@article {8098, title = {Analysis of smart biomaterial containing umbilical cord blood serum protein conjugated with P-(NIPAAM) using spectroscopy [Bio-incubation Services]}, journal = {Materials Today: Proceedings}, year = {2023}, abstract = {

Human Umbilical Cord Blood Serum (HUCBS) is a complex and evolving collection of proteins that promote fetal development. In the realm of regenerative medicine, the important proteins found in HUCBS are of great interest. The smart biomaterial generated from HUCBS is described in this paper. To characterize this novel biomaterial, human umbilical cord blood was obtained in sterile vacutainers from mothers and left to clot for 24\ h at 37 {\textdegree}C. The supernatant serum was collected, centrifuged and lyophilized. The lyophilized HUCBS was homogenized with smart polymer. This sample was subjected to physico-chemical characterization using Attenuated Total Reflectance-Fourier-Transform Infrared (ATR-FTIR) Spectroscopy and Nuclear Magnetic Resonance (NMR). The quantification of protein-polymer conjugate using ATR-FTIR revealed peaks ranging between 3264 and 531\ cm-1 and that of NMR showed wide resonances in the region 0{\textendash}5\ ppm. ATR-FTIR and NMR investigations were used to determine the structural stability of protein molecules in protein-polymer complex which helps in understanding the possible clinical effectiveness of the smart biomaterial in drug delivery.

}, keywords = {ATR-FTIR, H NMR, HUCBS, P-NIPAAM, Protein-polymer conjugate}, issn = {2214-7853}, doi = {https://doi.org/10.1016/j.matpr.2023.01.285}, url = {https://www.sciencedirect.com/science/article/pii/S2214785323003759}, author = {Manasa Biligowda Latha and Ashmitha Kishan Shetty and Rajamanickam Deveswaran and Ashish Jagannath Rai and Serene Joy and Hadonahalli Munegowda Shashanka and Siddique Sha Muhammad Hussain and Suraksha Shetty} } @article {5253, title = {Analysis of water soluble fractions of crude oil by gas chromatography: Mass spectroscopy [Mass Spectrometry - Metabolomics Facility]}, journal = {The Pharma Innovation Journal}, volume = {SP-11(4)}, year = {2022}, month = {06/2022}, chapter = {1119}, abstract = {

Crude oil is the major source of energy in the modern society to meet the global energy demand by which exploitation of crude oil and its transportation increasing rapidly leading to frequent catastrophic oil spills. When oil spill occurs or when oil is discharged into aquatic environment the components of the crude oil present in it are mostly volatile, evaporates into the environment and the fraction of the oil soluble in water (i.e., WSF) is available to the organisms directly which is the main determinant of crude oil toxicity to aquatic organisms. Although this fraction is present only in relatively low concentrations, it is this fraction which is in most intimate contact with fish and other pelagic organisms with carcinogenic and mutagenic potential. So, the aim of the study was to determine different hydrocarbons dissolved in water soluble fraction (WSF) of crude oil qualitatively and quantitatively. The determination of different hydrocarbons present in water soluble fraction of crude oil can be used as reference point for many studies in future for determining the toxicity of water soluble fraction of crude oil to fishes.

}, keywords = {crude oil, gas chromatography, Water soluble fraction}, url = {https://www.thepharmajournal.com/archives/2022/vol11issue4S/PartP/S-11-4-135-968.pdf}, author = {Rishika, V. and Lakshmipathi, M.T. and Sampath Kumar, B.} } @article {1866, title = {Alterations of Primary Metabolites in Root Exudates of Intercropped Cajanus cajan{\textendash}Zea mays Modulate the Adaptation and Proteome of Ensifer (Sinorhizobium) fredii NGR234 [Mass Spectrometry - Proteomics Facility]}, journal = {Microbial Ecology}, year = {2021}, month = {08/2021}, pages = {1{\textendash}18}, type = {Journal Article}, abstract = {

