01700nas a2200169 4500008004100000022001400041245014200055210006900197260000900266300000900275490000600284520105700290100002401347700001901371700001901390856012101409 2019 eng d a2045-232200aEvidence of nitrite acting as a stable and robust inducer of non-cultivability in Mycobacterium tuberculosis with physiological relevance0 aEvidence of nitrite acting as a stable and robust inducer of non cJUN  a92610 v93 a

Tuberculosis (TB) is the ninth leading cause of death worldwide, ranking above human immunodeficiency virus. Latency is the major obstacle in the eradication of this disease. How the physiology of the pathogen changes in transition to the latent stage needs to be understood. The latent bacteria extracted from animal hosts exist in a nonculturable (NC) phase, whereas bacteria extracted from most in vitro models are culture-positive. In the present study, we observed that nitrite, up to a concentration of 5 mM, shows the growth of Mycobacterium tuberculosis (MTB) in liquid media, but this effect starts reversing at higher concentrations. At a concentration of 10 mM, nitrite induces rapid nonculturability of MTB at the aerobic stage. This noncultivable dormancy was confirmed by analyzing the characteristics of NC bacteria. Further differential gene expression analyses clearly supported the formation of a dormancy phenotype. This study will be helpful for the use of this bacillus as a dormancy model in future studies on TB latency.

1 aGample, Suwarna, P.1 aAgrawal, Sonia1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/evidence-nitrite-acting-stable-and-robust-inducer-non-cultivability-mycobacterium01839nas a2200145 4500008004100000245013400041210006900175260000800244490000800252520125400260100001701514700001901531700001901550856012401569 2019 eng d00aHigh content screening assay for identifying inhibitors against active and dormant state intracellular Mycobacterium tuberculosis0 aHigh content screening assay for identifying inhibitors against  cSEP0 v1643 a

The antitubercular drug development pipeline could start with an in vitro investigation of several compounds to examine their effect on active and dormant state Mycobacterium tuberculosis (Mtb). However, in vitro screening of dormant state bacilli cannot provide enough information on the simultaneous effect of a compound on the host. Therefore, we developed a live cell fluorescence based screening protocol by utilizing the high content system for determining the effect of inhibitors against active and dormant state intracellular mycobacteria. THP-1 macrophages infected with an actively growing and hypoxia derived dormant Mtb culture were standardized to develop the screening protocol. The signal to noise ratio and the Z' factor of this assay were found to be 7.5-29 and 0.6-0.8, respectively, which confirm the robustness of the protocol. The protocol was then validated with standard inhibitors. This newly developed drug screening assay offers an easy, safe, image based high content screening tool to search for novel antitubercular inhibitors against both active and dormant state intracellular mycobacteria. Therefore, this assay could fill in the gap between in vitro and in vivo latent tuberculosis drug screening programs.

1 aYeware, Amar1 aAgrawal, Sonia1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/high-content-screening-assay-identifying-inhibitors-against-active-and-dormant-state02012nas a2200181 4500008004100000022001400041245013800055210006900193260000800262300001300270490000700283520133400290100001701624700002001641700001901661700001901680856013101699 2019 eng d a1932-620300aUsing diphenyleneiodonium to induce a viable but non-culturable phenotype in Mycobacterium tuberculosis and its metabolomics analysis0 aUsing diphenyleneiodonium to induce a viable but nonculturable p cAUG ae02206280 v143 a

