02190nas a2200193 4500008004100000245010600041210006900147260000800216300001200224490000600236520148400242100002901726700002401755700001901779700002801798700001901826700002101845856013001866 2015 eng d00aLeucas mollissima, a source of bioactive compounds with antimalarial and antimycobacterium activities0 aLeucas mollissima a source of bioactive compounds with antimalar cJAN ae35-e380 v23 a

A phytochemical investigation of the acetone extract from the aerial parts of Leucas mollissima afforded one new (−)epi-marmelo lactone, (2 S, 4R, 6 S)-2,6-dimethyl-6 hydroxy-7-ene-4-olide (1), along with five known compounds, schensianol A (2), vanillin (3), β-hydroxy propiovanillone (4), lanost-9(11),25-diene-3β,24β-diol (5), and lanost-9(11),23E(24)-diene-3β,25-diol (6). Similarly, an investigation of the methanol extract of the aerial parts of L. mollissima resulted in the isolation of three known compounds, (+)-syringaresinol (7), anisofolin A (8), and apigenin 7-O-β-D(− 6′′-p-E-coumaroyl)-glucoside (9). Structure elucidation of the isolated compounds was carried out using detailed analysis of 1D and 2D nuclear magnetic resonance. All compounds were evaluated for antimalarial activity against Plasmodium falciparum (3D7) and for antimycobacterium activity against Mycobacterium tuberculosis H37Ra and Mycobacterium bovis. Compound 8 was found to have promising antimalarial activity (IC50 4.39 ± 0.25 µM), promising antimycobacterium activity [IC50 4.50 ± 0.75 µM (3.31 µg/mL)] against M. tuberculosis H37Ra and at 100 µg/mL, showed 55.6 % inhibition of M. bovis. Compound 9 showed moderate inhibition of P. falciparum growth (35 % inhibition at 10 µM) with respect to the positive control atovaquone and 67.4 % inhibition against M. bovis at 100 µg/mL with respect to the positive control rifampicin.

