The tyrosine kinase FER is responsible for the capacitation-associated increase in tyrosine phosphorylation in murine sperm

Sperm capacitation is required for fertilization. At the molecular level, this process is associated with fast activation of protein kinase A. Downstream of this event, capacitating conditions lead to an increase in tyrosine phosphorylation. The identity of the tyrosine kinase(s) mediating this process has not been conclusively demonstrated. Recent experiments using stallion and human sperm have suggested a role for PYK2 based on the use of small molecule inhibitors directed against this kinase. However, crucially, loss-of-function experiments have not been reported. Here, we used both pharmacological inhibitors and genetically modified mice models to investigate the identity of the tyrosine kinase(s) mediating the increase in tyrosine phosphorylation in mouse sperm. Similar to stallion and human, PF431396 blocks the capacitation-associated increase in tyrosine phosphorylation. Yet, sperm from Pyk2(-/-) mice displayed a normal increase in tyrosine phosphorylation, implying that PYK2 is not responsible for this phosphorylation process. Here, we show that PF431396 can also inhibit FER, a tyrosine kinase known to be present in sperm. Sperm from mice targeted with a kinase-inactivating mutation in Fer failed to undergo capacitation-associated increases in tyrosine phosphorylation. Although these mice are fertile, their sperm displayed a reduced ability to fertilize metaphase II-arrested eggs in vitro.Fil: Alvau, Antonio. University of Massachussets; Estados UnidosFil: Battistone, Maria Agustina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Gervasi, Maria Gracia. University of Massachussets; Estados UnidosFil: Navarrete, Felipe A.. University of Massachussets; Estados UnidosFil: Xu, Xinran. State University of Colorado - Fort Collins; Estados UnidosFil: Sánchez Cárdenas, Claudia. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: De la Vega Beltran, José Luis. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Da Ros, Vanina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Greer, Peter. Queens University; CanadáFil: Darszon, Alberto. Universidad Nacional Autónoma de México. Instituto de Biotecnología; MéxicoFil: Krapf, Diego. State University of Colorado - Fort Collins; Estados UnidosFil: Salicioni, Ana María. University of Massachussets; Estados UnidosFil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Visconti, Pablo E.. University of Massachussets; Estados Unido

Avaliação da associação do biosilicato® ao laser de Nd:YAG para o tratamento de cárie/ Nd:YAG biosilicate® association assessment for the treatment of cárie

A cárie é uma doença infecciosa, transmissível, crônica, multifatorial e de lenta progressão. Ultimamente, há a busca pelo desenvolvimento de tratamentos minimamente invasivos das lesões cariosas com preservação estética. Uma estratégia interessante é o tratamento com Biomateriais e Laserterapia, pois permitiria uma melhor absorção e aproveitamento do Biosilicato® (BS) pelo dente lesionado com intuito de regeneração. Este estudo objetivou a investigação de métodos de aderência de BS ao dente cariado, assim como o desenvolvimento de metodologia para melhor interação do BS com o laser de Nd:YAG. O estudo foi conduzido em 2 fases experimentais. Na primeira, adotou-se o modelo de cárie química em dentina radicular bovina e, após, foi feito o tratamento com as partículas de BS em diferentes veículos de aplicação (água destilada, silicone e gel dental), com posterior avaliação composicional e morfológica, que mostraram a efetividade do tratamento de cárie com o BS veiculado em água destilada. Na segunda fase, a dentina cariada foi tratada com BS, associado ou não à irradiação laser e diferentes fotoabsorvedores, e avaliaram-se a morfologia e temperatura intrapulpar decorrentes dos tratamentos. A irradiação laser após a aplicação do BS, com utilização de carvão como fotoabsorvedor, possibilitou o recobrimento dos túbulos dentinários de forma uniforme, com aspecto de derretimento e recristalização do BS, proveniente do aquecimento promovido pelo laser. Ademais, a temperatura intrapulpar, monitorada durante a etapa de irradiação não apresentou variação superior a 5,6 ºC, que é considerada crítica para a vitalidade da polpa. Em conclusão, o tratamento de BS combinado ao laser de Nd:YAG se mostrou promissor ao tratamento da cárie

