92 research outputs found
Relationship between apoptosis and the BH2 domain sequence of the VP5 peptide of infectious pancreatic necrosis virus
Objective. To determine whether the level of apoptosis induced by infectious pancreatic necrosis
virus (IPNV) is related to the amino acid sequence of the BH2 domain of the VP5 protein and the
level of infectivity. Materials and methods. Three IPNV strains were used, the VP2 protein gene was
amplified for genotyping and the VP5 sequence was also obtained. The infectivity of the strains was
calculated using the viral titer obtained at 12, 24, 36 and 45 hpi in CHSE-214 cells. The percentage
of apoptosis in infected cells was visualized by TUNEL assay and immunohistochemistry (caspase 3
detection). Results. The V70/06 and V33/98 strains corresponded to genotype Sp, while V112/06 to
VR-299; the amino acid analysis of the V70/06 strain allows its classification as middle virulent strain
and V33/98 and V112/06 strains as low virulent ones; infection with the V112/06 strain produced a
lower viral titer (p<0.05). The VP5 gene of the 3 strains showed four homologous domains to Bcl-2,
however, the BH2 domain was truncated in V70/06 and V33/98 (12 kDa), being complete (15kDa) in
V112/06, which also showed the Trp155 residue, equivalent to Trp188 considered as a critical factor
for the function of Bcl-2. The average apoptosis was below 12%, showing no differences between
strains (p>0.05). Conclusions. The results showed that the differences in the BH2 sequence of the
VP5 protein, infectivity and the VP2 sequence are not associated with the modulation of apoptosis
Inhibition of Non-flux-Controlling Enzymes Deters Cancer Glycolysis by Accumulation of Regulatory Metabolites of Controlling Steps
Glycolysis provides precursors for the synthesis of macromolecules and may contribute to the ATP supply required for the constant and accelerated cellular duplication in cancer cells. In consequence, inhibition of glycolysis has been reiteratively considered as an anti-cancer therapeutic option. In previous studies, kinetic modeling of glycolysis in cancer cells allowed the identification of the main steps that control the glycolytic flux: glucose transporter, hexokinase (HK), hexose phosphate isomerase (HPI) and glycogen degradation in human cervix HeLa cancer cells and rat AS-30D ascites hepatocarcinoma. It was also previously experimentally determined that simultaneous inhibition of the non-controlling enzymes lactate dehydrogenase (LDH), pyruvate kinase (PYK) and enolase (ENO) brings about significant decrease in the glycolytic flux of cancer cells and accumulation of intermediate metabolites, mainly fructose-1,6-bisphosphate (Fru1,6BP) and dihydroxyacetone phosphate (DHAP), which are inhibitors of HK and HPI, respectively. Here it was found by kinetic modeling that inhibition of cancer glycolysis can be attained by blocking downstream non flux-controlling steps as long as Fru1,6BP and DHAP, regulatory metabolites of flux-controlling enzymes, are accumulated. Furthermore, experimental results and further modeling showed that oxamate and iodoacetate inhibitions of PYK, ENO and glyceraldehyde3-phosphate dehydrogenase (GAPDH), but not of LDH and phosphoglycerate kinase, induced accumulation of Fru1,6BP and DHAP in AS-30D hepatoma cells. Indeed, PYK, ENO and GAPDH exerted the highest control on the Fru1,6BP and DHAP concentrations. The high levels of these metabolites inhibited HK and HPI and led to glycolytic flux inhibition, ATP diminution and accumulation of toxic methylglyoxal. Hence, the anticancer effects of downstream glycolytic inhibitors are very likely mediated by this mechanism. In parallel, it was also found that uncompetitive inhibition of the flux-controlling steps is a more potent mechanism than competitive and mixed-type inhibition to efficiently perturb cancer glycolysis
Glycoprotein Ib activation by thrombin stimulates the energy metabolism in human platelets
<div><p>Thrombin-induced platelet activation requires substantial amounts of ATP. However, the specific contribution of each ATP-generating pathway <i>i</i>.<i>e</i>., oxidative phosphorylation (OxPhos) versus glycolysis and the biochemical mechanisms involved in the thrombin-induced activation of energy metabolism remain unclear. Here we report an integral analysis on the role of both energy pathways in human platelets activated by several agonists, and the signal transducing mechanisms associated with such activation. We found that thrombin, Trap-6, arachidonic acid, collagen, A23187, epinephrine and ADP significantly increased glycolytic flux (3â38 times <i>vs</i>. non-activated platelets) whereas ristocetin was ineffective. OxPhos (33 times) and mitochondrial transmembrane potential (88%) were increased only by thrombin. OxPhos was the main source of ATP in thrombin-activated platelets, whereas in platelets activated by any of the other agonists, glycolysis was the principal ATP supplier. In order to establish the biochemical mechanisms involved in the thrombin-induced OxPhos activation in platelets, several signaling pathways associated with mitochondrial activation were analyzed. Wortmannin and LY294002 (PI3K/Akt pathway inhibitors), ristocetin and heparin (GPIb inhibitors) as well as resveratrol, ATP (calcium-release inhibitors) and PP1 (Tyr-phosphorylation inhibitor) prevented the thrombin-induced platelet activation. These results suggest that thrombin activates OxPhos and glycolysis through GPIb-dependent signaling involving PI3K and Akt activation, calcium mobilization and protein phosphorylation.</p></div
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(The features of state control over the activities of small businesses are analyzed. The authorâs definition of the method of monitoring in the state financial control over tax payments is given, and the expedience for its use in the prospects is grounded.
