00757nas a2200181 4500008004100000022001400041245011100055210006900166260010300235300000800338490000600346100002500352700001800377700002300395700001500418700001200433856013000445 2006 eng d a1535-947600aIntact cell MALDI mass spectrometry as a tool to screen drugs in vivo for regulation of protein expression0 aIntact cell MALDI mass spectrometry as a tool to screen drugs in a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcOCT aS620 v51 aKulkarni, Mahesh, J.1 aVinod, V., P.1 aUmashankar, P., K.1 aPatole, M.1 aRao, M. uhttps://library.ncl.res.in/content/intact-cell-maldi-mass-spectrometry-tool-screen-drugs-vivo-regulation-protein-expression-002113nas a2200253 4500008004100000022001400041245010900055210006900164260006100233300001200294490000700306520124400313653001101557653001101568653002301579653001401602653001401616100001901630700002501649700002401674700002101698700001801719856012201737 2007 eng d a0959-399300aAttempt at taxonomical characterization of some Rhizobial species by intact cell MALDI mass spectrometry0 aAttempt at taxonomical characterization of some Rhizobial specie a233 SPRING STREET, NEW YORK, NY 10013 USAbSPRINGERcFEB a177-1850 v233 a

In the present study an attempt was made to exploit the benefit of intact cell MALDI mass spectrometry (ICM-MS) in bringing out similarities and differences among some Rhizobium species and a species of Agrobacterium based on specific mass:charge (m/z) values. Rhizobium species isolated from the root nodules of selected leguminous plants were analysed by ICM-MS. The spectra were acquired in the range of 500-10,000 Da yielding several peaks specific to each species. The peaks obtained corresponded to the respective bacterial cell surface molecules which were desorbed during matrix-assisted laser desorption ionization. The number of peaks were more in the range of 500-1200 Da. Dice similarity coefficient analysis of m/z values indicated that Rhizobium species isolated from Trigonella foenum-graecum and Pisum arvense showed more similarity than any other species. Agrobacterium species did show a few common m/z values in comparison with other Rhizobium species. This clearly shows that Agrobacterium is closely related to Rhizobium. Eventually, ICM-MS technique offers clear, distinct, and consistent results for replicates, in less than an hour's time, therefore this technique has high potential in molecular taxonomy.

10aICM-MS10alegume10amolecular taxonomy10aRhizobium10asymbiotic1 aPrabhu, R., R.1 aKulkarni, Mahesh, J.1 aParasharami, V., A.1 aSanthakumari, B.1 aParanjape, S. uhttps://library.ncl.res.in/content/attempt-taxonomical-characterization-some-rhizobial-species-intact-cell-maldi-mass01963nas a2200229 4500008004100000022001400041245014700055210006900202260007300271300001200344490000800356520107500364653001201439653002501451653002201476100001901498700002001517700001901537700002501556700002201581856013001603 2007 eng d a0168-165600aCharacterization of the proteins of bacterial strain isolated from contaminated site involved in heavy metal resistance - a proteomic approach0 aCharacterization of the proteins of bacterial strain isolated fr aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcFEB a444-4510 v1283 a

The present study describes response of a bacterial strain isolated from a polluted river to heavy metal toxicity. The bacterium was identified to be Klebsiella pneumoniae by biochemical tests using API 20E strips and 16S ribotyping. The isolate was studied for its tolerance to two heavy metals, i.e., cobalt (Co2+) and lead (Pb2+) by growing it in citrate mineral medium (CMM). Proteomic approach involving two-dimensional polyacrylamide gel electrophoresis (2D PAGE) and mass spectrometry (MS) was used to identify the differentially expressed proteins under heavy metal stress. Two of the differentially expressed proteins were identified to be L-isoaspartate protein carboxymethyltransferase type II and DNA gyrase A. To our knowledge, this is for the first time that K pneumoniae has been reported to be present in metal contaminated site and L-isoaspartate protein carboxymethyltransferase type II protein to be over expressed under heavy metal stress. The role of these proteins in metal tolerance is discussed. (c) 2006 Elsevier B.V. All rights reserved.

10a2D PAGE10aheavy metal toxicity10aMass spectrometry1 aBar, Chinmayee1 aPatil, Rajendra1 aDoshi, Jignesh1 aKulkarni, Mahesh, J.1 aGade, Wasudeo, N. uhttps://library.ncl.res.in/content/characterization-proteins-bacterial-strain-isolated-contaminated-site-involved-heavy-metal01416nas a2200253 4500008004100000022001400041245007100055210006900126260011100195300001200306490000700318520052500325653001300850653001200863653001200875653001700887653002600904100002500930700002500955700002600980700002601006700002501032856010501057 2008 eng d a0929-866500aAlbumin competitively inhibits glycation of less abundant proteins0 aAlbumin competitively inhibits glycation of less abundant protei aEXECUTIVE STE Y26, PO BOX 7917, SAIF ZONE, 1200 BR SHARJAH, U ARAB EMIRATESbBENTHAM SCIENCE PUBL LTDcJUL a663-6670 v153 a

Glycation, a non-enzymatic reaction between glucose and protein is the primary cause of diabetic complications. Albumin, the most abundant plasma protein undergoes glycation both in vivo and in vitro. The influence of albumin on glycation of less abundant proteins has not been addressed. For the first time, we show that albumin competitively inhibits the glycation of less abundant proteins. This study suggests that at least in the initial stages of diabetes, albumin may protect other proteins from glycation.

10aDiabetes10aglucose10ainsulin10aMALDI-TOF-MS10avascular complication1 aBhonsle, Hemangi, S.1 aSingh, Sameer, Kumar1 aSrivastava, Ghanshyam1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/albumin-competitively-inhibits-glycation-less-abundant-proteins-000554nas a2200145 4500008004100000245009000041210006900131260000800200300001000208490000600218100002300224700002100247700002500268856011500293 2008 eng d00aIdentification of ripening specific proteins in tomato by intact tissue MALDI-TOF-MS.0 aIdentification of ripening specific proteins in tomato by intact cJAN a10-130 v31 aKorwar, Arvind, M.1 aSanthakumari, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/identification-ripening-specific-proteins-tomato-intact-tissue-maldi-tof-ms01627nas a2200229 4500008004100000022001400041245007400055210006900129260006100198300000900259490000700268520085000275653002101125653001001146653002001156653003101176653001901207100001501226700002501241700002601266856010501292 2008 eng d a1572-388700aStudy of papain-cystatin interaction by intensity fading MALDI-TOF-MS0 aStudy of papaincystatin interaction by intensity fading MALDITOF a233 SPRING STREET, NEW YORK, NY 10013 USAbSPRINGERcJAN a7-120 v273 a

Intensity fading (IF) matrix assisted laser desorption ionization (MALDI) time of flight (TOF) mass spectrometry (MS ) has become an alternative screening approach for the affinity-binding analysis of proteins and peptides with molecular ligands. In this investigation an attempt has been made to study the protein ligand interaction by intensity fading (IF) MALDI-MS using papain and cystatin as model system for protein-ligand interactions. The intensity fading of cystatin was monitored using various concentration of cystatin ranging from (1 to 8.6 mu M) in presence of target protein, papain. The results indeed indicate that the intensity of cystatin decreases upon addition of papain. Furthermore, for the first time we have used IF-MALDI-MS for determining the number of binding sites for cystatin on papain by Scatchard analysis.

10aintensity fading10aMALDI10apapain-cystatin10aprotein-ligand interaction10aScatchard plot1 aShabab, M.1 aKulkarni, Mahesh, J.1 aKhan, Mohammad, Islam uhttps://library.ncl.res.in/content/study-papain-cystatin-interaction-intensity-fading-maldi-tof-ms-002218nas a2200157 4500008004100000245016200041210006900203260000800272300001200280490000700292520155300299100002601852700002501878700002501903856013201928 2009 eng d00aColumn liquid chromatography-ultraviolet and column liquid chromatography/mass spectrometry evaluation of stress degradation behavior of escitalopram oxalate0 aColumn liquid chromatographyultraviolet and column liquid chroma cJAN a138-1470 v923 a

{The objective of this work was to study the degradation behavior of escitalopram oxalate under different International Conference on Harmonization (ICH)-recommended stress conditions by column liquid chromatography (LC)-UV and LC/mass spectrometry (LC/MS) and to establish a validated stability-indicating LC assay method. Escitalopram oxalate was subjected to stress conditions of hydrolysis, oxidation, photolysis, and thermal decomposition. Extensive degradation was found to occur in alkaline medium. Mild degradation was observed in acidic and oxidative conditions. Escitalopram oxalate was stable to neutral, photolytic, and thermal stress. Successful separation of the drug from degradation products formed under stress conditions was achieved on a PerfectSil-100 ODS-3 column [C(18) (5 mu m, 25 cm x 4.6 mm id)] using methanol-0.01 M acetate buffer pH 3.8 adjusted with acetic acid (45 + 55) as the mobile phase. The flow rate was 1 mL/min, and the detection wavelength was 239 nm. The method was validated according to ICH guidelines. Major degradation products formed in hydrolysis and oxidative conditions were isolated, and structural elucidation of degradation products was done by LC/MS and infrared spectrometry studies. The major hydrolysis degradation product was confirmed as 1-(3-dimethylaminopropyl)-1-(4-fluoro-phenyl)-1,3dihydroisobenzofuran-5- carboxylic acid, and the major oxidative degradation product was confirmed as 1-{[3-dimethylamino(oxide)-propyl]-1(4-flurophenyl)}-1,3-dihydro-iso benzofuran-5-carbonitrile.

1 aDhaneshwar, Sunil, R.1 aMahadik, Mahadeo, V.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/column-liquid-chromatography-ultraviolet-and-column-liquid-chromatographymass-spectrometry-002207nas a2200241 4500008004100000022001400041245008000055210006900135260010600204300001200310490000700322520133200329653000901661653003201670653001901702653001801721653001501739100002501754700002401779700002501803700002601828856011101854 2009 eng d a0731-708500aLC-UV and LC-MS evaluation of stress degradation behaviour of tenatoprazole0 aLCUV and LCMS evaluation of stress degradation behaviour of tena aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcDEC a787-7930 v503 a

In the present study, comprehensive stress testing of tenatoprazole was carried out according to ICH guideline Q1A (R2). Tenatoprazole was subjected to stress conditions of hydrolysis, oxidation, photolysis and neutral decomposition. Additionally, the solid drug was subjected to 50 degrees C for 60 days in dry-bath, and to the combined effect of temperature and humidity at 40 degrees C/75% RH. Extensive degradation was found to occur in acidic, neutral and oxidative conditions. Mild degradation was observed in basic conditions. The drug is relatively stable in the solid-state. The products formed under different stress conditions were investigated by LC and LC-MS. Successful separation of drug from degradation products formed under stress conditions was achieved on a Chemito ODS-3 column [C(18) (5 mu m, 250 mm x 4.6 mm, i.d.)] using methanol: 0.01 M acetate buffer pH 4.5 adjusted with glacial acetic acid (55:45) as the mobile phase at flow rate of 1 mL/min and the peak was detected using a UV detector set at 306 nm. The LC-MS m/z values and fragmentation patterns of degradation products formed under different stress conditions were studied and characterized through LC-MS fragmentation. Based on the results, degradation pathway for drug has been proposed. (C) 2009 Elsevier B.V. All rights reserved.

10aHPLC10aStability-indicating method10aStress studies10aTenatoprazole10aValidation1 aMahadik, Mahadeo, V.1 aBhusari, Vidhya, K.1 aKulkarni, Mahesh, J.1 aDhaneshwar, Sunil, R. uhttps://library.ncl.res.in/content/lc-uv-and-lc-ms-evaluation-stress-degradation-behaviour-tenatoprazole-001656nas a2200253 4500008004100000022001400041245015600055210006900211260009300280300001200373490000700385520069100392653000801083653001901091653001501110653002601125653002301151653001101174100002901185700002501214700002601239700002501265856011201290 2010 eng d a1469-066700aDiscovery of rifampicin as a new anti-glycating compound by matrix-assisted laser desorption/ionization mass spectrometry-based insulin glycation assay0 aDiscovery of rifampicin as a new antiglycating compound by matri a6 CHARLTON MILL, CHARLTON, CHICHESTER,, W SUSSEX PO18 0HY, ENGLANDbIM PUBLICATIONScFEB a221-2260 v163 a

An in vitro insulin glycation assay was developed for screening glycation inhibitors. The assay involves the use of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for monitoring the formation of glycated insulin. The assay is simple, rapid and amenable for high throughput screening. Using this assay we have discovered a strong anti-glycation activity for the anti-tuberculosis drug rifampicin. These results were compared with bovine serum albumin glucose fluorescence assay. In addition, the IC(50) of rifampicin was lower than that of aminoguanidine, a known anti-glycating agent, suggesting that rifampicin is a more potent glycation inhibitor.

10aAGE10aaminoguanidine10aantibiotic10adiabetic complication10adrug repositioning10aIC(50)1 aGolegaonkar, Sandeep, B.1 aBhonsle, Hemangi, S.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/discovery-rifampicin-new-anti-glycating-compound-matrix-assisted-laser-003017nas a2200277 4500008004100000022001400041245013800055210006900193260007700262300001400339490000700353520197800360653001002338653003002348653003402378653001102412653002102423653002902444100001902473700002802492700002102520700002502541700002102566700002002587856013202607 2010 eng d a1615-985300aInteraction of recombinant CanPIs with helicoverpa armigera gut proteases reveals their processing patterns, stability and efficiency0 aInteraction of recombinant CanPIs with helicoverpa armigera gut  aPO BOX 10 11 61, D-69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJUL a2845-28570 v103 a

Six diverse representative Capsicum annuum (common name: hot pepper; Solanaceae) protease inhibitor genes, viz CanPI-5, -7, -13, -15, -19, and 22 comprising 1-4 inhibitory repeat domains (IRDs), were cloned and expressed in Pichia pastoris. The recombinant proteins were evaluated for their interactions with bovine trypsin, chymotrypsin, and Helicoverpa armigera gut proteases (HGP) using electrophoretic (native and denaturing) and mass spectrometric (MALDI-TOF-MS in combination with intensity fading assays) techniques. These techniques allow qualitative and semiquantitative analysis of multiple and processed IRDs of purified recombinant Capsicum annuum proteinase inhibitor (rCanPI) proteins. rCanPIs showed over 90% trypsin inhibition, varying chymotrypsin inhibition depending on the number of respective IRDs and over 60% inhibition of total HGP. rCanPI-15 that has only one IRD showed exceptionally low inhibition of these proteases. Interaction studies of rCanPIs with proteases using intensity fading-MALDI-TOF-MS revealed gradual processing of multi-IRD rCanPIs into single IRD forms by the action of HGP at the linker region, unlike their interactions with trypsin and chymotrypsin. Intensity fading-MALDI-TOF-MS assay showed that CanPI-13 and -15, possessing single IRD and expressed predominantly in stem tissue are degraded by HGP; indicating their function other than defense. In vitro and in vivo studies on rCanPI-5 and -7 showed maximum inhibition of HGP isoforms and their processed IRDs were also found to be stable in the presence of HGP. Even single amino acid variations in IRDs were found to change the HGP specificity like in the case of HGP-8 inhibited only by IRD-12. The presence of active PI in insect gut might be responsible for changed HGP profile. rCanPI-5 and -7 enhanced HGP-7, reduced HGP-4, -5, -10 expression and new protease isoforms were induced. These results signify isoform complexity in plant PIs and insect proteases.

10aCanPI10aH. armigera gut proteases10aIntensity fading MALDI-TOF-MS10aPin-II10aPlant proteomics10aPlant-insect interaction1 aMishra, Manasi1 aTamhane, Vaijayanti, A.1 aKhandelwal, Neha1 aKulkarni, Mahesh, J.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/interaction-recombinant-canpis-helicoverpa-armigera-gut-proteases-reveals-their-processing-001852nas a2200241 4500008004100000022001400041245012100055210006900176260016500245300001200410490000700422520084600429653001601275653003601291653002001327653001501347653001001362100002401372700002501396700003701421700002501458856012701483 2011 eng d a0388-135000aComparative and chemical proteomic approaches reveal gatifloxacin deregulates enzymes involved in glucose metabolism0 aComparative and chemical proteomic approaches reveal gatifloxaci aINTERNATIONAL MEDICAL INFORMATION CENTER, SHINANOMACHI RENGAKAN, 35 SHINANO-MACHI, SHINJUKU-KU, TOKYO, 160-0016, JAPANbJAPANESE SOC TOXICOLOGICAL SCIENCEScDEC a787-7960 v363 a

Gatifloxacin has been associated with increased risks of hypoglycemic and hyperglycemic side effects. In order to understand the molecular mechanism of gatifloxacin induced deregulation of glucose metabolism, a Combination of comparative and chemical proteomic approaches were employed using yeast as a model system. Differential protein expression studies using two dimensional electrophoresis and mass spectrometry reveal that gatifloxacin deregulates the expression of key enzymes involved in glucose metabolism. Furthermore, affinity chromatography and LC-MSE analysis led to identification of enolase, as one of the key gatifloxacin binding proteins. Fluorescence spectrometric studies confirmed that the gatifloxacin indeed binds to enolase. Role of enolase in regulation of gatifloxacin induced dysglycemic effect is discussed.

10aantibiotics10aDifferential protein expression10aFluoroquinolone10aGlycolysis10aYeast1 aSuresh, Kesavan, K.1 aBhosale, Santosh, D.1 aThulasiram, Hirekodathakallu, V.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/comparative-and-chemical-proteomic-approaches-reveal-gatifloxacin-deregulates-enzymes-001657nas a2200265 4500008004100000022001400041245010500055210006900160260009500229300001200324490000800336520066400344653003601008653001801044653000901062100002301071700002501094700002401119700002101143700002201164700002201186700003301208700002501241856012501266 2012 eng d a0006-291X00aAnalysis of AGE modified proteins and RAGE expression in HER2/neu negative invasive ductal carcinoma0 aAnalysis of AGE modified proteins and RAGE expression in HER2neu a525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USAbACADEMIC PRESS INC ELSEVIER SCIENCEcMAR a490-4940 v4193 a

Cancer is associated with increased glycolysis and carbonyl stress. In view of this, AGE modified proteins were identified from clinical breast cancer tissue using 2DE-immunoblot and mass-spectrometry. These proteins were identified to be serotransferrin, fibrinogen gamma chain, glycerol-3-phosphate dehydrogenase, lactate dehydrogenase, annexin II, prohibitin and peroxiredoxin 6, which have established role in cancer. Further, RAGE expression and its downstream signaling proteins NADPH oxidase and NF-kB were studied. Role of these AGE modified proteins and RAGE signaling in breast cancer is discussed. (C) 2012 Elsevier Inc. All rights reserved.

10aadvanced glycation end products10aBreast cancer10aRAGE1 aKorwar, Arvind, M.1 aBhonsle, Hemangi, S.1 aChougale, Ashok, D.1 aKote, Sachin, S.1 aGawai, Kachru, R.1 aGhole, Vikram, S.1 aKoppikar, Chaitanyananda, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/analysis-age-modified-proteins-and-rage-expression-her2neu-negative-invasive-ductal-000684nas a2200181 4500008004100000245011100041210006900152260000800221300001200229490000600241100001900247700002100266700001900287700001900306700001700325700002500342856013500367 2012 eng d00aBioanalytical method development and validation of levalbuterol a β2-adrenergic agonist by RP-HPLC method0 aBioanalytical method development and validation of levalbuterol  cMAR a249-2530 v41 aGhemud, A., S.1 aSanthakumari, B.1 aPharne, A., B.1 aJadhav, M., M.1 aJain, K., S.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/bioanalytical-method-development-and-validation-levalbuterol-%CE%B22-adrenergic-agonist-rp-hplc01823nas a2200181 4500008004100000245012800041210006900169260000800238300001200246490000600258520114600264100001901410700002101429700001901450700001701469700002501486856013001511 2012 eng d00aBioanalytical method development and validation of vildagliptin a novel dipeptidyl peptidase IV inhibitor by RP-HPLC method0 aBioanalytical method development and validation of vildagliptin  cJUN a119-1230 v43 a

A simple, selective, rapid, precise and economical Reverse-Phase HPLC method has been developed and validated for quantitative determination of vildagliptin in plasma. Vildagliptin is a potent dipeptidyl peptidase IV inhibitor used for treatment of diabetes. Tolbutamide is used as an internal standard. The method was carried out with UV Spectrophotometric detection using a Perkin Elmer Series 200 HPLC system equipped with XBridge Shield C18 column (3.5 μm, 4.6x150mm) and a guard column of the same type, at a flow rate of 1.0mL/min. Detection was carried out at 210 nm. The mobile phase consisted of 50mM ammonium bicarbonate (pH 7.8) (solvent A) and acetonitrile (solvent B). The retention times of vildagliptin and tolbutamide were 11.2 min and 13.4 min respectively. The method was developed and tested for linearity range of 10μg/ml to 120μg/ml. The developed method was validated in terms of selectivity, accuracy, precision, linearity, and stability study. The proposed method uses less biological material and the method is MS compatible also. Method can be applicable for pharmacokinetic studies using HPLC or LC-MS.

1 aPharne, A., B.1 aSanthakumari, B.1 aGhemud, A., S.1 aJain, H., K.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/bioanalytical-method-development-and-validation-vildagliptin-novel-dipeptidyl-peptidase-iv01672nas a2200265 4500008004100000022001400041245010700055210006900162260007500231300001000306490000600316520072300322653001701045653002101062653002701083653001601110100001601126700002101142700002501163700002501188700002301213700002601236700002001262856012401282 2012 eng d a1862-834600aDynamic proteome in enigmatic preeclampsia: an account of molecular mechanisms and biomarker discovery0 aDynamic proteome in enigmatic preeclampsia an account of molecul aPOSTFACH 101161, 69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJAN a79-900 v63 a

The coevolution of genomics and proteomics has led to advancements in the field of diagnosis and molecular mechanisms of disease. Proteomics is now stepping into the field of obstetrics, where early diagnosis of pregnancy complication such as preeclampsia (PE) is imperative. PE is a multifactorial disease characterized by hypertension with proteinuria, which is a leading cause of maternal and neonatal morbidity and mortality occurring in 5-7% of pregnancies worldwide. This review discusses the probable molecular mechanisms that lead to PE and summarizes the proteomics research carried out in understanding the pathogenicity of PE, and for identifying the candidate biomarker for diagnosis of the disease.

10aHypertension10aOxidative stress10aPregnancy complication10aProteinuria1 aMary, Sheon1 aPatil, Gouri, V.1 aKulkarni, Asmita, V.1 aKulkarni, Mahesh, J.1 aJoshi, Sadhana, R.1 aMehendale, Savita, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/dynamic-proteome-enigmatic-preeclampsia-account-molecular-mechanisms-and-biomarker-002315nas a2200277 4500008004100000022001400041245007900055210006900134260007300203300000900276490000800285520137500293653001201668653002001680653002601700653001801726653004001744100002001784700002401804700002001828700001701848700001801865700002501883700002201908856010701930 2012 eng d a0168-165600aEffects of arsenite stress on growth and proteome of Klebsiella pneumoniae0 aEffects of arsenite stress on growth and proteome of Klebsiella  aPO BOX 211, 1000 AE AMSTERDAM, NETHERLANDSbELSEVIER SCIENCE BVcMAR a8-160 v1583 a

In the present study an arsenite, As(III), tolerating bacterium, MR4, was isolated from Mulla River Pune, India, capable of reducing arsenate to arsenite and identified as Klebsiella pneumoniae (HQ857583). Comparative proteomic analysis using two-dimensional gel electrophoresis (2-DGE) and matrix assisted laser desorption ionization-time of flight-time of flight (MALDI-TOF/ TOF) was used to monitor the proteins undergoing changes in expression levels under 2.5 mM As(III) stress. The 2-DGE proteome map has shown that 60 proteins were differentially expressed under As(III) stress, of which 39 proteins were successfully identified with a MASCOT score greater than 70 (p < 0.05). Among the identified proteins, membrane transport/ binding proteins, porins, and amino acid metabolism enzymes were down-regulated while stress responsive proteins and antioxidant enzymes were up-regulated. Proteins involved in carbohydrate metabolism, particularly those in pentose phosphate pathway were also up-regulated while those involved in pyruvate metabolism were down-regulated. However, proteins involved in glycolysis and tricarboxylic acid cycle showed a mixed regulation response. These findings provide new insights into the probable mechanisms by which K. pneumoniae (HQ857583) could be adapting to As(III) stress. (C) 2012 Elsevier B.V. All rights reserved.