TPCS/PBAT blown extruded films added with curcumin as a technological approach for active packaging materials

The development of active packaging is a relevant topic demanding the development of films with diverse properties to preserve specific foodstuff. The objective of this work was to obtain extruded TPCS/PBAT films containing curcumin and evaluate it as an active antimicrobial and antioxidant packaging to protect chia oil from oxidative degradation. Morphology, thermal, mechanical, antimicrobial, and antioxidant evaluation of the films were conducted to determine whether the presence of curcumin affected the film’s properties. Infrared Spectroscopy indicated that curcumin addition affected the crosslinking reaction between citric acid and starch, which explains the changes in hydrophilicity and mechanical strength of the films. The incorporation of curcumin conferred antimicrobial activity against Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa and Escherichia coli) bacteria, as well as antioxidant activity. Films were tested as chia oil packaging, being verified that they successfully prevented oil degradation under accelerated stability test (60 °C for 7 days), demonstrating the feasibility of using TPCS/PBAT biodegradable films containing curcumin to obtain active packaging materials.Authors thank to CNPq (Chamada Universal– MCTI/CNPq Nº 14/2014, Processo 447768/2014-0), CAPES (Coordenação de Pessoal de Nível Superior Master's scholarship) and Fundação Araucária (Programa Universal/Pesquisa Básica e Aplicada 24/2012, protocolo 7334133700514041013) for the finantial support. This work was financially supported by Associate Laboratory LSRE-LCM (UID/EQU/50020/2019) funded by national funds through FCT/MCTES (PIDDAC), and Foundation for Science and Technology (FCT, Portugal). CIMO (UID/AGR/00690/2019) through FEDER under Program PT2020. To the national funding by FCT, P.I., through the institutional scientific employment program-contract for I.P. Fernandes contract.info:eu-repo/semantics/publishedVersio

Antiparasitic Activity of Natural and Semi-Synthetic Tirucallane Triterpenoids from Schinus terebinthifolius (Anacardiaceae): Structure/Activity Relationships

Leishmaniasis and Chagas are diseases caused by parasitic protozoans that affect the poorest population in the World, causing a high mortality and morbidity. As a result of highly toxic and long-term treatments, the discovery of novel, safe and more efficacious drugs is essential. in this work, the in vitro antiparasitic activity and mammalian cytotoxicity of three natural tirucallane triterpenoids, isolated from leaves of Schinus terebinthifolius (Anacardiaceae), and nine semi-synthetic derivatives were investigated against Leishmania (L.) infantum and Trypanosoma cruzi. Trypomastigotes of T. cruzi were the most susceptible parasites and seven compounds demonstrated a trypanocidal activity with IC50 values in the range between 15 and 58 mu g/mL. Four compounds demonstrated selectivity towards the intracellular amastigotes of Leishmania, with IC50 values in the range between 28 and 97 mu g/mL. the complete characterization of triterpenoids was afforded after thorough analysis of nuclear magnetic resonance (NMR) data as well as electrospray ionization mass spectrometry (ESI-MS). Additionally, structure-activity relationships were performed using Decision Trees.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Universidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09972270 Diadema, SP, BrazilAdolfo Lutz Inst, Ctr Parasitol, BR-01246902 São Paulo, BrazilUniv Fed Paraiba, Ctr Ciencias Aplicadas & Educ, BR-58297000 Rio Tinto, BrazilUniv Fed Uberlandia, Inst Quim, BR-38400902 Uberlandia, MG, BrazilUniversidade Federal de São Paulo, Inst Ciencias Ambientais Quim & Farmaceut, BR-09972270 Diadema, SP, BrazilCNPq: 300546/2012-2CNPq: 471458/2012-0FAPESP: 2011/51739-0FAPESP: 2012/18756-1FAPESP: 2013/16320-4Web of Scienc

Versatility in the biological behavior of two aminobenzoate oxidovanadium (V, IV) compounds. Inhibition or simulation of enzymes