Defining the Molecular Basis of Tumor Metabolism: a Continuing Challenge Since Warburg's Discovery
Cancer cells are the product of genetic disorders that alter crucial intracellular signaling pathways associated with the regulation of cell survival, proliferation, differentiation and death mechanisms. the role of oncogene activation and tumor suppressor inhibition in the onset of cancer is well established. Traditional antitumor therapies target specific molecules, the action/expression of which is altered in cancer cells. However, since the physiology of normal cells involves the same signaling pathways that are disturbed in cancer cells, targeted therapies have to deal with side effects and multidrug resistance, the main causes of therapy failure. Since the pioneering work of Otto Warburg, over 80 years ago, the subversion of normal metabolism displayed by cancer cells has been highlighted by many studies. Recently, the study of tumor metabolism has received much attention because metabolic transformation is a crucial cancer hallmark and a direct consequence of disturbances in the activities of oncogenes and tumor suppressors. in this review we discuss tumor metabolism from the molecular perspective of oncogenes, tumor suppressors and protein signaling pathways relevant to metabolic transformation and tumorigenesis. We also identify the principal unanswered questions surrounding this issue and the attempts to relate these to their potential for future cancer treatment. As will be made clear, tumor metabolism is still only partly understood and the metabolic aspects of transformation constitute a major challenge for science. Nevertheless, cancer metabolism can be exploited to devise novel avenues for the rational treatment of this disease. Copyright (C) 2011 S. Karger AG, BaselFundação de Amparo Ă Pesquisa do Estado de SĂŁo Paulo (FAPESP)Conselho Nacional de Desenvolvimento CientĂfico e TecnolĂłgico (CNPq)Univ Fed ABC UFABC, CCNH, Santo Andre, BrazilUniversidade Federal de SĂŁo Paulo UNIFESP, Dept Ciencias Biol, SĂŁo Paulo, BrazilUniversidade Federal de SĂŁo Paulo UNIFESP, Dept Bioquim, SĂŁo Paulo, BrazilUniv Fed Sao Carlos UFSCar, DFQM, Sorocaba, BrazilUniversidade Federal de SĂŁo Paulo UNIFESP, Dept Ciencias Biol, SĂŁo Paulo, BrazilUniversidade Federal de SĂŁo Paulo UNIFESP, Dept Bioquim, SĂŁo Paulo, BrazilFAPESP: 10/16050-9FAPESP: 10/11475-1FAPESP: 08/51116-0Web of Scienc
Development of the supply chain oriented quality assurance system for aerospace manufacturing SMEs and its implementation perspectives
Aerospace manufacturing SMEs are continuously facing the challenge on managing their supply chain and complying with the aerospace manufacturing quality standard requirement due to their lack of resources and the nature of business. In this paper, the ERP system based approach is presented to quality control and assurance work in light of seamless integration of in-process production data and information internally and therefore managing suppliers more effectively and efficiently. The Aerospace Manufacturing Quality Assurance Standard (BS/EN9100) is one of the most recognised and essential protocols for developing the industry-operated-and-driven quality assurance systems. The research investigates using the ERP based system as an enabler to implement BS/EN9100 quality management system at manufacturing SMEs and the associated implementation and application perspectives. An application case study on a manufacturing SME is presented by using the SAP based implementation, which helps further evaluate and validate the approach and application system development
Polymorphisms in the Mitochondrial Ribosome Recycling Factor EF-G2mt/MEF2 Compromise Cell Respiratory Function and Increase Atorvastatin Toxicity
