the gene NAT2. Single nucleotide polymorphism (SNP) changes from the wild type NAT2 *4 allele result in allelic variants *5, *6 and *7. Homozygotes for the NAT2 *4 wild type are fast acetylators; heterozygotes with one wild-type allele and a variant NAT2 *5, *6 or *7 allele have reduced enzyme activity and individuals with two variant alleles are slow acetylators. Previous studies have implicated NAT2 as a susceptibility factor in endometriosis. This study investigated the NAT2 allele frequencies and genotype distributions in 252 unrelated women with endometriosis and 264 controls of South Indian origin. No differences were found between the frequencies of fast and slow acetylators in cases (34.9% and 65.1%) and controls (33.3% and 66.7%). Two NAT2 genotypes *7/*7 (1.2%) and *5/*6/*7 (1.6%) were detected in endometriosis cases only. Four new combinations, 6D (481 + 590 mutation), 7C (590 + 857), 7D (590 + 803 + 857) and 7E (481 + 590 + 803 + 857) were detected, which have not been reported earlier. Similar genotype and phenotype results were obtained in 33 affected sister-pairs. The case-control data from this study suggest there is no association between endometriosis and NAT2 in South Indian women; however, two new variant genotypes and seven SNP combinations were also identified in cases only, which suggests that the gene may still have some as yet undetermined role in the disease.

Adolescent↗

Pseudonocardia antarctica sp. nov. an Actinomycetes from McMurdo Dry Valleys, Antarctica.

Strain DVS 5a1 was isolated from a moraine sample from the McMurdo Dry Valleys region of Antarctica. The strain is aerobic, Gram-positive, with white aerial mycelia and brown substrate mycelia, sporulating, has meso-diaminopimelic acid, arabinose and galactose in the cell wall, MK-8 (H4) as the major menaquinone and a mol% G+C content of DNA of 71% thus confirming to the description of the genus Pseudonocardia. Phylogenetic analysis based on the 16S rRNA gene sequence analysis further confirms that DVS 5al which forms a robust clade with P. alni, P. compacta, P. autotrophica and P. kongjuensis is closely related to the genus Pseudonocardia and exhibits maximum similarity of 99.7% with Pseudonocardia alni. However, at whole genome level as determined by DNA-DNA hybridisation DVS 5al exhibits a similarity of only 50% with Pseudonocardia alni. Further, DVS 5al differs from Pseudonocardia alni in that it does not produce acid from D-arabinose, meso-erythritol, melizitose, sorbitol, sucrose, D-trehalose; but produces acid from D-mannitol, D-galactose, D-maltose, D-mannose, inulin, D-ribose and D-xylose. Further, compared to Pseudonocardia alni, it has two additional fatty acids namely Me-C(18:0) and Me-C(19:0) and also possesses one additional unidentified lipid. It also shows distinct differences with P. compacta, P. autotrophica and P. kongjuensis and the other species of Pseudonocardia. It is proposed to assign DVS 5a1 the status of a new species for which the name Pseudonocardia antarctica sp. nov. is suggested.

Antarctic Regions↗

Psychrobacter salsus sp. nov. and Psychrobacter adeliensis sp. nov. isolated from fast ice from Adelie Land, Antarctica.

Nine psychrotolerant bacteria were isolated from fast ice in the middle of Geologie Archipelago, Adelie Land, Antarctica and were categorized into two groups, based on their SDS-PAGE profiles. Representatives from each of the two groups, namely strains DD 48T and SJ 14T exhibited phenotypic and chemotaxonomic characteristics confirming to the genus Psychrobacter. The 16S rRNA gene sequence indicated that the two isolates are closely related to each other and to the already reported fifteen species of Psychrobacter. Detailed studies on the phenotypic characteristics, chemotaxonomic properties and phylogenetic analysis of strains DD 48T and SJ 14T indicated that they are distinctly different from each other and the reported species of Psychrobacter. At the DNA-DNA hybridisation level, the two species exhibit less than 70% similarity. Thus, strains DD 48T and SJ 14T are identified as new species of the genus Psychrobacter for which the names Psychrobacter salsus sp. nov. and Psychrobacter adeliensis sp. nov. respectively are proposed.

Antarctic Regions↗

Deinococcus indicus sp. nov., an arsenic-resistant bacterium from an aquifer in West Bengal, India.