10a(2-DGE)10aArsenite stress10aKlebsiella pneumoniae10aMALDI-TOF-TOF10aTwo dimensional gel electrophoresis1 aDaware, Vandana1 aKesavan, Suresh, K.1 aPatil, Rajendra1 aNatu, Arvind1 aKumar, Ameeta1 aKulkarni, Mahesh, J.1 aGade, Wasudeo, N. uhttps://library.ncl.res.in/content/effects-arsenite-stress-growth-and-proteome-klebsiella-pneumoniae-002403nas a2200313 4500008004100000022001400041245010700055210006900162260007100231300001400302490000700316520127100323653004301594653001201637653001201649653003601661100002501697700002301722700002101745700002901766700002401795700002301819700002201842700002001864700002701884700002601911700002501937856012701962 2012 eng d a1535-389300aLow plasma albumin levels are associated with increased plasma protein glycation and HbA1c in diabetes0 aLow plasma albumin levels are associated with increased plasma p a1155 16TH ST, NW, WASHINGTON, DC 20036 USAbAMER CHEMICAL SOCcFEB a1391-13960 v113 a

Albumin is one of the most abundant plasma proteins and is heavily glycated in diabetes. In this study, we have addressed whether variation in the albumin levels influence glycation of plasma proteins and HbA1c. The study was performed in three systems: (1) streptozotocin (STZ)-induced diabetic mice plasma, (2) diabetic clinical plasma, and (3) in vitro glycated plasma. Diabetic mice and clinical plasma samples were categorized as diabetic high albumin plasma (DHAP) and diabetic low albumin plasma (DLAP) on the basis of their albumin levels. For the in vitro experiment, two albumin levels, high albumin plasma (HAP) and low albumin plasma (LAP), were created by differential depletion of plasma albumin. Protein glycation was studied by using a combination of two-dimensional electrophoresis (2DE), Western blotting, and LC-MSE. In both mice and clinical experiments, an increased plasma protein glycation was observed in DLAP than in DHAP. Additionally, plasma albumin levels were negatively correlated with HbA1c. The in vitro experiment with differential depletion of albumin mechanistically showed that the low albumin levels are associated with increased plasma protein glycation and that albumin competes for glycation with other plasma proteins.

10aadvanced glycation end products (AGEs)10aAlbumin10aglucose10apost translational modification1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aKote, Sachin, S.1 aGolegaonkar, Sandeep, B.1 aChougale, Ashok, D.1 aShaik, Mahemud, L.1 aDhande, Nitin, L.1 aGiri, Ashok, P.1 aShelgikar, Kishore, M.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/low-plasma-albumin-levels-are-associated-increased-plasma-protein-glycation-and-hba1c-002107nas a2200301 4500008004100000022001400041245009200055210006900147260007900216300001000295490000700305520106700312653000801379653001301387653002101400653001601421653003401437100002401471700002101495700002501516700002401541700002601565700002201591700002601613700002001639700002501659856012101684 2012 eng d a1073-608500aProteomic analysis of glycated proteins from streptozotocin-induced diabetic rat kidney0 aProteomic analysis of glycated proteins from streptozotocininduc a999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USAbHUMANA PRESS INCcJAN a28-380 v503 a

Glycation of proteins leading to formation of advanced glycation end products (AGEs) has been considered as one of the important causes of diabetic nephropathy. Therefore, in this study, glycated proteins were detected by anti-AGE antibodies from kidney of streptozotocin-induced diabetic rat showing nephropathic symptoms, by using two dimensional electrophoresis and western blot analysis. These glycated proteins were identified and characterized by using combination of peptide mass finger printing and tandem mass spectrometric approaches. Glycated proteins identified included proteins from metabolic pathways, oxidative stress, cell signaling, and transport. Several of the proteins modified by glycation were involved in glucose metabolism. The extent of glycation was higher in diabetes compared to control, in the glycated proteins that were common to both control and diabetic kidney. Two dimensional electrophoresis proteins profiling of glycated proteins suggest that four of the glycated proteins were significantly up regulated in diabetes.

10aAGE10aDiabetes10aGlucose toxicity10aNephropathy10aPost-translation modification1 aChougale, Ashok, D.1 aBhat, Shweta, P.1 aBhujbal, Swapnil, V.1 aZambare, Mandar, R.1 aPuntambekar, Shraddha1 aSomani, Rahul, S.1 aBoppana, Ramanamurthy1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-analysis-glycated-proteins-streptozotocin-induced-diabetic-rat-kidney-002099nas a2200253 4500008004100000022001400041245011400055210006900169260009900238300001200337490000800349520107900357653004201436653005301478653002101531653001701552653002901569100002501598700001901623700002101642700003001663700002501693856012701718 2012 eng d a0308-814600aPurification and characterization of an antioxidant protein (similar to 16 kDa) from Terminalia chebula fruit0 aPurification and characterization of an antioxidant protein simi aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLANDbELSEVIER SCI LTDcMAR a141-1480 v1313 a

Terminalia chebula fruit is used as folk medicine in India and Southeast Asia. An antioxidant protein was isolated by bioassay guided fractionation of T. chebula fruit by homogenizing in the citrate phosphate buffer. The isolated protein (TCP-III) obtained from fruit was purified by gel chromatography and preparative HPLC, showed apparent molecular weight of 16 kDa by SDS-PAGE and MALDI-TOF/MS analyses. Amino acid sequence obtained by LC-MSE analysis showed homology with the predicted protein fragments of Populus trichocarpa, putative uncharacterized protein fragments from Oryza sativa and with fragments of 17 kDa thylakoid lumenal protein from Spinacia oleracea. TCP-III exhibited significant radical scavenging in DPPH, NO, H2O2 and ABTS assays. In addition, TCP-III inhibited oxidation of linoleic acid in beta-carotene bleaching assay, reduced ferric ions and chelated ferrous ions. The present finding demonstrates uniquely, for the first time, characterization of an antioxidant protein from T. chebula fruit. (C) 2011 Elsevier Ltd. All rights reserved.

10a16 kDa protein molecule of T. chebula10aAntioxidant and free radical scavenging activity10aCharacterization10aPurification10aTerminalia chebula fruit1 aSrivastava, Pratibha1 aRaut, Hema, N.1 aWagh, Renuka, S.1 aPuntambekar, Hemalata, M.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/purification-and-characterization-antioxidant-protein-similar-16-kda-terminalia-chebula03296nas a2200277 4500008004100000022001400041245007100055210006900126260008100195300000800276490000700284520235500291653001002646653001402656653002002670653003802690653002902728100001902757700002202776700002802798700002502826700002102851700002102872700002002893856010502913 2012 eng d a1471-222900aStress inducible proteinase inhibitor diversity in capsicum annuum0 aStress inducible proteinase inhibitor diversity in capsicum annu a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcNOV a2170 v123 a

Background: Wound-inducible Pin-II Proteinase inhibitors (PIs) are one of the important plant serine PIs which have been studied extensively for their structural and functional diversity and relevance in plant defense against insect pests. To explore the functional specialization of an array of Capsicum annuum (L.) proteinase inhibitor (CanPIs) genes, we studied their expression, processing and tissue-specific distribution under steady-state and induced conditions. Inductions were performed by subjecting C. annuum leaves to various treatments, namely aphid infestation or mechanical wounding followed by treatment with either oral secretion (OS) of Helicoverpa armigera or water. Results: The elicitation treatments regulated the accumulation of CanPIs corresponding to 4-, 3-, and 2-inhibitory repeat domains (IRDs). Fourty seven different CanPI genes composed of 28 unique IRDs were identified in total along with those reported earlier. The CanPI gene pool either from uninduced or induced leaves was dominated by 3-IRD PIs and trypsin inhibitory domains. Also a major contribution by 4-IRD CanPI genes possessing trypsin and chymotrypsin inhibitor domains was specifically revealed in wounded leaves treated with OS. Wounding displayed the highest number of unique CanPIs while wounding with OS treatment resulted in the high accumulation of specifically CanPI-4, -7 and -10. Characterization of the PI protein activity through two dimensional gel electrophoresis revealed tissue and induction specific patterns. Consistent with transcript abundance, wound plus OS or water treated C. annuum leaves exhibited significantly higher PI activity and isoform diversity contributed by 3- and 4-IRD CanPIs. CanPI accumulation and activity was weakly elicited by aphid infestation yet resulted in the higher expression of CanPI-26, -41 and -43. Conclusions: Plants can differentially perceive various kinds of insect attacks and respond appropriately through activating plant defenses including regulation of PIs at transcriptional and post-translational levels. Based on the differentially elicited CanPI accumulation patterns, it is intriguing to speculate that generating sequence diversity in the form of multi-IRD PIs is a part of elaborative plant defense strategy to obtain a diverse pool of functional units to confine insect attack.

10aCanPI10aHerbivory10aOral secretions10aPin-II type proteinase inhibitors10aPlant-insect interaction1 aMishra, Manasi1 aMahajan, Neha, S.1 aTamhane, Vaijayanti, A.1 aKulkarni, Mahesh, J.1 aBaldwin, Ian, T.1 aGupta, Vidya, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/stress-inducible-proteinase-inhibitor-diversity-capsicum-annuum-002064nas a2200253 4500008004100000022001400041245013500055210006900190260009300259300001200352490000700364520103800371653004301409653003301452653001301485653003701498100002501535700002301560700002401583700002501607700002301632700002501655856013001680 2012 eng d a1469-066700aZoom-in - a targeted database search for identification of glycation modifications analyzed by untargeted tandem mass spectrometry0 aZoomin a targeted database search for identification of glycatio a6 CHARLTON MILL, CHARLTON, CHICHESTER,, W SUSSEX PO18 0HY, ENGLANDbIM PUBLICATIONScJAN a475-4810 v183 a

Post-translational modifications (PTMs) are very important to biological function, however their identification and characterization is technically challenging. In this study, we have identified glycation modifications by nano LC-MSE, a data independent acquisition work flow, followed by database search using the Protein Lynx Global Server (PLGS). PLGS search with a complete human protein database hardly identified glycation modifications in a glycated human serum albumin (HSA), which was detected to be glycated by western blotting with advanced glycation end products (AGE) antibody and fluorescence spectroscopy. To overcome this difficulty, ``Zoom-In'' approach, a targeted database search was used to identify glycation modifications in a glycated HSA, which were further manually validated. This approach was useful for identification of glycation modifications from untargeted tandem mass spectrometry workflow such as MSE, but may require the development of a new algorithm or an upgrade of the existing software.

10aadvanced glycation end products (AGEs)10adata independent acquisition10aDiabetes10apost-translational modifications1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aKesavan, Suresh, K.1 aBhosale, Santosh, D.1 aBansode, Sneha, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/zoom-targeted-database-search-identification-glycation-modifications-analyzed-untargeted-002342nas a2200205 4500008004100000022001400041245005200055210005000107260011000157300001400267490000600281520163200287100002001919700002501939700002001964700002501984700002102009700002202030856008402052 2013 eng d a2046-206900aArsenomics of exiguobacterium sp PS (NCIM 5463)0 aArsenomics of exiguobacterium sp PS NCIM 5463 aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcAPR a9705-97130 v33 a

In the present study, arsenate [As(V)] tolerant organisms were isolated from a metal contaminated site, and identified by 16S rDNA sequencing as Exiguobacterium sp. Arsenomics (omics of arsenic response) in Exiguobacterium sp. PS via two-dimensional gel electrophoresis (2-DGE) coupled with identification of proteins using MALDI-TOF MS, MALDI-TOF MS/MS and LC/MSE are studied vide this paper. Out of 2460 Coomassie stained proteins, 270 were differentially expressed (p < 0.05). Considering the resolution and abundance level, 45 spots (proteins) were subjected to MALDI-TOF MS/MS, MALDI-TOF/TOF and LC/MSE. Out of these, 33 were identified and categorized into several functional categories (including both known and novel/putative stress responsive proteins) viz., carbohydrate and energy metabolism, stress response, motility, amino acid, and purine and protein synthesis. The identifications of unique proteins, phosphate ABC transporter ATPase subunit and pyridine nucleotide disulfide oxidoreductase, are the significant findings of the present study which may be used as markers of As(V) stress in Exiguobacterium sp. PS. The absence of As(V) reductase activity and non-amplification of the arsC gene of the ars operon suggests that the classical mechanism of As(V) resistance may not be operating in Exiguobacterium sp. PS. Lastly, considering the fact that proteomic studies under As(V) stress were performed only in Gram negative organisms, this study is the first comprehensive endeavour to explore the mechanism of As(V) response in Gram positive organisms, Exiguobacterium sp. PS at the proteomic level.

1 aSacheti, Poonam1 aBhonsle, Hemangi, S.1 aPatil, Rajendra1 aKulkarni, Mahesh, J.1 aSrikanth, Rapole1 aGade, Wasudeo, N. uhttps://library.ncl.res.in/content/arsenomics-exiguobacterium-sp-ps-ncim-5463-001301nas a2200241 4500008004100000022001400041245005300055210005200108260007500160300001200235490000600247520057000253653001000823653001300833653001400846653000900860100002500869700002300894700001600917700002500933700002000958856008100978 2013 eng d a1862-834600aGlycated proteome: from reaction to intervention0 aGlycated proteome from reaction to intervention aPOSTFACH 101161, 69451 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcJAN a155-1700 v73 a

Glycation, a nonenzymatic reaction between reducing sugars and proteins, is a proteome wide phenomenon, predominantly observed in diabetes due to hyperglycemia. Glycated proteome of plasma, kidney, lens, and brain are implicated in the pathogenesis of various diseases, including diabetic complications, neurodegenerative diseases, cancer, and aging. This review discusses the strategies to characterize protein glycation, its functional implications in different diseases, and intervention strategies to protect the deleterious effects of protein glycation.

10aaging10aDiabetes10aGlycation10aRAGE1 aKulkarni, Mahesh, J.1 aKorwar, Arvind, M.1 aMary, Sheon1 aBhonsle, Hemangi, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/glycated-proteome-reaction-intervention-002538nas a2200193 4500008004100000022001400041245010300055210006900158260011000227300001400337490000600351520172700357100002502084700002702109700002402136700002502160700002902185856013002214 2013 eng d a1742-206X00aNovel mechanism for antiglycative action of limonene through stabilization of protein conformation0 aNovel mechanism for antiglycative action of limonene through sta aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcJUN a2463-24720 v93 a

Inhibition of protein glycation is known to ameliorate secondary complications in diabetes. In the present study antiglycative properties of limonene, a natural product, were evaluated using BSA as a model protein. AMG (aminoguanidine) was used as a positive control. Measurement of total AGEs (Advanced Glycation End-products) and specific AGEs revealed that limonene could inhibit protein glycation to the extent of 56.3% and 75.1% respectively at 50 mu M concentration as against 54.4% and 82.2% by AMG at 1 mM. Congo red binding and CD (Circular Dichroism) analysis revealed inhibition of alpha-helix to beta-sheet transition wherein 18.5% beta-sheet structures were observed in glycated BSA (bovine serum albumin) as against 4.9% with limonene. Glycation of protein in the presence of urea was enhanced by 18%, while in the presence of limonene it was reduced by 23% revealing the stabilizing effect of limonene. Electrophoretic mobility was similar to the normal control and a zeta potential value of -12.1 mV as against -15.1 mV in diabetic control was observed. Inhibition of glycation in limonene treated samples was confirmed through LC-MS analysis wherein AGEs such as pentosidine, CML (N-epsilon-(carboxymethyl)lysine), CEL (N-epsilon-(carboxyethyl) lysine), MOLD (methylglyoxal-lysine dimer) and imidazolone observed in glycated samples were absent in limonene treated samples. PatchDock studies revealed that limonene could bind to the major glycation sites IB, IIA and IIB sub domains and AMG to the IIIA sub domain. Thus limonene is a potent protein glycation inhibitor that prevents protein glycation through a novel mechanism of stabilization of protein structure through hydrophobic interactions.

1 aJoglekar, Madhav, M.1 aPanaskar, Shrimant, N.1 aChougale, Ashok, D.1 aKulkarni, Mahesh, J.1 aArvindekar, Akalpita, U. uhttps://library.ncl.res.in/content/novel-mechanism-antiglycative-action-limonene-through-stabilization-protein-conformation-001910nas a2200265 4500008004100000022001400041245012700055210006900182260009000251300000900341490000600350520086400356100002401220700001701244700002901261700003901290700002401329700002301353700002501376700002401401700003701425700002601462700002501488856013101513 2013 eng d a2045-232200aProteome wide reduction in AGE modification in streptozotocin induced diabetic mice by hydralazine mediated transglycation0 aProteome wide reduction in AGE modification in streptozotocin in aMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLANDbNATURE PUBLISHING GROUPcOCT a29410 v33 a

The non-enzymatic reaction between glucose and protein can be chemically reversed by transglycation. Here we report the transglycation activity of hydralazine using a newly developed MALDI-TOF-MS based assay. Hydralazine mediated transglycation of HbA1c, plasma proteins and kidney proteins was demonstrated in streptozotocin (STZ) induced diabetic mice, as evidenced by decrease in protein glycation, as well as presence of hydralazine-glucose conjugate in urine of diabetic mice treated with hydralazine. Hydralazine down regulated the expression of Receptor for Advanced Glycation End products (RAGE), NADPH oxidase (NOX), and super oxide dismutase (SOD). These findings will provide a new dimension for developing intervention strategies for the treatment of glycation associated diseases such as diabetes complications, atherosclerosis, and aging.

1 aKesavan, Suresh, K.1 aBhat, Shweta1 aGolegaonkar, Sandeep, B.1 aJagadeeshaprasad, Mashanipalya, G.1 aDeshmukh, Arati, B.1 aPatil, Harshal, S.1 aBhosale, Santosh, D.1 aShaikh, Mahemud, L.1 aThulasiram, Hirekodathakallu, V.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteome-wide-reduction-age-modification-streptozotocin-induced-diabetic-mice-hydralazine-002124nas a2200217 4500008004100000022001400041245008100055210006900136260010300205300001200308490000700320520130500327100002301632700002401655700002201679700002001701700002701721700002501748700002501773856010801798 2013 eng d a1535-947600aProteomic analysis of protease resistant proteins in the diabetic rat kidney0 aProteomic analysis of protease resistant proteins in the diabeti a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcJAN a228-2360 v123 a

Glycation induced protein aggregation has been implicated in the development of diabetic complications and neurodegenerative diseases. These aggregates are known to be resistant to proteolytic digestion. Here we report the identification of protease resistant proteins from the streptozotocin induced diabetic rat kidney, which included enzymes in glucose metabolism and stress response proteins. These protease resistant proteins were characterized to be advanced glycation end products modified and ubiquitinated by immunological and mass spectrometry analysis. Further, diabetic rat kidney exhibited significantly impaired proteasomal activity. The functional analysis of identified physiologically important enzymes showed that their activity was reduced in diabetic condition. Loss of functional activity of these proteins was compensated by enhanced gene expression. Aggregation prone regions were predicted by in silico analysis and compared with advanced glycation end products modification sites. These findings suggested that the accumulation of protein aggregates is an inevitable consequence of impaired proteasomal activity and protease resistance due to advanced glycation end products modification. Molecular & Cellular Proteomics 12: 10.1074/mcp.M112.020651, 228-236, 2013.

1 aBansode, Sneha, B.1 aChougale, Ashok, D.1 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aBodhankar, Subhash, L.1 aHarsulkar, Abhay, M.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-analysis-protease-resistant-proteins-diabetic-rat-kidney-002352nas a2200205 4500008004100000022001400041245015300055210006900208260008800277300001000365490000700375520148400382100002301866700002501889700002201914700002201936700003301958700002501991856013002016 2013 eng d a1536-231000aProteomic profiling and interactome analysis of ER-positive/HER2/neu negative invasive ductal carcinoma of the breast: towards proteomics biomarkers0 aProteomic profiling and interactome analysis of ERpositiveHER2ne a140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USAbMARY ANN LIEBERT, INCcJAN a27-400 v173 a

Breast cancer, especially ER positive/HER2/neu negative IDC, is the predominant subtype of invasive ductal carcinoma. Although proteomic approaches have been used towards biomarker discovery in clinical breast cancer, ER positive/HER2/neu negative IDC is the least studied subtype. To discover biomarkers, as well as to understand the molecular events associated with disease progression of estrogen receptor positive/HER2/neu negative subtype of invasive ductal carcinoma, differential protein expression profiling was performed by using LC-MSE (MS at elevated energy). A total of 118 proteins were identified, of which 26 were differentially expressed. These identified proteins were functionally classified and their interactions and coexpression were analyzed by using bioinformatic tools PANTHER (Protein Analysis THrough Evolutionary Relationships) and STRING (Search Tool for the Retrieval of Interacting Genes). These proteins were found to be upregulated and were involved in cytoskeletal organization, calcium binding, and stress response. Interactions of annexin A5, actin, S100 A10, glyceraldehyde 3 phosphate dehydrogenase, superoxide dismutase 1, apolipoprotein, fibrinogen, and heat shock proteins were prominent. Differential expression of these proteins was validated by two-dimensional gel electrophoresis and Western blot analysis. The cluster of these proteins may serve as a signature profile for estrogen receptor positive/ HER2/neu negative subtype.

1 aKorwar, Arvind, M.1 aBhonsle, Hemangi, S.1 aGhole, Vikram, S.1 aGawai, Kachru, R.1 aKoppikar, Chaitanyananda, B.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-profiling-and-interactome-analysis-er-positiveher2neu-negative-invasive-ductal-001879nas a2200253 4500008004100000022001400041245011900055210006900174260006300243300001200306490000600318520096300324653001401287653001001301653001501311100002501326700002301351700002401374700002101398700002201419700002701441700002501468856013201493 2013 eng d a1791-299700aProteomic study reveals downregulation of apolipoprotein A1 in plasma of poorly controlled diabetes: a pilot study0 aProteomic study reveals downregulation of apolipoprotein A1 in p aPOB 18179, ATHENS, 116 10, GREECEbSPANDIDOS PUBL LTDcFEB a495-4980 v73 a

Proteomic approaches aid in gaining a better understanding of the pathophysiology of diabetic complications. In view of this, differential protein expression in diabetic plasma samples was studied by a combination of proteomic and western blot analyses. Diabetic plasma samples were categorized based on glycated haemoglobin levels as controlled diabetes (CD; 7-8%), poorly controlled diabetes (PCD; >8%) and non-diabetic control (ND;<6.4%). Two-dimensional electrophoresis and liquid chromatography-mass spectrometry revealed differential expression of proteins including upregulation of fibrinogen and haptoglobin and downregulation of vitamin D binding protein, alpha-1-antitrypsin, transthyretin and apolipoprotein A1 (Apo A1) in diabetic compared with non-diabetic plasma samples. Amongst these proteins, Apo A1 downregulation was prominent in PCD. Downregulation of Apo A1 may serve as an early predictive marker of diabetic complications.