The pharmacological potential of vanadium compounds is of great interest to researchers in treatments of various diseases (diabetes, cancer, tropical endemic diseases, etc.). On the basis of the potential biological/pharmacological applications, in this work we have synthesized and physico-chemically characterized, two new complexes containing vanadium (IV) and (V) with 4-aminobenzoic acid as the ligand (L)

Detecting spatio-temporal mortality clusters of European countries by sex and ag

[EN] Background: Mortality decreased in European Union (EU) countries during the last century. Despite these similar trends, there are still considerable differences in the levels of mortality between Eastern and Western European countries. Sub-group analysis of mortality in Europe for different age and sex groups is common, however to our knowledge a spatio-temporal methodology as in this study has not been applied to detect significant spatial dependence and interaction with time. Thus, the objective of this paper is to quantify the dynamics of mortality in Europe and detect significant clusters of mortality between European countries, applying spatio-temporal methodology. In addition, the joint evolution between the mortality of European countries and their neighbours over time was studied. Methods: The spatio-temporal methodology used in this study takes into account two factors: time and the geographical location of countries and, consequently, the neighbourhood relationships between them. This methodology was applied to 26 European countries for the period 1990-2012. Results: Principally, for people older than 64 years two significant clusters were obtained: one of high mortality formed by Eastern European countries and the other of low mortality composed of Western countries. In contrast, for ages below or equal to 64 years only the significant cluster of high mortality formed by Eastern European countries was observed. In addition, the joint evolution between the 26 European countries and their neighbours during the period 1990-2012 was confirmed. For this reason, it can be said that mortality in EU not only depends on differences in the health systems, which are a subject to national discretion, but also on supra-national developments. Conclusions: This paper proposes statistical tools which provide a clear framework for the successful implementation of development public policies to help the UE meet the challenge of rethinking its social model (Social Security and health care) and make it sustainable in the medium term.The authors are grateful for the financial support provided by the Ministry of Economy and Competitiveness, project MTM2013-45381-P. Adina Iftimi gratefully acknowledges financial support from the MECyD (Ministerio de Educacion, Cultura y Deporte, Spain) Grant FPU12/04531. Francisco Montes is grateful for the financial support provided by the Spanish Ministry of Economy and Competitiveness, project MTM2016-78917-R. The research by Patricia Carracedo and Ana Debon has been supported by a grant from the Mapfre Foundation.Carracedo-Garnateo, P.; Debón Aucejo, AM.; Iftimi, A.; Montes-Suay, F. (2018). Detecting spatio-temporal mortality clusters of European countries by sex and ag. International Journal for Equity in Health. 17:1-19. https://doi.org/10.1186/s12939-018-0750-zS11917Anderson TW, Goodman LA. Statistical Inference about Markov Chains. Ann Math Stat. 1957; 28(1):89–110.Anselin L. Local Indicators of Spatial Association–LISA. Geographical Anal. 1995; 27(2):93–115.Bilbao-Ubillos J. Is there still such a thing as the ‘European social model’?. Int J Soc Welf. 2016; 25:110–25.Bivand R. spdep: Spatial Dependence:Weighting Schemes, Statistics and Models. 2012. R package version 0.5-53. http://CRAN.R-project.org/package=spdep .Bivand R, Hauke J, Kossowski T. Computing the Jacobian in Gaussian Spatial Autoregressive Models: An Illustrated Comparison of Available Methods. Geographical Anal. 2013; 45(2):150–79.Bivand R, Keitt T, Rowlingson B. rgdal: Bindings for the Geospatial Data Abstraction Library. 