An arsenic- and radiation-resistant bacterium, strain Wt/1a(T), was isolated from water from an arsenic-contaminated aquifer located in the Chakdah district of West Bengal, India. The bacterium stains Gram-negative and is rod-shaped, non-motile, non-sporulating and red-pigmented. Cell-wall peptidoglycan contains ornithine as the diamino acid, MK-8 is the major menaquinone, C(15 : 1) and C(16 : 1) are the major fatty acids and the DNA G+C content of the organism is 65.8 mol%. Based on these phenotypic and chemotaxonomic characteristics, strain Wt/1a(T) was identified as a member of the genus Deinococcus. Strain Wt/1a(T) exhibited maximum 16S rRNA gene sequence similarity (95 %) with Deinococcus grandis; however, strain Wt/1a(T) exhibited only 14 % similarity to D. grandis IAM 13005(T) at the DNA-DNA level. Furthermore, strain Wt/1a(T) (compared to D. grandis IAM 13005(T)) is more resistant to arsenate and arsenite, is positive for arginine dihydrolase, utilizes a number of carbon sources and exhibits quantitative differences in fatty acid composition and qualitative differences in lipid composition. Strain Wt/1a(T) is identified as a novel species of the genus Deinococcus, for which the name Deinococcus indicus sp. nov. is proposed. The type strain of Deinococcus indicus is Wt/1a(T) (=MTCC 4913(T)=DSM 15307(T)).

Arsenic↗

Reclassification of Amycolatopsis mediterranei DSM 46095 as Amycolatopsis rifamycinica sp. nov.

Previous experiments have suggested that the rifamycin-producing strain DSM 46095 might not belong to Amycolatopsis mediterranei. Analysis of its 16S rRNA gene sequence and construction of a phylogenetic tree showed most similarity to Amycolatopsis kentuckyensis NRRL B-24129T, Amycolatopsis lexingtonensis NRRL B-24129T and Amycolatopsis pretoriensis NRRL B-24133T, but the strain was probably not a member of any of these species. Results from DNA-DNA hybridization experiments and comparison of DNA profiling patterns using pulsed-field gel electrophoresis also supported the assignment of strain DSM 46095 to a novel species. Analyses of phospholipids, fatty acid methyl esters and physiological characteristics also showed that the differences between different isolates of A. mediterranei and A. mediterranei DSM 46095 were as large as those between Amycolatopsis species. Strain DSM 46095 represents a novel species of the genus Amycolatopsis for which the name Amycolatopsis rifamycinica sp. nov. is proposed, with the type strain NT 19T (=DSM 46095T=ATCC 27643T).

Actinomycetales↗

Bacillus indicus sp. nov., an arsenic-resistant bacterium isolated from an aquifer in West Bengal, India.

Strain Sd/3T (=MTCC 4374T=DSM 15820T), an arsenic-resistant bacterium, was isolated from a sand sample obtained from an arsenic-contaminated aquifer in Chakdah district in West Bengal, India (23 degrees 3' N 88 degrees 35' E). The bacterium was Gram-positive, rod-shaped, non-motile, endospore-forming and yellowish-orange pigmented. It possessed all the characteristics that conform to the genus Bacillus, such as it had A4beta murein type (L-orn-D-Asp) peptidoglycan variant, MK-7 as the major menaquinone and iso-C15 : 0 and anteiso-C15 : 0 as the major fatty acids. Based on its chemotaxonomic and phylogenetic characteristics, strain Sd/3T was identified as a species of the genus Bacillus. It exhibited maximum similarity (95%) at the 16S rRNA gene level with Bacillus cohnii; however, DNA-DNA similarity with B. cohnii was 60.7%. Strain Sd/3T also exhibited a number of phenotypic differences from B. cohnii (DSM 6307T). These data suggest that Sd/3T represents a novel species of the genus Bacillus. The name Bacillus indicus sp. nov. is proposed.

Anti-Bacterial Agents↗

Arthrobacter gangotriensis sp. nov. and Arthrobacter kerguelensis sp. nov. from Antarctica.