10abiomarker10aHbA1c10aproteomics1 aBhonsle, Hemangi, S.1 aKorwar, Arvind, M.1 aChougale, Ashok, D.1 aKote, Sachin, S.1 aDhande, Nitin, L.1 aShelgikar, Kishore, M.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-study-reveals-downregulation-apolipoprotein-a1-plasma-poorly-controlled-diabetes-002040nas a2200193 4500008004100000022001400041245013600055210006900191260011000260300001400370490000600384520121700390100002401607700002201631700002501653700002001678700002201698856012601720 2013 eng d a2046-206900aSynergistic action of flavin containing NADH dependant azoreductase and cytochrome P450 monooxygenase in azoaromatic mineralization0 aSynergistic action of flavin containing NADH dependant azoreduct aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcDEC a3062-30700 v33 a

An alkaliphilic strain Bacillus lentus Bl377 was isolated from contaminated soil of the textile area of Solapur, India. The strain was able to degrade almost 98% of recalcitrant azoic compounds by a mutually regulated process of azoreductase and a monooxygenase system. An enzyme activity study and a periodical carbon monoxide (CO) binding spectra study on a UV-visible spectrophotometer revealed that the intermediate amines formed by typical azoreduction (N=N cleavage), subsequently underwent hydroxylation by the cytochrome P450 monooxygenase (CYP450) system. Azoreductase was purified by chromatographic techniques and characterization by MALDI-TOF substantiated its identity as FMN containing NADH dependent azoreductase of 32 kDa in size. Surprisingly, purified azoreductase showed the highest activity at 80 degrees C and pH 8.0. An increase in the activity of superoxide dismutase after decolorization confirmed the signature of oxidative stress and its involvement in the dismutation of reactive metabolites. Intermediate metabolite analysis by HPLC, GC-MS and FTIR and the removal of total organic carbon (TOC) suggested the azoaromatics' degradation leads to mineralization via a TCA cycle.

1 aOturkar, Chetan, C.1 aOthman, Munif, A.1 aKulkarni, Mahesh, J.1 aMadamwar, Datta1 aGawai, Kachru, R. uhttps://library.ncl.res.in/content/synergistic-action-flavin-containing-nadh-dependant-azoreductase-and-cytochrome-p450-002581nas a2200277 4500008004100000022001400041245012800055210006900183260008100252300002000333490000700353520154700360653001501907653001601922653002901938653001401967100002201981700002102003700002402024700002902048700002302077700002902100700002502129700002302154856012602177 2014 eng d a1476-459800aCholesterol depletion by methyl-beta-cyclodextrin augments tamoxifen induced cell death by enhancing its uptake in melanoma0 aCholesterol depletion by methylbetacyclodextrin augments tamoxif a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcSEP aArticle No. 2040 v133 a

Background: Despite modern advances in treatment, skin cancer is still one of the most common causes of death in the western countries. Chemotherapy plays an important role in melanoma management. Tamoxifen has been used either alone or in-combination with other chemotherapeutic agents to treat melanoma. However, response rate of tamoxifen as a single agent has been comparatively low. In the present study, we investigated whether treatment with methyl-beta-cyclodextrin (MCD), a cholesterol depleting agent, increases the efficacy of tamoxifen in melanoma cells. Methods: This was a two-part study that incorporated in vitro effects of tamoxifen and MCD combination by analyzing cell survival, apoptosis and cell cycle analysis and in vivo antitumor efficacy on tumor isografts in C57BL/6J mice. Results: MCD potentiated tamoxifen induced anticancer effects by causing cell cycle arrest and induction of apoptosis. Sensitization to tamoxifen was associated with down regulation of antiapoptotic protein Bcl-2, up-regulation of proapoptotic protein Bax, reduced caveolin-1 (Cav-1) and decreased pAkt/pERK levels. Co-administration of tamoxifen and MCD caused significant reduction in tumor volume and tumor weight in mice due to enhancement of drug uptake in the tumor. Supplementation with cholesterol abrogated combined effect of tamoxifen and MCD. Conclusion: Our results emphasize a potential synergistic effect of tamoxifen with MCD, and therefore, may provide a unique therapeutic window for improvement in melanoma treatment.

10aCaveolin-110aCholesterol10aMethyl beta-cyclodextrin10aTamoxifen1 aMohammad, Naoshad1 aMalvi, Parmanand1 aMeena, Avtar, Singh1 aSingh, Shivendra, Vikram1 aChaube, Balkrishna1 aVannuruswamy, Garikapati1 aKulkarni, Mahesh, J.1 aBhat, Manoj, Kumar uhttps://library.ncl.res.in/content/cholesterol-depletion-methyl-beta-cyclodextrin-augments-tamoxifen-induced-cell-death-001848nas a2200253 4500008004100000022001400041245005200055210005200107260007700159300001000236490000600246520106800252653001701320653002401337653001901361653002101380653000901401100001701410700001601427700002001443700002001463700002501483856008601508 2014 eng d a1862-834600aImmune response to chemically modified proteome0 aImmune response to chemically modified proteome aBOSCHSTRASSE 12, D-69469 WEINHEIM, GERMANYbWILEY-V C H VERLAG GMBHcFEB a19-340 v83 a

Both enzymatic and nonenzymatic PTMs of proteins involve chemical modifications. Some of these modifications are prerequisite for the normal functioning of cell, while other chemical modifications render the proteins as neo-self antigens, which are recognized as non-self leading to aberrant cellular and humoral immune responses. However, these modifications could be a secondary effect of autoimmune diseases, as in the case of type I diabetes, hyperglycemia leads to protein glycation. The enigma of chemical modifications and immune response is akin to the chick-and-egg paradox. Nevertheless, chemical modifications regulate immune response. In some of the well-known autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, and multiple sclerosis, chemically modified proteins act as autoantigens forming immune complexes. In some instances, chemical modifications are also involved in regulating immune response during pathogen infection. Further, the usefulness of proteomic analysis of immune complexes is briefly discussed.

10aAutoantibody10aAutoimmune diseases10aImmune complex10aImmunoproteomics10aPTMs1 aBhat, Shweta1 aMary, Sheon1 aBanarjee, Reema1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/immune-response-chemically-modified-proteome-002477nas a2200205 4500008004100000022001400041245010900055210006900164260009100233300001200324490000600336520165100342100002001993700002902013700001802042700003702060700002502097700002002122856012902142 2014 eng d a1932-620300aInsecticidal potential of defense metabolites from ocimum kilimandscharicum against helicoverpa armigera0 aInsecticidal potential of defense metabolites from ocimum kilima a1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USAbPUBLIC LIBRARY SCIENCEcAUG ae1043770 v93 a

Genus Ocimum contains a reservoir of diverse secondary metabolites, which are known for their defense and medicinal value. However, the defense-related metabolites from this genus have not been studied in depth. To gain deeper insight into inducible defense metabolites, we examined the overall biochemical and metabolic changes in Ocimum kilimandscharicum that occurred in response to the feeding of Helicoverpa armigera larvae. Metabolic analysis revealed that the primary and secondary metabolism of local and systemic tissues in O. kilimandscharicum was severely affected following larval infestation. Moreover, levels of specific secondary metabolites like camphor, limonene and beta-caryophyllene (known to be involved in defense) significantly increased in leaves upon insect attack. Choice assays conducted by exposing H. armigera larvae on O. kilimandscharicum and tomato leaves, demonstrated that O. kilimandscharicum significantly deters larval feeding. Further, when larvae were fed on O. kilimandscharicum leaves, average body weight decreased and mortality the larvae increased. Larvae fed on artificial diet supplemented with O. kilimandscharicum leaf extract, camphor, limonene and beta-caryophyllene showed growth retardation, increased mortality rates and pupal deformities. Digestive enzymes of H. armigera - namely, amylase, protease and lipase- showed variable patterns after feeding on O. kilimandscharicum, which implies striving of the larvae to attain required nutrition for growth, development and metamorphosis. Evidently, selected metabolites from O. kilimandscharicum possess significant insecticidal activity.

1 aSingh, Priyanka1 aJayaramaiah, Ramesha, H.1 aSarate, Priya1 aThulasiram, Hirekodathakallu, V.1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/insecticidal-potential-defense-metabolites-ocimum-kilimandscharicum-against-helicoverpa-002257nas a2200217 4500008004100000022001400041245010300055210006900158260009100227300001200318490000600330520141100336100002301747700001901770700002501789700002501814700002201839700002501861700002501886856012801911 2014 eng d a1932-620300aMolecular investigations of protriptyline as a multi-target directed ligand in alzheimer's disease0 aMolecular investigations of protriptyline as a multitarget direc a1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USAbPUBLIC LIBRARY SCIENCEcAUG ae1051960 v93 a

Alzheimer's disease (AD) is a complex neurodegenerative disorder involving multiple cellular and molecular processes. The discovery of drug molecules capable of targeting multiple factors involved in AD pathogenesis would greatly facilitate in improving therapeutic strategies. The repositioning of existing non-toxic drugs could dramatically reduce the time and costs involved in developmental and clinical trial stages. In this study, preliminary screening of 140 FDA approved nervous system drugs by docking suggested the viability of the tricyclic group of antidepressants against three major AD targets, viz. Acetylcholinesterase (AChE), beta-secretase (BACE-1), and amyloid beta (A beta) aggregation, with one member, protriptyline, showing highest inhibitory activity. Detailed biophysical assays, together with isothermal calorimetry, fluorescence quenching experiments, kinetic studies and atomic force microscopy established the strong inhibitory activity of protriptyline against all three major targets. The molecular basis of inhibition was supported with comprehensive molecular dynamics simulations. Further, the drug inhibited glycation induced amyloid aggregation, another important causal factor in AD progression. This study has led to the discovery of protriptyline as a potent multi target directed ligand and established its viability as a promising candidate for AD treatment.

1 aBansode, Sneha, B.1 aJana, Asis, K.1 aBatkulwar, Kedar, B.1 aWarkad, Shrikant, D.1 aJoshi, Rakesh, S.1 aSengupta, Neelanjana1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/molecular-investigations-protriptyline-multi-target-directed-ligand-alzheimers-disease-002190nas a2200229 4500008004100000245014400041210006900185260000800254300002300262490000700285520132200292100002501614700003901639700002101678700002101699700002101720700001901741700002501760700002001785700002401805856013101829 2014 eng d00aProteomic analysis of human plasma in chronic rheumatic mitral stenosis reveals proteins involved in the complement and coagulation cascade0 aProteomic analysis of human plasma in chronic rheumatic mitral s cSEP aArticle Number: 350 v113 aBackground: Rheumatic fever in childhood is the most common cause of Mitral Stenosis in developing countries. The disease is characterized by damaged and deformed mitral valves predisposing them to scarring and narrowing (stenosis) that results in left atrial hypertrophy followed by heart failure. Presently, echocardiography is the main imaging technique used to diagnose Mitral Stenosis. Despite the high prevalence and increased morbidity, no biochemical indicators are available for prediction, diagnosis and management of the disease. Adopting a proteomic approach to study Rheumatic Mitral Stenosis may therefore throw some light in this direction. In our study, we undertook plasma proteomics of human subjects suffering from Rheumatic Mitral Stenosis (n = 6) and Control subjects (n = 6). Six plasma samples, three each from the control and patient groups were pooled and subjected to low abundance protein enrichment. Pooled plasma samples (crude and equalized) were then subjected to in-solution trypsin digestion separately. Digests were analyzed using nano LC-MSE. Data was acquired with the Protein Lynx Global Server v2.5.2 software and searches made against reviewed Homo sapiens database (UniProtKB) for protein identification. Label-free protein quantification was performed in crude plasma only. 1 aMukherjee, Somaditya1 aJagadeeshaprasad, Mashanipalya, G.1 aBanerjee, Tanima1 aGhosh, Sudip, K.1 aBiswas, Monodeep1 aDutta, Santanu1 aKulkarni, Mahesh, J.1 aPattari, Sanjib1 aBandyopadhyay, Arun uhttps://library.ncl.res.in/content/proteomic-analysis-human-plasma-chronic-rheumatic-mitral-stenosis-reveals-proteins-involved02477nas a2200325 4500008004100000022001400041245009500055210006900150260005700219300001400276490000700290520142400297653001201721653002001733653001501753653001101768653002001779100002001799700002001819700001701839700002501856700002101881700002001902700002201922700001901944700002501963700002101988700002202009856012002031 2014 eng d a0175-759800aProteomics of arsenic stress in the gram-positive organism Exiguobacterium sp PS NCIM 54630 aProteomics of arsenic stress in the grampositive organism Exiguo a233 SPRING ST, NEW YORK, NY 10013 USAbSPRINGERcAUG a6761-67730 v983 a

The general responses of microorganisms to environmental onslaughts are modulated by altering the gene expression pattern to reduce damage in the cell and produce compensating stress responses. The present study attempts to unravel the response of the Gram-positive Exiguobacterium sp. PS NCIM 5463 in the presence of [As(III)] and arsenate [As(V)] using comparative proteomics via two-dimension gel electrophoresis (2-DE) coupled with identification of proteins using matrix-assisted laser desorption/ionisation (MALDI-TOF/MALDI-TOF/TOF). Out of 926 Coomassie-stained proteins, 45 were differentially expressed (p < 0.05). Considering the resolution and abundance level, 24 spots (peptides) were subjected to MALDI analysis, identified and categorised into several functional categories, viz., nitrogen metabolism, energy and stress regulators, carbohydrate metabolism, protein synthesis components and others. A functional role of each protein is discussed in Exiguobacterium sp. PS 5463 under arsenic stress and validated at their transcript level using a quantitative real-time polymerase chain reaction. Unlike previous reports that unravel the responses toward arsenic stress in Gram-negative organisms, the present study identified new proteins under arsenic stress in a Gram-positive organism, Exiguobacterium sp. PS NCIM 5463, which could elucidate the physiology of organisms under arsenic stress.

10aArsenic10aExiguobacterium10aproteomics10aStress10aTranscriptomics1 aSacheti, Poonam1 aPatil, Rajendra1 aDube, Ankita1 aBhonsle, Hemangi, S.1 aThombre, Dipalee1 aMarathe, Sayali1 aVidhate, Ravindra1 aWagh, Priyanka1 aKulkarni, Mahesh, J.1 aRapole, Srikanth1 aGade, Wasudeo, N. uhttps://library.ncl.res.in/content/proteomics-arsenic-stress-gram-positive-organism-exiguobacterium-sp-ps-ncim-546302439nas a2200205 4500008004100000245008700041210006900128260000800197490000700205520173800212100002001950700002001970700001801990700002502008700002402033700001502057700001702072700002502089856011902114 2015 eng d00aAssociation between serum albumin and glycated hemoglobin in Asian Indian subjects0 aAssociation between serum albumin and glycated hemoglobin in Asi cJAN0 v193 a

Protein glycation plays a significant role in diabetic complications. Glycated hemoglobin (HbA1c) is a known predictor of diabetes and its complications. Albumin, found to be profoundly glycated in diabetes, and its level could regulate plasma protein as well as hemoglobin glycation. We aimed to evaluate the association between variations in albumin level with HbA1c in the Asian Indian population. We screened data of 929 subjects who have had a simultaneous measurement of fasting plasma glucose (FPG), HbA1c and albumin levels via the same blood collection. Data were analyzed by SPSS for 610 subjects who met the study criteria. There was a significant negative correlation between HbA1c and albumin concentration (r = -0.284; P < 0.001). Univariate analysis showed the statistically significant decrease of average HbA1c but not for fasting plasma glucose (FPG) across increasing tertiles of albumin. Stepwise multiple regression model showed a significant correlation between HbA1c and serum albumin (P < 0.05), FPG (P < 0.001), hemoglobin (Hb) (P < 0.001) and serum globulin (P < 0.05). FPG was the strongest predictor (63.4%) of variation of HbA1c. The albumin concentration (r = -0.114) accounted for 0.3% (P < 0.05) of the total variance in HbA1c independent of age, body mass index, FPG, Hb, creatinine, total protein and globulin. It was also observed that HbA1c decreases with increasing albumin concentration in those having FPG between 100 to <126 mg/dl. Serum albumin negatively correlates with HbA1c in Asian Indians independent of other variables. This study suggests that predicting diabetes and its complication based on the HbA1c needs to be further investigated in Indian subjects.

1 aTiwari, Shalbha1 aBotahle, Manish1 aHasan, Imtiaz1 aKulkarni, Mahesh, J.1 aSayyad, Mehmood, G.1 aBasu, Rita1 aBasu, Ananda1 aG., Unnikrishnan, A. uhttps://library.ncl.res.in/content/association-between-serum-albumin-and-glycated-hemoglobin-asian-indian-subjects02633nas a2200265 4500008004100000022001400041245022300055210006900278260010300347300001400450490000700464520147500471100002301946700002901969700003901998700002902037700001702066700002502083700002702108700002002135700002302155700003202178700002502210856013202235 2015 eng d a1535-947600aDevelopment of diagnostic fragment ion library for glycated peptides of human serum albumin: targeted quantification in prediabetic, diabetic, and microalbuminuria plasma by parallel reaction monitoring, SWATH, and MSE0 aDevelopment of diagnostic fragment ion library for glycated pept a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcAUG a2150-21590 v143 a

Human serum albumin is one of the most abundant plasma proteins that readily undergoes glycation, thus glycated albumin has been suggested as an additional marker for monitoring glycemic status. Hitherto, only Amadori-modified peptides of albumin were quantified. In this study, we report the construction of fragment ion library for Amadori-modified lysine (AML), N(epsilon)-(carboxymethyl) lysine (CML)-, and N(epsilon)-(carboxyethyl) lysine (CEL)-modified peptides of the corresponding synthetically modified albumin using high resolution accurate mass spectrometry (HR/AM). The glycated peptides were manually inspected and validated for their modification. Further, the fragment ion library was used for quantification of glycated peptides of albumin in the context of diabetes. Targeted Sequential Window Acquisition of all THeoretical Mass Spectra (SWATH) analysis in pooled plasma samples of control, prediabetes, diabetes, and microalbuminuria, has led to identification and quantification of 13 glycated peptides comprised of four AML, seven CML, and two CEL modifications, representing nine lysine sites of albumin. Five lysine sites namely K549, K438, K490, K88, and K375, were observed to be highly sensitive for glycation modification as their respective m/z showed maximum fold change and had both AML and CML modifications. Thus, peptides involving these lysine sites could be potential novel markers to assess the degree of glycation in diabetes.

1 aKorwar, Arvind, M.1 aVannuruswamy, Garikapati1 aJagadeeshaprasad, Mashanipalya, G.1 aJayaramaiah, Ramesha, H.1 aBhat, Shweta1 aRegin, Bhaskaran, S.1 aRamaswamy, Sureshkumar1 aGiri, Ashok, P.1 aMohan, Viswanathan1 aBalasubramanyam, Muthuswamy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/development-diagnostic-fragment-ion-library-glycated-peptides-human-serum-albumin-targeted-001713nas a2200205 4500008004100000022001400041245006000055210006000115260011000175300001600285490000600301520095000307100002401257700002901281700002301310700002101333700003701354700002501391856009101416 2015 eng d a2046-206900aInvestigation of antiglycation activity of isoprenaline0 aInvestigation of antiglycation activity of isoprenaline aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcMAR a25051-250580 v53 a

Advanced glycation end products (AGEs) are implicated in pathogenesis of diabetes and its complications. In this study, we report the ability of isoprenaline to inhibit the AGE modification of protein by fluorescence spectroscopy and western blotting. Isoprenaline was more effective in inhibiting AGE modification than aminoguanidine, a well known glycation inhibitor. Further, we show that isoprenaline inhibits at Amadori product formation during glycation reaction by various techniques such as MALDI-TOF-MS, LC-MS/MS, and fructosamine assay. Mass spectrometric analysis of the glycation reaction mixture incubated with isoprenaline suggested that it forms adducts with glucose and thus inhibits glycation. The finding of the additional property of isoprenaline of inhibiting glycation suggests that it is a potential candidate for drug repositioning for the treatment of diabetes and its complications, as it is an FDA approved drug.

1 aKolekar, Yogesh, M.1 aVannuruswamy, Garikapati1 aBansode, Sneha, B.1 aSanthakumari, B.1 aThulasiram, Hirekodathakallu, V.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/investigation-antiglycation-activity-isoprenaline-002643nas a2200301 4500008004100000022001400041245005500055210005500110260006700165300001200232490000700244520171800251653001701969653001201986653001401998653001102012653002002023653000902043100002502052700002302077700002102100700002402121700002102145700003602166700002802202700002502230856008602255 2015 eng d a1615-985300aInvestigation of phosphoproteome in RAGE signaling0 aInvestigation of phosphoproteome in RAGE signaling a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcJAN a245-2590 v153 a

The receptor for advanced glycation end products (RAGE) is one of the most important proteins implicated in diabetes, cardiovascular diseases, neurodegenerative diseases, and cancer. It is a pattern recognition receptor by virtue of its ability to interact with multiple ligands, RAGE activates several signal transduction pathways through involvement of various kinases that phosphorylate their respective substrates. Only few substrates have been known to be phosphorylated in response to activation by RAGE (e.g., nuclear factor kappa B); however, it is possible that these kinases can phosphorylate multiple substrates depending upon their expression and localization, leading to altered cellular responses in different cell types and conditions. One such example is, glycogen synthase kinase 3 beta which is known to phosphorylate glycogen synthase, acts downstream to RAGE, and hyperphosphorylates microtubule-associated protein tau causing neuronal damage. Thus, it is important to understand the role of various RAGE-activated kinases and their substrates. Therefore, we have reviewed here the details of RAGE-activated kinases in response to different ligands and their respective phosphoproteome. Furthermore, we discuss the analysis of the data mined for known substrates of these kinases from the PhosphoSitePlus (http://www.phosphosite.org) database, and the role of some of the important substrates involved in cancer, diabetes, cardiovascular diseases, and neurodegenerative diseases. In summary, this review provides information on RAGE-activated kinases and their phosphoproteome, which will be helpful in understanding the possible role of RAGE and its ligands in progression of diseases.

10aCell biology10aDisease10aGlycation10aKinase10aPhosphoproteome10aRAGE1 aBatkulwar, Kedar, B.1 aBansode, Sneha, B.1 aPatil, Gouri, V.1 aGodbole, Rashmi, K.1 aKazi, Rubina, S.1 aChinnathambi, Subashchandrabose1 aShanmugam, Dhanasekaran1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/investigation-phosphoproteome-rage-signaling-002140nas a2200217 4500008004100000022001400041245007400055210006900129260011000198300001600308490000600324520132500330100002201655700002401677700002301701700002401724700002101748700002501769700002401794856010401818 2015 eng d a2046-206900aN-(3-Aminoalkyl)proline derivatives with potent antigycation activity0 aN3Aminoalkylproline derivatives with potent antigycation activit aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcSEP a77332-773400 v53 a

The importance of amino acids in the therapy of conditions such as renal failure, neurological disorders and congenital defects has been documented. Some amino acids such as lysine and glycine have also been reported to have antiglycating activity. Herein we report the synthesis of a new series of N-(3-aminoalkyl)proline derivatives which are non-natural in nature. The compounds were unambiguously characterized by NMR, mass and IR spectroscopy. Their in vitro antiglycation activity was studied by circular dichroism and fluorescence spectrometry. The mechanism of action was also studied and found to take place by inhibition of Amadori product formation. The inhibition of AGE formation was further confirmed by western blot and LC-MS/MS analyses and the IC50 values of the potent compounds were determined. Compounds containing hydroxyl substituents at C4 were found to have superior antiglycation properties than those containing azide substituents at the same position. The compounds were additionally found to possess good anti-oxidant properties, which could lead to further reduction in AGE formation. Moreover, the title compounds were found to have low cytotoxicity in mammalian cells, another important attribute. Thus, the title compounds represent a novel promising class of antiglycating agents.

