03062nas a2200241 4500008004100000022001400041245011700055210006900172260008500241300000900326490000600335520215300341100002002494700002302514700002402537700002402561700002102585700001902606700002002625700002302645700002402668856012802692 2007 eng d a1932-620300aAnalysis of mitochondrial DNA sequences in childhood encephalomyopathies reveals new disease-associated variants0 aAnalysis of mitochondrial DNA sequences in childhood encephalomy a185 BERRY ST, STE 1300, SAN FRANCISCO, CA 94107 USAbPUBLIC LIBRARY SCIENCEcSEP ae9420 v23 a

Background. Mitochondrial encephalomyopathies are a heterogeneous group of clinical disorders generally caused due to mutations in either mitochondrial DNA (mtDNA) or nuclear genes encoding oxidative phosphorylation (OXPHOS). We analyzed the mtDNA sequences from a group of 23 pediatric patients with clinical and morphological features of mitochondrial encephalopathies and tried to establish a relationship of identified variants with the disease. Methodology/Principle Findings. Complete mitochondrial genomes were amplified by PCR and sequenced by automated DNA sequencing. Sequencing data was analyzed by SeqScape software and also confirmed by BLASTn program. Nucleotide sequences were compared with the revised Cambridge reference sequence (CRS) and sequences present in mitochondrial databases. The data obtained shows that a number of known and novel mtDNA variants were associated with the disease. Most of the non-synonymous variants were heteroplasmic (A4136G, A9194G and T11916A) suggesting their possibility of being pathogenic in nature. Some of the missense variants although homoplasmic were showing changes in highly conserved amino acids (T3394C, T3866C, and G9804A) and were previously identified with diseased conditions. Similarly, two other variants found in tRNA genes (G5783A and C8309T) could alter the secondary structure of Cys-tRNA and Lys-tRNA. Most of the variants occurred in single cases; however, a few occurred in more than one case (e. g. G5783A and A10149T). Conclusions and Significance. The mtDNA variants identified in this study could be the possible cause of mitochondrial encephalomyopathies with childhood onset in the patient group. Our study further strengthens the pathogenic score of known variants previously reported as provisionally pathogenic in mitochondrial diseases. The novel variants found in the present study can be potential candidates for further investigations to establish the relationship between their incidence and role in expressing the disease phenotype. This study will be useful in genetic diagnosis and counseling of mitochondrial diseases in India as well as worldwide.

1 aWani, Aijaz, A.1 aAhanger, Sajad, H.1 aBapat, Sharmila, A.1 aRangrez, Ashraf, Y.1 aHingankar, Nitin1 aSuresh, C., G.1 aBarnabas, Shama1 aPatole, Milind, S.1 aShouche, Yogesh, S. uhttps://library.ncl.res.in/content/analysis-mitochondrial-dna-sequences-childhood-encephalomyopathies-reveals-new-disease-002357nas a2200277 4500008004100000022001400041245015900055210006900214260009000283300001200373490000700385520127400392653002401666653002801690653002901718653003401747100002001781700002401801700002001825700002401845700001901869700002001888700002301908700002001931856012801951 2008 eng d a1071-576200aAnalysis of reactive oxygen species and antioxidant defenses in complex I deficient patients revealed a specific increase in superoxide dismutase activity0 aAnalysis of reactive oxygen species and antioxidant defenses in  aTELEPHONE HOUSE, 69-77 PAUL STREET, LONDON EC2A 4LQ, ENGLANDbINFORMA HEALTHCAREcMAY a415-4270 v423 a

The mechanism of free radical production by complex I deficiency is ill-defined, although it is of significant contemporary interest. This study studied the ROS production and antioxidant defenses in children with mitochondrial NADH dehydrogenase deficiency. ROS production has remained significantly elevated in patients compared to controls. The expression of all antioxidant enzymes significantly increased at mRNA level. However, the enzyme activities did not correlate with high mRNA or protein expression. Only the activity of superoxide dismutase (SOD) was found to correlate with higher mRNA expression in patient derived cell lines. The activities of the enzymes such as glutathione peroxidase (GPx), Catalase (CAT) and glutathione-S-transferase (GST) were significantly reduced in patients (p < 0.05 or p < 0.01). Glutathione reductase (GR) activity and intracellular glutathione (GSH) levels were not changed. Decreased enzyme activities could be due to post-translational or oxidative modification of ROS scavenging enzymes. The information on the status of ROS and marking the alteration of ROS scavenging enzymes in peripheral lymphocytes or lymphoblast cell lines will provide a better way to design antioxidant therapies for such disorders.

10aantioxidant enzymes10amitochondrial complex I10amitochondrial myopathies10aReactive Oxygen Species (ROS)1 aWani, Aijaz, A.1 aRangrez, Ashraf, Y.1 aKumar, Himanshu1 aBapat, Sharmila, A.1 aSuresh, C., G.1 aBarnabas, Shama1 aPatole, Milind, S.1 aShouche, Yogesh uhttps://library.ncl.res.in/content/analysis-reactive-oxygen-species-and-antioxidant-defenses-complex-i-deficient-patients-001757nas a2200277 4500008004100000022001400041245006100055210006000116260009200176300001400268490000600282520086800288653002401156653001801180653000801198653000801206653000801214100002201222700001801244700002301262700002301285700002301308700003001331700002401361856009401385 2008 eng d a1533-488000aBacterial synthesis of copper/copper oxide nanoparticles0 aBacterial synthesis of coppercopper oxide nanoparticles a26650 THE OLD RD, STE 208, VALENCIA, CA 91381-0751 USAbAMER SCIENTIFIC PUBLISHERScJUN a3191-31960 v83 a

A bacterial mediated synthesis of copper/copper oxide nanoparticle composite is reported. A Gram-negative bacterium belonging to the genus Serratia was isolated from the midgut of Stibara sp., an insect of the Cerambycidae family of beetles found in the Northwestern Ghats of India. This is a unique bacterium that is quite specific for the synthesis of copper oxide nanoparticles as several other strains isolated from the same insect and common Indian mosquitoes did not result in nanoparticle formation. By following the reaction systematically, we could delineate that the nanoparticle formation occurs intracellularly. However, the process results in the killing of bacterial cells. Subsequently the nanoparticles leak out as the cell wall disintegrates. The nanoparticles formed are thoroughly characterized by UV-Vis, TEM, XRD, XPS and FTIR studies.