Two coryneform bacteria were isolated from a penguin rookery soil sample collected in Antarctica, near the Indian station Dakshin Gangotri (strain Lz1y(T)), and from sea water from Kerguelen island, Antarctica (strain KGN15(T)). They have morphological and chemotaxonomic properties (peptidoglycan A4alpha type; major menaquinones MK-8, MK-9 and MK-10; predominant fatty acids anteiso-C(15 : 0) and anteiso-C(17 : 0)) that are characteristic of members of the genus Arthrobacter. The isolates shared 97.8 % 16S rRNA gene sequence similarity to each other and were most closely related to Arthrobacter sulfureus (about 98.5 % sequence similarity). DNA-DNA hybridization experiments revealed 50 % relatedness between the isolates, while the levels of DNA-DNA relatedness between strains Lz1y(T) and KGN15 (T) and their phylogenetic relative, A. sulfureus, were respectively 54 and 12 %. Based on the above data and distinct phenotypic differences between the isolates and A. sulfureus, two novel species are proposed, Arthrobacter gangotriensis sp. nov. (type strain Lz1y(T)=DSM 15796(T)=JCM 12166(T)) and Arthrobacter kerguelensis sp. nov. (type strain KGN15(T)=DSM 15797(T)=JCM 12165(T)).

Antarctic Regions↗

Bacterial diversity of a soil sample from Schirmacher Oasis, Antarctica.

The bacterial diversity of a soil sample collected in the vicinity of Lake Zub, Schirmacher Oasis, Antarctica, was determined both by establishing pure colonies of culturable bacteria and by cloning the total 16S rDNA of the soil and establishing the phylogeny of the clones. Analysis of the 16S rRNA gene clones indicated that the bacteria belonged to the classes alpha-proteobacteria, beta-proteobacteria, gamma-proteobacteria, Gemmatimonas, Bacteriodetes, Actinobacteria, Chloroflexi and Chlamydiae. In addition, seven clones were categorized as unidentified and unculturable in the classes of beta-Proteobacteria, Actinobacteria, Chloroflexi and Chlamydiae. Further, the culturable bacteria from the same site were identified as belonging to the genera Pseudomonas, Sphingobacterium, Arthrobacter, Micrococcus, Brevondimonas, Rhodococcus and Microbacterium. These results identify for the first time the presence of bacteria belonging to the genera Brevundimonas, Microbacterium, Rhodococcus, Serratia, Enterobacter, Rhodopseudomonas, Sphingomonas, Acidovorax, Burkholderia, Nevskia, Gemmatimonas, Xanthomonas and Flexibacter in Antarctica. Further, comparison of the Antarctic soil bacterial diversity with other cold habitats of Antarctica like from sediments, ice and cyanobacterial mat samples indicated that the bacterial diversity in soil was similar to the diversity observed in the continental shelf sediment sample. The Antarctic soil clones also resembled the bacterial diversity of soils from other geographical regions, but were unique in that none of the clones from the soil belonged to the uncultured Y, O, G, A and B groups common to all soil samples.

Antarctic Regions↗

Role of membrane lipid fatty acids in cold adaptation.

Psychrophilic and psychrotolerant bacteria have evolved various strategies to adapt to low temperature. One important strategy, which is crucial to the survival of the cell at low temperature, relates to the ability of the cell to modulate the fluidity of the membrane. Bacteria in general modulate membrane fluidity by altering their fatty acid composition. But, bacteria could also achieve the same by various other strategies such as by altering the lipid head group, the protein content of the membrane, the type of carotenoids synthesized, the fatty acid chain length and the proportion of cis to trans fatty acids. In addition bacteria have a two-component signal transduction pathway consisting of a membrane-bound sensor and a soluble cytoplasmic response regulator involved in the perception and transduction of low temperature signals. This review on cold adaptation highlights the various strategies by which bacteria modulate the fluidity of the membrane and the process by which it senses and transduces the low temperature signal.

Adaptation, Physiological↗

Novel tyrosine-phosphorylated post-pyruvate metabolic enzyme, dihydrolipoamide dehydrogenase, involved in capacitation of hamster spermatozoa.

Capacitation is a process that confers fertilizing ability to spermatozoa and this critical event occurs in the development of mammalian spermatozoa during their transit through the female reproductive tract and precedes fertilization. Because spermatozoa are relatively silent in transcription and translation, posttranslational modifications perform the regulatory functions in these cells during capacitation. In this report, we identify a candidate protein, dihydrolipoamide dehydrogenase, which is a post-pyruvate metabolic enzyme, exhibiting tyrosine phosphorylation during hamster spermatozoal capacitation. This is the first report showing dihydrolipoamide dehydrogenase as a phosphoprotein. The cDNA sequence of hamster testes dihydrolipoamide dehydrogenase does not show any variation from the already reported mammalian dihydrolipoamide dehydrogenases. Downregulation of the activity of the hamster spermatozoal enzyme by its specific inhibitor, 5-methoxyindole-2-carboxylic acid, blocks acrosome reaction completely and hyperactivation partially, confirming the role of dihydrolipoamide dehydrogenase in hamster spermatozoal capacitation. We also delineate the temporal involvement of glucose and pyruvate-lactate, showing that the former is required in the earlier stages and the latter for the later stages of hamster spermatozoal capacitation. The essentiality of pyruvate-lactate during hyperactivation and acrosome reaction necessitates the involvement of the post-pyruvate-lactate enzyme, dihydrolipoamide dehydrogenase.