1 aChilukuri, Harsha1 aKolekar, Yogesh, M.1 aBhosle, Govind, S.1 aGodbole, Rashmi, K.1 aKazi, Rubina, S.1 aKulkarni, Mahesh, J.1 aFernandes, Moneesha uhttps://library.ncl.res.in/content/n-3-aminoalkylproline-derivatives-potent-antigycation-activity-002083nas a2200181 4500008004100000022001400041245008000055210006900135260011000204300001600314490000600330520136100336100002901697700002301726700002501749700002101774856010601795 2015 eng d a2046-206900aRecent development of plant products with anti-glycation activity: a review0 aRecent development of plant products with antiglycation activity aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcMAR a31113-311380 v53 a

Diabetes mellitus (DM) is an endocrine disorder characterized by chronic hyperglycemia, which results from an absolute or a relative deficiency of insulin or resistance to insulin. Hyperglycemia is increasingly linked to the pathogenesis of diabetic complications in individuals with long-duration diabetes. One of the inevitable consequences of hyperglycemia is the enhanced accumulation of advanced glycation end-products (AGEs), which are implicated in the pathogenesis of diabetes. Various natural products and their active constituents have reportedly been used for the treatment of diabetes and its complications. Some of these molecules are known to have anti-glycation activity. The search for novel anti-glycation agents from various sources is gaining a lot of importance. Attention has especially been focused on plants with an ethnopharmacological background and also on plants rich in triterpenoids and phenolics, which generally exhibit antioxidant and anti-glycation effects. Plant extracts or compounds obtained from them that possess both antioxidant and anti-glycation activities might have great therapeutic potential for treating diabetic complications. This review highlights the anti-glycation activities of phytochemicals, which will aid in the identification of lead molecules for the development of new anti-glycation drugs.

1 aChinchansure, Ashish, A.1 aKorwar, Arvind, M.1 aKulkarni, Mahesh, J.1 aJoshi, Swati, P. uhttps://library.ncl.res.in/content/recent-development-plant-products-anti-glycation-activity-review-002873nas a2200325 4500008004100000022001400041245012300055210006900178260006700247300001200314490000700326520171600333653003602049653001002085653002702095653001102122653001402133653001302147653001502160100002502175700002102200700002102221700002702242700003902269700002302308700003602331700002502367700002402392856013102416 2015 eng d a1474-971800aRifampicin reduces advanced glycation end products and activates DAF-16 to increase lifespan in Caenorhabditis elegans0 aRifampicin reduces advanced glycation end products and activates a111 RIVER ST, HOBOKEN 07030-5774, NJ USAbWILEY-BLACKWELLcJUN a463-4730 v143 a

Advanced glycation end products (AGEs) are formed when glucose reacts nonenzymatically with proteins; these modifications are implicated in aging and pathogenesis of many age-related diseases including type II diabetes, atherosclerosis, and neurodegenerative disorders. Thus, pharmaceutical interventions that can reduce AGEs may delay age-onset diseases and extend lifespan. Using LC-MSE, we show that rifampicin (RIF) reduces glycation of important cellular proteins in vivo and consequently increases lifespan in Caenorhabditis elegans by up to 60%. RIF analog rifamycin SV (RSV) possesses similar properties, while rifaximin (RMN) lacks antiglycation activity and therefore fails to affect lifespan positively. The efficacy of RIF and RSV as potent antiglycating agents may be attributed to the presence of a p-dihydroxyl moiety that can potentially undergo spontaneous oxidation to yield highly reactive p-quinone structures, a feature absent in RMN. We also show that supplementing rifampicin late in adulthood is sufficient to increase lifespan. For its effect on longevity, rifampicin requires DAF-18 (nematode PTEN) as well as JNK-1 and activates DAF-16, the FOXO homolog. Interestingly, the drug treatment modulates transcription of a different subset of DAF-16 target genes, those not controlled by the conserved Insulin-IGF-1-like signaling pathway. RIF failed to increase the lifespan of daf-16 null mutant despite reducing glycation, showing thereby that DAF-16 may not directly affect AGE formation. Together, our data suggest that the dual ability to reduce glycation in vivo and activate prolongevity processes through DAF-16 makes RIF and RSV effective lifespan-extending interventions.

10aadvanced glycation end products10aaging10aCaenorhabditis elegans10aDAF-1610aGlycation10alifespan10arifampicin1 aGolegaonkar, Sandeep1 aTabrez, Syed, S.1 aPandit, Awadhesh1 aSethurathinam, Shalini1 aJagadeeshaprasad, Mashanipalya, G.1 aBansode, Sneha, B.1 aSampathkumar, Srinivasa-Gopalan1 aKulkarni, Mahesh, J.1 aMukhopadhyay, Arnab uhttps://library.ncl.res.in/content/rifampicin-reduces-advanced-glycation-end-products-and-activates-daf-16-increase-lifespan-001692nas a2200205 4500008004100000022001400041245007700055210006900132260011000201300001600311490000600327520089700333100002301230700002501253700002501278700001901303700002501322700002501347856011401372 2015 eng d a2046-206900aTolbutamide induces conformational change and promotes albumin glycation0 aTolbutamide induces conformational change and promotes albumin g aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcAPR a40070-400750 v53 a

Tolbutamide is a first generation sulfonylurea drug used for the treatment of type II diabetes. In the present study, the effect of tolbutamide on albumin conformation was investigated using BSA as a model protein. The binding of tolbutamide induced a conformational change in albumin, which was confirmed by thioflavin T assay, ANS assay, and CD spectroscopy. Computer simulations suggested that the binding of tolbutamide increases the solvent accessibility of lysine residues, the hotspots of glycation. Furthermore, the change in conformation of albumin facilitates increased glycation, which was observed by AGE specific fluorescence and the results were corroborated by mass spectrometric analysis. This study suggested that tolbutamide enhances albumin glycation by inducing conformational change in BSA and hence could be a potential risk factor if used for a prolonged period.

1 aBansode, Sneha, B.1 aBatkulwar, Kedar, B.1 aWarkad, Shrikant, D.1 aJana, Asis, K.1 aSengupta, Neelanjana1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/tolbutamide-induces-conformational-change-and-promotes-albumin-glycation-002342nas a2200265 4500008004100000022001400041245016300055210006900218260012700287300001200414490000700426520130200433653001301735653001301748653001701761653001401778653001101792653001901803100002601822700002501848700002301873700002501896700002501921856013001946 2016 eng d a1138-754800aDietary flaxseed oil and fish oil ameliorates renal oxidative stress, protein glycation, and inflammation in streptozotocin-nicotinamide-induced diabetic rats0 aDietary flaxseed oil and fish oil ameliorates renal oxidative st aCAMPUS UNIV, CARRETERA DEL SADAR S-N, APARTADO 177, 31080 PAMPLONA, SPAINbSERVICIO PUBLICACIONES UNIVERSIDAD NAVARRAcJUN a327-3360 v723 a

Protective and prophylactic effects of omega-3 fatty acids on oxidative stress and inflammation are well known. We assessed beneficial effects of flaxseed oil and fish oil on streptozotocin (65 mg/kg; i.p.)-nicotinamide (110 mg/kg; i.p.) induced diabetic rats by studying renal expression of antioxidant and inflammatory genes. Diabetic rats given 10 % flaxseed oil or 10 % fish oil diet for 35 days showed significant decrease in renal lipid peroxidation. Flaxseed oil diet resulted in up-regulation of renal superoxide dismutase-1 (SOD-1) (activity and expression) and glutathione peroxidase-1 (GPx-1) expression. Furthermore, both diets up-regulated catalase (CAT) (activity and expression) and down-regulated heme oxygenase-1 (HO-1) expression. Both diets were able to limit the renal advanced glycation end products (AGEs) formation and reduced receptor of AGE (RAGE) protein expression significantly. Expressions of interleukin-6 (IL-6) and NF-kappa B p65 subunit were down-regulated significantly by flaxseed oil or fish oil diet. The histological tubular injuries were also lowered by both diets. These results suggest that dietary omega-3 fatty acids may slow the progression of diabetic nephropathy (DN) associated with oxidative stress, glycation, and inflammation in the kidney.

10aDiabetes10aFish oil10aFlaxseed oil10aGlycation10aKidney10aStreptozotocin1 aJangale, Nivedita, M.1 aDevarshi, Prasad, P.1 aBansode, Sneha, B.1 aKulkarni, Mahesh, J.1 aHarsulkar, Abhay, M. uhttps://library.ncl.res.in/content/dietary-flaxseed-oil-and-fish-oil-ameliorates-renal-oxidative-stress-protein-glycation-and00644nas a2200157 4500008004100000245012600041210006900167260000800236300001600244490000700260100001900267700002500286700002500311700002500336856012500361 2016 eng d00aGlycation induces conformational changes in Amyloid-? peptide and enhances its aggregation propensity: molecular insights0 aGlycation induces conformational changes in Amyloid peptide and  cOCT a31446-314580 v181 aJana, Asis, K.1 aBatkulwar, Kedar, B.1 aKulkarni, Mahesh, J.1 aSengupta, Neelanjana uhttps://library.ncl.res.in/content/glycation-induces-conformational-changes-amyloid-peptide-and-enhances-its-aggregation00727nas a2200181 4500008004100000245011800041210007000159260000800229300001800237490000600255100002500261700001900286700002400305700002300329700002500352700002500377856014300402 2016 eng d00aHydralazine inhibits amyloid beta (Aβ) aggregation and glycation and ameliorates Aβ1–42 induced neurotoxicity0 aHydralazine inhibits amyloid beta Aβ aggregation and glycation a cNOV a108768-1087760 v61 aBatkulwar, Kedar, B.1 aJana, Asis, K.1 aGodbole, Rashmi, K.1 aKhandelwal, Puneet1 aSengupta, Neelanjana1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/hydralazine-inhibits-amyloid-beta-a%CE%B2-aggregation-and-glycation-and-ameliorates-a%CE%B21%E2%80%934202204nas a2200253 4500008004100000022001400041245009100055210006900146260011000215300001600325490000600341520123900347100002901586700003901615700001801654700002401672700001701696700002301713700002401736700002601760700002101786700002501807856011801832 2016 eng d a2046-206900aMolecules with O-acetyl group protect protein glycation by acetylating lysine residues0 aMolecules with Oacetyl group protect protein glycation by acetyl aTHOMAS GRAHAM HOUSE, SCIENCE PARK, MILTON RD, CAMBRIDGE CB4 0WF, CAMBS, ENGLANDbROYAL SOC CHEMISTRYcJUL a65572-655780 v63 a

Pharmaceutical intervention for reduction of advanced glycation end products (AGEs) is considered as a therapeutic strategy to attenuate the pathogenesis of diabetes. Many molecules have been reported to possess antiglycation activity, one such example is acetylsalicylic acid (aspirin). It protects proteins from glycation by acetylating the lysine residues. Therefore, in this study we have synthesized and screened molecules containing free N-acetyl, O-acetyl and acetophenone groups. All the selected molecules in this study showed glycation inhibition but interestingly, only molecules with O-acetyl but not N-acetyl and acetophenone groups were capable of acetylating lysine residue. Furthermore, we have demonstrated that pre-acetylation or aspirin treatment prior to the induction of diabetes helps in reducing HbA1c and AGE formation in the streptozotocin induced diabetic mice. Hence pre-acetylation may have an additional therapeutic efficacy of reducing AGE levels in vivo. Incorporation of O-acetyl group into anti-diabetic molecules could be a useful strategy, as it may have an additive effect in reducing AGEs. Identification of such novel acetylating agents represents a new area in the drug discovery process.

1 aVannuruswamy, Garikapati1 aJagadeeshaprasad, Mashanipalya, G.1 aKashinath, K.1 aKesavan, Suresh, K.1 aBhat, Shweta1 aKorwar, Arvind, M.1 aChougale, Ashok, D.1 aBoppana, Ramanamurthy1 aReddy, Srinivasa1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/molecules-o-acetyl-group-protect-protein-glycation-acetylating-lysine-residues02556nas a2200277 4500008004100000022001400041245012900055210006900184260009000253490000600343520147700349100002001826700002901846700002401875700002901899700002301928700001601951700002601967700002401993700002202017700002602039700002502065700003702090700002002127856013102147 2016 eng d a2045-232200aPotential dual role of eugenol in inhibiting advanced glycation end products in diabetes: proteomic and mechanistic insights0 aPotential dual role of eugenol in inhibiting advanced glycation  aMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLANDbNATURE PUBLISHING GROUPcJAN0 v63 a

Medicinally important genus Ocimum harbors a vast pool of chemically diverse metabolites. Current study aims at identifying anti-diabetic candidate compounds from Ocimum species. Major metabolites in O. kilimandscharicum, O. tenuiflorum, O. gratissimum were purified, characterized and evaluated for anti-glycation activity. In vitro inhibition of advanced glycation end products (AGEs) by eugenol was found to be highest. Preliminary biophysical analysis and blind docking studies to understand eugenol-albumin interaction indicated eugenol to possess strong binding affinity for surface exposed lysines. However, binding of eugenol to bovine serum albumin (BSA) did not result in significant change in secondary structure of protein. In vivo diabetic mice model studies with eugenol showed reduction in blood glucose levels by 38% likely due to inhibition of alpha-glucosidase while insulin and glycated hemoglobin levels remain unchanged. Western blotting using anti-AGE antibody and mass spectrometry detected notably fewer AGE modified peptides upon eugenol treatment both in vivo and in vitro. Histopathological examination revealed comparatively lesser lesions in eugenol-treated mice. Thus, we propose eugenol has dual mode of action in combating diabetes; it lowers blood glucose by inhibiting a-glucosidase and prevents AGE formation by binding to epsilon-amine group on lysine, protecting it from glycation, offering potential use in diabetic management.

1 aSingh, Priyanka1 aJayaramaiah, Ramesha, H.1 aAgawane, Sachin, B.1 aVannuruswamy, Garikapati1 aKorwar, Arvind, M.1 aAnand, Atul1 aDhaygude, Vitthal, S.1 aShaikh, Mahemud, L.1 aJoshi, Rakesh, S.1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J.1 aThulasiram, Hirekodathakallu, V.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/potential-dual-role-eugenol-inhibiting-advanced-glycation-end-products-diabetes-proteomic-002505nas a2200253 4500008004100000022001400041245010700055210006900162260010300231300001400334490000700348520151500355100001701870700003901887700002401926700002401950700002501974700002301999700002002022700003202042700002602074700002502100856012602125 2016 eng d a1535-947600aProteomic insight reveals elevated levels of albumin in circulating immunecomplexes in diabetic plasma0 aProteomic insight reveals elevated levels of albumin in circulat a9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USAbAMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INCcJUN a2011-20200 v153 a

A Hyperglycemic condition in diabetes promotes formation of advanced glycation end products, which are known to elicit immune response and form complexes with immunoglobulins called circulating immune complexes. To investigate the involvement of advanced glycation end product (AGE)-modified proteins in the elicitation of an immune response, circulating immune complexes were isolated and proteins associated were identified and characterized. Label-free-based mass spectrometric analysis of circulating immune complexes in clinical plasma of prediabetic, newly diagnosed diabetes, and diabetic microalbuminurea revealed elevated levels of serum albumin in the circulating immune complexes, which were also observed to be AGE modified. Further, to examine the role of glycation, circulating immune complexeswere analyzed in the streptozotocin-induced diabetic mice treated with or without aminoguanidine, a prototype glycation inhibitor. Mass spectrometric analysis of circulating immune complexes showed elevated levels of serum albumin in plasma from diabetic mice over that of control animals. Aminoguanidine-treated diabetic mice displayed decreased AGE modification of plasma albumin, accompanied by a reduced level of albumin in the circulating immune complexes. In addition, elevated levels of proinflammatory cytokines such as IL-1b, IL-2, and TNF-alpha were observed in diabetes, which were reduced with aminoguanidine treatment, suggesting the involvement of glycation in the immune response.

1 aBhat, Shweta1 aJagadeeshaprasad, Mashanipalya, G.1 aPatil, Yugendra, R.1 aShaikh, Mahemud, L.1 aRegin, Bhaskaran, S.1 aMohan, Viswanathan1 aGiri, Ashok, P.1 aBalasubramanyam, Muthuswamy1 aBoppana, Ramanamurthy1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-insight-reveals-elevated-levels-albumin-circulating-immunecomplexes-diabetic02720nas a2200289 4500008004100000022001400041245015100055210006900206260008100275300002200356490000700378520163500385653001302020653001402033653001402047653002102061653001002082653002202092100003902114700002502153700002502178700002002203700002302223700002902246700002502275856013002300 2016 eng d a1542-641600aTargeted quantification of N-1-(carboxymethyl) valine and N-1-(carboxyethyl) valine peptides of beta-hemoglobin for better diagnostics in diabetes0 aTargeted quantification of N1carboxymethyl valine and N1carboxye a236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLANDbBIOMED CENTRAL LTDcMAR aArticle Number: 70 v133 a

Background: N-1-(Deoxyfructosyl) valine (DFV) beta-hemoglobin (beta-Hb), commonly referred as HbA1c, is widely used diagnostic marker in diabetes, believed to provide glycemic status of preceding 90-120 days. However, the turnover of hemoglobin is about 120 days, the DFV-beta-Hb, an early and reversible glycation product eventually may undergo irreversible advanced glycation modifications such as carboxymethylation or carboxyethylation. Hence quantification of N-1-(carboxymethyl) valine (CMV) and N-1-(carboxyethyl) valine (CEV) peptides of beta-Hb would be useful in assessing actual glycemic status. Results: Fragment ion library for synthetically glycated peptides of hemoglobin was generated by using high resolution-accurate mass spectrometry (HR/AM). Using parallel reaction monitoring, deoxyfructosylated, carboxymethylated and carboxyethylated peptides of hemoglobin were quantified in clinical samples from healthy control, pre-diabetes, diabetes and poorly controlled diabetes. For the first time, we report N-1-beta-valine undergoes carboxyethylation and mass spectrometric quantification of CMV and CEV peptides of beta-hemoglobin. Carboxymethylation was found to be the most abundant modification of N-1-beta-valine. Both CMV-beta-Hb and CEV-beta-Hb peptides showed better correlation with severity of diabetes in terms of fasting glucose, postprandial glucose and microalbuminuria. Conclusions: This study reports carboxymethylation as a predominant modification of N-1-beta-valine of Hb, and quantification of CMV-beta-Hb and CEV-beta-Hb could be useful parameter for assessing the severity of diabetes.

10aDiabetes10aDiagnosis10aGlycation10aGlycemic control10aHbA1c10aMass spectrometry1 aJagadeeshaprasad, Mashanipalya, G.1 aBatkulwar, Kedar, B.1 aMeshram, Nishita, N.1 aTiwari, Shalbha1 aKorwar, Arvind, M.1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/targeted-quantification-n-1-carboxymethyl-valine-and-n-1-carboxyethyl-valine-peptides-beta00717nas a2200193 4500008004100000022001400041245012400055210006900179260000800248300001200256490000800268100002000276700001900296700002000315700002000335700001700355700002500372856012600397 2016 eng d a1355-603700aUnderstanding endothelial dysfunction in diabetic cardiovascular complications using mass spectrometry-based proteomics0 aUnderstanding endothelial dysfunction in diabetic cardiovascular cOCT aA16-A170 v1021 aBanarjee, Reema1 aSharma, Akshay1 aBai, Shakuntala1 aDeshmukh, Arati1 aKazi, Rubina1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/understanding-endothelial-dysfunction-diabetic-cardiovascular-complications-using-mass02013nas a2200157 4500008004100000245007700041210006900118260000800187490000700195520143900202100001701641700003901658700003101697700002501728856010201753 2017 eng d00aAbundance matters: role of albumin in diabetes, a proteomics perspective0 aAbundance matters role of albumin in diabetes a proteomics persp cJUL0 v143 aIntroduction: Human serum albumin (HSA) is a multifaceted protein with vital physiological functions. It is the most abundant plasma protein with inherent capability to bind to diverse ligands, and thus susceptible to various post-translational modifications (PTMs) which alter its structure and functions. One such PTM is glycation, a non-enzymatic reaction between reducing sugar and protein leading to formation of heterogeneous advanced glycation end products (AGEs). Glycated albumin (GA) concentration increases significantly in diabetes and is implicated in development of secondary complications. Areas covered: In this review, we discuss in depth formation of GA and its consequences, approaches used for characterization and quantification of GA, milestones in GA proteomics, clinical relevance of GA as a biomarker, significance of maintaining abundant levels of albumin and future perspectives. Expert commentary: Elevated GA levels are associated with development of insulin resistance as well as secondary complications, in healthy and diabetic individuals respectively. Mass spectrometry (MS) based approaches aid in precise characterization and quantification of GA including early and advanced glycated peptides, which can be useful in prediction of the disease status. Thus GA has evolved to be one of the best candidates in the pursuit of diagnostic markers for prediction of prediabetes and diabetic complications.1 aBhat, Shweta1 aJagadeeshaprasad, Mashanipalya, G.1 aVenkatasubramani, Vinashya1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/abundance-matters-role-albumin-diabetes-proteomics-perspective01559nas a2200181 4500008004100000245007800041210006900119260000800188300001200196490000700208520094600215100001601161700002501177700002601202700002301228700002001251856010601271 2017 eng d00a Differential accumulation of vimentin fragments in preeclamptic placenta0 aDifferential accumulation of vimentin fragments in preeclamptic  cNOV a420-4250 v743 aPreeclampsia is a pregnancy complication that is the result of abnormal placentation because of inadequate trophoblast invasion into spiral arteries that prevent normal blood flow to the placenta. We report the alteration in vimentin protein proteolysis in placenta of normotensive and preeclamptic women, which is known to have a role in many physiological functions other than its major function in the structural integrity of the cell. Placental proteome from normotensive (n=25) and preeclamptic pregnancies (n=25) showed eight differentially accumulated protein spots of vimentin (proteolytic fragments) by two-dimensional electrophoresis. Immunoblots of normotensive and preeclamptic placenta revealed a difference in proteolytic processing of vimentin. In particular, lower molecular weight vimentin fragments of 32 and 20 kDa were 3.3 and 2.6-fold (p<0.0001) higher, respectively, in preeclampsia compared with normotensive placenta.1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMehendale, Savita, S.1 aJoshi, Sadhana, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/differential-accumulation-vimentin-fragments-preeclamptic-placenta02880nas a2200193 4500008004100000245017800041210006900219260000800288300001200296490000800308520208800316100002102404700002402425700002402449700002102473700003902494700002502533856012802558 2017 eng d00aGlycation inhibitors extend yeast chronological lifespan by reducing advanced glycation end products and by back regulation of proteins involved in mitochondrial respiration0 aGlycation inhibitors extend yeast chronological lifespan by redu cMAR a104-1120 v1563 aAdvanced Glycation End products (AGEs) are implicated in aging process. Thus, reducing AGEs by using glycation inhibitors may help in attenuating the aging process. In this study using Saccharomyces cerevisiae yeast system, we show that Aminoguanidine (AMG), a well-known glycation inhibitor, decreases the AGE modification of proteins in non-calorie restriction (NR) (2% glucose) and extends chronological lifespan (CLS) similar to that of calorie restriction (CR) condition (0.5% glucose). Proteomic analysis revealed that AMG back regulates the expression of differentially expressed proteins especially those involved in mitochondrial respiration in NR condition, suggesting that it switches metabolism from fermentation to respiration, mimicking CR. AMG induced back regulation of differentially expressed proteins could be possibly due to its chemical effect or indirectly by glycation inhibition. To delineate this, Metformin (MET), a structural analog of AMG and a mild glycation inhibitor and Hydralazine (HYD), another potent glycation inhibitor but not structural analog of AMG were used. HYD was more effective than MET in mimicking AMG suggesting that glycation inhibition was responsible for restoration of differentially expressed proteins. Thus glycation inhibitors particularly AMG, HYD and MET extend yeast CLS by reducing AGEs, modulating the expression of proteins involved in mitochondrial respiration and possibly by scavenging glucose. Significance: This study reports the role of glycation in aging process. In the non-caloric restriction condition, carbohydrates such as glucose promote protein glycation and reduce CLS. While, the inhibitors of glycation such as AMC, HYD, MET mimic the caloric restriction condition by back regulating deregulated proteins involved in mitochondria( respiration which could facilitate shift of metabolism from fermentation to respiration and extend yeast CLS. These findings suggest that glycation inhibitors can be potential molecules that can be used in management of aging. (C) 2017 Elsevier B.V. All rights reserved.1 aKazi, Rubina, S.1 aBanarjee, Reema, M.1 aDeshmukh, Arati, B.1 aPatil, Gouri, V.1 aJagadeeshaprasad, Mashanipalya, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/glycation-inhibitors-extend-yeast-chronological-lifespan-reducing-advanced-glycation-end01776nas a2200205 4500008004100000245012700041210006900168260000800237300001400245490000700259520101100266100002301277700002501300700002401325700002001349700002101369700002501390700002401415856013101439 2017 eng d00aMass spectrometry based identification of galectin-3 interacting proteins potentially involved in lung melanoma metastasis0 aMass spectrometry based identification of galectin3 interacting  cNOV a2303-23090 v133 aAdhesive interactions between molecules on tumor cells and those on target organs play a key role in organ specific metastasis. Poly-N-acetyl-lactosamine (polyLacNAc) substituted N-oligosaccharides on melanoma cell surface glycoproteins promote lung specific metastasis via galectin-3 by facilitating their arrest and extravasation. This study reports the identification and characterization of galectin-3 interacting proteins using a combination of galectin-3 sepharose affinity and leucoagglutinating phytohemagglutinin (L-PHA) columns. A total of 83 proteins were identified as galectin-3 interacting glycoproteins, of which 35 were constituents of the L-PHA bound fraction, suggesting that these proteins carry polyLacNAc substituted beta 1,6 branched N-glycans. The identities of some of these proteins, like LAMP-1, LAMP-3, basigin, embigin, and alpha 5 and beta 1 Integrin, have been confirmed by western blotting, and functional relevance with respect to metastatic properties has been established.1 aDange, Manohar, C.1 aBhonsle, Hemangi, S.1 aGodbole, Rashmi, K.1 aMore, Shyam, K.1 aBane, Sanjay, M.1 aKulkarni, Mahesh, J.1 aKalraiya, Rajiv, D. uhttps://library.ncl.res.in/content/mass-spectrometry-based-identification-galectin-3-interacting-proteins-potentially-involved02820nas a2200241 4500008004100000245011400041210006900155260000800224300001400232490000700246520197800253100002402231700002002255700001502275700002102290700002002311700002402331700002402355700002202379700002302401700002502424856012902449 2017 eng d00aMethylglyoxal attenuates insulin signaling and downregulates the enzymes involved in cholesterol biosynthesis0 aMethylglyoxal attenuates insulin signaling and downregulates the cNOV a2338-23490 v133 a