Depletion of oxygen levels is a well-accepted model for induction of non-replicating, persistent states in mycobacteria. Increasing the stress levels in mycobacterium bacilli facilitates their entry into a non-cultivable, dormant state. In this study, it was shown that diphenyleneiodonium, an inhibitor of NADH oxidase, induced a viable, but non-culturable state in mycobacteria, having similar features to dormant bacilli, like loss of acid-fastness, upregulation of stress-regulated genes and decreased superoxide levels as compared to actively growing bacilli. Comprehensive, untargeted metabolic profiling also confirmed a decrease in biogenesis of amino acids, NAD, unsaturated fatty acids and nucleotides. Additionally, an increase in the level of lactate, fumarate, succinate and pentose phosphate pathways along with increased mycothiol and sulfate metabolites, similar to dormant bacilli, was observed in the granuloma. These non-cultivable bacilli were resuscitated by supplementation of fetal bovine serum, regaining their culturability in liquid as well as on agar medium. This study focused on the effect of diphenyleneiodonium treatment in causing mycobacteria to rapidly transition from an active state into a viable, but non-cultivable state, and comparing their characteristics with dormant phenotypes.

1 aYeware, Amar1 aGample, Suwarna1 aAgrawal, Sonia1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/using-diphenyleneiodonium-induce-viable-non-culturable-phenotype-mycobacterium-tuberculosis02851nas a2200277 4500008004100000022001400041245013700055210006900192260000800261300001100269490000700280520193000287653002202217653001002239653001502249653002202264653001502286653001002301100002402311700002502335700001902360700002202379700001902401700002202420856013102442 2020 eng d a0946-672X00aAnticancer potential of AgNPs synthesized using acinetobacter sp. and curcuma aromatica against HeLa cell lines: a comparative study0 aAnticancer potential of AgNPs synthesized using acinetobacter sp cDEC a1266300 v623 a

Background: Biogenic nanoparticles are gaining attention due to their low toxicity and numerous biomedical applications. Present study aimed to compare the potential anticancer activity of two biogenic silver nanoparticles (bAgNPs and pAgNPs) against human cervical cancer cell lines (HeLa). Methods: bAgNPs were synthesized using Acinetobacter sp. whereas pAgNPs were synthesized using aqueous root extract of Curcuma aromatica. Effect of these nanoparticles on HeLa cells viability was studied using MTT assay and colony formation assay. Anticancer potential was determined using fluorescence microscopy and flow cytometry studies. Bio-compatibility studies were performed against peripheral blood mononuclear cells (PBMCs). Results: Both the nanoparticles showed 50 % viability of peripheral blood mononuclear cells (PBMCs) when used at high concentration (200 mu g/mL). IC50 for bAgNPs and pAgNPs against HeLa cells were 17.4 and 14 mu g/mL respectively. Colony formation ability of Hela cells was reduced on treatment with both nanoparticles. Acridine orange and ethidium bromide staining demonstrated that bAgNPs were cytostatic whereas pAgNPs were apoptotic. JC-1 dye staining revealed that the mitochondrial membrane potential was affected on treatment with pAgNPs while it remained unchanged on bAgNPs treatment. Flow cytometry confirmed cell cycle arrest in HeLa cells on treatment with nanoparticles further leading to apoptosis in case of pAgNPs. About 77 and 58 % HeLa cells were found in subG1 phase on treatment with bAgNPs and pAgNPs respectively. bAgNPs showed cytostatic effect on HeLa cells arresting the cell growth in subG1 phase, whereas, pAgNPs triggered death of HeLa cells through mitochondrial membrane potential impairment and apoptosis. Conclusion: Overall, bAgNPs and pAgNPs could be safe and showed potential to be used as anticancer nanoantibiotics against human cervical cancer cells.