Mitochondrial translation, essential for synthesis of the electron transport chain complexes in the mitochondria, is governed by nuclear encoded genes. Polymorphisms within these genes are increasingly being implicated in disease and may also trigger adverse drug reactions. Statins, a class of HMG-CoA reductase inhibitors used to treat hypercholesterolemia, are among the most widely prescribed drugs in the world. However, a significant proportion of users suffer side effects of varying severity that commonly affect skeletal muscle. The mitochondria are one of the molecular targets of statins, and these drugs have been known to uncover otherwise silent mitochondrial mutations. Based on yeast genetic studies, we identify the mitochondrial translation factor MEF2 as a mediator of atorvastatin toxicity. The human ortholog of MEF2 is the Elongation Factor Gene (EF-G) 2, which has previously been shown to play a specific role in mitochondrial ribosome recycling. Using small interfering RNA (siRNA) silencing of expression in human cell lines, we demonstrate that the EF-G2mt gene is required for cell growth on galactose medium, signifying an essential role for this gene in aerobic respiration. Furthermore, EF-G2mt silenced cell lines have increased susceptibility to cell death in the presence of atorvastatin. Using yeast as a model, conserved amino acid variants, which arise from non-synonymous single nucleotide polymorphisms (SNPs) in the EF-G2mt gene, were generated in the yeast MEF2 gene. Although these mutations do not produce an obvious growth phenotype, three mutations reveal an atorvastatin-sensitive phenotype and further analysis uncovers a decreased respiratory capacity. These findings constitute the first reported phenotype associated with SNPs in the EF-G2mt gene and implicate the human EF-G2mt gene as a pharmacogenetic candidate gene for statin toxicity in humans
The CARMENES search for exoplanets around M dwarfs. Two temperate Earth-mass planet candidates around Teegardenâs Star
Context.Teegardenâs Star is the brightest and one of the nearest ultra-cool dwarfs in the solar neighbourhood. For its late spectral type (M7.0 V),the star shows relatively little activity and is a prime target for near-infrared radial velocity surveys such as CARMENES.Aims.As part of the CARMENES search for exoplanets around M dwarfs, we obtained more than 200 radial-velocity measurements of TeegardenâsStar and analysed them for planetary signals.Methods.We find periodic variability in the radial velocities of Teegardenâs Star. We also studied photometric measurements to rule out stellarbrightness variations mimicking planetary signals.Results.We find evidence for two planet candidates, each with 1.1Mâminimum mass, orbiting at periods of 4.91 and 11.4 d, respectively. Noevidence for planetary transits could be found in archival and follow-up photometry. Small photometric variability is suggestive of slow rotationand old age.Conclusions.The two planets are among the lowest-mass planets discovered so far, and they are the first Earth-mass planets around an ultra-cooldwarf for which the masses have been determined using radial velocities.We thank the referee Rodrigo DĂaz for a careful review andhelpful comments. M.Z. acknowledges support from the Deutsche Forschungs-gemeinschaft under DFG RE 1664/12-1 and Research Unit FOR2544 âBluePlanets around Red Starsâ, project no. RE 1664/14-1. CARMENES isan instrument for the Centro AstronĂłmico Hispano-AlemĂĄn de Calar Alto(CAHA, AlmerĂa, Spain). CARMENES is funded by the German Max-Planck-Gesellschaft (MPG), the Spanish Consejo Superior de InvestigacionesCientĂficas (CSIC), the European Union through FEDER/ERF FICTS-2011-02 funds, and the members of the CARMENES Consortium (Max-Planck-Institut fĂŒr Astronomie, Instituto de AstrofĂsica de AndalucĂa, LandessternwarteKönigstuhl, Institut de CiĂšncies de lâEspai, Institut fĂŒr Astrophysik Göttingen,Universidad Complutense de Madrid, ThĂŒringer Landessternwarte Tautenburg,Instituto de AstrofĂsica de Canarias, Hamburger Sternwarte, Centro de Astro-biologĂa and Centro AstronĂłmico Hispano-AlemĂĄn), with additional contribu-tions by the Spanish Ministry of Economy, the German Science Foundationthrough the Major Research Instrumentation Programme and DFG ResearchUnit FOR2544 âBlue Planets around Red Starsâ, the