Methylglyoxal (MG) is a highly reactive dicarbonyl known to be elevated under the hyperglycemic conditions of diabetes and is implicated in the development of diabetic complications. Therefore, the current study investigates the role of MG in exacerbating insulin resistance at the insulin signaling level, as well as its effect on the global proteomic level. By using insulin sensitive rat muscle cells (L6) and Chinese hamster ovary (CHO) cells stably expressing the insulin receptor (IR) and a glucose transporter fused with green fluorescent protein (GLUT4-GFP), we have observed that MG impairs insulin signaling, inhibits GLUT4 translocation and reduces glucose uptake. SWATH MS analysis, a label-free quantitative mass spectrometric approach, showed altered expression of 99 proteins out of 2404 identified in response to MG treatment. These proteins are mainly involved in stress response, protein folding and proteolysis. Some of the deregulated proteins such as thioredoxin 2, glutathione S transferase, T complex protein 1 subunit beta (tcbp1), heat shock protein 90 and E3 ubiquitin ligase were previously reported to be associated with either diabetes or insulin resistance. Interestingly, aminoguanidine (AMG), a potent dicarbonyl scavenger, restored the deleterious effects of MG. For the first time, we report that MG induces downregulation of enzymes involved in cholesterol biosynthesis such as acetyl-CoA acetyltransferase, hydroxymethylglutaryl-CoA synthase, farnesyl pyrophosphate synthetase, squalene monooxygenase, and lanosterol synthase. GC MS analysis for sterol metabolites corroborated the proteomic results; MG significantly reduced cholesterol production whereas AMG treatment restored cholesterol production to levels similar to the control. Thus, MG leads to primary defects in insulin signaling and cellular abnormalities at the proteomic and metabolic levels, both of which may contribute to the development of insulin resistance.

1 aDeshmukh, Arati, B.1 aBai, Shakuntala1 aAarthy, T.1 aKazi, Rubina, S.1 aBanarjee, Reema1 aRathore, Rajeshwari1 aVijayakumar, M., V.1 aThulasiram, H. V.1 aBhat, Manoj, Kumar1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/methylglyoxal-attenuates-insulin-signaling-and-downregulates-enzymes-involved-cholesterol02108nas a2200193 4500008004100000245012100041210006900162260000800231300001400239490000700253520138400260100001601644700002501660700002201685700003101707700002601738700002001764856013001784 2017 eng d00aPlacental proteomics provides insights into pathophysiology of pre-eclampsia and predicts possible markers in plasma0 aPlacental proteomics provides insights into pathophysiology of p cFEB a1050-10600 v163 a

Pre-eclampsia is a hypertensive disorder characterized by the new onset of hypertension >140/90 mmHg and proteinuria after the 20th week of gestation. The disorder is multi-factorial and originates with abnormal placentation. Comparison of the placental proteome of normotensive (n=25) and pre-eclamptic (n=25) patients by gel-free proteomic techniques, identified a total of 2145 proteins in the placenta of which 180 were differentially expressed (>1.3 fold, p<0.05). Gene ontology enrichment analysis of biological process suggested that the differentially expressed proteins belonged to various physiological processes such as angiogenesis, apoptosis, oxidative stress, hypoxia, placental development, which are implicated in the pathophysiology of pre-eclampsia. Some of the differentially expressed proteins were monitored in the plasma by multiple reaction monitoring (MRM) analysis, which showed an increase in Apolipoprotein AI and AII in gestational week 26-30th (2-fold, p<0.01) while haptoglobin and hemopexin decreased in gestational week 26-30th and 40th week/at delivery (1.8 fold, p<0.01) in pre-eclamptic patients. This study provides a proteomic insight into the pathophysiology of pre-eclampsia. Identified candidate proteins can be evaluated further for the development of potential biomarkers associated with pre-eclampsia pathogenesis.

1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMalakar, Dipankar1 aJoshi, Sadhana, Ramchandra1 aMehendale, Savita, S.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/placental-proteomics-provides-insights-pathophysiology-pre-eclampsia-and-predicts-possible01499nas a2200145 4500008004100000245007600041210006900117300001200186520093800198100002901136700002301165700003901188700002501227856010101252 2017 eng d00aTargeted quantification of the glycated peptides of human serum albumin0 aTargeted quantification of the glycated peptides of human serum  a403-4163 a

Glycated human serum albumin (HSA) serves as an important marker for monitoring the glycemic status. Developing methods for unambiguous identification and quantification of glycated peptides of HSA using high-throughput technologies such as mass spectrometry has a great clinical significance. The following protocol describes the construction of reference spectral libraries for Amadori-modified lysine (AML), N(ε)-(carboxymethyl) lysine (CML)-, and N(ε)-(carboxyethyl)lysine (CEL)-modified peptides of synthetically modified HSA using high-resolution mass spectrometers. The protocol also describes work flows, for unambiguous identification and quantification of glycated modified peptides of HSA in clinical plasma using standard spectral libraries by various mass spectrometry approaches such as parallel reaction monitoring (PRM), sequential window acquisition of all theoretical fragment ion spectra (SWATH), and MSE.

1 aGarikapati, Vannuruswamy1 aKorwar, Arvind, M.1 aJagadeeshaprasad, Mashanipalya, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/targeted-quantification-glycated-peptides-human-serum-albumin01944nas a2200181 4500008004100000245011200041210006900153260000800222300002200230490000700252520126400259100001601523700002501539700002601564700002301590700002001613856012901633 2017 eng d00aTubulointerstitial nephritis antigen-like 1 protein is downregulated in the placenta of pre-eclamptic women0 aTubulointerstitial nephritis antigenlike 1 protein is downregula cMAR aArticle Number: 80 v143 aBackground: Tubulointerstitial nephritis antigen-like 1 protein (TINAGL1), is a matricellular protein, known to play role in cell adhesion and cell receptor interaction. Research related to TINAGL1 is limited to cell culture and animal models. Demonstration of TINAGL1 as a positive regulator of angiogenesis and its expression in the decidua of postimplantation mouse uterus, prompted us to validate its expression in human placenta during impaired angiogenesis in pre-eclamptic condition. Methods: Placental tissue from normotensive (n = 25) and pre-eclamptic (n = 25) pregnancies were used to study the differentially expressed proteins by two-dimensional gel electrophoresis and TINAGL1 protein was validated with Western blotting. Results: A total of 55 protein spots were differentially expressed (fold change > 1.5, p < 0.05), of which 27 were upregulated and 28 were downregulated in the pre-eclamptic placenta. TINAGL1 was found to be downregulated in pre-eclamptic compared to normotensive pregnant women. Conclusion: This is the first study reporting TINAGL1 to be present in human placenta and differentially expressed in pre-eclamptic condition. The functional role of TINAGL1 in association to human pregnancy needs to be explored further.1 aMary, Sheon1 aKulkarni, Mahesh, J.1 aMehendale, Savita, S.1 aJoshi, Sadhana, R.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/tubulointerstitial-nephritis-antigen-1-protein-downregulated-placenta-pre-eclamptic-women01873nas a2200193 4500008004100000245017900041210006900220260000800289300001300297490000600310520111700316100002101433700002001454700002001474700001801494700002501512700002501537856011701562 2018 eng d00aAdvanced glycation end products modulate amyloidogenic APP processing and tau phosphorylation: a mechanistic link between glycation and the development of alzheimer's disease0 aAdvanced glycation end products modulate amyloidogenic APP proce cMAY a988-10000 v93 aAdvanced glycation end products (AGEs) are implicated in the pathology of Alzheimer's disease (AD), as they induce neurodegeneration following interaction with the receptor for AGE (RAGE). This study aimed to establish a mechanistic link between AGE-RAGE signaling and AD pathology. AGE-induced changes in the neuro2a proteome were monitored by SWATH-MS. Western blotting and cell-based reporter assays were used to investigate AGE-RAGE regulated APP processing and tau phosphorylation in primary cortical neurons. Selected protein expression was validated in brain samples affected by AD. The AGE-RAGE axis altered proteome included increased expression of cathepsin B and asparagine endopeptidase (AEP), which mediated an increase in A beta(1-)(42) formation and tau phosphorylation, respectively. Elevated cathepsin B, AEP, RAGE, and pTau levels were found in human AD brain, coincident with enhanced AGEs. This study demonstrates that the AGE-RAGE axis regulates A beta(1-)(42) formation and tau phosphorylation via increased cathepsin B and AEP, providing a new molecular link between AGEs and AD pathology.1 aBatkulwar, Kedar1 aGodbole, Rashmi1 aBanarjee, Reema1 aKassaar, Omar1 aWilliams, Robert, J.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/advanced-glycation-end-products-modulate-amyloidogenic-app-processing-and-tau02404nas a2200169 4500008004100000245007900041210006900120260000800189300001600197490000600213520177800219100003901997700003102036700002902067700002502096856011302121 2018 eng d00a Albumin abundance and its glycation Status determine hemoglobin glycation0 aAlbumin abundance and its glycation Status determine hemoglobin  cOCT a12999-130080 v33 aDiabetes diagnosis and management majorly depend upon the measurement of glycated hemoglobin (HbAlc) levels. Various factors influence HbAlc levels such as the use of various analytical methods and the presence of various clinical conditions. Plasma albumin levels were known to be negatively associated with HbAlc. However, the precise mechanism by which they affect HbAlc is not well' understood. Therefore, we have studied the influence of albumin levels and its glycation status on hemoglobin glycation using erythrocyte culture experiments. Erythrocytes maintained at low albumin concentration exhibited relatively increased albumin and hemoglobin glycation as compared to that in those maintained at higher albumin concentration. Increase in albumin glycation may decrease its ability to protect hemoglobin glycation. This was demonstrated by treatment of erythrocytes with N(epsilon-(carboxymethyl)lysine-modified serum albumin (CMSA), which failed to protect hemoglobin glycation; instead, it increased hemoglobin glycation. The inability of CMSA to reduce hemoglobin glycation was due to the lack of free lysine residues of albumin, which was corroborated by using N(epsilon-(acetyI)lysine serum albumin (AcSA) and clinical diabetic plasma. This is the first study which demonstrates that the modification of lysine residues of albumin impairs its ability to inhibit hemoglobin glycation. Furthermore, correlation studies between HbAlc and albumin levels or relative albumin fructosamine from clinical subjects supported our experimental finding that albumin abundance and its glycation status influence hemoglobin glycation. Therefore, we propose albumin level and its glycation status to be quantified in conjunction with HbAlc for better management of diabetes.1 aJagadeeshaprasad, Mashanipalya, G.1 aVenkatasubramani, Vinashya1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/albumin-abundance-and-its-glycation-status-determine-hemoglobin-glycation02086nas a2200157 4500008004100000245014300041210006900184260000800253520141500261100002401676700002101700700002501721700002801746700002201774856013201796 2018 eng d00aChemo-biological evaluation of antidiabetic activity of M entha arvensis L. and it's role in inhibition of advanced glycation end products0 aChemobiological evaluation of antidiabetic activity of M entha a cFEB3 a

Background: There has been enormous curiosity in the development of alternative plant based medicines to control diabetes, oxidative stress and related disorders. One of the therapeutic approaches is to reduce postprandial release of glucose in the blood. Two key enzymes that are involved in reducing postprandial glucose are α-amylase and α-glucosidase. Mentha arvensis L. has been traditionally used by several tribes as a medicinal plant to treat various disorders. Objective: The present study was undertaken to test M. arvenisis L. for inhibition of postprandial hyperglycemia. Material and method: We performed various in vitro and in vivo tests to evaluate efficacy of M. arvenisis L. for antidiabetic activity (postprandial hyperglycemia). Results: Methanolic extract of M. arvensis L. leaves showed DPPH free radical scavenging activity (more than 78% μg/μl) and high antiglycation potential (more than 90% inhibition of AGE formation). Methanolic extract also showed remarkable inhibitory effects on α-amylase (more than 50% μg/μl) and α-glucosidase (68% μg/μl) and significant inhibition of postprandial hyperglycemia in starch induced diabetic Wistar rats. Conclusion: The non-insulin dependent antidiabetic or inhibition of postprandial hyperglycemic activity of methanolic extract of M. arvensis L. leaves was shown by using in vitro and in vivo approaches in the present study.

1 aAgawane, Sachin, B.1 aGupta, Vidya, S.1 aKulkarni, Mahesh, J.1 aBhattacharya, Asish, K.1 aKoratkar, Santosh uhttps://library.ncl.res.in/content/chemo-biological-evaluation-antidiabetic-activity-m-entha-arvensis-l-and-its-role-inhibition02639nas a2200265 4500008004100000022001400041245006800055210006800123260000800191300001300199490000700212520181400219100002002033700001802053700002102071700001702092700002102109700002002130700001902150700002902169700002502198700001802223700003202241856010002273 2018 eng d a1369-378600aLimonene inhibits Candida albicans growth by inducing apoptosis0 aLimonene inhibits Candida albicans growth by inducing apoptosis cJUL a 565-5780 v563 a

Anti-Candida potential of limonene was evaluated against planktonic growth, biofilm (adhesion, development and maturation) and morphogenesis of Candida albicans in this study. Limonene is a major constituent of citrus oil and most frequently used terpene in food and beverage industry due to its pleasant fragrance, nontoxic, and is generally recognized as safe (GRAS) flavoring agent as well as treatment option in many gastrointestinal diseases. Limonene exhibited excellent anti-Candida activity and was equally effective against planktonic growth of C. albicans isolates differentially susceptible to FLC (N = 35). Limonene inhibited morphogenesis significantly at low concentration. However, it showed stage dependent activity against biofilm formation, that is, it was more effective against adhesion followed by development and maturation. Limonene also exhibited excellent synergy with FLC against planktonic and biofilm growth. SWATH-MS analysis led to identification of limonene responsive proteins that provided molecular insight of its anti-Candida activity. Proteomic analysis revealed upregulation of proteins involved in cell wall glucan synthesis (Kre6); oxidative stress (Rhr2, Adh7 and Ebp1); DNA damage stress (Mbf1 and Npl3); nucleolar stress (Rpl11, Rpl7, Rpl29, Rpl15) and down regulation of cytoskeleton organization (Crn1, Pin3, Cct8, Rbl2), and so forth, in response to limonene. Limonene mediated down regulation of Tps3 indicates activation of caspase (CaMca1) and induction of apoptosis in C. albicans. These results suggest that limonene inhibits C. albicans growth by cell wall/membrane damage induced oxidative stress that leads to DNA damage resulting into modulation of cell cycle and induction of apoptosis through nucleolar stress and metacaspase dependent pathway.

1 aThakre, Archana1 aZore, Gajanan1 aKodgire, Santosh1 aKazi, Rubina1 aMulange, Shradha1 aPatil, Rajendra1 aShelar, Amruta1 aSanthakumari, Bayitigeri1 aKulkarni, Mahesh, J.1 aKharat, Kiran1 aKaruppayil, Sankunny, Mohan uhttps://library.ncl.res.in/content/limonene-inhibits-candida-albicans-growth-inducing-apoptosis02887nas a2200169 4500008004100000245008700041210006900128260000800197520226400205100002002469700001802489700002502507700002502532700002202557700003002579856010802609 2018 eng d00aPatho-physiological evaluation of Duranta erecta for the treatment of urolithiasis0 aPathophysiological evaluation of Duranta erecta for the treatmen cMAY3 a

Background: Urolithiasis is the third common disorder of the urinary system affecting 10-15% of the general population. In recent years, search for new antilithiatic drugs from natural sources has assumed greater importance. Objectives: This study was performed to investigate the anti-urolithiatic activity of methanolic extract of Duranta erecta leaves by in vitro and in vivo analysis. Materials and methods: The study was designed to determine presence of phytochemicals in D. erecta, its yield in percentage, antioxidant activity against 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and anti-microbial property against few bacteria. In vitro analysis was carried out study anti-urolithiatic property of D. erecta by nucleation assay and synthetic urine assay for inhibition of calcium oxalate and calcium oxalate monohydrate crystals formation. An in vivo experiment was performed on Wistar rats for confirmation of anti-urolithiatic property of D. erecta in animal model. Results: D. erecta has the presence of primary and secondary metabolites like glycoside, saponins, sterols, flavonoids, phenols, tannins, alkaloids, carbohydrates and proteins. Methanolic extract of D. erecta gave a very good yield (60%). D. erecta proved its antioxidant potential by 93.51% inhibition of DPPH radical at a concentration of 1000 μg/mL where ascorbic showed 94.71% of DPPH radical at the same concentration. In vitro tests like nucleation assay and synthetic urine assay showed that D. erecta inhibits formation of calcium oxalate and calcium oxalate monohydrate crystals. It also showed the anti-microbial property by formation of zone of inhibition against few bacteria. An in vivo experiment on Wistar rat animal model confirmed the anti-urolithiatic property of D. erecta L. leaves extract. Conclusions: Based on the results, we reported that D. erecta may treat calcium oxalate crystal deposition in the kidney by preventing hyperoxaluria-induced peroxidative damage to the renal tubular membrane surface (lipid peroxidation). It has anti-microbial potential so it may also inhibit the secondary bacterial infection in kidney. Based on the data, it can be concluded that this herb can be used as a potential anti-urolithiasis agent for kidney stone removal.

1 aAgawane, S., B.1 aGupta, V., S.1 aKulkarni, Mahesh, J.1 aBhattacharya, A., K.1 aKoratkar, Santosh1 aRao, Vankudoth, Koteswara uhttps://library.ncl.res.in/content/patho-physiological-evaluation-duranta-erecta-treatment-urolithiasis02974nas a2200181 4500008004100000245012600041210006900167260000800236300001000244490000800254520230300262100002002565700001902585700002002604700002002624700002502644856012302669 2018 eng d00aProteomic study of endothelial dysfunction induced by Ages and its possible role in diabetic cardiovascular complications0 aProteomic study of endothelial dysfunction induced by Ages and i cSEP a69-790 v1873 a

Endothelial dysfunction is one of the primary steps in the development of diabetes associated cardiovascular diseases. Hyperglycemic condition in diabetes promotes accumulation of advanced glycation end products (AGEs) in the plasma, that interact with the receptor for AGEs (RAGE) present on the endothelial cells and negatively affect their function. Using Human umbilical vascular endothelial cells (HUVECs) in culture, the effect of glycated human serum albumin on global proteomic changes was studied by SWATH-MS, a label free quantitative proteomic approach. Out of the 1860 proteins identified, 161 showed higher abundance while 123 showed lesser abundance in cells treated with glycated HSA. Bioinformatic analysis revealed that the differentially regulated proteins were involved in various processes such as apoptosis, oxidative stress etc. that are associated with endothelial dysfunction. Furthermore, the iRegulon analysis and immunofuorescence studies indicated that several of the differentially regulated proteins were transcriptionally regulated by NF-kappa B, that is downstream to AGE-RAGE axis. Some of the important differentially regulated proteins include ICAM1, vWF, PAI-1 that affect important endothelial functions like cell adhesion and blood coagulation. qPCR analysis showed an increase in expression of the AGE receptor RAGE along with other genes involved in endothelial function. AGE treatment to HUVEC cells led to increased oxidative stress and apoptosis. This is the first proteomics study that provides insight into proteomic changes downstream to AGE-RAGE axis leading to endothelial dysfunction and predisposing to cardiovascular complications. Significance: Cardiovascular disease (CVD) is a major pathological outcome in diabetic patients and it is important to address ways that target its development before the onset. Elevated plasma AGEs in diabetes can affect endothelial function and can continue to show their effects even after blood glucose levels are back to normal. Since endothelial dysfunction acts as one of the initiating factors for the development of CVD, understanding how AGEs affect the endothelial cell proteome to cause dysfunction will provide insight into the mechanisms involved and aid designing new therapeutic approaches.

1 aBanarjee, Reema1 aSharma, Akshay1 aBai, Shakuntala1 aDeshmukh, Arati1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteomic-study-endothelial-dysfunction-induced-ages-and-its-possible-role-diabetic01523nas a2200169 4500008004100000022001400041245006500055210006400120260000800184300001200192490000600204520098500210100001801195700002501213700001801238856009701256 2018 eng d a2040-250300aRevisiting amino acids and peptides as anti-glycation agents0 aRevisiting amino acids and peptides as antiglycation agents cAPR a614-6240 v93 a

The importance of controlling or preventing protein glycation cannot be overstated and is of prime importance in the treatment of diabetes and associated complications including Alzheimer's disease, cataracts, atherosclerosis, kidney aliments among others. In this respect, simple molecules such as amino acids and peptides hold much promise both in terms of ease and scale-up of synthesis as well as in relation to negligible/low associated toxicity. In view of this, a comprehensive account of literature reports is presented, that documents the anti-glycation activity of natural and non-natural amino acids and peptides. This review also discusses the chemical reactions involved in glycation and the formation of advanced glycation end-products (AGEs) and possible/probable intervention sites and mechanism of action of the reported amino acids/peptides. This aspect of amino acids/peptides adds to their growing importance in medicinal and therapeutic applications.