1 aChinchansure, Ashish, A.1 aArkile, Manisha, A.1 aShukla, Anurag1 aShanmugam, Dhanasekaran1 aSarkar, Dhiman1 aJoshi, Swati, P. uhttps://library.ncl.res.in/content/leucas-mollissima-source-bioactive-compounds-antimalarial-and-antimycobacterium-activities02124nas a2200157 4500008004100000245016700041210006900208260000800277300002500285490000600310520145000316100003101766700001901797700002501816856012501841 2018 eng d00aComprehensive metabolic and transcriptomic profiling of various tissues provide insights for saponin biosynthesis in the medicinally important Asparagus racemosus0 aComprehensive metabolic and transcriptomic profiling of various  cJUN aArticle Number: 90980 v83 aAsparagus racemosus (Shatavari), belongs to the family Asparagaceae and is known as a "curer of hundred diseases" since ancient time. This plant has been exploited as a food supplement to enhance immune system and regarded as a highly valued medicinal plant in Ayurvedic medicine system for the treatment of various ailments such as gastric ulcers, dyspepsia, cardiovascular diseases, neurodegenerative diseases, cancer, as a galactogogue and against several other diseases. In depth metabolic fingerprinting of various parts of the plant led to the identification of 13 monoterpenoids exclusively present in roots. LC-MS profiling led to the identification of a significant number of steroidal saponins (33). However, we have also identified 16 triterpene saponins for the first time in A. racemosus. In order to understand the molecular basis of biosynthesis of major components, transcriptome sequencing from three different tissues (root, leaf and fruit) was carried out. Functional annotation of A. racemosus transcriptome resulted in the identification of 153 transcripts involved in steroidal saponin biosynthesis, 45 transcripts in triterpene saponin biosynthesis, 44 transcripts in monoterpenoid biosynthesis and 79 transcripts in flavonoid biosynthesis. These findings will pave the way for better understanding of the molecular basis of steroidal saponin, triterpene saponin, monoterpenoids and flavonoid biosynthesis in A. racemosus.1 aSrivastava, Prabhakar, Lal1 aShukla, Anurag1 aKalunke, Raviraj, M. uhttps://library.ncl.res.in/content/comprehensive-metabolic-and-transcriptomic-profiling-various-tissues-provide-insights01008nas a2200157 4500008004100000245015600041210006900197260000800266520033300274100002500607700001900632700001900651700002800670700002100698856013100719 2018 eng d00aEfforts to access the potent antitrypanosomal marine natural product janadolide: synthesis of des- tert -butyl janadolide and its biological evaluation0 aEfforts to access the potent antitrypanosomal marine natural pro cFEB3 aTo identify novel antitrypanosomal agents based on Janadolide, a potent macrocyclic polyketide–peptide hybrid, a macrolactonization strategy was explored. We prepared des-tert-butyl Janadolide and evaluated its antitrypanosomal activity. Our findings suggest that the tert-butyl group is necessary for the desired bioactivity.1 aAthawale, Paresh, R.1 aJachak, Gorakh1 aShukla, Anurag1 aShanmugam, Dhanasekaran1 aReddy, Srinivasa uhttps://library.ncl.res.in/content/efforts-access-potent-antitrypanosomal-marine-natural-product-janadolide-synthesis-des-tert02706nas a2200265 4500008004100000022001500041245011400056210006900170260000800239300001200247490000700259520181200266100001902078700002602097700001902123700002402142700002102166700002002187700001402207700002302221700002002244700002002264700002802284856012802312 2018 eng d a 0020-751900a Glycolysis is important for optimal asexual growth and formation of mature tissue cysts by Toxoplasma gondii0 aGlycolysis is important for optimal asexual growth and formation cOCT a955-9680 v483 aToxoplasma gondii can grow and replicate using either glucose or glutamine as the major carbon source. Here, we have studied the essentiality of glycolysis in the tachyzoite and bradyzoite stages of T. gondii, using transgenic parasites that lack a functional hexokinase gene (Delta hk) in RH (Type-1) and Prugniaud (Type-II) strain parasites. Tachyzoite stage 411k parasites exhibit a fitness defect similar to that reported previously for the major glucose transporter mutant, and remain virulent in mice. However, although Prugniaud strain Delta hk tachyzoites were capable of transforming into bradyzoites in vitro, they were severely compromised in their ability to make mature bradyzoite cysts in the brain tissue of mice. Isotopic labelling studies reveal that glucose-deprived tacyzoites utilise glutamine to replenish glycolytic and pentose phosphate pathway intermediates via gluconeogenesis. Interestingly, while glutaminedeprived intracellular Delta hk tachyzoites continued to replicate, extracellular parasites were unable to efficiently invade host cells. Further, studies on mutant tachyzoites lacking a functional phosphoenolpyruvate carboxykinase (Delta pepck1) revealed that glutaminolysis is the sole source of gluconeogenic flux in glucose-deprived parasites. In addition, glutaminolysis is essential for sustaining oxidative phosphorylation in zihk parasites, while wild type (wt) and Delta pepck1 parasites can obtain ATP from either glycolysis or oxidative phosphorylation. This study provides insights into the role of nutrient metabolism during asexual propagation and development of T. gondii, and validates the versatile nature of central carbon and energy metabolism in this parasite. (C) 2018 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.1 aShukla, Anurag1 aOlszewski, Kellen, L.1 aLlinas, Manuel1 aRommereim, Leah, M.1 aFox, Barbara, A.1 aBzik, David, J.1 aXia, Dong1 aWastling, Jonathan1 aBeiting, Daniel1 aRoos, David, S.1 aShanmugam, Dhanasekaran uhttps://library.ncl.res.in/content/glycolysis-important-optimal-asexual-growth-and-formation-mature-tissue-cysts-toxoplasma02881nas a2200277 4500008004100000022001300041245016600054210006900220260000800289300001500297490000600312520188200318100002502200700002702225700001902252700001902271700001702290700002202307700002502329700002302354700002102377700002102398700002802419700002902447856012702476 2018 eng d a2379504200aTargeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules0 aTargeted phenotypic screening in plasmodium falciparum and toxop cJAN a e00534-170 v33 aThe Malaria Box collection includes 400 chemically diverse small molecules with documented potency against malaria parasite growth, but the underlying modes of action are largely unknown. Using complementary phenotypic screens against Plasmodium falciparum and Toxoplasma gondii, we report phenotype-specific hits based on inhibition of overall parasite growth, apicoplast segregation, and egress or host invasion, providing hitherto unavailable insights into the possible mechanisms affected. First, the Malaria Box library was screened against tachyzoite stage T. gondii and the half-maximal effective concentrations (EC50s) of molecules showing ≥80% growth inhibition at 10 µM were determined. Comparison of the EC50s for T. gondii and P. falciparum identified a subset of 24 molecules with nanomolar potency against both parasites. Thirty molecules that failed to induce acute growth inhibition in T. gondii tachyzoites in a 2-day assay caused delayed parasite death upon extended exposure, with at least three molecules interfering with apicoplast segregation during daughter cell formation. Using flow cytometry and microscopy-based examinations, we prioritized 26 molecules with the potential to inhibit host cell egress/invasion during asexual developmental stages of P. falciparum. None of the inhibitors affected digestive vacuole integrity, ruling out a mechanism mediated by broadly specific protease inhibitor activity. Interestingly, five of the plasmodial egress inhibitors inhibited ionophore-induced egress of T. gondii tachyzoites. These findings highlight the advantage of comparative and targeted phenotypic screens in related species as a means to identify lead molecules with a conserved mode of action. Further work on target identification and mechanism analysis will facilitate the development of antiparasitic compounds with cross-species efficacy.1 aSubramanian, Gowtham1 aBelekar, Meenakshi, A.1 aShukla, Anurag1 aTong, Jie, Xin1 aSinha, Ameya1 aChu, Trang, T. T.1 aKulkarni, Akshay, S.1 aPreiser, Peter, R.1 aReddy, Srinivasa1 aTan, Kevin, S. W1 aShanmugam, Dhanasekaran1 aChandramohanadas, Rajesh uhttps://library.ncl.res.in/content/targeted-phenotypic-screening-plasmodium-falciparum-and-toxoplasma-gondii-reveals-novel03253nas a2200301 4500008004100000022001400041245015900055210006900214260000800283300001100291490000800302520224400310653002002554653000902574653001402583653000802597653001702605653001402622100002702636700002302663700002102686700001902707700002502726700002402751700002802775700002202803856012602825 2019 eng d a0014-489400aMitochondrial cytochrome oxidase C subunit III (cox3) gene as a sensitive and specific target for molecular detection of Babesia gibsoni infection in dogs0 aMitochondrial cytochrome oxidase C subunit III cox3 gene as a se cNOV a1077710 v2063 a