10aAcinetobacter sp.10aAgNPs10aAnticancer10aCurcuma aromatica10aHeLa cells10aPBMCs1 aNadhe, Shradhda, B.1 aTawre, Madhumita, S.1 aAgrawal, Sonia1 aChopade, Balu, A.1 aSarkar, Dhiman1 aPardesi, Karishma uhttps://library.ncl.res.in/content/anticancer-potential-agnps-synthesized-using-acinetobacter-sp-and-curcuma-aromatica-against01959nas a2200181 4500008004100000022001400041245012100055210006900176260000900245300001000254490000700264520130300271100001901574700002001593700001701613700001901630856012801649 2021 eng d a2045-232200aNovel gene similar to nitrite reductase (NO forming) plays potentially important role in the latency of tuberculosis0 aNovel gene similar to nitrite reductase NO forming plays potenti cOCT  a198130 v113 aThe development of the latent phenotype of Mycobacterium tuberculosis (Mtb) in the human lungs is the major hurdle to eradicate Tuberculosis. We recently reported that exposure to nitrite (10 mM) for six days under in vitro aerobic conditions completely transforms the bacilli into a viable but non-cultivable phenotype. Herein, we show that nitrite (beyond 5 mM) treated Mtb produces nitric oxide (NO) within the cell in a dose-dependent manner. Our search for the conserved sequence of NO synthesizing enzyme in the bacterial system identified MRA2164 and MRA0854 genes, of which the former was found to be significantly up regulated after nitrite exposure. In addition, the purified recombinant MRA2164 protein shows significant nitrite dependent NO synthesizing activity. The knockdown of the MRA2164 gene at mRNA level expression resulted in a significantly reduced NO level compared to the wild type bacilli with a simultaneous return of its replicative capability. Therefore, this study first time reports that nitrite induces dormancy in Mtb cells through induced expression of the MRA2164 gene and productions of NO as a mechanism for maintaining non-replicative stage in Mtb. This observation could help to control the Tuberculosis disease, especially the latent phenotype of the bacilli.1 aAgrawal, Sonia1 aGample, Suwarna1 aYeware, Amar1 aSarkar, Dhiman uhttps://library.ncl.res.in/content/novel-gene-similar-nitrite-reductase-no-forming-plays-potentially-important-role-latency02593nas a2200229 4500008004100000022001400041245012400055210006900179260000900248300001600257490000700273520179100280100002002071700001602091700001902107700002302126700003002149700001602179700001902195700001902214856013002233 2022 eng d a1477-922600aDNA binding, antitubercular, antibacterial and anticancer studies of newly designed piano-stool ruthenium(ii) complexes0 aDNA binding antitubercular antibacterial and anticancer studies  cNOV  a16371-163820 v513 a

The chemotherapeutic potential of ruthenium(ii) complexes has recently attracted researchers' interest as antibacterial and anticancer agents. In this study, two novel half-sandwich imine-based Ru complexes ([Ru(p-cymene)Cl(L-1)][PF6] (Ru-1) and [Ru(p-cymene)Cl(L-2)][PF6] (Ru-2)) were reported for their deoxyribonucleic acid (DNA) binding and antitubercular, antibacterial, and anticancer activities. The molecular structure of Ru-2 was obtained by single-crystal X-ray crystallography. DNA interaction studies were conducted by UV-Vis absorbance and fluorescence spectral titration which gave rise to DNA binding constants (K-b) of 1.32 x 10(6) and 1.82 x 10(6) for Ru-1 and Ru-2, respectively and the Stern-Volmer binding constant (K-SV) values for Ru-1 and Ru-2 were 1.7763 x 10(4) M-1 and 7.6 x 10(3) M-1, respectively. The in vitro antitubercular activity was evaluated against Mycobacterium tuberculosis H37Ra. The antibacterial potential of both the Ru-complexes was examined against Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacteria. The half-maximal inhibitory concentration (IC50) values for the antitubercular activity of Ru-1 and Ru-2 were 4.87 +/- 1.32 mu M and 5.78 +/- 0.54 mu M, respectively. A cytotoxic study of these complexes was performed against the human breast cancer cell line (MCF-7) and the human embryonic kidney cell line (HEK293) (normal cells). The study revealed meaningful activity of the Ru-1 complex against (cancer) MCF-7 cells, while the viability of HEK293 (normal) cells in the presence of Ru-2 was higher as compared to a reference drug 5FU. We suggest that these kinds of Ru-complexes could have potential for application in metallopharmaceuticals.