Acrosome Reaction↗

Psychrophilic Planococcus maitriensis sp.nov. from Antarctica.

An orange pigmented bacterium, S1, was isolated from a cyanobacterial mat sample collected in the vicinity of Schirmacher Oasis, Maitri, the Indian station, in Antarctica. The bacterium is Gram-positive and possesses all the characteristics of the genus Planococcus. It is non-sporulating, motile and has A4alpha type peptidoglycan, MK-7 and MK-8 as the major menaquinones and anteiso-C(15:0) as the major fatty acid. Based on the phylogenetic characteristics, the bacterium S1 is identified as a close relative of Planococcus citreus with which it shares 98.12% similarity at the 16S rRNA gene level but exhibits a low similarity of 52% at the whole genome level. Apart from the above major differences, S1 also exhibits phenotypic differences with Planococcus citreus and other members of the genus Planococcus. Based on these differences, the bacterium S1 is identified as a new species of the genus Planococcus for which the name Planococcus maitriensis is proposed. The type strain of Planococcus maitriensis is S1(T) (= MTCC 4827; DSM 15305).

Antarctic Regions↗

Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov., psychrophiles from a pond in Antarctica.

Two unique psychrophilic strains (CMS 76rT and CMS 81yT) were isolated from a cyanobacterial mat sample from a pond in Wright Valley, McMurdo, Antarctica. Both isolates were assigned to the genus Leifsonia, since they were gram-positive, curved rods, non-motile, catalase-positive, contained DL-2,4-diaminobutyric acid, menaquinone MK-11, phosphatidylglycerol and diphosphatidylglycerol, had a high content of anteiso- and iso-branched fatty acids and had a DNA G + C content of 64-66 mol%. In addition, both isolates were related to the five reported species of Leifsonia at a level of about 95-96% 16S rDNA sequence similarity and differed from one another by 2.5%. Strains CMS 76rT and CMS 81yT also differed from one another in many other phenotypic characteristics and exhibited only 30% relatedness at the DNA-DNA level, thus indicating that they represent two different species. Furthermore, these two isolates also showed many distinct differences with respect to the reported species of Leifsonia in terms of their phenotypic characteristics, biochemical properties, chemotaxonomic features, sensitivity to various antibiotics and 16S rDNA similarity, clearly indicating that strains CMS 76rT (= MTCC 4210T = DSM 15304T = CIP 107783T) and CMS 81yT (= MTCC 4657T = DSM 15303T = CIP 107785T) represent the type strains of two novel species of Leifsonia, for which the names Leifsonia rubra sp. nov. and Leifsonia aurea sp. nov. are proposed.

Actinomycetales↗

Sporosarcina macmurdoensis sp. nov., from a cyanobacterial mat sample from a pond in the McMurdo Dry Valleys, Antarctica.

Strain CMS 21w(T) was isolated from a cyanobacterial mat sample taken from a pond located in the McMurdo Dry Valleys, Antarctica. Based on its phenotypic, chemotaxonomic and phylogenetic properties, strain CMS 21w(T) was identified as a member of the genus SPOROSARCINA: At the 16S rRNA gene level, CMS 21w(T) exhibited about 93-96 % similarity to all reported species of Sporosarcina and exhibited a maximum similarity of 96 % to both Sporosarcina globispora and Sporosarcina psychrophila. Based on more than 3 % difference at the 16S rRNA gene sequence level and the presence of distinct differences with respect to phenotypic, biochemical and chemotaxonomic features, strain CMS 21w(T) (=MTCC 4670(T)=DSM 15428(T)=CIP 107784(T)) is proposed as the type strain of a novel species of Sporosarcina, Sporosarcina macmurdoensis sp. nov.

Antarctic Regions↗

Role of signaling pathways in regulating the capacitation of mammalian spermatozoa.