1 aChilukuri, H.1 aKulkarni, Mahesh, J.1 aFernandes, M. uhttps://library.ncl.res.in/content/revisiting-amino-acids-and-peptides-anti-glycation-agents02763nas a2200289 4500008004100000022001400041245013200055210006900187260000800256300001000264490000800274520177900282653001302061653002102074653002202095653004402117653001802161653003002179653001802209100001802227700002102245700002102266700002002287700002502307700001802332856012302350 2018 eng d a1874-391900aSerum responsive proteome reveals correlation between oxidative phosphorylation and morphogenesis in Candida albicans ATCC102310 aSerum responsive proteome reveals correlation between oxidative  cAUG a25-380 v1853 a

To understand the impact of fetal bovine serum (FBS) on metabolism and cellular architecture in addition to morphogenesis, we have identified FBS responsive proteome of Candida albicans. FBS induced 34% hyphae and 60% pseudohyphae in C. albicans at 30 C while 98% hyphae at 37 C. LC-MS/MS analysis revealed that 285 proteins modulated significantly in response to FBS at 30 degrees C and 37 degrees C. Out of which 152 were upregulated and 62 were downregulated at 30 degrees C while 18 were up and 53 were downregulated at 37 degrees C. Functional annotation suggests that FBS may inhibit glycolysis and fermentative pathway and enhance oxidative phosphorylation (OxPhos), TCA cycle, amino acid and fatty acid metabolism indicating a use of alternative energy source by C. albicans. OxPhos inhibition assay using sodium azide corroborated the correlation between inhibition of glycolysis and enhanced OxPhos with pseudohyphae formation. C. albicans induced hyphae in response to FBS irrespective of down regulation of Rasl,Asrl/Asr2, indicates the possible involvement of MAPK and cAMP-PKA independent pathway. The Cell wall of cells grown in presence of FBS at 30 degrees C was rich in mannan, Beta 1,3glucan and chitin while membranes were rich in ergosterol compared to those grown at 37 degrees C. Significance of the study: This is the first study suggesting a correlation between OxPhos and morphogenesis especially pseudohyphae formation in C. albicans. Our data also indicate that fetal bovine serum (FBS) induced morphogenesis is multifactorial and may involve MAPK and cAMP-PKA independent pathway. In addition to morphogenesis, our study provides an insight in to the modulation of metabolism and cellular architecture of C. albicans in response to FBS.

10aanalysis10aCandida albicans10aFetal bovin serum10aLiquid chromatography mass spectrometry10amorphogenesis10aOxidative phosphorylation10apathogenicity1 aAhmed, Radfan1 aKodgire, Santosh1 aSanthakumari, B.1 aPatil, Rajendra1 aKulkarni, Mahesh, J.1 aZore, Gajanan uhttps://library.ncl.res.in/content/serum-responsive-proteome-reveals-correlation-between-oxidative-phosphorylation-and02850nas a2200181 4500008004100000245011200041210006900153260000800222520215600230100001602386700002002402700002902422700001402451700002102465700002502486700002502511856013202536 2019 eng d00aGlycation of glucose sensitive lysine residues K36, K438 and K549 of albumin is associated with prediabetes0 aGlycation of glucose sensitive lysine residues K36 K438 and K549 cAUG3 a

Prediabetes is a risk factor for the development of diabetes. Early diagnosis of prediabetes may prevent the onset and progression of diabetes and its associated complications. Therefore, this study aimed at the identification of novel markers for efficient prediction of prediabetes. In this pursuit, we have evaluated the ability of glycated peptides of albumin in predicting prediabetes. Glycated peptides of in vitro glycated albumin were characterized by data dependent acquisition and parallel reaction monitoring using LC-HRMS. Amongst 14 glycated peptides characterized in vitro, four peptides, particularly, FK(CML)DLGEENFK, K(AML)VPQVSTPTLVEVSR, K(CML)VPQVSTPTLVEVSR, and K(AML)QTALVELVK, corresponding to 3 glucose sensitive lysine residues K36, K438, and K549, respectively showed significantly higher abundance in prediabetes than control. Additionally, the abundance of three of these peptides, namely K(AML)QTALVELVK, K(CML)VPQVSTPTLVEVSR and FK(CML)DLGEENFK was >1.8-fold in prediabetes, which was significantly higher than the differences observed for FBG, PPG, and HbA1c. Further, the four glycated peptides showed a significant correlation with FBG, PPG, HbA1c, triglycerides, VLDL, and HDL. This study supports that glycated peptides of glucose sensitive lysine residues K36, K438 and K549 of albumin could be potentially useful markers for prediction of prediabetes. SIGNIFICANCE: Undiagnosed prediabetes may lead to diabetes and associated complications. This study reports targeted quantification of four glycated peptides particulary FK(CML)DLGEENFK, K(AML)VPQVSTPTLVEVSR, K(CML)VPQVSTPTLVEVSR, and K(AML)QTALVELVK, corresponding to 3 glucose sensitive lysine residues K36, K438 and K549 respectively by parallel reaction monitoring in healthy and prediabetic subjects. These peptides showed significantly higher abundance in prediabetes than healthy subjects, and showed significant correlation with various clinical parameters including FBG, PPG, HbA1c, and altered lipid profile. Therefore, together these four peptides constitute a panel of markers that can be useful for prediction of prediabetes.

1 aRathore, R.1 aSonwane, B., P.1 aJagadeeshaprasad, M., G.1 aKahar, S.1 aSanthakumari, B.1 aUnnikrishnan, A., G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/glycation-glucose-sensitive-lysine-residues-k36-k438-and-k549-albumin-associated-prediabetes02264nas a2200217 4500008004100000022001400041245011500055210006900170260000900239300001600248490000600264520148300270100002001753700003201773700002001805700002201825700001801847700002901865700002501894856012701919 2020 eng d a2470-134300aCD44, a predominant protein in methylglyoxal-induced secretome of muscle cells, is elevated in diabetic plasma0 aCD44 a predominant protein in methylglyoxalinduced secretome of  cOCT  a25016-250280 v53 a

Methylglyoxal (MG), a glycolytic intermediate and reactive dicarbonyl, is responsible for exacerbation of insulin resistance and diabetic complication. In this study, NIG-induced secretome of rat muscle cells was identified and relatively quantified by SWATH-MS. A total of 643 proteins were identified in MG induced' secretome, of which 82 proteins were upregulated and 99 proteins were downregulated by more than 1.3-fold in SWATH analysis. Further, secretory proteins from the dassical secretory pathway and nonclassical secretory pathway were identified using SignalP and SecretomeP, respectively. A total of 180 proteins were identified with SignalP, and 113 proteins were identified with SecretomeP. The differentially expressed proteins were functionallyannotated by KEGG pathway analysis using Cytoscape soft ware with plugin clusterMaker. The differentially expressed proteins were'sfound to he involved m various pathways like extracellular matrix (ECM) receptor interaction, leukocyte transendothelial migration, fluid shear tress and atherosclerosis, complement and coagulation cascades, and lysosomal pathway. Since the MG levels are high in diabetic conditions, the presence of MG-induced isecreted proteins was inspected `by profiling human plasma of healthy and diabetic subjects (n = 10 each). CD44, a predominant MG -induced secreted protein, was found to be elevated in the diabetic plasma and to have a role in the development of insulin resistance.

1 aBai, Shakuntala1 aChaurasiya, Arvindkumar, H.1 aBanarjee, Reema1 aWalke, Prachi, B.1 aRashid, Faraz1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/cd44-predominant-protein-methylglyoxal-induced-secretome-muscle-cells-elevated-diabetic02148nas a2200193 4500008004100000245010200041210006900143260000900212300001200221490000700233520143900240100002701679700002301706700002401729700002501753700002701778700002501805856012401830 2020 eng d00aDevelopment of multiple reaction monitoring assay for quantification of carnosine in human plasma0 aDevelopment of multiple reaction monitoring assay for quantifica cJAN  a763-7690 v103 a

Carnosine, a histidine containing dipeptide, exerts beneficial effects by scavenging reactive carbonyl compounds (RCCs) that are implicated in pathogenesis of diabetes. However, the reduced carnosine levels may aggravate the severity of diabetes. The precise quantification of carnosine levels may serve as an indicator of pathophysiological state of diabetes. Therefore, we have developed a highly sensitive targeted multiple reaction monitoring (MRM) method for quantification of carnosine in human plasma samples. Various mass spectrometry parameters such as ionization of precursor, fragment abundance and stability, collision energy, tube lens offset voltage were optimized to develop a sensitive and robust assay. Using the optimized MRM assay, the lower limit of detection (LOD) and limit of quantification (LOQ) for carnosine were found to be 0.4 nM and 1.0 nM respectively. Standard curves were constructed ranging from 1.0 nM to 15.0 mu M and the levels of carnosine in mice and human plasma were determined. Further, the MRM assay was extended to study carnosine hydrolyzing activity of human carnosinases, the serum carnosinase (CN1) and the cytosolic carnosinase (CN2). CN1 showed three folds higher activity than CN2. The MRM assay developed in this study is highly sensitive and can be used for basal plasma carnosine quantification, which can be developed as a novel marker for scavenging of RCCs in diabetes.

1 aPandya, Vaibhav, Kumar1 aSonwane, Babasaheb1 aRathore, Rajeshwari1 aUnnikrishnan, A., G.1 aKumaran, Sangaralingam1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/development-multiple-reaction-monitoring-assay-quantification-carnosine-human-plasma02932nas a2200313 4500008004100000022001400041245013200055210006900187260000800256520189100264653001602155653001302171653001102184653000902195653003302204653002302237653000902260653002102269100002202290700002502312700002302337700001902360700002602379700002702405700001702432700002002449700002502469856012402494 2020 eng d a0739-110200aDiscovery of potential multi-target-directed ligands by targeting host-specific SARS-CoV-2 structurally conserved main protease0 aDiscovery of potential multitargetdirected ligands by targeting  cMAY3 a

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has resulted in the current COVID-19 pandemic. Worldwide this disease has infected over 2.5 million individuals with a mortality rate ranging from 5 to 10%. There are several efforts going on in the drug discovery to control the SARS-CoV-2 viral infection. The main protease (M-Pro) plays a critical role in viral replication and maturation, thus can serve as the primary drug target. To understand the structural evolution of M-Pro, we have performed phylogenetic and Sequence Similarity Network analysis, that depicted divergence of Coronaviridae M-Pro in five clusters specific to viral hosts. This clustering was corroborated with the comparison of M-Pro structures. Furthermore, it has been observed that backbone and binding site conformations are conserved despite variation in some of the residues. These attributes can be exploited to repurpose available viral protease inhibitors against SARS-CoV-2 M-Pro. In agreement with this, we performed screening of similar to 7100 molecules including active ingredients present in the Ayurvedic anti-tussive medicines, anti-viral phytochemicals and synthetic anti-virals against SARS-CoV-2 M-Pro as the primary target. We identified several natural molecules like delta-viniferin, myricitrin, taiwanhomoflavone A, lactucopicrin 15-oxalate, nympholide A, afzelin, biorobin, hesperidin and phyllaemblicin B that strongly binds to SARS-CoV-2 M-Pro. Intrestingly, these molecules also showed strong binding with other potential targets of SARS-CoV-2 infection like viral receptor human angiotensin-converting enzyme 2 (hACE-2) and RNA dependent RNA polymerase (RdRp). We anticipate that our approach for identification of multi-target-directed ligand will provide new avenues for drug discovery against SARS-CoV-2 infection. Communicated by Ramaswamy H. Sarma

10aCoronavirus10aCOVID-1910ahACE-210aMPro10amulti-target-directed ligand10aprotease inhibitor10aRdRp10aSARS-CoV-2 virus1 aJoshi, Rakesh, S.1 aJagdale, Shounak, S.1 aBansode, Sneha, B.1 aShankar, Shiva1 aTellis, Meenakshi, B.1 aPandya, Vaibhav, Kumar1 aChugh, Anita1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/discovery-potential-multi-target-directed-ligands-targeting-host-specific-sars-cov-202789nas a2200289 4500008004100000022001400041245015800055210006900213260000800282300001400290490000700304520175600311653002102067653001702088653003302105653002102138653002002159100002502179700002102204700003202225700002202257700002002279700002502299700002202324700002402346856012902370 2020 eng d a0721-759500aEnzymatic and non-enzymatic detoxification of reactive carbonyl compounds improves the oxidative stress tolerance in cucumber, tobacco and rice seedlings0 aEnzymatic and nonenzymatic detoxification of reactive carbonyl c cSEP a1359-13720 v393 a

Detoxification of reactive carbonyl compounds (RCC) is crucial to sustain cellular activity to improve plant growth and development. Seedling growth is highly affected by accumulation of RCC under stress. We report non-enzymatic, enzymatic mechanisms of detoxification of RCC in the cucumber, tobacco and rice seedling systems exposed to glucose, NaCl, methyl viologen (MV) induced oxidative stress. The cucumber seedlings exposed to carbonyl stress had higher levels of malondialdehyde (MDA), protein carbonyls (PCs) and advanced glycation end-product N-carboxymethyl-lysine (AGE-CML) that negatively affected the seedling growth. The overexpression of enzyme encoding aldo-keto reductase-1 (AKR1) in tobacco and rice showed detoxification of RCC, MDA and methylglyoxal (MG) with improved seedling growth under glucose, NaCl and MV-induced oxidative stress. Further, small molecules like acetylsalicylic acid (ASA), aminoguanidine (AG), carnosine (Car), curcumin (Cur) and pyridoxamine (PM) showed detoxification of RCC non-enzymatically and rescued the cucumber seedling growth from glucose, NaCl and MV-stress. In autotrophically grown rice seedlings these molecules substantially improved seedling growth under MV-induced oxidative stress. Seedlings treated with the small molecules sustained higher guaiacol peroxidase (GPX) enzyme activity signifying the role of small molecules in reducing carbonyl stress-induced protein inactivation and AGE-CML protein modifications. The results showed that besides enzymatic detoxification of RCC, the small molecules also could reduce cytotoxic effect of RCC under stress. The study demonstrates that small molecules are attractive compounds to improve the seedling growth under stress conditions.

10aOxidative stress10aplant growth10aReactive carbonyl scavengers10aSeed germination10asmall molecules1 aNareshkumar, Ambekar1 aSubbarao, Sindhu1 aVennapusa, Amarnatha, Reddy1 aAshwin, Vargheese1 aBanarjee, Reema1 aKulkarni, Mahesh, J.1 aRamu, Vemanna, S.1 aUdayakumar, Makarla uhttps://library.ncl.res.in/content/enzymatic-and-non-enzymatic-detoxification-reactive-carbonyl-compounds-improves-oxidative02148nas a2200301 4500008004100000022001400041245013100055210006900186260000800255300001100263490000800274520114300282653002001425653002501445653001701470653003501487653001901522653001301541653001201554653001401566653002001580100002101600700002201621700002101643700002501664700002501689856013201714 2020 eng d a0306-987700aPossible role of glycation in the regulation of amyloid beta precursor protein processing leading to amyloid beta accumulation0 aPossible role of glycation in the regulation of amyloid beta pre cSEP a1097990 v1423 a

Alzheimer's disease (AD) is one of the most common forms of neurodegenerative diseases amongst the aged population. The disease is multifactorial, and diabetes has been considered as one of the major risk factors for the development of AD. Chronic hyperglycemic condition in diabetes promotes non-enzymatic protein modification by glucose termed as glycation, which affects protein structure and function. Previous studies have shown that many of the enzymes, including proteases, are affected by glycation. Conversely, glycated proteins are known to become resistant to protease action. In these hypotheses, we have extended these two concepts to the regulation of amyloid-beta protein precursor (A beta PP) by secretases leading to amyloid-beta (Af3) accumulation. The first hypothesis deals with the glycation of alpha-secretases leading to its reduced activity, while in the second hypothesis, A beta PP glycation may prevent a-secretases action, rendering its processing by beta secretase. As diabetes is a risk factor for the development of AD, either or both these pathways may operate, leading to the manifestation of AD.

10aAlpha-secretase10aAlzheimer `s disease10aAmyloid-beta10aAmyloid-beta protein precursor10aBeta-secretase10aDiabetes10aglucose10aGlycation10aType-3 diabetes1 aPatil, Gouri, V.1 aJoshi, Rakesh, S.1 aKazi, Rubina, S.1 aKulsange, Shabda, E.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/possible-role-glycation-regulation-amyloid-beta-precursor-protein-processing-leading-amyloid00794nas a2200193 4500008004100000022001400041245016300055210006900218260000800287300001100295490000800306100002400314700002900338700002900367700002500396700002900421700002400450856012600474 2020 eng d a1874-391900aProteomic analysis detects deregulated reverse cholesterol transport in human subjects with ST-segment elevation myocardial infarction (vol 222, 103796, 2020)0 aProteomic analysis detects deregulated reverse cholesterol trans cJUL a1038280 v2241 aDas, Apabrita, Ayan1 aChoudhury, Kamalika, Roy1 aJagadeeshaprasad, M., G.1 aKulkarni, Mahesh, J.1 aMondal, Prakash, Chandra1 aBandyopadhyay, Arun uhttps://library.ncl.res.in/content/proteomic-analysis-detects-deregulated-reverse-cholesterol-transport-human-subjects-st03322nas a2200277 4500008004100000022001400041245013900055210006900194260000800263300001100271490000800282520236100290653002002651653001002671653000902681653002202690653003402712653001002746100002402756700002902780700002902809700002502838700002902863700002402892856012802916 2020 eng d a1874-391900aProteomic analysis detects deregulated reverse cholesterol transport in human subjects with ST-segment elevation myocardial infarction0 aProteomic analysis detects deregulated reverse cholesterol trans cJUN a1037960 v2223 a

Reverse cholesterol transport (RCT) plays a critical role in removing cholesterol from the arterial wall. However, very few reports directly relate chronic inflammation and RCT with atherosclerosis. The present study was undertaken to investigate clinical implications of significantly altered circulating proteins in subjects with ST-segment elevation myocardial infarction (STEMI) in the manifestation of atherosclerotic events. Using a casecontrol design, more than 2500 proteins in both STEMI and healthy control subjects were identified by Orbitrap mass spectrometer. Quantitative proteomics study revealed downregulation of 26 proteins while expression of 38 proteins increased significantly in STEMI subjects compared to healthy controls. Pathway enrichment analyses indicated that most of the identified proteins were related to chronic inflammation, atherosclerosis, and RCT. Altered proteins such as AZGP1, ABCA5, Calicin, PGLYRP2, HAVCR2 and C17ORF57 were further validated by Western blotting analysis of human plasma. Pathophysiological significance was studied using macrophage derived foam cell for their critical role in RCT which indicated the imbalance of RCT via the interaction of AZGP1 with CD36. In summary, this study revealed a unique relationship of some novel proteins apparently responsible for impaired RCT and chronic inflammation leading to atherothrombosis and myocardial infarction. Significance: In the present study we identified >= 2500 unique circulating proteins in healthy control and clinically diagnosed STEMI subjects among which 423 proteins were found to be common in both the groups. We further show 64 proteins significantly different between healthy control and STEMI subjects. Proteomic analyses reveal a panel of proteins associated with atherosclerosis and STEMI. One of the proteins, AZGP1, an adipokine, is likely to act as the missing link between chronic inflammation and cholesterol transport. Deregulation of reverse cholesterol transport might be orchestrated by AZGP1, CD36, ABCA5, and PPARy in STEMI subjects. The present study employs shotgun and quantitative proteomics followed by in vitro validations demonstrating a biochemical basis for reverse cholesterol transport in the local milieu of the luminal wall of the artery which are critical for plaque build-up and atherosclerosis.

10aAtherosclerosis10aAZGP110aCD3610aPlasma proteomics10aReverse cholesterol transport10aSTEMI1 aDas, Apabrita, Ayan1 aChoudhury, Kamalika, Roy1 aJagadeeshaprasad, M., G.1 aKulkarni, Mahesh, J.1 aMondal, Prakash, Chandra1 aBandyopadhyay, Arun uhttps://library.ncl.res.in/content/proteomic-analysis-detects-deregulated-reverse-cholesterol-transport-human-subjects-st-002068nas a2200253 4500008004100000022001400041245018500055210006900240260000900309300001100318490000800329520112700337100002801464700001701492700002001509700002801529700002001557700002601577700001801603700002101621700002501642700001701667856013001684 2020 eng d a1874-391900aSWATH-MS based quantitative proteomics analysis to evaluate the antileishmanial effect of Commiphora wightii- Guggul and Amphotericin B on a clinical isolate of Leishmania donovani0 aSWATHMS based quantitative proteomics analysis to evaluate the a cJUL  a1038000 v2233 a

The present study provides comprehensive proteomics analyses of the response of L. donovani parasite to pharamacological stress in vitro. Identification of differentially expressed proteins with associated molecular functions and metabolic pathways, clearly provides an insight into the potential mechanism of the antileishmanial effects as well as a comparative response of the parasite to Guggul and AmB. Treatment of parasite with AmB results in an enhanced modulatory mechanism to counteract the drug induced stress which may have contributed to relapse. In the case of Guggul treatment, an effective antipromastigote activity was observed, which is being reported for the first time. Thus, a deeper understanding of the molecular pathways in the Leishmania parasite in response to pharmacological stress would help in designing novel and effective strategies in targeting the key molecules essential for parasite survival. It will also help in screening of new lead molecules targeting these vital pathways which could be used as an adjunct therapy along with the limited repertoire of antileishmanial drugs.

1 aBar Routaray, Chinmayee1 aBhor, Renuka1 aBai, Shakuntala1 aKadam, Nitin, Suryakant1 aJagtap, Surabhi1 aDoshi, Pooja, Jignesh1 aSundar, Shyam1 aSawant, Sangeeta1 aKulkarni, Mahesh, J.1 aPai, Kalpana uhttps://library.ncl.res.in/content/swath-ms-based-quantitative-proteomics-analysis-evaluate-antileishmanial-effect-commiphora02681nas a2200349 4500008004100000022001400041245017000055210006900225260000800294300001200302490000700314520143400321653004401755653001501799653002701814653002901841653001701870100002201887700002801909700001801937700002801955700002901983700001902012700001802031700002402049700002602073700002502099700002102124700002702145700003302172856012602205 2020 eng d a1615-985300aTriton X-114 fractionated subcellular proteome ofleptospira interrogansshows selective enrichment of pathogenic and outer membrane proteins in the detergent fraction0 aTriton X114 fractionated subcellular proteome ofleptospira inter cOCT a20001700 v203 a

The Triton X-114-based solubilization and temperature-dependent phase separation of proteins is used for subcellular fractionation where, aqueous, detergent, and pellet fractions represents cytoplasmic, outer membrane (OM), and inner membrane proteins, respectively. Mass spectrometry-based proteomic analysis of Triton X-114 fractions of proteomic analysis ofLeptospira interrogansidentified 2957 unique proteins distributed across the fractions. The results are compared with bioinformatics predictions on their subcellular localization and pathogenic nature. Analysis of the distribution of proteins across the Triton X-114 fractions with the predicted characteristics is performed based on ``number'' of unique type of proteins, and ``quantity'' which represents the amount of unique protein. The highest number of predicted outer membrane proteins (OMPs) and pathogenic proteins are found in aqueous and pellet fractions, whereas detergent fraction representing the OM has the highest quantity of OMPs and pathogenic proteins though lower in number than the aqueous and pellet fractions. This leaves the possibility of an upsurge in pathogenic proteins and OMPs on the OM under pathogenic conditions suggesting their potential use to combat leptospirosis. Further, the Triton X-114 subcellular fractions are more correlated to enrichment of pathogenic proteins predicted by MP3 software than predicted localization.

10aintensity-based absolute quantification10aLeptospira10aouter membrane protein10asubcellular localization10aTriton X-1141 aThoduvayil, Sikha1 aDhandapani, Gunasekaran1 aBrahma, Rahul1 aBalaya, Rex, Devasahaya1 aMangalaparthi, Kiran, K.1 aPatel, Krishna1 aKumar, Manish1 aTennyson, Jebasingh1 aSatheeshkumar, P., K.1 aKulkarni, Mahesh, J.1 aPinto, Sneha, M.1 aPrasad, T., S. Keshava1 aMadanan, Madathiparambil, G. uhttps://library.ncl.res.in/content/triton-x-114-fractionated-subcellular-proteome-ofleptospira-interrogansshows-selective02341nas a2200289 4500008004100000022001400041245008700055210006900142260000900211300001400220490000800234520143200242653002101674653001101695653001801706653003301724653001501757100002201772700002001794700002501814700001801839700001801857700002101875700002301896700001701919856011501936 2021 eng d a0011-389100aAntiviral drugs prioritization for COVID-19 management based on rational selection0 aAntiviral drugs prioritization for COVID19 management based on r cMAY  a1464-14700 v1203 a

The SARS-CoV-2 infection has resulted in COVID-19 pandemic worldwide. It has infected around 0.1 billion individuals and caused 2 million fatalities across the globe till mid-January 2021. Drug repurposing has been utilized as the most preferred therapeutic intervention for COVID-19 mitigation due to its necessity and feasibility. To prioritize therapeutic regime against COVID-19, we used 61 antiviral drugs and their combinations. Selected molecules were subjected to virtual screening against: (i) human angiotensin-converting enzyme 2 receptor binding domain (hACE-2) which serves as an anchor for virus attachment and entry, (ii) SARS-CoV-2 RNA dependent RNA polymerase (RdRp) responsible for viral RNA replication, and (iii) SARS-CoV-2 main protease (M-Pro) needed for viral polyprotein slab proteolytic processing. Based on docking score, pharmacodynamic and pharmacokinetic parameters, combinations of Daclatasvir, Elbasvir, Indinavir, Ledipasvir, Paritaprevir and Rilpivirine were analysed further. Our analysis suggested Sofosbuvir in combination with Ledipasvir and Daclatasvir as potential therapeutic agents for SARS-CoV-2. The combined score suggests that these combinations have superior anti-SARS-CoV-2 potential than Remdesivir and other investigational drugs. The present work provides a rationale-based approach to select drugs with possible anti-SARS-CoV-2 activity for further clinical evaluation.