Legume-cereal intercropping systems, in the context of diversity, ecological function, and better yield have been widely studied. Such systems enhance nutrient phytoavailability by balancing root-rhizosphere interactions. Root exudates (RE) play an important role in the rhizospheric interactions of plant-plant and/or plant-microbiome interaction. However, the influence of the primary metabolites of RE on plant-rhizobia interactions in a legume-cereal intercrop system is not known. To understand the plant communication with rhizobia, Cajanus cajan-Zea mays intercropped plants and the broad host range legume nodulating Ensifer fredii NGR234 as the model plants and rhizobium used respectively. A metabolomics-based approach revealed a clear separation between intercropped and monocropped RE of the two plants. Intercropped C. cajan showed an increase in the myo-inositol, and proline, while intercropped Z. mays showed enhanced galactose, D-glucopyranoside, and arginine in the RE. Physiological assays of NGR234 with the RE of intercropped C. cajan exhibited a significant enhancement in biofilm formation, while intercropped Z. mays RE accelerated the bacterial growth in the late log phase. Further, using label-free proteomics, we identified a total of 2570 proteins of NGR234 covering 50\% annotated protein sequences upon exposure to Z. mays RE. Furthermore, intercropped Z. mays RE upregulated bacterioferritin comigratory protein (BCP), putative nitroreductase, IlvD, LeuC, D (branched-chain amino acid proteins), and chaperonin proteins GroEL2. Identification offered new insights into the metabolome of the legume-cereal intercrop and proteome of NGR234-Z. mays interactions that underline the new molecular candidates likely to be involved in the fitness of rhizobium in the intercropping system.

}, keywords = {Cajanus cajan Zea mays, Ensifer fredii NGR234, Metabolomics, Proteomics, Root exudates (RE)}, doi = {https://doi.org/10.1007/s00248-021-01818-4}, url = {https://link.springer.com/article/10.1007/s00248-021-01818-4}, author = {Vora, Siddhi M and Ankati, Sravani and Patole, Chhaya and Podile, Appa Rao and Archana, G} } @article {1863, title = {Antimicrobial and antibiofilm activity of GNP-Tannic Acid-Ag nanocomposite and their epoxy-based coatings [Log9 Materials, a C-CAMP startup]}, journal = {Progress in Organic Coatings}, volume = {159}, year = {2021}, month = {07/2021}, pages = {106421}, type = {Journal Article}, abstract = {

In order to deal with the challenge of biofilms and its associated infections, it is important to develop some efficient methodology which prevents the formation of biofilms over the surfaces. Herein, we have successfully developed an antibacterial and antibiofilm composite using non-toxic and hydrophobic nanomaterial, graphene nanoplatelets, via facile two step methodology including functionalization and subsequent hydrothermal treatment. The as-prepared composites were characterized using FE-SEM, FTIR and Raman spectroscopy to obtain morphological, compositional and structural details. In addition, the antibacterial efficiency of these composites was investigated against antibiotic resistant S. aureus and E. coli. Afterwards, graphene composite based epoxy coatings were prepared on glass substrate and tested for their antibiofilm efficiency against methicillin resistant S. aureus strain. The abundant presence of hydroxyl groups over the surface due to tannic acid, hydrophobicity of graphene and epoxy and remarkable antibacterial efficiency of tannic acid (TA), silver (Ag) and graphene synergistically enhance the antibiofilm efficiency of such coatings. This work presents a new strategy to develop a multifunctional coating and subsequently lessen the risk of biofilm assisted infections.