Mammalian testicular spermatozoa are immotile and incompetent for fertilization. They acquire motility during epididymal maturation and fertilizing ability during a second phase of maturation in the female reproductive tract, termed as capacitation. Capacitation was discovered independently by Austin and Cang in early 1950s and was defined as the obligate period of residency of spermatozoa in the female reproductive tract, which confers on the spermatozoa the ability to fertilize an oocyte. Over the years, the definition of capacitation has changed and it has been recognized as a complex phenomenon, which is correlated with changes associated with the spermatozoa in the female tract. These alterations in metabolism, intracellular ion concentration, membrane fluidity, intracellular pH, cAMP concentration and concentration of reactive oxygen species, ultimately make the spermatozoa fertilization-competent. The molecular basis of capacitation is poorly understood despite the fact that it is an important event preceding fertilization. This review presents our current understanding of the signaling events involved in the process of capacitation.

Acrosome↗

Inhibition of in vitro capacitation of hamster spermatozoa by nitric oxide synthase inhibitors.

In an attempt to understand the role of nitric oxide(NO) in sperm capacitation, in the present study, hamster spermatozoa were used to evaluate the effects of NO on motility, viability, hyperactivation, capacitation and protein tyrosine and serine phosphorylation using specific inhibitors of nitric oxide synthase (NOS); namely L-NAME (N-nito-L-aginine methyl ester) and 7-Ni (7-nitroindazole). The results indicated that L-NAME inhibits sperm motility, hyperactivation and acrosome reaction where as 7-Ni inhibits only hyperactivation and acrosome reaction thus implying that NOS inhibitors exhibit subtle differences with respect to their effects on sperm functions. This study also provides evidence that NOS inhibitors inhibit sperm capacitation by their ability to modulate protein tyrosine phosphorylation. However, the inhibitors had no effect on the protein serine phosphorylation of hamster spermatozoa during capacitation. Thus, these results indicate that NO is required

Acrosome Reaction↗

Conservation of wild animals by assisted reproduction and molecular marker technology.

Wild animals are an integral component of the ecosystem. Their decimation due to abrupt natural calamities or due to gradual human intervention would be disastrous to the ecosystem and would alter the balance in nature between various biotic components. Such an imbalance could have an adverse effect on the ecosystem. Therefore, there is an urgent need to put an end to the ever increasing list of endangered species by undertaking both in situ and ex situ conservation using tools of modern biology, to ascertain the degree of genetic variation and reproductive competence in these animals. This review highlights the development and use of molecular markers such as microsatellites, minisatellites, mitochondrial control region, cytochrome b and MHC loci to assess the genetic variation in various Indian wild animals such as the lion, tiger, leopard and deer. The review also presents data on the semen profile of the big cats of India. Reproductive technologies such as cryopreservation of semen and artificial insemination in big cats are also highlighted.

Animals↗

Halomonas glaciei sp. nov. isolated from fast ice of Adelie Land, Antarctica.

Eleven psychrophilic bacteria were isolated from a solid layer of fast ice in the middle of Pointe-Geologie Archipelago, Adelie Land, Antarctica. The 11 isolates based on the phenotypic characteristics, chemotaxonomic and phylogenetic analysis have been identified as members of the genus Halomonas. All the isolates at the 16S rDNA sequence level were identical, possessed the 15 conserved nucleotides of the family Halomonadaceae and four nucleotides of the genus Halomonas. Therefore, the 16S rDNA sequence of DD 39 was used for calculating the evolutionary distances and for phylogenetic analysis. It was observed that DD 39 formed a robust cluster with H. variabilis, from which it differed by 0.7%. Further DNA-DNA hybridization studies indicated low DNA-DNA homology (15%) between H. variabilis and DD 39. Between the 11 Antarctic isolates the homology was >85%. In addition it was observed that DD 39 was different from H. variabilis in that it was psychrophilic, could tolerate only up to 15% sodium chloride, could not hydrolyse esculin, could not reduce nitrate, was urease negative, could not utilize glycerol as a carbon source, and was resistant to ampicillin and erythromycin and sensitive to nalidixic acid. In addition, it also exhibited distinct differences with respect to high content of C(16:1) and low levels of cyclo-C(17:0) and cyclo-C(19:0). DD 39 also differed from all the other reported species of Halomonas with respect to many phenotypic characteristics. It is proposed therefore that DD 39 should be placed in the genus Halomonas as a new species that is Halomonas glaciei. The type strain of H. glaciei is DD 39(T) (MTCC 4321; JCM 11692).

Antarctic Regions↗