1 aNavale, Govinda1 aSingh, Sain1 aAgrawal, Sonia1 aGhosh, Chandrachur1 aChoudhury, Angshuman, Roy1 aRoy, Partha1 aSarkar, Dhiman1 aGhosh, Kaushik uhttps://library.ncl.res.in/content/dna-binding-antitubercular-antibacterial-and-anticancer-studies-newly-designed-piano-stool03072nas a2200313 4500008004100000022001400041245014000055210006900195260000900264300001100273490000800284520201300292653001602305653002802321653002002349653001902369653002202388653002402410100001602434700002402450700001902474700002502493700002002518700001902538700002202557700002902579700001902608856013102627 2023 eng d a0141-813000aDesign and synthesis of piano-stool ruthenium(II) complexes and their studies on the inhibition of amyloid ? (1-42) peptide aggregation0 aDesign and synthesis of pianostool rutheniumII complexes and the cJUN  a1241970 v2393 a

Misfolding , protein aggregation have been linked to numerous human neurodegenerative disorders such as Alzheimer's, prion , Parkinson's diseases. Ruthenium (Ru) complexes have received considerable attention in studying protein aggregation due to their interesting photophysical and photo properties. In this study, we have synthesized the novel Ru complexes ([Ru(p-cymene)Cl(L-1)][PF6](Ru-1), and [Ru(p-cymene)Cl(L-2)][PF6](Ru-2)) and investigated their inhibitory activity against the bovine serum albumin (BSA) aggregation and the A beta 1-42 peptides amyloid formation. Several spectroscopic methods were used to characterize these complexes, and the molecular structure of the complex was determined by X-ray crystallography. Amyloid aggregation and inhi-bition activities were examined using the Thioflavin-T (ThT) assay, and the secondary structures of the protein were analyzed by circular dichroism (CD) spectroscopy and transmission electron microscopy (TEM). The cell viability assay was carried out on the neuroblastoma cell line, revealing that the complex Ru-2 showed better protective effects against A beta 1-42 peptide toxicity on neuro-2a cells than the complex Ru-1. Molecular docking studies elucidate the binding sites and interactions between the Ru-complexes and A beta 1-42 peptides. The exper-imental studies revealed that these complexes significantly inhibited the BSA aggregation and A beta 1-42 amyloid fibril formation at 1:3 and 1:1 molar concentrations, respectively. Antioxidant assays demonstrated that these complexes act as antioxidants, protecting from amyloid-induced oxidative stress. Molecular docking studies with the monomeric A beta 1-42 (PDB: 1IYT) show hydrophobic interaction, and both complexes bind preferably in the central region of the peptide and coordinate with two binding sites of the peptide. Hence, we suggest that the Ru-based complexes could be applied as a potential agent in metallopharmaceutical research against Alzheimer's disease.

10aAggregation10aAmyloid ?(1-42) peptide10aDFT computation10aFibrillization10aMolecular docking10aRuthenium complexes1 aSingh, Sain1 aNavale, Govinda, R.1 aAgrawal, Sonia1 aSingh, Haobam, Kisan1 aSingla, Labhini1 aSarkar, Dhiman1 aSarma, Manabendra1 aChoudhury, Anghuman, Roy1 aGhosh, Kaushik uhttps://library.ncl.res.in/content/design-and-synthesis-piano-stool-rutheniumii-complexes-and-their-studies-inhibition-amyloid02379nas a2200277 4500008004100000022001400041245010700055210007000162260000900232300001100241490000800252520144600260653002501706653003801731653002201769100001701791700002401808700001601832700002501848700001901873700001901892700001901911700002201930700001901952856013001971 2023 eng d a0141-813000aInhibition of amyloid β1-42 peptide aggregation by newly designed cyclometallated palladium complexes0 aInhibition of amyloid β142 peptide aggregation by newly designed cSEP  a1258470 v2483 a