}, keywords = {Antibiofilm, Antimicrobial, Composite, Cytotoxicity, Graphene nanoplatelets}, doi = {https://doi.org/10.1016/j.porgcoat.2021.106421}, url = {https://www.sciencedirect.com/science/article/pii/S0300944021002927$\#$}, author = {Singhal, Akshay V and Malwal, Deepika and Thiyagarajan, Shankar and Lahiri, Indranil} } @article {1858, title = {Azaindole Based Potentiator of Antibiotics against Gram-Negative Bacteria [C-CAMP Startup Bugworks]}, journal = {ACS Infectious Diseases}, year = {2021}, month = {10/2021}, type = {Journal Article}, abstract = {
We discovered azaindole-based compounds with weak innate activity that exhibit substantial potentiation of antibacterial activities of different antibiotics, viz., rifampicin, erythromycin, solithromycin, and novobiocin in Gram-negative bacteria. In the presence of the azaindole derivatives, these antibiotics exhibited submicromolar minimum inhibitory concentrations (MICs) against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. The fold improvements in MIC of these antibiotics that were otherwise weak or inactive on their own against these bacteria were also observed against drug-resistant clinical isolates. Our studies indicate that this selective potentiation is probably through destabilization of the outer membrane{\textquoteright}s integrity, known to be regulated by the lipopolysaccharides (LPS). Thus, the azaindole based compounds described here open opportunities for those antibiotics that are otherwise ineffective due to LPS mediated entry barriers in Gram-negative bacteria.
}, keywords = {Azaindole, Bacterial Permeability, Lipopolysachhardies (LPS), Polymyxin, Synergy}, doi = {10.1021/acsinfecdis.1c00171}, url = {https://pubs.acs.org/doi/abs/10.1021/acsinfecdis.1c00171$\#$}, author = {Sharma, Sreevalli and Rao, Ranga and Reeve, Stephanie M and Phelps, Gregory A and Bharatham, Nagakumar and Katagihallimath, Nainesh and Ramachandran, Vasanthi and Raveendran, Savitha and Sarma, Maitrayee and Nath, Anubha} } @article {1145, title = {Aspartic protease from Aspergillus niger: Molecular characterization and interaction with pepstatin A [Mass Spectrometry Facility - Proteomics].}, journal = {Int J Biol Macromol}, year = {2019}, month = {2019 Jul 30}, abstract = {

In the pursuit of industrial aspartic proteases, aspergillopepsin A-like endopeptidase from the fungi Aspergillus niger, was identified and cultured by solid state fermentation. Conventional chromatographic techniques were employed to purify the extracellular aspartic protease to apparent homogeneity. The enzyme was found to have single polypeptide chain with a molecular mass of 50 {\textpm} 0.5 kDa. The optimum pH and temperature for the purified aspartic protease was found to be 3.5 and 60 {\textdegree}C respectively. The enzyme was stable for 60 min at 50 {\textdegree}C. The purified enzyme had specific activity of 40,000 {\textpm} 1800 U/mg. The enzyme had 85\% homology with the reported aspergillopepsin A-like aspartic endopeptidase from Aspergillus niger CBS 513.88, based on tryptic digestion and peptide analysis. Pepstatin A reversibly inhibited the enzyme with a K value of 0.045 μM. Based on homology modeling and predicted secondary structure, it was inferred that the aspartic protease is rich in β-structures, which was also confirmed by CD measurements. Interaction of pepstatin A with the enzyme did not affect the conformation of the enzyme as evidenced by CD and fluorescence measurements. Degree of hydrolysis of commercial substrates indicated the order of cleaving ability of the enzyme to be hemoglobin \> defatted soya flour \> gluten \> gelatin \> skim milk powder. The enzyme also improved the functional characteristics of defatted soya flour. This aspartic protease was found to be an excellent candidate for genetic manipulation for biotechnological application in food and feed industries, due to its high catalytic turn over number and thermostability.

}, issn = {1879-0003}, doi = {10.1016/j.ijbiomac.2019.07.133}, author = {Purushothaman, Kavya and Bhat, Sagar Krishna and Singh, Sridevi Annapurna and Marathe, Gopal Kedihithlu and Appu Rao, Appu Rao G} } @article {690, title = {Automation aided optimization of cloning, expression and purification of enzymes of the bacterial sialic acid catabolic and sialylation pathways enzymes for structural studies. [Protein Technology (INT)]}, year = {2018}, author = {Sneha Bairy and Lakshmi Narayanan Gopalan and Thanuja Gangi Setty and Sathya Srinivasachari and Lavanyaa Manjunath and Jay Prakash Kumar and Sai R Guntupalli and Sucharita Bose and Vinod Nayak and Swagatha Ghosh and Nitish Sathyanarayanan and Rhawnie Caing-Carlsson and Weixiao Yuan Wahlgren and Rosmarie Friemann and S. Ramaswamy and Muniasamy Neerathilingam} } @article {713, title = {Active remodeling of cortical actin regulates spatiotemporal organization of cell surface molecules.}, journal = {Cell}, volume = {149}, year = {2012}, month = {2012 Jun 08}, pages = {1353-67}, abstract = {

