Identification of Escherichia coli strains from water vending machines of Kelantan, Malaysia using 16S rRNA gene sequence analysis

Water vending machines provide an alternative source of clean and safe drinking water to the consumers. However, the quality of drinking water may alter due to contamination from lack of hygienic practices and maintenance of the machines. Hence, this study was conducted to determine the microbiological quality of water from vending machines and associated contact surfaces. Seventeen water samples and 85 swab samples (nozzles, drip trays, coin slots, buttons and doors) from 3 locations in Kelantan were collected. Polymerase chain reaction amplification and 16S ribosomal ribonucleic acid (rRNA) sequencing were carried out and sequences obtained were compared against the sequences available in the National Centre for Biotechnology Information database using the basic local alignment search tool programme. Coliform counts were observed in 94.12 % of water samples, 76.47 % of nozzles and 82.35 % of drip tray swabs. Furthermore, results of 16S rRNA sequence analysis indicated that two gram-negative isolates were identified as Escherichia coli U 5/41 (Accession no. NR_024570.1) and E. coli O157:H7 EDL933 (Accession no. CP008957.1) with similarity value of 100 %, respectively. The results from this study further improve our understanding of the potential microorganisms in drinking water. Regular maintenance and cleaning of water vending machines are important to reduce bacterial growth and the presence of waterborne pathogens

Measurement of the viscoelastic properties of blood plasma clot formation in response to tissue factor concentration-dependent activation

© 2016, The Author(s). The coagulation of blood plasma in response to activation with a range of tissue factor (TF) concentrations was studied with a quartz crystal microbalance (QCM), where frequency and half width at half maximum (bandwidth) values measured from the conductance spectrum near resonant frequency were used. Continuous measurement of bandwidth along with the frequency allows for an understanding of the dissipative nature of the forming viscoelastic clot, thus providing information on the complex kinetics of the viscoelastic changes occurring during the clot formation process. Using a mathematical model, these changes in frequency and bandwidth have been used to derive novel QCM parameters of effective elasticity, effective mass density and rigidity factor of the viscoelastic layer. The responses of QCM were compared with those from thromboelastography (TEG) under identical conditions. It was demonstrated that the nature of the clot formed, as determined from the QCM parameters, was highly dependent on the rate of clot formation resulting from the TF concentration used for activation. These parameters could also be related to physical clot characteristics such as fibrin fibre diameter and fibre density, as determined by scanning electron microscopic image analysis. The maximum amplitude (MA) as measured by TEG, which purports to relate to clot strength, was unable to detect these differences

New structural insights into the role of TROVE2 complexes in the on-set and pathogenesis of systemic lupus eythematosus determined by a combiantion of QCM-D and DPI

