Thioredoxin promotes survival signaling events under nitrosative/oxidative stress associated with cancer development

Accumulating mutations may drive cells into the acquisition of abnormal phenotypes that are characteristic of cancer cells. Cancer cells feature profound alterations in proliferation programs that result in a new population of cells that overrides normal tissue construction and maintenance programs. To achieve this goal, cancer cells are endowed with up regulated survival signaling pathways. They also must counteract the cytotoxic effects of high levels of nitric oxide (NO) and of reactive oxygen species (ROS), which are by products of cancer cell growth. Accumulating experimental evidence associates cancer cell survival with their capacity to up-regulate antioxidant systems. Elevated expression of the antioxidant protein thioredoxin-1 (Trx1) has been correlated with cancer development. Trx1 has been characterized as a multifunctional protein, playing different roles in different cell compartments. Trx1 migrates to the nucleus in cells exposed to nitrosative/oxidative stress conditions. Trx1 nuclear migration has been related to the activation of transcription factors associated with cell survival and cell proliferation. There is a direct association between the p21Ras-ERK1/2 MAP Kinases survival signaling pathway and Trx1 nuclear migration under nitrosative stress. The expression of the cytoplasmic protein, the thioredoxin-interacting protein (Txnip), determines the change in Trx1 cellular compartmentalization. The anti-apoptotic actions of Trx1 and its denitrosylase activity occur in the cytoplasm and serve as important regulators of cell survival. Within this context, this review focuses on the participation of Trx1 in cells under nitrosative/oxidative stress in survival signaling pathways associated with cancer development.Brazilian Funding Institutions: Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed São Paulo, Paulista Med Sch, Ctr Cellular & Mol Therapy CTCMol, Dept Biochem, São Paulo, SP, BrazilKarolinska Inst, Div Biochem Med Biochem & Biophys, Stockholm, SwedenNYU, Sch Med, New York, NY 10016 USAUniv Fed São Paulo, Paulista Med Sch, Ctr Cellular & Mol Therapy CTCMol, Dept Biochem, São Paulo, SP, BrazilFAPESP: 2007/59617-6FAPESP: 2009/52730-7FAPESP: 2012/10470-1CNPq: 481154/2013-2Web of Scienc

Inhibition of protein tyrosine phosphatase activity by diamide is reversed by epidermal growth factor in fibroblasts

AbstractDiamide (azodicarboxylic acid bis(dimethylamide)) inhibits protein tyrosine phosphatase activity in fibroblasts without altering protein tyrosine kinase activity associated with the epidermal growth factor receptor. The loss of protein tyrosine phosphatase activity caused by diamide is reversed by 2-mercaptoethanol or epidermal growth factor

The Venoarteriolar Reflex Significantly Reduces Contralateral Perfusion as Part of the Lower Limb Circulatory Homeostasis in vivo

Perfusion at microvascular level involves the contribution of both local and central regulators, under a complex vascular signaling frame. The venoarteriolar reflex (VAR) is one of such regulatory responses, of particular relevance in the lower limb to prevent edema. Although known for quite some time, many of the complex interactions involving all of these regulatory mechanisms still need clarification. Our objective was to look deeper into VAR through modern photoplethymography (PPG). Twelve healthy subjects (both sexes, 26.0 ± 5.0 y.o.) were enrolled in this study after informed written consent. Subjects were submitted to a leg lowering maneuver while lying supine to evoke the VAR, involving three phases–10 min baseline register, both legs extended, 10 min challenge, with one randomly chosen leg (test) pending 50 cm below heart level, while the contralateral (control) remained in place, and 10 minutes recovery, resuming the initial position. PPG signals were collected from both feet and treated by the wavelet transform (WT) revealing six spectral bands in frequency intervals comprising the cardiac [1.6–0.7 Hz], respiratory [0.4–0.26 Hz], myogenic [0.26–0.1 Hz], neurogenic/sympathetic [0.1–0.045 Hz], endothelial NO-dependent (NOd) [0.045–0.015 Hz], and NO-independent (NOi) [0.015–0.007 Hz] activities. For the first time, this approach revealed that, with VAR, perfusion significantly decreased in both limbs, although the change was more pronounced in the test foot. Here, a significant decrease in myogenic, neurogenic and NOd, were noted, while the control foot recorded a decrease in neurogenic and an increase in NOd. These results confirm the utility of WT spectral analysis for flowmotion. Further, it strongly suggests that VAR results from a complex cooperation between local myogenic-endothelial responses, where a central neurogenic reflex might also be involved

