02578nas a2200229 4500008004100000022001400041245019000055210006900245260007300314300001000387490000700397520165500404653002102059653002902080653002002109653002102129653001702150100001702167700001802184700001902202856012702221 2008 eng d a0924-857900aBactericidal activity of 2-nitroimidazole against the active replicating stage of mycobacterium bovis BCG and mycobacterium tuberculosis with intracellular efficacy in THP-1 macrophages0 aBactericidal activity of 2nitroimidazole against the active repl aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcJUL a40-450 v323 a

This study evaluated the antituberculous potential of 2-nitroimidazole under in vitro conditions. Minimal bactericidal concentrations of the compound against actively replicating Mycobacterium bovis BCG and Mycobacterium tuberculosis H37Ra were found to be 0.226 mu g/mL and 0.556 mu g/mL in enriched and minimal medium, respectively. Minimal inhibitory concentrations were > 100 times lower than reported antituberculous nitroimidazoles such as nitrofurantoin and furaltadone, indicating the greater potential of 2-nitroimidazole. No discernible effect of 2-nitroimidazole was seen on saprophytic Mycobacterium smegmatis and the representative bacterial strain Escherichia coli DH5 alpha, indicating the specificity of the molecule against tuberculous mycobacteria. The compound was also found to be effective against M. tuberculosis in the intracellular environment of the human monocytic cell line THP-1, with a reduction in viability of bacilli by 2.5 log after 144 h of incubation at a concentration of 0.113 mu g/mL. A five-fold higher concentration (0.565 mu g/mL) of 2-nitroimidazole sterilised the macrophages of intracellular pathogens within 192 h, without affecting the host. However, 2-nitroimidazole was unable to affect significantly the viability of dormant non-replicating bacilli of M. bovis BCG and M. tuberculosis in Wayne's in vitro model. Overall, the results indicate that 2-nitroimidazole is a potent antituberculous agent active against the organism's active replicating stage, with promising intracellular efficacy as well. (c) 2008 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

10a2-nitroimidazole10aantimycobacterial agents10anitroimidazoles10aTHP-1 macrophage10atuberculosis1 aKhan, Arshad1 aSarkar, Sampa1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/bactericidal-activity-2-nitroimidazole-against-active-replicating-stage-mycobacterium-001275nas a2200253 4500008004100000022001400041245006000055210006000115260009500175300001200270490000800282520047900290653001200769653001200781653001700793653001800810653001600828653001100844100001900855700001800874700002100892700001900913856008900932 2011 eng d a0003-269700aMethod to extract intact and pure RNA from mycobacteria0 aMethod to extract intact and pure RNA from mycobacteria a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcOCT a286-2880 v4173 a

We describe a high-yielding, simple, and aerosol-free protocol for the isolation of RNA from mycobacteria that does not require sophisticated instruments. The method yielded 50 mu g of RNA from 10(7) cells, 50 times more than a recently reported method. Our method can extract total RNA from aerobically grown bacteria and from in vitro hypoxia-induced dormant bacilli and mycobacteria residing within infected macrophages. (C) 2011 Elsevier Inc. All rights reserved.

10aDormant10aHypoxia10aMycobacteria10aRNA isolation10aSpheroplast10aTrizol1 aAkhtar, Shamim1 aSarkar, Sampa1 aMishra, Abhishek1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/method-extract-intact-and-pure-rna-mycobacteria-002223nas a2200217 4500008004100000022001400041245016600055210006900221260007600290300001200366490000700378520134900385653003001734653001401764653003101778653002201809653000901831100001801840700001901858856012801877 2012 eng d a1087-057100aPotential use of nitrate reductase as a biomarker for the identification of active and dormant inhibitors of mycobacterium tuberculosis in a THP1 infection model0 aPotential use of nitrate reductase as a biomarker for the identi a2455 TELLER RD, THOUSAND OAKS, CA 91320 USAbSAGE PUBLICATIONS INCcAUG a966-9730 v173 a