The final publication is available at link.springer.com.[EN] The mechanism of self-recognition of the autoantigen TROVE2, a common biomarker in autoimmune diseases, has been studied with a quartz crystal microbalance with dissipation monitoring (QCM-D) and dual polarization interferometry (DPI). The complementarity and remarkable analytical features of both techniques has allowed new insights into the onset of systemic lupus erythematosus (SLE) to be achieved at the molecular level. The in vitro study for SLE patients and healthy subjects suggests that anti-TROVE2 autoantibodies may undergo an antibody bipolar bridging. An epitope-paratope-specific binding initially occurs to activate a hidden Fc receptor in the TROVE2 tertiary structure. This bipolar mechanism may contribute to the pathogenic accumulation of anti-TROVE2 autoantibody immune complex in autoimmune disease. Furthermore, the specific calcium-dependent protein-protein bridges point out at how the TRIM21/TROVE2 association might occur, suggesting that the TROVE2 protein could stimulate the intracellular immune signaling via the TRIM21 PRY-SPRY domain. These findings may help to better understand the origins of the specificity and affinity of TROVE2 interactions, which might play a key role in the SLE pathogenesis. This manuscript gives one of the first practical applications of two novel functions (-df/dD and Delta h/molec) for the analysis of the data provided by QCM-D and DPI. In addition, it is the first time that QCM-D has been used for mapping hidden Fc receptors as well as linear epitopes in a protein tertiary structure.We would like to thank Sylvia Daunert for her invaluable help with the discussion of the paper. Furthermore, we acknowledge financial support from the Generalitat Valenciana (GVA-PROMETEOII/2014/040) as well as the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund under award numbers CTQ2013-45875-R and CTQ2013-42914-RJuste-Dolz, AM.; Do Nascimento, NM.; Monzó, IS.; Grau-García, E.; Roman-Ivorra, JA.; López-Paz, JL.; Escorihuela Fuentes, J.... (2019). New structural insights into the role of TROVE2 complexes in the on-set and pathogenesis of systemic lupus eythematosus determined by a combiantion of QCM-D and DPI. Analytical and Bioanalytical Chemistry. 411(19):4709-4720. https://doi.org/10.1007/s00216-018-1407-xS4709472041119Kakatia S, Teronpia R, Barmanb B. Frequency, pattern and determinants of flare in systemic lupus erythematosus: a study from North East India. Egypt Rheumatol. 2015;37:S55–9.Kuhn A, Wenzel J, Weyd H. Photosensitivity, apoptosis, and cytokines in the pathogenesis of lupus erythematosus: a critical review. 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An RNA degradation machine sculpted by Ro autoantigen and noncoding RNA. Cell. 2013;153:166–77.Stein AJ, Fuchs G, Fu C, Wolin SL, Reinisch KM. Structural insights into RNA quality control: the Ro autoantigen binds misfolded RNAs via its central cavity. Cell. 2005;121:529–39.Reed JH, Gordon TP. Autoimmunity: Ro60-associated RNA takes its toll on disease pathogenesis. Nat Rev Rheumatol. 2016;12:136–8.Sim S, Weinberg DE, Fuchs G, Choi K, Chung J, Wolin SL. The subcellular distribution of an RNA quality control protein, the Ro autoantigen, is regulated by noncoding Y RNA binding. Mol Biol Cell. 2009;20:1555–64.Reed JH, Jackson MW, Gordon TP. A B cell apotope of Ro 60 in systemic lupus erythematosus. Arthritis Rheum. 2008;58:1125–9.Wolin SL, Reinisch KM. The Ro 60 kDa autoantigen comes into focus: interpreting epitope mapping experiments on the basis of structure. Autoimmun Rev. 2006;5:367–72.Routsias JG, Tzioufas AG. B-cell epitopes of the intracellular autoantigens Ro/SSA and La/SSB: tools to study the regulation of the autoimmune response. J Autoimmun. 2010;35:256–64.Whittaker CA, Hynes RO. Distribution and evolution of von Willebrand/integrin a domains: widely dispersed domains with roles in cell adhesion and elsewere. Mol Bio Cell. 2002;13:3369–87.Lacy DB, Wigelsworth DJ, Scobie HM, Young JA, Collier RJ. Crystal structure of the von Willebrand factor a domain of human capillary morphogenesis protein 2: an anthrax toxin receptor. Proc Natl Acad Sci U S A. 2004;101:6367–72.O’Brien CA, Wolin SL. A possible role for the 60-kD Ro autoantigen in a discard pathway for defective 5S rRNA precursors. Genes Dev. 1994;8:2891–903.Chen X, Wolin SL. The Ro 60 autoantigen : insights into cellular function and role in autoimmunity. J Mol Med (Berl). 2004;82:232–9.Escorihuela J, González-Martínez MA, López-Paz JL, Puchades R, Maquieira A, Gimenez-Romero D. Dual-polarization interferometry: a novel technique to light up the nanomolecular world. Chem Rev. 2014;115:265–94.do Nascimento NM, Juste-Dolz A, Bueno PR, Monzó I, Tejero R, Lopez-Paz JL, et al. Mapping molecular binding by means of conformational dynamics measurements. RSC Adv. 2018;8:867–76.do Nascimento NM, Juste-Dolz A, Grau-García E, Román-Ivorra J, Puchades R, Maquieira A, et al. Label-free piezoelectric biosensor for prognosis and diagnosis of systemic lupus erythematosus. Biosens. Bioelectron. 2016;90:166–73.Seo MH, Park J, Kim E, Hohng S, Kim HS. Protein conformational dynamics dictate the binding affinity for a ligand. Nat Commun. 2014;5:3724.Lakshmanan RS, Efremov V, O’Donnell JS, Killard AJ. Measurement of the viscoelastic properties of blood plasma clot formation in response to tissue factor concentration-dependent activation. Anal Bioanal Chem. 2016;408:6581–8.Fakhrullin RF, Vinter VG, Zamaleeva AI, Matveeva MV, Kourbanov RA, Temesgen BK, et al. Quartz crystal microbalance immunosensor for the detection of antibodies to double-stranded DNA. Anal Bioanl Chem. 2007;388:367–75.Shen F, Rojas OJ, Genzer J, Gurgel PV, Carbonell RG. Affinity interactions of human immunoglobulin G with short peptides: role of ligand spacer on binding, kinetics, and mass transfer. Anal Bioanl Chem. 2015;408:1829–41.Fogarty AC, Laage D. Water dynamics in protein hydration shells: the molecular origins of the dynamical perturbation. J Phys Chem B. 2014;118:7715–29.Born B, Kim SJ, Ebbinghaus S, Gruebelebc M, Havenith M. The terahertz dance of water with the proteins: the effect of protein flexibility on the dynamical hydration shell of ubiquitin. Faraday Discuss. 2009;141:161–73.Yoshimi R, Ueda A, Ozato K, Ishigatsubo Y. Clinical and pathological roles of Ro/SSA autoantibody system. Clin Dev Immunol. 2012;2012:606195.Boire G, Gendron M, Monast N, Bastin B, Ménard HA. Purification of antigenically intact Ro ribonucleoproteins; biochemical and immunological evidence that the 52-kD protein is not a Ro protein. Clin Exp Immunol. 1995;100:489–98.Gazzaruso C, Montecucco CM, Geroldi D, Garzaniti A, Finardi G. Severe hypercalcemia and systemic lupus erythematosus. Joint Bone Spine. 2000;67:485–8.Hassan AB, Lundberg IE, Isenberg D, Wahren-Herlenius M. Serial analysis of Ro/SSA and La/SSB antibody levels and correlation with clinical disease activity in patients with systemic lupus erythematosus. Scand J Rheumatol. 2002;31:133–9.Huang RY, Chen G. Higher order structure characterization of protein therapeutics by hydrogen/deuterium exchange mass spectrometry. Anal Bioanal Chem. 2014;406:6541–58.Yu F, Roy S, Arevalo E, Schaeck J, Wang J, Holte K, et al. Characterization of heparin-protein interaction by saturation transfer difference (STD) NMR. Anal Bioanal Chem. 2014;406:3079–89.Rizzuto R, Pozzan T. Microdomains of intracellular Ca2+: molecular determinants and functional consequences. Physiol Rev. 2006;86:369–408.Gaipl US, Kuhn A, Sheriff A, Munoz LE, Franz S, Voll RE, et al. Clearance of apoptotic cells in human SLE. Curr Dir Autoimmun. 2006;9:173–87.Falati S, Edmead CE, Poole AW. Glycoprotein Ib-V-IX, a receptor for Von Willebrand factor, couples physically and functionally to the Fc receptor gamma-chain, Fyn, and Lyn to activate human platelets. Blood. 1999;94:1648–56.Muñoz LE, Lauber K, Schiller M, Manfredi AA, Herrmann M. The role of defective clearance of apoptotic cells in systemic autoimmunity. Nat Rev Rheumatol. 2010;6:280–9