Characterization of thimet oligopeptidase and neurolysin activities in B16F10-Nex2 tumor cells and their involvement in angiogenesis and tumor growth

<p>Abstract</p> <p>Background</p> <p>Angiogenesis is a fundamental process that allows tumor growth by providing nutrients and oxygen to the tumor cells. Beyond the oxygen diffusion limit from a capillary blood vessel, tumor cells become apoptotic. Angiogenesis results from a balance of pro- and anti-angiogenic stimuli. Endogenous inhibitors regulate enzyme activities that promote angiogenesis. Tumor cells may express pro-angiogenic factors and hydrolytic enzymes but also kinin-degrading oligopeptidases which have been investigated.</p> <p>Results</p> <p>Angiogenesis induced by B16F10-Nex2 melanoma cells was studied in a co-culture with HUVEC on Matrigel. A stimulating effect on angiogenesis was observed in the presence of B16F10-Nex2 lysate and plasma membrane. In contrast, the B16F10-Nex2 culture supernatant inhibited angiogenesis in a dose-dependent manner. This effect was abolished by the endo-oligopeptidase inhibitor, JA-2. Thimet oligopeptidase (TOP) and neurolysin activities were then investigated in B16F10-Nex2 melanoma cells aiming at gene sequencing, enzyme distribution and activity, influence on tumor development, substrate specificity, hydrolytic products and susceptibility to inhibitors. Fluorescence resonance energy transfer (FRET) peptides as well as neurotensin and bradykinin were used as substrates. The hydrolytic activities in B16F10-Nex2 culture supernatant were totally inhibited by <it>o</it>-phenanthrolin, JA-2 and partially by Pro-Ile. Leupeptin, PMSF, E-64, Z-Pro-Prolinal and captopril failed to inhibit these hydrolytic activities. Genes encoding M3A enzymes in melanoma cells were cloned and sequenced being highly similar to mouse genes. A decreased proliferation of B16F10-Nex2 cells was observed in vitro with specific inhibitors of these oligopeptidases. Active rTOP but not the inactive protein inhibited melanoma cell development in vivo increasing significantly the survival of mice challenged with the tumor cells. On Matrigel, rTOP inhibited the bradykinin – induced angiogenesis. A possible regulation of the homologous tumor enzyme in the perivascular microenvironment is suggested based on the observed rTOP inhibition by an S-nitrosothiol NO donor.</p> <p>Conclusion</p> <p>Data show that melanoma cells secrete endo-oligopeptidases which have an important role in tumor proliferation in vitro and in vivo. rTOP inhibited growth of subcutaneously injected B16F10-Nex2 cells in mice. TOP from tumor cells and bradykinin in endothelial cells are two antagonist factors that may control angiogenesis essential for melanoma growth. A regulatory role of NO or S-nitrosothiols is suggested.</p

3-D Photoionization Structure and Distances of Planetary Nebulae II. Menzel 1

We present the results of a spatio-kinematic study of the planetary nebula Menzel 1 using spectro-photometric mapping and a 3-D photoionization code. We create several 2-D emission line images from our long-slit spectra, and use these to derive the line fluxes for 15 lines, the Halpha/Hbeta extinction map, and the [SII] line ratio density map of the nebula. We use our photoionization code constrained by these data to derive the three-dimensional nebular structure and ionizing star parameters of Menzel 1 by simultaneously fitting the integrated line intensities, the density map, and the observed morphologies in several lines, as well as the velocity structure. Using theoretical evolutionary tracks of intermediate and low mass stars, we derive a mass for the central star of 0.63+-0.05 Msolar. We also derive a distance of 1050+_150 pc to Menzel 1.Comment: To be published in ApJ of 10th February 2005. 12 figure