The development of a macrophage-based, antitubercular high-throughput screening system could expedite discovery programs for identifying novel inhibitors. In this study, the kinetics of nitrate reduction (NR) by Mycobacterium tuberculosis during growth in Thp1 macrophages was found to be almost parallel to viable bacilli count. NR in the culture medium containing 50 mM of nitrate was found to be optimum on the fifth day after infection with M. tuberculosis. The signal-to-noise (S/N) ratio and Z-factor obtained from this macrophage-based assay were 5.4 and 0.965, respectively, which confirms the robustness of the assay protocol. The protocol was further validated by using standard antitubercular inhibitors such as rifampicin, isoniazid, streptomycin, ethambutol, and pyrazinamide, added at their IC90 value, on the day of infection. These inhibitors were not able to kill the bacilli when added to the culture on the fifth day after infection. Interestingly, pentachlorophenol and rifampicin killed the bacilli immediately after addition on the fifth day of infection. Altogether, this assay protocol using M. tuberculosis-infected Thp-1 macrophages provides a novel, cost-efficient, robust, and easy-to-perform screening platform for the identification of both active and hypoxic stage-specific inhibitors against tuberculosis.

10ahigh-throughput screening10ainhibitor10aMycobacterium tuberculosis10anitrate reductase10aTHP11 aSarkar, Sampa1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/potential-use-nitrate-reductase-biomarker-identification-active-and-dormant-inhibitors-003028nas a2200169 4500008004100000245016800041210007100209260000800280520230800288100002502596700001802621700001902639700002102658700002802679700001802707856013302725 2012 eng d00aSelf–assembled histidine acid phosphate nanocapsules as templates for multifunctional hollow platinum nanospheres in ionic liquid [BMIM][BF 4 ] for drug delivery0 aSelf–assembled histidine acid phosphate nanocapsules as template cJAN3 a

During the past few decades, the design and fabrication of nano drug delivery vehicle has received considerable attention for incorporating multi-functionality, emphasizing transportation of anticancer drugs to tumour sites. Protein-based nanoparticles hold significant promise in the development of anticancer drug delivery systems since they have remarkable superiority in biocompatibility, biodegradability and natural metabolism in physiologic systems. Ionic liquids (ILs) are a class of solvents that are composed solely of ions (ILs – commonly referred as room temperature molten salts or room temperature ionic liquids). They have recently emerged as green solvents or green reaction media, because of their reported unique physico-chemical properties. We report for the first time the controlled self-assembly of Histidine Acid phosphatase (HAP) enzyme in the IL 1-butyl-3-methylimidazolium tetrafluoroborate [BMIM][BF 4 ], leading to the formation of HAP nanocapsules as template for synthesis of platinum nanospheres. These HAP nano-containers were loaded with an anticancer drug curcumin. We further demonstrated in vitro drug release and synergistic anticancer effect of these systems on three different cell lines viz. hepatocellular carcinoma (HepG2), breast cancer (MCF-7) and human acute monocytes (THP-1). The present work can extend our theoretical understanding on nano-bio interfacial molecular self assembly and will facilitate the motivated insights of inorganic/organic hybrid nano-materials/ nano-particles/ nano-objects and ionic liquids based multi functional drug delivery vehicles with synergistic combination chemotherapy for potential application in tumor targeting. INTRODUCTION During the past few decades, the design and fabrication of drug delivery vehicle has received considerable attention for incorporating multi-functionality emphasizing transportation of anticancer drugs to tumour sites. These nano drug delivery vehicles have many advantages including prolonged circulation time of the drugs (Duncan, 2003, Kiick, 2007) , improved drug targeting and solubility (Duncan, 2003, Kiick, 2007), higher payloads (Duncan, 2003) and controlled release of the therapeutics into the blood stream or the targeted tumour tissues (Uhrich et al. 1999, Duncan, 2003).

1 aSoni, Sarvesh, Kumar1 aSarkar, Sampa1 aSarkar, Dhiman1 aColoe, Peter, J.1 aBhargava, Suresh, Kumar1 aBansal, Vipul uhttps://library.ncl.res.in/content/self%E2%80%93assembled-histidine-acid-phosphate-nanocapsules-templates-multifunctional-hollow02293nas a2200265 4500008004100000022001400041245016000055210006900215260009900284300001400383490000700397520126900404653001301673653001301686653001201699653002901711653002701740100002401767700002501791700002201816700001701838700001801855700002601873856012801899 2013 eng d a1359-511300aPhytase isozymes from aspergillus niger NCIM 563 under solid state fermentation: biochemical characterization and their correlation with submerged phytases0 aPhytase isozymes from aspergillus niger NCIM 563 under solid sta aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcNOV a1618-16250 v483 a