10aDrug repurposing10ahACE-210amain protease10aRNA dependent RNA polymerase10aSARS-CoV-21 aJoshi, Rakesh, S.1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J.1 aGupta, Mahesh1 aVerma, Savita1 aChaudhry, Dhruva1 aDeshmukh, Narendra1 aChugh, Anita uhttps://library.ncl.res.in/content/antiviral-drugs-prioritization-covid-19-management-based-rational-selection03472nas a2200181 4500008004100000245013100041210006900172260000800241300001100249490000800260520278500268100001903053700002803072700002403100700002503124700001403149856012703163 2021 eng d00aComparison of glycoprofiles of rituximab versions licensed for sale in India and an analytical approach for quality assessment0 aComparison of glycoprofiles of rituximab versions licensed for s cJUL a1042670 v2443 a

Glycosylation affects clinical efficacy and safety; therefore, is a critical quality attribute of therapeutic monoclonal antibodies. Glycans are often labile and complex in patterns, giving rise to macro- and micro-heterogeneity. Recombinant production, diverse geographical locations, associated transportation and storage conditions further compound the problem. Two-way studies comparing glycoprofile of the originator and its given biosimilar are aplenty. However, the extent of analytical variation and similarity in glycoprofile across all approved versions of a drug is hardly explored. Using UHPLC and mass spectrometry, we compared the glycoprofiles of eight rituximab drug samples licensed for sale in India. While the types of glycans were found identical, the abundance of some glycans varied significantly within the tested population. The quality range of glycosylation parameters of the tested sample population differed significantly from the previously established values for US/EU licensed rituximab. As the mean abundance of the 90% of identified glycans falls within ±3SD, the extent of mutual variations amongst tested lots is less significant compared to the extreme deviation from previously established QR limits. Thus, we propose this approach as an orthogonal method to capture glycan variations in licensed versions of mAbs for quality surveillance and in cases where originator samples' are limiting. SIGNIFICANCE: As fluctuation in glycosylation may be of clinical significance, we identify that a one-to-one comparison with originator alone is insufficient in sensing the extent of variations in glycosylation parameters in licensed biosimilars of a given therapeutic mAb. Here we propose that future biosimilarity analysis may include an orthogonal approach of generating an additional combined QR range representing variations across the originator and its biosimilars. The glycosylation profiles of eight rituximab drug samples of different make obtained from the point of sale in India were found identical amongst the tested rituximab versions. However, the QR limits corresponding to important glycosylation parameters differed significantly across all tested samples from the previously established QR limits of US- and EU-licensed rituximab in statistical terms. Such an approach may be useful in defining the true range of glycan variations in licensed versions of therapeutic mAbs.

1 aKaur, Tejinder1 aShukla, Bhupendra, Nath1 aYadav, Vinay, Kumar1 aKulkarni, Mahesh, J.1 aRao, Alka uhttps://library.ncl.res.in/content/comparison-glycoprofiles-rituximab-versions-licensed-sale-india-and-analytical-approach02638nas a2200277 4500008004100000022001400041245011500055210006900170260000900239300001100248490000900259520174400268653000802012653001302020653001402033653001802047653001202065653000902077100002202086700002302108700002102131700003202152700002202184700002502206856012902231 2021 eng d a0925-443900aMolecular investigation of glycated insulin-induced insulin resistance via insulin signaling and AGE-RAGE axis0 aMolecular investigation of glycated insulininduced insulin resis cFEB  a1660290 v18673 a

Hyperglycemic condition in diabetes promotes glycation of various plasma proteins including insulin. Glycation of insulin has been reported to reduce its biological activity. Reduced biological activity of glycated insulin could be either due to reduced affinity for the insulin receptor and impaired insulin signaling, or it can act as a ligand for the receptor for advanced glycation end products (RAGE) and activates oxidative stress and pro-inflammatory pathways leading to insulin resistance. This study investigates the effect of glycated insulin on both insulin and RAGE signaling. Glycated insulin treatment to Chinese hamster ovary (CHO-IR-GLUT4) cells stably expressing insulin receptor (IR) and glucose transporter fused with a green fluorescent protein (GLUT4-GFP) resulted in the impairment of insulin signaling, as the phosphorylation of IR and AKT significantly reduced, which affected GLUT4 translocation and glucose uptake. Moreover, it also activated RAGE signaling as observed by increased expression of NADPH oxidase accompanied by an increase in reactive oxygen species (ROS). Immunofluorescence study indicated the translocation of NF-kappa B to the nucleus upon treatment of glycated insulin. This was associated with increased RAGE expression, Caspase 3, and cell death. Downregulation of RAGE with the losartan treatment restored the impaired insulin signaling and glucose uptake. Additionally, in silico study demonstrated that glycated insulin has reduced binding affinity to insulin receptor and increased binding affinity to RAGE. Overall, this study demonstrates the role of glycated insulin in exacerbating insulin resistance by impairing insulin signaling as well as stimulating AGE-RAGE signaling.

10aAGE10aDiabetes10aGlycation10aHyperglycemia10ainsulin10aRAGE1 aWalke, Prachi, B.1 aBansode, Sneha, B.1 aMore, Nikita, P.1 aChaurasiya, Arvindkumar, H.1 aJoshi, Rakesh, S.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/molecular-investigation-glycated-insulin-induced-insulin-resistance-insulin-signaling-and02995nas a2200265 4500008004100000022001400041245015900055210006900214260000800283300001100291490000800302520207900310653001302389653000802402653001402410653003802424653001602462100001902478700001802497700002702515700002102542700001402563700002502577856012702602 2021 eng d a1874-391900aMultiple-parallel-protease digestion coupled with high-resolution mass spectrometry: an approach towards comprehensive peptide mapping of therapeutic mAbs0 aMultipleparallelprotease digestion coupled with highresolution m cFEB a1040530 v2323 a

Therapeutic monoclonal antibodies (mAbs) are structurally large and complex molecules. To be safe and efficacious, a biosimilar mAb must show high similarity to its reference product in Critical Quality Attributes (CQA). mAbs are highly sensitive to protein expression, production, manufacturing, supply chain, and storage conditions. All these factors make biosimilar mAbs intrinsically susceptible for variability during production. Accordingly, several lots of references and tests are required to establish the biosimilarity of a test mAb. The primary structure is a CQA of a mAb affecting its safety and efficacy. Here, we apply peptide mapping as an analytical method to decipher the primary structure and associated modifications for a quick quality assessment of TrastuzumAb and RituximAb innovator and biosimilar. A multiple-parallel-protease digestion strategy followed by high-resolution mass spectrometric analysis consistently achieved 100% sequence coverage along with reliable detection of post-translational modifications. Additionally, the use of supporting methods such as intact mass analysis and circular dichroism helped us to decipher the primary and higher order structures of these mAbs. We identify discernible variations in the profile of the innovator and biosimilar mAbs and validate the method for quick yet deep comparability analysis of the primary structure of biosimilar mAbs sold in the market. Significance: Peptide mapping using bottom-up approach is one of the most common methods for the characterization of therapeutic monoclonal antibodies. Herein, we describe a multi-parallel-protease digestion strategy using a combination of five different proteases followed by high-resolution mass spectrometric analysis with TrastuzumAb and RituximAb as an example. This resulted in a comprehensive identification of peptides with increased reliability and identification of different PTMs. Additional supporting orthogonal methods like intact mass and higher-order structure analysis helped evaluate broader conformational properties.

10aLC-MS/MS10aPTM10aRituximAb10aTherapeutic monoclonal antibodies10aTrastuzumAb1 aPradhan, Gauri1 aSneha, J., M.1 aSonwane, Babasaheb, P.1 aSanthakumari, B.1 aRao, Alka1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/multiple-parallel-protease-digestion-coupled-high-resolution-mass-spectrometry-approach02781nas a2200313 4500008004100000022001400041245018000055210006900235260000900304300001100313490000900324520172200333653002402055653001902079653000902098653001102107653002102118653001802139100002102157700002102178700001602199700002702215700002402242700001002266700002502276700001902301700001902320856012802339 2021 eng d a0006-899300aProtriptyline improves spatial memory and reduces oxidative damage by regulating NF kappa B-BDNF/CREB signaling axis in streptozotocin-induced rat model of Alzheimer's disease0 aProtriptyline improves spatial memory and reduces oxidative dama cMAR  a1472610 v17543 a

Antidepressants are well known to exert their role via upregulation of brain derived neurotrophic factor (BDNF). BDNF has been reported to exerts its neuroprotective effect in rodent and primate models as well as in patients of Alzheimer's disease (AD). The aim of our study was to evaluate the effect of protriptyline (PRT), a tricyclic antidepressant, in streptozotocin (STZ)- induced rat model of AD. Total 10 mu l of STZ was injected into each ventricle (1 mg/kg). PRT (10 mg/kg, i.p.) treatment was started 3-day post STZ administration and continued till 21 days. We found that STZ treatment significantly increased pTau, A beta 42 and BACE-1 expression, oxidative stress and neurodegeneration in hippocampus and cortex of adult rats. STZ induced impairment in spatial learning and retention memory was associated with increased WO and reduced CREB and BDNF expression in cortex and hippocampus. Interestingly, PRT treatment significantly reduced pTau, A beta 42 and BACE-1 levels, neurodegeneration, oxidative stress and glial activation, contributing to the improved spatial learning and retention memory in STZ treated rats. Moreover, PRT treatment significantly improved p-ERK/ERK ratio and enhanced BDNF and CREB levels by reducing NFXB and GFAP expression in STZ treated rats. Our data suggest that impaired NFXB and CREB signaling potentially contribute in AD pathogenesis by elevating oxidative stress and neuroinflammation mediated neurodegeneration. Our study has established protriptyline as a multi target molecule in pre-clinical model of AD and further investigations on PRT like molecules could pave way for further development of effective new treatments in neurodegenerative disorders.

10aAlzheimer's disease10aantidepressant10aBDNF10aMemory10aOxidative stress10aProtriptyline1 aTiwari, Virendra1 aMishra, Akanksha1 aSingh, Sonu1 aMishra, Sandeep, Kumar1 aSahu, Kiran, Kumari1 aParul1 aKulkarni, Mahesh, J.1 aShukla, Rakesh1 aShukla, Shubha uhttps://library.ncl.res.in/content/protriptyline-improves-spatial-memory-and-reduces-oxidative-damage-regulating-nf-kappa-b02392nas a2200241 4500008004100000022001400041245020900055210006900264260000900333490000800342520144800350100002801798700001701826700002001843700002801863700002001891700002601911700001801937700002101955700002501976700001702001856013202018 2021 eng d a1874-391900aSWATH-MS based quantitative proteomics analysis to evaluate the antileishmanial effect of Commiphora wightii- Guggul and amphotericin B on a clinical isolate of Leishmania donovani (vol 223, 103800, 2020)0 aSWATHMS based quantitative proteomics analysis to evaluate the a cFEB 0 v2323 a

Drug resistance and relapse after treatment of visceral leishmaniasis (VL) with the chemotherapeutic drugs has impeded the VL elimination programme especially, in the endemic region of Bihar, India. Currently, Antimonials (Sbv) have been rendered obsolete (Bihar) as frequent treatment failure and relapse in Sbv treated patient's warrants greater vigilance and attention to the limited drugs. A clinical isolate of L.donovani obtained from an Amphotericin B (AmB) relapse patient was evaluated for its susceptibility to AmB and a hyperlipidemic drug Guggul. The evaluation of susceptibility or resistance to any drug still relies on in vitro assay on promastigote and amastigote stages of Leishmania spp. as there are no validated markers which can ascertain drug resistance in Leishmania. The anti-promastigote effect of AmB and Guggul were demonstrated by significant cellular and morphological changes exhibiting apoptosis-mediated cell death. To further illustrate the molecular mechanism of the parasite's response upon exposure to either AmB and Guggul, sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS) for quantitative proteomics analysis was performed along with computational data analysis; revealing considerable differences in the proteome profiles which could be regarded as putative markers for resistance or drug targets for development of therapeutic antileishmanials.

1 aBar Routaray, Chinmayee1 aBhor, Renuka1 aBai, Shakuntala1 aKadam, Nitin, Suryakant1 aJagtap, Surabhi1 aDoshi, Pooja, Jignesh1 aSundar, Shyam1 aSawant, Sangeeta1 aKulkarni, Mahesh, J.1 aPai, Kalpana uhttps://library.ncl.res.in/content/swath-ms-based-quantitative-proteomics-analysis-evaluate-antileishmanial-effect-commiphora-002685nas a2200229 4500008004100000022001400041245016600055210006900221260000800290300001600298490000600314520183500320100001802155700002402173700002002197700002302217700002002240700002002260700002002280700002502300856013002325 2022 eng d a2470-134300aInvestigation of the captopril-insulin interaction by mass spectrometry and computational approaches reveals that captopril induces structural changes in insulin0 aInvestigation of the captoprilinsulin interaction by mass spectr cJUL a23115-231260 v73 a

ABSTRACT: Post-translational modifications remarkably regulate proteins' biological function. Small molecules such as reactive thiols, metabolites, and drugs may covalently modify the proteins and cause structural changes. This study reports the covalent modification and noncovalent interaction of insulin and captopril, an FDA-approved antihypertensive drug, through mass spectrometric and computation-based approaches. Mass spectrometric analysis shows that captopril modifies intact insulin, reduces it into its ``A'' and ``B'' chains, and covalently modifies them by forming adducts. Since captopril has a reactive thiol group, it might reduce the insulin dimer or modify it by reacting with cysteine residues. This was proven with dithiothreitol treatment, which reduced the abundance of captopril adducts of insulin A and B chains and intact Insulin. Liquid chromatography tandem mass spectrometric analysis identified the modification of a total of four cysteine residues, two in each of the A and B chains of insulin. These modifications were identified to be Cys6 and Cys7 of the A chain and Cys7 and Cys19 of the B chain. Mass spectrometric analysis indicated that captopril may simultaneously modify the cysteine residues of intact insulin or its subunits A and B chains. Biophysical studies involving light scattering and thioflavin T assay suggested that the binding of captopril to the protein leads to the formation of aggregates. Docking and molecular dynamics studies provided insights into the noncovalent interactions and associated structural changes in insulin. This work is a maiden attempt to understand the detailed molecular interactions between captopril and insulin. These findings suggest that further investigations are required to understand the long-term effect of drugs like captopril.

1 aGhosh, Amrita1 aPawar, Aiswarya, B.1 aChirmade, Tejas1 aJathar, Swaraj, M.1 aBhambure, Rahul1 aSengupta, Durba1 aGiri, Ashok, P.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/investigation-captopril-insulin-interaction-mass-spectrometry-and-computational-approaches02949nas a2200385 4500008004100000022001400041245017500055210006900230260000800299300001200307490000800319520168600327653001902013653001802032653001502050653001402065653001402079653002202093653001402115653000902129653001202138653001602150100003302166700002002199700002302219700002902242700002002271700003302291700002502324700002502349700002102374700001902395700002202414856012702436 2022 eng d a0981-942800aStacking herbicide detoxification and resistant genes improves glyphosate tolerance and reduces phytotoxicity in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.)0 aStacking herbicide detoxification and resistant genes improves g cOCT a126-1380 v1893 a

Glyphosate residues retained in the growing meristematic tissues or in grains of glyphosate-resistant crops affect the plants physiological functions and crop yield. Removing glyphosate residues in the plants is desirable with no penalty on crop yield and quality. We report a new combination of scientific strategy to detoxify glyphosate that reduces the residual levels and improve crop resistance. The glyphosate detoxifying enzymes Aldo-keto reductase (AKR1) and mutated glycine oxidase (mGO) with different modes of action were co-expressed with modified EPSPS, which is insensitive to glyphosate in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.). The transgenic tobacco plants expressing individual PsAKR1, mGO, CP4-EPSPS, combinations of PsAKR1:CP4EPSPS, PsAKR1:mGO, and multigene with PsAKR1: mGO: CP4EPSPS genes were developed. The bio-efficacy studies of in-vitro leaf regeneration on different concentrations of glyphosate, seedling bioassay, and spray on transgenic tobacco plants demonstrate that glyphosate detoxification with enhanced resistance. Comparative analysis of the transgenic tobacco plants reveals that double and multigene expressing transgenics had reduced accumulation of shikimic acid, glyphosate, and its primary residue AMPA, and increased levels of sarcosine were observed in all PsAKR1 expressing transgenics. The multigene expressing rice transgenics showed improved glyphosate resis-tance with yield maintenance. In summary, results suggest that stacking genes with two different detoxification mechanisms and insensitive EPSPS is a potential approach for developing glyphosate-resistant plants with less residual content.

10aDetoxification10aGene stacking10aGlyphosate10aHerbicide10aMultigene10aResidual toxicity10aResistant10arice10aTobacco10aTransgenics1 aVennapusa, Amaranatha, Reddy1 aAgarwal, Subham1 aHm, Hanumanth, Rao1 aAarthy, Thiagarayaselvam1 aBabitha, K., C.1 aThulasiram, Hirekodathakallu1 aKulkarni, Mahesh, J.1 aMelmaiee, Kalpalatha1 aSudhakar, Chinta1 aUdayakumar, M.1 aVemanna, Ramu, S. uhttps://library.ncl.res.in/content/stacking-herbicide-detoxification-and-resistant-genes-improves-glyphosate-tolerance-and01832nas a2200217 4500008004100000022001400041245011300055210006900168260000900237300001600246490000600262520104300268100003201311700002301343700002901366700001801395700002001413700002901433700002501462856012701487 2023 eng d a2470-134300aElevated level of glycated KQTALVELVK peptide of albumin is associated with the risk of diabetic nephropathy0 aElevated level of glycated KQTALVELVK peptide of albumin is asso cJUN  a20654-206600 v83 a

Diabetic nephropathyis a leading cause of end-stagerenal disease.Hence, early detection of diabetic nephropathy is essential to mitigatethe disease burden. Microalbuminuria, the currently used diagnosticmarker of diabetic nephropathy, is not efficient in detecting it atan early stage. Therefore, we explored the utility of glycated humanserum albumin (HSA) peptides for risk prediction of diabetic nephropathy.Three glycation-sensitive HSA peptides, namely, FKDLG-EENFK,KQTAL-VELVK, and KVPQVST-PTLVEVSR, with deoxyfructosyllysine(DFL) modification were quantified by targeted mass spectrometry (MS)in a study population comprising healthy and type II diabetes subjectswith and without nephropathy. Mass spectrometry, receiver operatingcharacteristic (ROC) curve, and correlation analysis revealed thatthe DFL-modified KQTALVELVK peptide was better than other glycatedHSA peptides and HbA(1c) for identifying diabetic nephropathy.DFL-modified KQTALVELVK could be a potential marker for risk predictionof diabetic nephropathy.

1 aChaurasiya, Arvindkumar, H.1 aJaiswal, Meera, R.1 aBayatigeri, Santhakumari1 aKahar, Shweta1 aTiwari, Shalbha1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/elevated-level-glycated-kqtalvelvk-peptide-albumin-associated-risk-diabetic-nephropathy02457nas a2200289 4500008004100000022001400041245008200055210006900137260000800206490000900214520156800223653001301791653001601804653001401820653001301834653001001847653000701857653003901864653001601903100002701919700001901946700001501965700002401980700002702004700002502031856011102056 2023 eng d a0304-416500aMechanistic insights on anserine hydrolyzing activities of human carnosinases0 aMechanistic insights on anserine hydrolyzing activities of human cMAR0 v18673 a

Anserine and carnosine represent histidine-containing dipeptides that exert a pluripotent protective effect on human physiology. Anserine is known to protect against oxidative stress in diabetes and cardiovascular diseases. Human carnosinases (CN1 and CN2) are dipeptidases involved in the homeostasis of carnosine. In poikilothermic vertebrates, the anserinase enzyme is responsible for hydrolyzing anserine. However, there is no specific anserine hydrolyzing enzyme present in humans. In this study, we have systematically investigated the anserine hydrolyzing activity of human CN1 and CN2. A targeted multiple reaction monitoring (MRM) based approach was employed for studying the enzyme kinetics of CN1 and CN2 using carnosine and anserine as substrates. Surprisingly, both CN1 and CN2 can hydrolyze anserine effectively. The observed catalytic turnover rate (Vmax/[E]t) was 21.6 s-1 and 2.8 s-1 for CN1 and CN2, respectively. CN1 is almost eight-fold more efficient in hydrolyzing anserine compared to CN2, which is comparable to the efficiency of the carnosine hydrolyzing activity of CN2. The Michaelis constant (Km) value for CN1 (1.96 mM) is almost three-fold lower compared to CN2 (6.33 mM), representing higher substrate affinity for anserine-CN1 interactions. Molecular docking studies showed that anserine binds at the catalytic site of the carnosinases with an affinity similar to carnosine. Overall, the present study elucidated the inherent promiscuity of human carnosinases in hydrolyzing anserine using a sensitive LCMS/MS approach.

10aAnserine10aCarnosinase10aCarnosine10aDiabetes10aLC-MS10aMS10aMultiple reaction monitoring (MRM)10aPromiscuity1 aPandya, Vaibhav, Kumar1 aShankar, Shiva1 aRajesh, S.1 aRathore, Rajeshwari1 aKumaran, Sangaralingam1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/mechanistic-insights-anserine-hydrolyzing-activities-human-carnosinases02193nas a2200241 4500008004100000022001400041245009400055210006900149260000800218300001600226490000600242520135800248100003201606700002501638700001701663700002301680700002201703700002401725700002801749700002801777700002501805856012101830 2023 eng d a2470-134300aNanopore sequencing of RAGE gene polymorphisms and their association with type 2 diabetes0 aNanopore sequencing of RAGE gene polymorphisms and their associa cJUL a25727-257380 v83 a

The receptor foradvanced glycation end products (RAGE)is a transmembraneprotein that interacts with its ligands, advanced glycation end products(AGEs). AGEs are elevated in diabetes and diabetic complications,leading to increased oxidative stress and activation of pro-inflammatorypathways facilitated by AGE-RAGE signaling. Polymorphisms inthe RAGE gene can potentially affect AGE-RAGEinteraction and its downstream signaling, which plays a crucial rolein the progression of diabetes and its complications. In this study,we used nanopore sequencing for genotyping of RAGE polymorphism and identified a maximum number of 33 polymorphisms,including two previously unreported novel mutations in a cohort ofhealthy, type 2 diabetics without nephropathy and type 2 diabeticswith nephropathy in order to identify associations. Two novel RAGE polymorphisms in the intron 8 and 3 & PRIME;UTR regionat genomic locations 32181834 and 32181132, respectively, were detectedwith a low frequency. For four previously reported polymorphisms,cross-validation by PCR-RFLP showed 99.75% concordance with nanoporesequencing. Analysis of genotype distribution and allele frequenciesrevealed that five single nucleotide polymorphisms, i.e., rs1800625,rs3131300, rs3134940, rs2070600, and rs9391855, were associated withan increased risk for type 2 diabetes.