A PCR targeting mitochondrial cytochrome oxidase subunit III (cox3) for molecular detection of Babesia gibsoni infection in dogs has been developed in this study. Fifty blood samples from suspected clinical cases from dogs, brought to the veterinary college clinics, were examined for presence of B. gibsoni using conventional diagnosis by microscopic examination of Giemsa stained thin blood smears. In addition, species specific PCRs targeting ITS-1 region (BgITS-1 PCR) and nested PCR targeting 18S ribosomal RNA gene (Bg18SnPCR) were carried out. A 634 bp PCR fragment of B. gibsoni cox3 gene was amplified in positive samples from three geographical locations of Satara, Wai and Pune in Maharashtra state of India. From analysis of the sequence of the B. gibsoni cox3 gene, we found that the Indian isolate had 96-98% similarity to the isolate from Japan and China. Post sequencing, de-novo diagnostic primer pair for species specific amplification of 164 bp fragment of B. gibsoni cox3 was designed and the PCR was standardized. The diagnostic results of de-novo Bgcox3 PCR were compared with BgITS-1 PCR and Bg18S nPCR. Thin blood smears detected 22% (11/50) samples positive for small form of Babesia species. The BgITS-1 PCR detected 25% samples (15/50) as positive and Bg18S nPCR detected 80% (40/50) B. gibsoni positive samples. The de-novo Bgcox3 PCR detected 66% (33/50) samples positive for B. gibsoni (at 95% CI). The analytical sensitivity of cox3 PCR was evaluated as 0.000003% parasitaemia or 09 parasites in 100 mu l of blood. The de-novo diagnostic cox3 PCR did not cross react with control positive DNA from other haemoprotozoa and rickettsia like B. vogeli, Hepatozoon canis, Trypanosoma evansi, Ehrlichia cants and Anaplasma platys. Statistically, cox3 PCR had better diagnostic efficiency than ITS-1 PCR in terms of sensitivity (p = 0.0006). No statistically significant difference between results of cox3 PCR and 18S nPCR was observed (p = 0.1760). Kappa values estimated for each test pair showed fair to moderate agreement between the observations. Specificity of Bgcox3 PCR was 100% when compared with microscopy or BgITS-1 PCR. Sensitivity of Bgcox3 PCR was 100% when compared with that of Bg18S nPCR.