Aspergillus niger NCIM 563 produces dissimilar phytase isozymes under solid state and submerged fermentation conditions. Biochemical characterization and applications of phytase Phy III and Phy IV in SSF and their comparison with submerged fermentation Phy I and Phy III were studied. SSF phytases have a higher metabolic potential as compared to SmF. Phy I is tetramer and Phy II, III and IV are monomers. Phy I and IV have pH optima of 2.5 and Phy II and III have pH optima of 5.0 and 5.6, respectively. Phy I, III and IV exhibited very broad substrate specificity while Phy II was more specific for sodium phytate. SSF phytase is less thermostable as compared to SmF phytase. Phy land II show homology with other known phytases while Phy III and IV show no homology with SmF phytases and any other known phytases from the literature suggesting their unique nature. This is the first report about differences among phytase produced under SSF and SmF by A. niger and this study provides basis for explanation of the stability and catalytic differences observed for these enzymes. Exclusive biochemical characteristics and multilevel application of SSF native phytases determine their efficacy and is exceptional. (C) 2013 Elsevier Ltd. All rights reserved.

10aA. niger10aIsozymes10aPhytase10aSolid state fermentation10aSubmerged fermentation1 aBhavsar, Kavita, P.1 aBuddhiwant, Priyanka1 aSoni, Sarvesh, K.1 aDepan, Dilip1 aSarkar, Sampa1 aKhire, Jayant, Malhar uhttps://library.ncl.res.in/content/phytase-isozymes-aspergillus-niger-ncim-563-under-solid-state-fermentation-biochemical-002692nas a2200193 4500008004100000022001400041245011800055210006900173260011000242300001600352490000600368520188100374100002202255700001802277700002402295700001902319700002802338856013202366 2015 eng d a2046-206900aIntrinsic therapeutic and biocatalytic roles of ionic liquid mediated self-assembled platinum-phytase nanospheres0 aIntrinsic therapeutic and biocatalytic roles of ionic liquid med aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcJUL a62871-628810 v53 a

We herein report the inherent antitumor efficiency of self-assembled phytase enzyme nanospheres and enhance their efficiency by decorating with platinum nanoparticles and with the anticancer drug curcumin. Firstly, controlled self-assembly of phytase enzyme in an Ionic Liquid 1-butyl-3-methylimidazolium tetrafluoroborate [Bmim][BF4], led to the formation of therapeutically active phytase nanospheres. These nanospheres were further decorated with platinum nanoparticles by adding the platinum ions to these spheres and the nanoparticles formation was mediated by the specific interaction between the histidine residue (in active site of phytase enzymes) and the platinum ions and subsequent reduction of the ions into nanoparticles. The enzyme spheres act as a functional soft template for the as-formed platinum nanoparticles. These Platinum decorated hybrid biomacromolecular phytase nanospheres were loaded with the anticancer drug curcumin and all the different kinds of nanospheres were subjected to in vitro cytotoxicity for their anticancer effect on three different kinds of cancer cell lines i.e. MCF-7, Hep-G2 and THP-1 derived human macrophages. We observed a gradual increase in the anticancer effect caused by only phytase nanospheres (25%), platinum-phytase nanospheres (37%), phytase-curcumin (78%) and platinum-phytase-curcumin nanospheres (90%) that establishes this protein based system as a robust combinatorial drug delivery vehicle. The platinum-phytase spheres also proved their usability as a highly efficient green and reusable biocatalytic system for phytate degradation. The present work facilitates our understanding of ionic liquid based synthesis for multifunctional protein based drug delivery vehicles incorporating combinatorial chemotherapy for potential application as biopharmaceutical agents for tumor treatment and bio-catalysis.