Klaus Tschira Stiftung, thestates of Baden-WĂŒrttemberg and Niedersachsen, and by the Junta de AndalucĂa.Based on data from the CARMENES data archive at CAB (INTA-CSIC). Thisarticle is based on observations made with the MuSCAT2 instrument, devel-oped by ABC, at Telescopio Carlos SĂĄnchez operated on the island of Tener-ife by the IAC in the Spanish Observatorio del Teide. Data were partly col-lected with the 150-cm and 90-cm telescopes at the Sierra Nevada Observa-tory (SNO) operated by the Instituto de AstrofĂsica de AndalucĂa (IAA-CSIC).Data were partly obtained with the MONET/South telescope of the MOnitoringNEtwork of Telescopes, funded by the Alfried Krupp von Bohlen und HalbachFoundation, Essen, and operated by the Georg-August-UniversitĂ€t Göttingen,the McDonald Observatory of the University of Texas at Austin, and the SouthAfrican Astronomical Observatory. We acknowledge financial support from theSpanish Agencia Estatal de InvestigaciĂłn of the Ministerio de Ciencia, Inno-vaciĂłn y Universidades and the European FEDER/ERF funds through projectsAYA2015-69350-C3-2-P, AYA2016-79425-C3-1/2/3-P, AYA2018-84089, BES-2017-080769, BES-2017-082610, ESP2015-65712-C5-5-R, ESP2016-80435-C2-1/2-R, ESP2017-87143-R, ESP2017-87676-2-2, ESP2017-87676-C5-1/2/5-R, FPU15/01476, RYC-2012-09913, the Centre of Excellence âSevero Ochoaâand âMarĂa de Maeztuâ awards to the Instituto de AstrofĂsica de Canarias (SEV-2015-0548), Instituto de AstrofĂsica de AndalucĂa (SEV-2017-0709), and Cen-tro de AstrobiologĂa (MDM-2017-0737), the Generalitat de Catalunya throughCERCA programmeâ, the Deutsches Zentrum fĂŒr Luft- und Raumfahrt throughgrants 50OW0204 and 50OO1501, the European Research Council through grant694513, the Italian Ministero dellâinstruzione, dellâuniversitĂ de della ricerca andUniversitĂ degli Studi di Roma Tor Vergata through FFABR 2017 and âMis-sion: Sustainability 2016â, the UK Science and Technology Facilities Council through grant ST/P000592/1, the Israel Science Foundation through grant848/16, the Chilean CONICYT-FONDECYT through grant 3180405, the Mexi-can CONACYT through grant CVU 448248, the JSPS KAKENHI through grantsJP18H01265 and 18H05439, and the JST PRESTO through grant JPMJPR1775
The CARMENES search for exoplanets around M dwarfs High-resolution optical and near-infrared spectroscopy of 324 survey stars
The CARMENES radial velocity (RV) survey is observing 324 M dwarfs to search for any orbiting planets. In this paper, we present the survey sample by publishing one CARMENES spectrum for each M dwarf. These spectra cover the wavelength range 520â1710 nm at a resolution of at least R >80 000, and we measure its RV, Hα emission, and projected rotation velocity. We present an atlas of high-resolution M-dwarf spectra and compare the spectra to atmospheric models. To quantify the RV precision that can be achieved in low-mass stars over the CARMENES wavelength range, we analyze our empirical information on the RV precision from more than 6500 observations. We compare our high-resolution M-dwarf spectra to atmospheric models where we determine the spectroscopic RV information content, Q, and signal-to-noise ratio. We find that for all M-type dwarfs, the highest RV precision can be reached in the wavelength range 700â900 nm. Observations at longer wavelengths are equally precise only at the very latest spectral types (M8 and M9). We demonstrate that in this spectroscopic range, the large amount of absorption features compensates for the intrinsic faintness of an M7 star. To reach an RV precision of 1 m sâ1 in very low mass M dwarfs at longer wavelengths likely requires the use of a 10 m class telescope. For spectral types M6 and earlier, the combination of a red visual and a near-infrared spectrograph is ideal to search for low-mass planets and to distinguish between planets and stellar variability. At a 4 m class telescope, an instrument like CARMENES has the potential to push the RV precision well below the typical jitter level of 3â4 m sâ1
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