10aBabesia gibsoni10acox310aDiagnosis10aDog10amitochondria10amolecular1 aBetgiri, Aishwarya, A.1 aJadhav, Sameer, N.1 aPawde, Mrunalini1 aShukla, Anurag1 aMote, Chandrashekhar1 aPawar, Prashant, D.1 aShanmugam, Dhanasekaran1 aKundu, Krishnendu uhttps://library.ncl.res.in/content/mitochondrial-cytochrome-oxidase-c-subunit-iii-cox3-gene-sensitive-and-specific-target01015nas a2200265 4500008004100000022001400041245019100055210006900246260000800315300001400323490000600337100002500343700002700368700001900395700001900414700001700433700002200450700002500472700002300497700002100520700002200541700002800563700002900591856012900620 2019 eng d a2379-504200aTargeted phenotypic screening in plasmodium falciparum and toxoplasma gondii reveals novel modes of action of medicines for malaria venture malaria box molecules (vol 3, e00534-17, 2018)0 aTargeted phenotypic screening in plasmodium falciparum and toxop cAPR ae00159-190 v41 aSubramanian, Gowtham1 aBelekar, Meenakshi, A.1 aShukla, Anurag1 aTong, Jie, Xin1 aSinha, Ameya1 aChu, Trang, T. T.1 aKulkarni, Akshay, S.1 aPreiser, Peter, R.1 aReddy, Srinivasa1 aTan, Kevin, S. W.1 aShanmugam, Dhanasekaran1 aChandramohanadas, Rajesh uhttps://library.ncl.res.in/content/targeted-phenotypic-screening-plasmodium-falciparum-and-toxoplasma-gondii-reveals-novel-002560nas a2200313 4500008004100000022001400041245010100055210006900156260000800225300001100233490000800244520153800252653002601790653001201816653001201828653003301840653001401873653002801887653001701915653002901932653002001961100002001981700002502001700001902026700001802045700003302063700002002096856013002116 2020 eng d a0031-942200aBiosynthesis and tissue-specific partitioning of camphor and eugenol in Ocimum kilimandscharicum0 aBiosynthesis and tissuespecific partitioning of camphor and euge cSEP a1124510 v1773 a

In Ocimum kilimandscharicum, the relative volatile composition of camphor in leaves was as high as 55%, while that of eugenol in roots was 57%. These metabolites were differentially partitioned between the aerial and root tissues. Global metabolomics revealed tissue-specific biochemical specialization, evident by the differential distribution of 2588 putative metabolites across nine tissues. Next-generation sequencing analysis indicated differential expression of 51 phenylpropanoid and 55 terpenoid pathway genes in aerial and root tissues. By integrating metabolomics with transcriptomics, the camphor biosynthesis pathway in O. kilimandscharicum was elucidated. In planta bioassays revealed the role of geranyl diphosphate synthase (gpps) and borneol dehydrogenase (bdh) in camphor biosynthesis. Further, the partitioning of camphor was attributed to tissue-specific gene expression of both the pathway entry point (gpps) and terminal (bdh) enzyme. Unlike camphor, eugenol accumulated more in roots; however, absence of the eugenol synthase gene in roots indicated long distance transport from aerial tissues. In silico co-expression analysis indicated the potential involvement of ATP-binding cassette, multidrug and toxic compound extrusion, and sugar transporters in eugenol transport. Similar partitioning was evident across five other Ocimum species. Overall, our work indicates that metabolite partitioning maybe a finely regulated process, which may have implications on plant growth, development, and defense.

10aBorneol dehydrogenase10aCamphor10aEugenol10aGeranyl diphosphate synthase10aLamiaceae10aMetabolite partitioning10aMetabolomics10aOcimum kilimandscharicum10aTranscriptomics1 aSingh, Priyanka1 aKalunke, Raviraj, M.1 aShukla, Anurag1 aTzfadia, Oren1 aThulasiram, Hirekodathakallu1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/biosynthesis-and-tissue-specific-partitioning-camphor-and-eugenol-ocimum-kilimandscharicum01926nas a2200253 4500008004100000022001400041245011500055210006900170260000800239300000900247490000700256520106700263100001801330700001901348700002301367700002201390700002301412700001901435700001801454700001901472700002801491700002401519856012901543 2020 eng d a2041-172300aDistinct metabolic states of a cell guide alternate fates of mutational buffering through altered proteostasis0 aDistinct metabolic states of a cell guide alternate fates of mut cJUN a29260 v113 a