1 aChaurasiya, Arvindkumar, H.1 aKhilari, Ajinkya, A.1 aKazi, Rubina1 aJaiswal, Meera, R.1 aBhoite, Gouri, M.1 aPadwal, Meghana, K.1 aMomin, Abdulrahaman, A.1 aShanmugam, Dhanasekaran1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/nanopore-sequencing-rage-gene-polymorphisms-and-their-association-type-2-diabetes02486nas a2200277 4500008004100000022001400041245017700055210006900232260000900301300001400310490000700324520149400331653001201825653001301837653001801850653002201868653003301890653002501923100001901948700002001967700002301987700001502010700002602025700002502051856013202076 2024 eng d a0739-110200aDe novo structure prediction of meteorin and meteorin-like protein for identification of domains, functional receptor binding regions, and their high-risk missense variants0 aDe novo structure prediction of meteorin and meteorinlike protei cJUN  a4522-45360 v423 a

Meteorin (Metrn) and Meteorin-like (Metrnl) are homologous secreted proteins involved in neural development and metabolic regulation. In this study, we have performed de novo structure prediction and analysis of both Metrn and Metrnl using Alphafold2 (AF2) and RoseTTAfold (RF). Based on the domain and structural homology analysis of the predicted structures, we have identified that these proteins are composed of two functional domains, a CUB domain and an NTR domain, connected by a hinge/loop region. We have identified the receptor binding regions of Metrn and Metrnl using the machine-learning tools ScanNet and Masif. These were further validated by docking Metrnl with its reported KIT receptor, thus establishing the role of each domain in the receptor interaction. Also, we have studied the effect of non-synonymous SNPs on the structure and function of these proteins using an array of bioinformatics tools and selected 16 missense variants in Metrn and 10 in Metrnl that can affect the protein stability. This is the first study to comprehensively characterize the functional domains of Metrn and Metrnl at their structural level and identify the functional domains, and protein binding regions. This study also highlights the interaction mechanism of the KIT receptor and Metrnl. The predicted deleterious SNPs will allow further understanding of the role of these variants in modulating the plasma levels of these proteins in disease conditions such as diabetes.

10adomains10aMeteorin10ameteorin-like10amissense variants10aprotein-protein interactions10astructure prediction1 aShankar, Shiva1 aBanarjee, Reema1 aJathar, Swaraj, M.1 aRajesh, S.1 aRamasamy, Sureshkumar1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/de-novo-structure-prediction-meteorin-and-meteorin-protein-identification-domains-functional02952nas a2200241 4500008004100000022001400041245011100055210006900166260000900235300001600244490000600260520209500266100002902361700002002390700002802410700002402438700002302462700002402485700002002509700002902529700002502558856012702583 2024 eng d a2470-134300aDiscovery of free glycated amines and glycated urea in diabetic plasma: potential implications in diabetes0 aDiscovery of free glycated amines and glycated urea in diabetic  cMAY  a24907-249150 v93 a

The role of protein glycation in the pathogenesis of diabetes has been well established. Akin to proteins, free amino acids and other small-molecule amines are also susceptible to glycation in hyperglycemic conditions and may have a role in the pathogenesis of the disease. However, information about glycation of free amino acids and other small-molecule amines is relatively obscure. In the quest to discover small-molecule glycated amines in the plasma, we have synthesized glycated amino acids, glycated creatine, and glycated urea, and by using a high-resolution accurate mass spectrometer, a mass spectral library was developed comprising the precursor and predominant fragment masses of glycated amines. Using this information, we report the discovery of the glycation of free lysine, arginine, and leucine/isoleucine from the plasma of diabetic patients. This has great physiological significance as glycation of these amino acids may create their deficiency and affect vital physiological processes such as protein synthesis, cell signaling, and insulin secretion. Also, these glycated amino acids could serve as potential markers of diabetes and its complications. While other amines, such as creatinine and urea, accumulate in the plasma and act as biomarkers of diabetic nephropathy. For the first time, we report the detection of glycated urea in diabetic plasma, which is confirmed by matching the precursor and fragment masses with the in vitro synthesized glycated urea by using C-12(6) and C-13(6)-glucose. Further, we quantified glycated urea detected in two forms, monoglycated urea (MGU) and diglycated urea (DGU), by a targeted mass spectrometric approach in the plasma of healthy, diabetic, and diabetic nephropathy subjects. Both MGU and DGU showed a positive correlation with clinical parameters, such as blood glucose and HbA1c. Given that urea gets converted to glycated urea in hyperglycemic conditions, it is crucial to quantify MGU and DGU along with the urea for the diagnosis of diabetic nephropathy and study their physiological role in diabetes.

1 aShaikh, Rashdajabeen, Q.1 aDas, Sancharini1 aChaurasiya, Arvindkumar1 aAshtamy, Murali, G.1 aSheikh, Amreen, B.1 aFernandes, Moneesha1 aTiwari, Shalbha1 aUnnikrishnan, Ambika, G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/discovery-free-glycated-amines-and-glycated-urea-diabetic-plasma-potential-implications06699nas a2200169 4500008004100000245010700041210006900148260000800217520600300225100003606228700002906264700002706293700003406320700002606354700002506380856012406405 2024 eng d00aDual modification of tau by pseudophosphorylation and glycation does not enhance amorphous aggregation0 aDual modification of tau by pseudophosphorylation and glycation  cAPR3 a

Background/Aims:

 The neurofibrillary tangles consisting of Tau protein are an important pathology in Alzheimer’s disease. The paired helical filaments of Tau form most of the NFTs. These PHFs of Tau are found to carry numerous post-translational modifications, which stabilize them and aid in aggregation. The mechanistic function of Tau is to bind and stabilize the axonal microtubules. Hyperphosphorylation of Tau causes it to compromise its physiological function and accumulate in the neurons in the form of aggregates. Such residue-specific phosphorylation has been studied by employing Tau pseudophosphorylation mutants. But in addition to phosphorylation, several other modifications also aid in stabilizing the Tau PHF. Glycation is one such non-enzymatic PTM caused by sugars and their reactive intermediates. In this study, we employed the pseudophosphorylated Tau double mutants (262/404D, 262/396D, and 231/262) for studying their modification by methyl glyoxal, a reactive intermediate of glucose metabolism. 

Methods:

 We studied various biophysical properties like aggregation propensity, Advanced glycation end-product formation, and global conformation of the Tau with dual modifications. Our study includes the use of in vitro techniques e.g., ThS fluorescence assay, electron microscopy, CD spectroscopy, SDS-PAGE. 

Results:

 The overall result of the study suggest that the MG-induced Tau aggregation is influenced by the residue-specific Tau phosphorylation. 

Conclusion:

 In conclusion, the combinatorial effect of discreet PTMs on Tau function could lead to a better understanding of Tauopathy.

1 aChinnathambi, Subashchandrabose1 aSonawane, Shweta, Kishor1 aChandrashekar, Madhura1 aChidambaram, Hariharakrishnan1 aThulasiram, Shrivatsa1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/dual-modification-tau-pseudophosphorylation-and-glycation-does-not-enhance-amorphous02333nas a2200289 4500008004100000022001400041245007800055210006900133260000800202300001100210490000800221520142300229653001401652653002401666653002401690653002201714653003301736653002201769100001801791700001601809700002401825700002201849700002001871700002501891700002001916856010701936 2024 eng d a0141-813000aMulti-dimensional impact of captopril modification on human serum albumin0 aMultidimensional impact of captopril modification on human serum cAUG a1332890 v2743 a

Captopril is a thiol drug, widely used for the management of hypertension and cardiovascular diseases. Reactive thiols are found to covalently modify the cysteines of plasma proteins and affect their structure and function. Human serum albumin (HSA) is prone to undergo modification by various low molecular weight compounds, including drugs. Cysteine34 (Cys34) in HSA has a free thiol group with antioxidant properties, considered to be the most redox-sensitive amino acid in plasma. Through mass-spectrometric analysis, we demonstrate for the first time that captopril forms a disulfide adduct at Cys34 residue and increases the protease susceptibility of HSA to trypsin. As evidenced by our biophysical and electron microscopy studies, HSA undergoes structural alteration, aggregation and morphological changes when treated with different captopril concentrations. Molecular dynamics studies further revealed the regions of secondary structural changes in HSA due to disulfide adduct formation by captopril at Cys34. It also elucidated the residues involved in the noncovalent interactions with captopril. It is envisaged that structural change in HSA may influence the efficacy of drug delivery as well as its own biological function. These findings may thus provide significant insights into the field of pharmacology intriguing further investigation into the effects of long-term captopril treatment.

10aCaptopril10aElectron microscopy10aHuman serum albumin10aMass spectrometry10aMolecular dynamic simulation10aSpectrophotometry1 aGhosh, Amrita1 aJani, Vinod1 aSonavane, Uddhavesh1 aNaphade, Anvi, N.1 aJoshi, Rajendra1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/multi-dimensional-impact-captopril-modification-human-serum-albumin00820nas a2200217 4500008004100000022001400041245014000055210006900195260001100264300001400275490000700289653003000296653002000326653001200346653001700358100002900375700002200404700002500426700002400451856012700475 2024 eng d a1477-052000aSupramolecular polyplexes from Janus peptide nucleic acids (bm-PNA-G5): self-assembled bm-PNA G-quadruplex and its tetraduplex with DNA0 aSupramolecular polyplexes from Janus peptide nucleic acids bmPNA cAUG 22 a6810-68210 v2210aDOUBLE-HEADED NUCLEOTIDES10aENERGY-TRANSFER10aGuanine10aMAJOR GROOVE1 aTodkari, Iranna, Annappa1 aChaudhary, Preeti1 aKulkarni, Mahesh, J.1 aGanesh, Krishna, N. uhttps://library.ncl.res.in/content/supramolecular-polyplexes-janus-peptide-nucleic-acids-bm-pna-g5-self-assembled-bm-pna-g02430nas a2200277 4500008004100000022001400041245014100055210006900196260000800265300001100273490000800284520148100292653001501773653001301788653001801801653003301819653002101852653001501873100002501888700001901913700002401932700002301956700002501979700002102004856012702025 2024 eng d a0278-691500aSWATH-MS reveals that bisphenol A and its analogs regulate pathways leading to disruption in insulin signaling and fatty acid metabolism0 aSWATHMS reveals that bisphenol A and its analogs regulate pathwa cJUN a1146670 v1883 a

Bisphenol A (BPA) is an endocrine-disrupting chemical (EDC), associated with obesity and insulin resistance. The FDA prohibited the use of BPA-based polycarbonate resins in infant formula packaging; thus, its analogs, viz. Bisphenol S (BPS) and Bisphenol F (BPF) were considered alternatives in epoxy resins, plastics, and food cans. As these analogs might evoke a similar response, we investigated the role of Bisphenols (BPA, BPF, and BPS), on insulin signaling in CHO-HIRc-myc-GLUT4eGFP cells at environmentally relevant concentrations of 2 nM and 200 nM. Insulin signaling demonstrated that Bisphenols reduced phosphorylation of IR and AKT2, GLUT4 translocation, and glucose uptake. This was accompanied by increased oxidative stress. Furthermore, SWATHMS-based proteomics of 3T3-L1 cells demonstrated that Bisphenol-treated cells regulate proteins in insulin resistance, adipogenesis, and fatty acid metabolism pathways differently. All three Bisphenols induced differentially expressed proteins enriched similar pathways, although their abundance differed for each Bisphenol. This might be due to their varying toxicity level, structural differences, and estrogen-mimetic activity. This study has important implications in addressing health concerns related to EDCs. Given that the analogs of BPA are considered alternatives to BPA, the findings of this study suggest they are equally potent in altering fatty acid metabolism and inducing insulin resistance.

10aBisphenols10aEstrogen10aLipid droplet10aMitochondrial beta-oxidation10aOxidative stress10aPPAR gamma1 aKulsange, Shabda, E.1 aSharma, Monika1 aSonawane, Babasaheb1 aJaiswal, Meera, R.1 aKulkarni, Mahesh, J.1 aSanthakumari, B. uhttps://library.ncl.res.in/content/swath-ms-reveals-bisphenol-and-its-analogs-regulate-pathways-leading-disruption-insulin03149nas a2200229 4500008004100000245010900041210006900150260000800219300001200227490000700239520235000246100002702596700001602623700002302639700001902662700001902681700001702700700002102717700002602738700002502764856013002789 2024 eng d00aYoga therapy attenuates the progression of diabetes - insights from proteomics and metabolomics analysis0 aYoga therapy attenuates the progression of diabetes insights fro cDEC a163-1740 v173 a

Objective: Diabetes management remains challenging despite advancements in therapeutics, with many subjects developing complications. Yoga has been shown to aid diabetes management. This study investigates the impact of yoga therapy on diabetes progression, utilizing proteomics and metabolomics analyses to explore underlying molecular mechanisms.

Methodology: A 3-month longitudinal study involving healthy subjects with prediabetes and diabetes was conducted. Blood glucose, glycated hemoglobin (HbA1c), lipid profile, and malondialdehyde (MDA) levels were measured before and after the yoga intervention.

Results and conclusion: Healthy subjects showed no significant changes in blood glucose, lipid profile, HbA1c, or MDA levels. However, subjects with prediabetes and diabetes experienced positive changes, with decreases in HbA1c and MDA levels. Proteomics and metabolomics analyses provided insights into the molecular mechanisms by which yoga attenuates diabetes progression in subjects with prediabetes and diabetes. This study is a pioneering effort to understand the molecular basis of yoga's beneficial effects on diabetes management.

1 aSonwane, Babasaheb, P.1 aRaut, Pooja1 aChitalkar, Jyotsna1 aGodbole, Smita1 aSabnis, Shanta1 aGupta, Jyoti1 aSanthakumari, B.1 aDeshpande, Mukund, V.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/yoga-therapy-attenuates-progression-diabetes-insights-proteomics-and-metabolomics-analysis01638nas a2200265 4500008004100000022001400041245014600055210007000201260000800271300001400279490000700293520073700300653000801037653002001045653001301065653001401078653002001092653000801112100002401120700002001144700002801164700002501192700002501217856013001242 2025 eng d a0973-393000aAutoantibodies against Nε-carboxymethyl lysine and methylglyoxal modified albumin are associated with cardiovascular risk in type 2 diabetes0 aAutoantibodies against Nεcarboxymethyl lysine and methylglyoxal  cDEC a1104-11100 v453 a

{Background Albumin is an abundant plasma protein which gets modified with advanced glycation end products (AGEs) predominantly in diabetic condition. AGE modification induces immune response and autoantibodies are generated which play an important role in disease pathology. Objective This study aimed to illustrate the role of autoantibodies against N epsilon-carboxymethyl lysine (CML) and methylglyoxal (MG) modified albumin in diabetic cardiovascular complications. MethodsType-2 diabetes subjects were enrolled and further grouped into stress test positive or stress test negative based on treadmill stress test (TMT). Autoantibody titer was quantified by ELISA assay for CML-modified albumin (stress test positive

10aAGE10aAtherosclerosis10aDiabetes10aGlycation10aImmune response10aROC1 aPatil, Yugendra, R.1 aTiwari, Shalbha1 aMomin, Abdulrahaman, A.1 aUnnikrishnan, A., G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/autoantibodies-against-n%CE%B5-carboxymethyl-lysine-and-methylglyoxal-modified-albumin-are02205nas a2200241 4500008004100000022001400041245009500055210006900150260000800219300001200227490000700239520141000246653002001656653002301676653001201699653002201711100002301733700002301756700002301779700001501802700002501817856012101842 2025 eng d a1572-388700aCharacterization of S-homocysteinylation of Human Insulin and Its Implications in Diabetes0 aCharacterization of Shomocysteinylation of Human Insulin and Its cOCT a598-6100 v443 a

Homocysteine thiolactone is a reactive thiol known for its interaction with various proteins. Nevertheless, there exists a paucity of information concerning the interaction between homocysteine thiolactone and human insulin, particularly regarding the mechanism by which homocysteine facilitates the reduction of disulfide bonds within insulin. In the present study, we have elucidated the binding sites of homocysteine to the cysteine residues (A6-B7 and A20-B19) that are implicated in the formation of intermolecular disulfide bonds in insulin through an in vitro reaction analyzed via LC-ESI MS/MS. This results in a reduction of disulfide bonds linking the A and B chains, which was corroborated by MALDI-TOF-MS and ESI-MS analysis. The secondary structure of insulin is affected by this modification, as evidenced by circular dichroism spectroscopy. In-silico studies also show that homocysteine affects the insulin structure. A glucose uptake assay conducted in Chinese hamster ovary (CHO) cells that stably express the insulin receptor revealed that HC-modified insulin is less effective in inducing glucose uptake compared to native insulin, suggesting that HC-induced structural modifications in insulin influence functional activity. This study provides insight into the HC-induced structural and functional changes in insulin and discusses the consequent implications for diabetes.

10aDisulfide bonds10aHomocysteinylation10ainsulin10aMass spectrometry1 aSheikh, Amreen, B.1 aJathar, Swaraj, M.1 aTammara, Vaishnavi1 aDas, Atanu1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/characterization-s-homocysteinylation-human-insulin-and-its-implications-diabetes02383nas a2200253 4500008004100000022001400041245009700055210006900152260000800221300001200229490000700241520151000248653002701758653001301785653003301798653001901831653002101850653003501871100002401906700002001930700002501950700002501975856012902000 2025 eng d a1937-538700aProteome profile of stress test assessed cardiovascular disease risk-prone diabetic subjects0 aProteome profile of stress test assessed cardiovascular disease  cAUG a960-9690 v183 a

Cardiovascular disease (CVD) is the leading cause of death in the diabetic population. There is a need for specific predictive markers to assess CVD risk. The present study explored the plasma proteome profile of treadmill test (TMT) assessed diabetic stress test positive (DSTP) and diabetic stress test negative (DSTN) subjects by performing a SWATH-MS-based label-free quantitative mass spectrometry approach to identify differentially expressed proteins (DEPs). CVD-relevant DEPs were further validated using a targeted mass spectrometry approach (MRM-HR). It was observed that CO4B, PON1 and LUM exhibited considerable differential expression in both the MS approaches, and ROC analysis showed significant AUC (0.97, 0.79 and 0.77, respectively). Overall, the present study reports these proteins as potential alternative markers for TMT in assessing CVD risk. These markers can possibly overcome the limitations of TMT with further validation in the large cohort.Graphical AbstractAn overview of experimental approaches used in the current study. The study design depicts diabetic subjects assessed for cardiovascular risk by TMT or stress test. The experimental design shows the use of the SWATH-MS approach to identify differentially expressed proteins and validate CVD-related proteins with targeted MS approaches such as MRM-HR. Finally, CO4B, PON1 and LUM exhibited significant AUC in ROC analysis, indicating their potential marker capabilities to predict CVD in diabetic subjects.

10acardiovascular disease10aDiabetes10aMultiple reaction monitoring10aPlasma markers10aProteome profile10aQuantitative mass spectrometry1 aPatil, Yugendra, R.1 aTiwari, Shalbha1 aUnnikrishnan, A., G.1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/proteome-profile-stress-test-assessed-cardiovascular-disease-risk-prone-diabetic-subjects02515nas a2200277 4500008004100000022001400041245011700055210006900172260000800241300001100249490000800260520162800268653001301896653002201909653001201931653001201943653002201955100001501977700001901992700002002011700001902031700002002050700001902070700002502089856012302114 2025 eng d a1046-592800aSimple freeze-thaw based method for efficient purification of recombinant human proinsulin from inclusion bodies0 aSimple freezethaw based method for efficient purification of rec cMAR a1066450 v2273 a

Insulin is a pivotal peptide hormone essential for regulating glucose homeostasis. It has been known for over 100 years, but its production and purification methods are still under improvement. Escherichia coli based bacterial expression system is primarily used for insulin production. The human insulin protein expressed in bacteria usually forms inclusion bodies, complicating the purification process. Traditionally, insulin purification is a timeconsuming process involving urea-based denaturation methods, and various refolding techniques, followed by extensive chromatographic methods. Here, we report an easy and efficient purification of human proinsulin involving freeze-thaw based solubilization method. The extracted proinsulin inclusion bodies are treated with different concentrations of urea, followed by a freeze-thaw based solubilization. The freezing was carried out at various temperatures, mainly -80 degrees C, -20 degrees C, and -196 degrees C to determine the optimum condition for solubilization. Highest solubilization of proinsulin from the inclusion body was achieved with 0.5M urea and -20 degrees C. Further Nickel NTA-based purification was performed, and the purified protein was characterized for disulfide mapping by high-resolution mass spectrometer (HRMS). We also performed glucose uptake assays to validate the functional properties of purified proinsulin. This freeze-thaw based mild solubilization approach is a fast and effective method for getting bioactive proinsulin, which will help further design better purification and processing strategies for insulin production.

10aDiabetes10aDisulfide mapping10aglucose10ainsulin10aMass spectrometry1 aRajesh, S.1 aJathar, Swaraj1 aBanarjee, Reema1 aSharma, Monika1 aPalkar, Shivani1 aShankar, Shiva1 aKulkarni, Mahesh, J. uhttps://library.ncl.res.in/content/simple-freeze-thaw-based-method-efficient-purification-recombinant-human-proinsulin02916nas a2200313 4500008004100000022001400041245012200055210006900177260000800246300001100254490000900265520183400274653002602108653002602134653003202160653003002192653002002222653003502242100002202277700001902299700002502318700002002343700001802363700002502381700002002406700002202426700002402448856013002472 2026 eng d a0022-286000aNovel prodrug-inspired eugenol derivatives with enhanced bioavailability, anti-diabetic and anti-glycation efficacies0 aNovel prodruginspired eugenol derivatives with enhanced bioavail cAPR a1451750 v13573 a

In this study, we present the design, synthesis, and evaluation of novel eugenol analogs aimed to overcome its limited bioavailability due to insolubility in aqueous media. Thus, we re-engineered eugenol using prodruginspired structural modifications to improve pharmacokinetic properties. First, we structurally modified eugenol and synthesized its natural amino acid conjugates as esters and carbamates. These were prepared in NBoc protected, free amine, and HCl salt forms. These modifications are expected to improve the polarity and solubility of eugenol congeners in biological systems. They can also release the parent eugenol through enzymatic hydrolysis, enhancing its therapeutic potential. Next, we comprehensively screened for these derivatives through in silico studies followed by in vitro and in vivo assays. These include DPPH radical scavenging (IC50 range: 37.7 to 103.7 mu M), inhibition of (i) alpha-amylase (IC50 23.1 to 67.3 mu M), (ii) alpha-glucosidase (IC50 43.6 to 50.4 mu M), (iii) glycation (IC50 31.9 to 110.3 mu M) along with pharmacokinetic profiling and toxicity assessments. These experiments collectively demonstrated improved activity of eugenol analogs for several important parameters. Specifically, six analogs-epoxy eugenol (39), hydroxy eugenol (43), aspartate eugenol (26), isoleucinate eugenol (24), glutamate eugenol (37), and glutamate-salt eugenol (27) exhibited superior bioavailability, absorption, and distribution over to the parent compound eugenol. These analogs were found to be non-toxic and safe for oral administration. Overall, the study establishes a mechanistic and rational framework for modifying eugenol to overcome its inherent biopharmaceutical limitations, positioning them as promising candidates for treating diabetes and glycation-related conditions.

10aAmino acid conjugates10aAntidiabetic activity10aBioavailability enhancement10aDrug design and synthesis10aEugenol analogs10aIn silico and in vitro studies1 aKataria, Priyanka1 aVairale, Shiva1 aMote, Chandrashekhar1 aJoshi, Kaustubh1 aJoshi, Rakesh1 aKulkarni, Mahesh, J.1 aGiri, Ashok, P.1 aKontham, Ravindar1 aAgawane, Sachin, B. uhttps://library.ncl.res.in/content/novel-prodrug-inspired-eugenol-derivatives-enhanced-bioavailability-anti-diabetic-and-anti