1 aSoni, Sarvesh, K.1 aSarkar, Sampa1 aSelvakannan, P., R.1 aSarkar, Dhiman1 aBhargava, Suresh, Kumar uhttps://library.ncl.res.in/content/intrinsic-therapeutic-and-biocatalytic-roles-ionic-liquid-mediated-self-assembled-platinum-002428nas a2200301 4500008004100000022001400041245009900055210006900154260006700223300001200290490000700302520136400309653002801673653003301701653001601734653002701750100002101777700002101798700002301819700003401842700001801876700001901894700002201913700002201935700002101957700002401978856012402002 2015 eng d a1747-027700aTargeting dormant tuberculosis bacilli: results for molecules with a novel pyrimidone scaffold0 aTargeting dormant tuberculosis bacilli results for molecules wit a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcFEB a201-2070 v853 a

Our inability to completely control TB has been due in part to the presence of dormant mycobacteria. This also renders drug regimens ineffective and is the prime cause of the appearance of drug-resistant strains. In continuation of our efforts to develop novel antitubercular agents that especially target dormant mycobacteria, a set of 55 new compounds belonging to the pyrimidone class were designed on the basis of CoMFA and CoMSIA studies, and these were synthesized and subsequently tested against both the dormant and virulent BCG strain of M.tuberculosis. Some novel compounds have been identified which selectively inhibit the dormant tuberculosis bacilli with significantly low IC50 values. This study reports the second molecule after TMC-207, having the ability to inhibit tuberculosis bacilli exclusively in its dormant phase. The synthesis was accomplished by a modified multicomponent Biginelli reaction. A classification model was generated using the binary QSAR approach - recursive partitioning (RP) to identify structural characteristics related to the activity. Physicochemical, structural, topological, connectivity indices, and E-state key descriptors were used for generation of the decision tree. The decision tree could provide insights into structure-activity relationships that will guide the design of more potent inhibitors.

10aAntitubercular activity10adormant tuberculosis bacilli10apyrimidones10arecursive partitioning1 aJoshi, Rohit, R.1 aBarchha, Avinash1 aKhedkar, Vijay, M.1 aPissurlenkar, Raghuvir, R. S.1 aSarkar, Sampa1 aSarkar, Dhiman1 aJoshi, Rohini, R.1 aJoshi, Ramesh, A.1 aShah, Anamik, K.1 aCoutinho, Evans, C. uhttps://library.ncl.res.in/content/targeting-dormant-tuberculosis-bacilli-results-molecules-novel-pyrimidone-scaffold-002277nas a2200337 4500008004100000022001400041245011600055210006900171260000800240300001400248490000700262520120700269653000601476653000601482653002101488653001101509653003001520653002801550653003101578100001801609700001701627700002501644700002401669700001701693700002301710700002401733700002201757700002201779700001901801856011901820 2022 eng d a1381-199100aDetection of a target protein (GroEl2) in Mycobacterium tuberculosis using a derivative of 1,2,4-triazolethiols0 aDetection of a target protein GroEl2 in Mycobacterium tuberculos cOCT a2535-25480 v263 a

Herein, we identified a potent lead compound RRA2, within a series of 54 derivatives of 1,2,4-triazolethiols (exhibit good potency as an anti-mycobacterial agents) against intracellular Mycobacterium tuberculosis (Mtb). Compound RRA2 showed significant mycobactericidal activity against active stage Mycobacterium bovis BCG and Mtb with minimum inhibitory concentration (MIC) values of 2.3 and 2.0 mu g/mL, respectively. At MIC value, RRA2 compound yielded 0.82 log reduction of colony-forming unit (cfu) against non-replicating Mtb. Furthermore, RRA2 compound was selected for further target identification due to the presence of alkyne group, showing higher selectivity index (> 66.66 +/- 0.22, in non-replicating stage). Using ``click'' chemistry, we synthesized the biotin linker-RRA2 conjugate, purified with HPLC method and confirmed the conjugation of biotin linker-RRA2 complex by HR-MS analysis. Furthermore, we successfully pulled down and identified a specific target protein GroEl2, from Mtb whole-cell extract. Furthermore, computational molecular modeling indicated RRA2 could interact with GroEl2, which explains the structure-activity relationship observed in this study.

10a110a210a4-triazolethiols10aGroEl210aMycobactericidal activity10aMycobacterium bovis BCG10aMycobacterium tuberculosis1 aSarkar, Sampa1 aSwami, Sagar1 aSoni, Sarvesh, Kumar1 aHolien, Jessica, K.1 aKhan, Arshad1 aKorwar, Arvind, M.1 aLikhite, Anjali, P.1 aJoshi, Ramesh, A.1 aJoshi, Rohini, R.1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/detection-target-protein-groel2-mycobacterium-tuberculosis-using-derivative-124