Metabolic changes alter the cellular milieu; can this also change intracellular protein folding? Since proteostasis can modulate mutational buffering, if change in metabolism has the ability to change protein folding, arguably, it should also alter mutational buffering. Here we find that altered cellular metabolic states in E. coli buffer distinct mutations on model proteins. Buffered-mutants have folding problems in vivo and are differently chaperoned in different metabolic states. Notably, this assistance is dependent upon the metabolites and not on the increase in canonical chaperone machineries. Being able to reconstitute the folding assistance afforded by metabolites in vitro, we propose that changes in metabolite concentrations have the potential to alter protein folding capacity. Collectively, we unravel that the metabolite pools are bona fide members of proteostasis and aid in mutational buffering. Given the plasticity in cellular metabolism, we posit that metabolic alterations may play an important role in cellular proteostasis.

1 aVerma, Kanika1 aSaxena, Kanika1 aDonaka, Rajashekar1 aChaphalkar, Aseem1 aRai, Manish, Kumar1 aShukla, Anurag1 aZaidi, Zainab1 aDandage, Rohan1 aShanmugam, Dhanasekaran1 aChakraborty, Kausik uhttps://library.ncl.res.in/content/distinct-metabolic-states-cell-guide-alternate-fates-mutational-buffering-through-altered01129nas a2200241 4500008004100000022001400041245007400055210006900129260000900198300001400207490000900221520036100230653001800591653001500609653002300624653001600647100002700663700002300690700001900713700002800732700002100760856010600781 2021 eng d a1434-193X00aSynthesis and biological evaluation of hoshionolactam-based compounds0 aSynthesis and biological evaluation of hoshionolactambased compo cAPR  a2212-22180 v20213 aIn search of novel antitrypanosomal agents based on hoshinolactam (IC50=3.9 nM), we disclose the synthesis and biological evaluations of 14 different analogues of the natural product using combinations of different acids and lactams. Antitrypanosomal activity assays revealed that the synthesized analogues were less potent than the parent natural product.10aHoshinolactam10aOzonolysis10aParasitic diseases10aTrypanosoma1 aJachak, Gorakhnath, R.1 aElizebath, Drishya1 aShukla, Anurag1 aShanmugam, Dhanasekaran1 aReddy, Srinivasa uhttps://library.ncl.res.in/content/synthesis-and-biological-evaluation-hoshionolactam-based-compounds02296nas a2200241 4500008004100000022001400041245011200055210006900167260000800236300001100244490000800255520148600263653001801749653002401767653001201791653001501803653001501818100002501833700001901858700002801877700001701905856013201922 2022 eng d a0166-685100aProfiling of metabolic alterations in mice infected with malaria parasites via high-resolution metabolomics0 aProfiling of metabolic alterations in mice infected with malaria cNOV a1115250 v2523 a

Background: Malaria infection can result in distinct clinical outcomes from asymptomatic to severe. The association between patho-physiological changes and molecular changes in the host, and their correlation with severity of malaria progression is not fully understood. Methods: In this study, we addressed mass spectrometry-based temporal profiling of serum metabolite levels from mice infected with Plasmodium berhgei (strain ANKA). Results: We show global perturbations and identify changes in specific metabolites in correlation with disease progression. While metabolome-wide changes were apparent in late-stage malaria, a subset of metabolites exhibited highly correlated changes with disease progression. These metabolites changed early on following infection and either continued or maintained the change as mice developed severe disease. Some of these have the potential to be sentinel metabolites for severe malaria. Moreover, glycolytic metabolites, purine nucleotide precursors, tryptophan and its bioactive derivatives were many fold decreased in late-stage disease. Interestingly, uric acid, a metabolic waste reported to be elevated in severe human malaria, increased with disease progression, and subsequently appears to be detoxified into allantoin. This detoxification mechanism is absent in humans as they lack the enzyme uricase. Conclusions: We have identified candidate marker metabolites that may be of relevance in the context of human malaria.

10aHost response10aInfectious diseases10amalaria10ametabolite10aPlasmodium1 aChhibber-Goel, Jyoti1 aShukla, Anurag1 aShanmugam, Dhanasekaran1 aSharma, Amit uhttps://library.ncl.res.in/content/profiling-metabolic-alterations-mice-infected-malaria-parasites-high-resolution-metabolomics