10abacterial synthesis10aNanoparticles10aTEM10aXPS10aXRD1 aHasan, Syed, Saif1 aSingh, Sanjay1 aParikh, Rasesh, Y.1 aDharne, Mahesh, S.1 aPatole, Milind, S.1 aPrasad, Bhagavatula L. V.1 aShouche, Yogesh, S. uhttps://library.ncl.res.in/content/bacterial-synthesis-coppercopper-oxide-nanoparticles-002718nas a2200265 4500008004100000022001400041245018300055210006900238260007700307300001400384490000600398520169400404653001702098653001902115653001802134653002402152653001102176100002302187700001802210700003002228700002302258700001902281700002402300856012802324 2008 eng d a1439-422700aExtracellular synthesis of crystalline silver nanoparticles and molecular evidence of silver resistance from morganella sp.: towards understanding biochemical synthesis mechanism0 aExtracellular synthesis of crystalline silver nanoparticles and  aPO BOX 10 11 61, D-69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJUN a1415-14220 v93 a

There has been significant progress in the biological synthesis of nanomaterials. However, the molecular mechanism of synthesis of such bio-nanomaterials remains largely unknown. Here, we report the extracellular synthesis of crystalline silver nanoparticles (AgNPs) by using Morganella sp., and show molecular evidence I of silver resistance by elucidating the synthesis mechanism. The AgNPs were 20 +/- 5 nm in diameter and were highly stable at 1 room temperature. The kinetics of AgNPs formation was investigated. Detectable particles were formed after on hour of reaction, and their production remained exponential up to 18 h, and saturated at 24 h. Morganella sp. was found to be highly resistant to silver cations and was able to grow in the presence of more than 0.5 mm AgNO3. Three gene homologues viz. silE, silP and silS were identified in silver-resistant Morganella sp. The homologue of silE from Morganella sp. showed 99% nucleotide sequence similarity with the previously reported gene, silE, which encodes a periplasmic silver-binding protein. The homologues of silP and silS were also highly similar to previously reported sequences. Similar activity was totally absent in closely related Escherichia coli; this suggests that a unique mechanism of extracellular AgNPs synthesis is associated with silver-resistant Morganella sp. The molecular mechanism of silver resistance and its gene products might have a key role to play in the overall synthesis process of AgNPs by Morganella sp. An understanding of such biochemical mechanisms at the molecular level might help in developing an ecologically friendly and cost-effective protocol for microbial AgNPs synthesis.

10aBiosynthesis10aMorganella sp.10aNanoparticles10asil gene homologues10aSilver1 aParikh, Rasesh, Y.1 aSingh, Sanjay1 aPrasad, Bhagavatula L. V.1 aPatole, Milind, S.1 aSastry, Murali1 aShouche, Yogesh, S. uhttps://library.ncl.res.in/content/extracellular-synthesis-crystalline-silver-nanoparticles-and-molecular-evidence-silver-002308nas a2200241 4500008004100000022001400041245015900055210006900214260007900283300001000362490000700372520137500379653002001754653001901774653001401793653001401807653001901821100002501840700003401865700002301899700001401922856013001936 2010 eng d a0196-978100aMolecular cloning, over expression, and activity studies of a peptidic HIV-1 protease inhibitor: designed synthetic gene to functional recombinant peptide0 aMolecular cloning over expression and activity studies of a pept a360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USAbELSEVIER SCIENCE INCcJAN a16-210 v313 a

The aspartic protease inhibitor (ATBI) purified from a Bacillus sp. is a potent inhibitor of several proteases including recombinant HIV-1 protease, pepsin, and fungal aspartic protease. In this study, we report the cloning, and over expression of a synthetic gene coding for ATBI in Escherichia coli and establish a purification Protocol. The ATBI molecule consists of eleven amino acids and is peptidic in nature. We used the peptide sequence data of ATBI to synthesize complementary oligonucleotides, which were annealed and subsequently cloned in-frame with the gene for glutathione-S-transferase (GST). The expression of the resulting fusion protein was induced in E. coli BL21-A1 cells using arabinose. The recombinant peptide was Purified using a reduced glutathione column, and cleaved with Factor Xa to remove the GST tag. The resultant product was further purified to homogeneity using RP-HPLC. Mass spectroscopy analysis revealed that the purified peptide had a molecular weight of 1186 Da which matches the theoretical molecular weight of the amino acids present in the synthetic gene. The recombinant peptide was found to be active in vitro against HIV-1 protease, pepsin, and fungal aspartic protease. The protocol described in this study may be used to clone pharmaceutically important peptide molecules. (C) 2009 Elsevier Inc. All rights reserved.

10aGateway cloning10aHIV-1 protease10ainhibitor10aProteases10aSynthetic gene1 aVathipadiekal, Vinod1 aUmasankar, Perunthottathu, K.1 aPatole, Milind, S.1 aRao, Mala uhttps://library.ncl.res.in/content/molecular-cloning-over-expression-and-activity-studies-peptidic-hiv-1-protease-inhibitor-0