Uncontrolled amyloid aggregation is a frequent cause of neurodegenerative disorders such as prions and Alzheimer's disease (AD). As a result, many drug development approaches focus on evaluating novel molecules that can alter self-recognition pathways. Herein, we designed and synthesized the cyclometallated pyrene (Pd-1 and Pd-3) and anthracene (Pd-2) based palladium complexes ([Pd((L-1)Cl] Pd-1, [Pd(L-2)Cl](Pd-2), and [Pd(L-3)Cl] (Pd-3)). This study explores the effect of these complexes on the aggregation, fibrillation, and amyloid formation of bovine serum albumin (BSA) and A beta(1-42) peptide. Several spectroscopic methods were used to characterize all the Pd-complexes, and the molecular structure of Pd -3 was determined by X-ray crystallography. The secondary structures were studied using circular dichroism (CD) and transmission electron microscopy (TEM), while am-yloid aggregation and inhibitory activities were investigated using the Thioflavin-T (ThT) fluorescence assay. Molecular docking of the Pd-complex (Pd-3) was done using fibril (PDB: 2BEG) and monomeric (PDB: 1IYT) peptides using Auto-dock Vina. As a result, the hydrogen bonding and hydrophobic interaction between the aromatic rings of the Pd-complexes and the amino acids of amyloid-beta peptides significantly reduced the pro-duction of ordered beta-sheets of amyloid fibrils and protein aggregation in the presence of Pd-2 and Pd-3 complexes.

10aA beta(1-42) peptide10aAggregation and molecular docking10aPalladium complex1 aSaini, Rahul1 aNavale, Govinda, R.1 aSingh, Sain1 aSingh, Haobam, Kisan1 aChauhan, Rahul1 aAgrawal, Sonia1 aSarkar, Dhiman1 aSarma, Manabendra1 aGhosh, Kaushik uhttps://library.ncl.res.in/content/inhibition-amyloid-%CE%B21-42-peptide-aggregation-newly-designed-cyclometallated-palladium02240nas a2200205 4500008004100000022001400041245015900055210006900214260000800283300001400291490000700305520147500312100001601787700001801803700001901821700001901840700003001859700002601889856011901915 2023 eng d a1477-052000aTotal synthesis, structure elucidation and expanded bioactivity of icosalide A: effect of lipophilicity and ester to amide substitution on its bioactivity0 aTotal synthesis structure elucidation and expanded bioactivity o cJUL a5725-57310 v213 a

The first total synthesis of icosalide A, an antibacterial depsipeptide that is unique in that it contains two lipophilic beta-hydroxy acids, has been achieved by following Fmoc solid-phase peptide synthesis in combination with solution-phase synthesis. The ambiguity in the absolute stereochemistry of icosalide A has been resolved by synthesizing the reported structures and other relevant diastereomers of icosalides and comparing their NMR data. NMR-based structure elucidation of icosalide A revealed a well-folded structure with cross-strand hydrogen bonds similar to the anti-parallel beta-sheet conformation in peptides and displayed a synergistic juxtaposition of the aliphatic sidechains. 12 analogues of icosalide A were synthesized by varying the constituent lipophilic beta-hydroxy acid residues, and their biological activities against Bacillus thuringiensis and Paenibacillus dendritiformis were explored. Most of these icosalide analogues showed an MIC of 12.5 & mu;g mL(-1) against both bacteria. Swarming inhibition by icosalides was least in B. thuringiensis (8.3%) compared to that in P. dendritiformis (33%). Furthermore, this is the first report of icosalides showing assured inhibitory action (MIC between 2 and 10 & mu;g mL(-1)) against the active stage of Mycobacterium tuberculosis and cancer cell lines such as HeLa and ThP1. This study could help optimize icosalides for anti-TB, antibacterial, and anti-cancer activities.

1 aDangi, Abha1 aPande, Bharat1 aAgrawal, Sonia1 aSarkar, Dhiman1 aVamkudoth, Koteswara, Rao1 aMarelli, Udaya, Kiran uhttps://library.ncl.res.in/content/total-synthesis-structure-elucidation-and-expanded-bioactivity-icosalide-effect