Many lipid-tethered proteins and glycolipids exist as monomers and nanoclusters on the surface of living cells. The spatial distribution and dynamics of formation and breakup of nanoclusters does not reflect thermal and chemical equilibrium and is controlled by active remodeling of the underlying cortical actin. We propose a model for nanoclustering based on active hydrodynamics, wherein cell surface molecules bound to dynamic actin are actively driven to form transient clusters. This consistently explains all of our experimental observations. Using FCS and TIRF microscopy, we provide evidence for the existence of short, dynamic, polymerizing actin filaments at the cortex, a key assumption of the theoretical framework. Our theory predicts that lipid-anchored proteins that interact with dynamic actin must exhibit anomalous concentration fluctuations, and a cell membrane protein capable of binding directly to actin can form nanoclusters. These we confirm experimentally, providing an active mechanism for molecular organization and its spatiotemporal regulation on the plasma membrane.

}, keywords = {Actins, Animals, Cell Line, Tumor, Cell Membrane, CHO Cells, Cricetinae, Cytoskeleton, Humans, Membrane Proteins, Models, Biological, Spectrometry, Fluorescence}, issn = {1097-4172}, doi = {10.1016/j.cell.2012.05.008}, author = {Gowrishankar, Kripa and Ghosh, Subhasri and Saha, Suvrajit and C, Rumamol and Mayor, Satyajit and Rao, Madan} } @article {720, title = {Antiproliferative property of n-hexane and chloroform extracts of Anisomeles malabarica (L). R. Br. in HPV16-positive human cervical cancer cells.}, journal = {J Pharmacol Pharmacother}, volume = {3}, year = {2012}, month = {2012 Jan}, pages = {26-34}, abstract = {

OBJECTIVES: To find the efficacy of serial extracts of Anisomeles malabarica in inhibiting proliferation of and inducing apoptosis in human cervical cancer cells, SiHa and ME 180, that are HPV 16-positive.

MATERIALS AND METHODS: The whole plant was extracted in n-hexane, chloroform, ethyl acetate, n-butanol, methanol, and water. The cells were treated with the extracts at increasing concentrations to find the IC(50), adopting MTT ([3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]) assay. Acridine orange (AO) and ethidium bromide (EB) and Hoechst 33258 staining were adopted to assess the mode of cell death, Annexin V-Cy3 staining to evaluate one of the early apoptotic features, JC-1 staining to assess the mitochondrial membrane depolarization, comet assay for DNA fragmentation, and cell cycle analysis for the distribution of cells after treatment.

RESULTS: n-Hexane and chloroform extracts were cytotoxic to the cervical cancer cells in dose- and duration-dependent manner. The cells that responded to the treatments revealed typical apoptotic features. Early features of apoptosis, phosphatidyl serine translocation and loss of mitochondrial trans-membrane potential, were observed in the treated cells, and comet assay revealed DNA damage. In the FACS analysis, the cells accumulated in the sub-G0/G1 phase of the cell cycle, except in n-hexane- and chloroform extract-treated SiHa cells at 24 h, which showed arrest in S- and G2/M phases.

CONCLUSIONS: n-Hexane and chloroform extracts of A. malabarica inhibit proliferation of and induce death in HPV16-positive cervical cancer cells, mostly by apoptosis and to some extent by necrosis.

}, issn = {0976-5018}, doi = {10.4103/0976-500X.92500}, author = {Preethy, Christo Paul and Padmapriya, Ramamoorthy and Periasamy, Vaiyapuri Subbarayan and Riyasdeen, Anvarbatcha and Srinag, Suresh and Krishnamurthy, Hanumanthappa and Alshatwi, Ali Abdullah and Akbarsha, Mohammad Abdulkader} }