S-D logic-informed customer engagement: Integrative framework, revised fundamental propositions, and application to CRM

Advance online in 2016</p

The Use of Trademarks Or Names O F Manufacturers I N

this report is for accurate reporting and does not constitute an official endorsement, e ither expressed or i mplied, o f such products or manufacturers by the Nationa l Aeronaut ics and Space Administration. This publication i s ava il able f rom the f ol lowing sources: NASA Center for AeroSpace Information National Technical Information Service (NTIS) 800 Elkridge Landing Road 5285 Port Royal Road Linthicum Heights, MD 21090-2934 Springfield, VA 22161-2171 (301) 621-0390 (703) 487-4650 Introduction Computational fluid dynamics (CFD) methods and advanced turbulence models are needed to predict propulsion aerodynamic effects in transonic and supersonic free-stream conditions. Analytical results are frequently used to supplement the experimental data in critical design decisions. Accurate prediction of the pressure distribution and the skin friction coefficient is paramount to the design of propulsion systems. In the area of propulsion integration, accurate predictions of boundary layer structure, skin friction, and flow separation by CFD methods are critical. Two-equation turbulence models (ref. 1) offer several advantages over other approaches that compute practical flow problems. For example, algebraic models lack turbulence history-dependent nonlocal effects (through the convection and viscous diffusion of the Reynolds stress models), effects which are known to be important in determining the turbulence structure in complex flows. The numerical calculations that use the more advanced Reynolds stress models (refs. 2 and 3) require the solution of transport equations for each component of the Reynolds stress tensor in addition to solution of the NavierStokes equations; this approach requires tremendous computational time for three-dimensional flow problems. The tran..

Extract of Penicillium sclerotiorum an endophytic fungus isolated from Cassia fistula L. induces cell cycle arrest leading to apoptosis through mitochondrial membrane depolarization in human cervical cancer cells

Seventeen endophytic fungi were isolated from various tissues of Cassia fistula and the ethyl acetate extracts obtained from 21-day cultures of all the endophytic fungal isolates were initially screened for their cytotoxicity against HeLa (human cervical carcinoma) cells using MIT assay. Of these, Penicillium sclerotiorum extract (PSE), significantly affected the viability of HeLa cells in a dose-dependent manner. The extract of P. Sclerotiorum was further analyzed by GC-MS, which showed three compounds, hexadecanoic acid, oleic acid and benzoic acid to be the major active principles in the extracts.The extract was further tested for invitro cytotoxicity against five cancer cell lines. Of the cell lines tested, HeLa cells showed maximum sensitivity followed by A549, while A431 and U251 were moderately sensitive and MCF-7 was insensitive to the treatment. In addition, normal human embryonic kidney cells, HEK293 remained insensitive to the treatment. Furthermore, the mechanism of cytotoxic activity exhibited by PSE was investigated by evaluating cell cycle progression and apoptotic induction in HeLa cells. Cell cycle analysis revealed that the PSE arrested cells at S and G2/M phase of the cell cycle in a dose-dependent manner. Annexin V- Propidium iodide double staining showed that, the extract potentiates apoptosis rather than necrosis in cells. This was supported by the down regulation in the proapoptotic protein BCL2 and up regulation of BAX (BCL2 Associated X), tumor suppressor protein, p53 and Apaf-1 Apoptotic Peptidase Activating Factor 1]. Loss of mitochondrial membrane potential and a distinct DNA fragmentation pattern observed following the treatment, suggest that the PSE treatment leads to activation of mitochondrial pathway of apoptosis. Further, the extract also exhibited both antioxidant and anti-angiogenic properties. These results indicate that endophytic fungi isolated from medicinal plants may serve as potential sources of the anti-cancerous compounds