Systematic Observation of Corner Kick Strategies in Portuguese Football Players

Set pieces are important for the success of football teams, with the corner kick being one of the most game defining events. The aim of this research was twofold: (1) to analyze the corner kicks of a senior football amateur team, and (2) to compare the corner kicks of successful and unsuccessful teams (of the 2020/21 sporting season). In total, 500 corners were observed using a bespoke notational analysis tool, using a specific observational instrument tool (8 criteria; 25 categories). Out of the 500 corner kicks, 6% resulted in a goal. A greater number of direct corners using inswing trajectories were performed (n = 54%). Corners were delivered to central and front post areas most frequently (n = 79%). Five attackers were most predominantly used for offensive corners (n = 58%), but defenders won the ball more frequently (n = 44%). Attempts at goal occurred following a corner most commonly from outside of the box (n = 7%). Goals were scored most frequently with the foot (n = 16%) and head (n = 15%). Successful teams are more effective at reaching the attackers and score more goals directly from corners. Unsuccessful teams deliver more corner kicks out of play, the first touch is more frequently from the opposition defenders, and fewe goals are scored from corner kicks. The study provides an insight into the determining factors and patterns that influence corner kicks and success in football matches. This information should be used by coaches to prepare teams for both offensive and defensive corner kicks to increase team success and match outcomes.info:eu-repo/semantics/publishedVersio

Atuação dos enfermeiros de saúde materna e obstetrícia nas situações de risco e perigo para recém-nascidos: Um estudo qualitativo

This study aimed to explore the perception of a team of maternal health nurses and obstetrics concerning the experience in working in situations of risk and danger to newborns. For this, a qualitative and cross-sectional research was conducted. Nineteen nurses from a team of a maternal health and obstetrics service were interviewed. Nurses were aged between 29 and 56 years, and had professional experience in the area between six months and 26 years. Data collection was performed through a sociodemographic questionnaire and semi-structured interview guide with open questions. The results suggest the criteria by which the interviewed nurses assess risk and danger situations, the experienced emotions, as well as the role attributed to training in this area. It is concluded that it may be important to invest in the specific qualification of these professionals to improve intervention in this domain.O presente estudo teve o propósito de explorar a perceção de uma equipa de enfermeiros de saúde materna e obstetrícia relativamente à experiência na atuação em situações de risco e perigo em recém-nascidos. Para tal, realizou-se uma investigação de caráter qualitativo e desenho transversal. Foram entrevistados 19 enfermeiros de um serviço de saúde materna e obstetrícia com idades compreendidas entre os 29 e os 56 anos, com experiência profissional na área entre os seis meses e 26 anos. A recolha de dados foi feita através de um questionário sociodemográfico e um guião de entrevista semiestruturado composto por questões abertas. Os resultados sugerem os critérios pelos quais os enfermeiros entrevistados avaliam as situações de risco e perigo, bem como as emoções experienciadas e o papel atribuído à formação neste âmbito. Conclui-se que pode ser importante o investimento na qualificação específica destes profissionais de modo a contribuir para aprimorar a intervenção neste domínio

Insights into milk-clotting activity of latex peptidases from <i>Calotropis procera</i> and <i>Cryptostegia grandiflora</i>

Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl2. However, pre-incubation of both samples at 75 °C for 10 min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16 kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.Centro de Investigación de Proteínas Vegetale

Insights into milk-clotting activity of latex peptidases from <i>Calotropis procera</i> and <i>Cryptostegia grandiflora</i>

Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl2. However, pre-incubation of both samples at 75 °C for 10 min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16 kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.Centro de Investigación de Proteínas Vegetale

Insights into milk-clotting activity of latex peptidases from <i>Calotropis procera</i> and <i>Cryptostegia grandiflora</i>

Latex fractions from Calotropis procera, Cryptostegia grandiflora, Plumeria rubra, and Himatanthus drasticus were assayed in order to prospect for new plant peptidases with milk-clotting activities, for use as rennet alternatives. Only C. procera and C. grandiflora latex fractions exhibited proteolytic and milk-clotting activities, which were not affected by high concentrations of NaCl and CaCl2. However, pre-incubation of both samples at 75 °C for 10 min eliminated completely their activities. Both proteolytic fractions were able to hydrolyze k-casein and to produce peptides of 16 kDa, a similar SDS-PAGE profile to commercial chymosin. RP-HPLC and mass spectrometry analyses of the k-casein peptides showed that the peptidases from C. procera or C. grandiflora hydrolyzed k-casein similar to commercial chymosin. The cheeses made with both latex peptidases exhibited yields, dry masses, and soluble proteins similar to cheeses prepared with commercial chymosin. In conclusion, C. procera and C. grandiflora latex peptidases with the ability to coagulate milk can be used as alternatives to commercial animal chymosin in the cheese manufacturing process.Centro de Investigación de Proteínas Vegetale