2016. R package version 1.1-10. https://CRAN.R-project.org/package=rgdal .Bivand R, Lewin-Koh N. maptools: Tools for Reading and Handling Spatial Objects. 2016. R package version 0.8-39 https://CRAN.R-project.org/package=maptools .Bonneux L, Huisman C. de Beer J. Mortality in 272 European regions, 2002-2004: an update. Eur J Epidemiol. 2010; 25(1):77–85. Reporting year: 2010.Charpentier A. Computational Actuarial Science with R. Chapman y Hall/CRC. 2014.Cliff AD, Ord JK. Spatial autocorrelation. London: Pion; 1973.Cutler D, Deaton A, Lleras-Muney A. The Determinants of Mortality. J Econ Perspect. 2006; 20(3):97–120.Debón A, Chaves L, Haberman S, Villa F. Characterization of between-group inequality of longevity in European Union countries. Insur Math Econ. 2017; 75:151–65.Fleiss J, Levin B, Paik M. Statistical Methods for Rates and Proportions: Wiley; 2013.Gordon M. Gmisc: Descriptive Statistics, Transition Plots, and More. 2016. R package version 1.3.1. https://CRAN.R-project.org/package=Gmisc .Hinde A. Demographic methods. Routledge: Routledge; 1998.Hyndman RJ, Booth H, Tickle L, Maindonald J. demography: Forecasting mortality, fertility, migration and population data. 2014. package version 1.18. https://CRAN.R-project.org/package=demography .Human Mortality Database. University of California, Berkeley (USA), and Max Planck Institute for Demographic Research (Germany). 2016. Available at www.mortality.org or www.humanmortality.de (data downloaded on 12th July 2016).Hatzopoulos P, Haberman S. Common mortality modeling and coherent forecasts. An empirical analysis of worldwide mortality data. Insurance Math Econ. 2013; 52(2):320–37.Iftimi A, Montes F, Santiyán AM, Martínez-Ruiz F. Space–time airborne disease mapping applied to detect specific behaviour of varicella in Valencia, Spain Spatial Spatio-Temporal Epidemiol. 2015; 14:33–44.Julious S, Nicholl J, George S. Why do we continue to use standardized mortality ratios for small area comparisons?. J Public Health. 2001; 23(1):40–6.Laurent T, Ruiz-Gazen A, Thomas-Agnan C. GeoXp: An R package for exploratory spatial data analysis. J Stat Softw. 2012; 47(2):1–23.Leon DA. Trends in European life expectancy: a salutary view. Int J Epidemiol. 2011; 40:271–7.Li H, Li L, Wu B, Xiong Y. The End of Cheap Chinese Labor. J Econ Perspect. 2013; 26(4):57–74.Mackenbach JP, Karanikolos M, McKee M. The unequal health of Europeans: successes and failures of policies. The Lancet. 2013; 381(9872):1125–34.Meslé F. Mortality in Central and Eastern Europe: Long-term trends and recent upturns. Demographic Res. 2004; 2:45–70.Meslé F, Vallin J. Mortality in Europe: The divergence between East and West. Population (English Edition). 2002; 57(1):157–97.Moran PAP. Notes on continuous stochastic phenomena. Biometrika. 1950; 37(1-2):17–23.Moran PAP. A Test for the Serial Independence of Residuals. Biometrika. 1950; 37(1/2):178–81.Neuwirth E. RColorBrewer: ColorBrewer Palettes. R package version. 2014; 1:1–2. https://CRAN.R-project.org/package=RColorBrewer .Oleckno WA. Epidemiology: concepts and methods: Waveland Press, Inc.; 2008.Quah D. Galton’s Fallacy and Tests of the Convergence Hypothesis. Scand J Econ. 1993; 95(4):427–43.R Core Team. R: A Language and Environment for Statistical Computing. Vienna: R Foundation for Statistical Computing. 2015. https://www.R-project.org/ .Rey S. In: Fischer MM, Nijkamp P, (eds).Spatial Dynamics and Space-Time Data Analysis. Berlin, Heidelberg: Springer: Handbook of Regional Science; 2014, pp. 1365–83.Rey SJ. Spatial Empirics for Economic Growth and Convergence. Geogr Anal. 2001; 33(3):195–214.Riffe T. Reading Human Fertility Database and Human Mortality Database data into R. Technical Report TR-2015-004, MPIDR. 2015.Schofield R, Reher D, Bideau A. The Decline of Mortality in Europe. International studies in demography. Oxford: Clarendon Press; 1991.Shaw M, Orford S, Brimblecombe N, Dorling D. Widening inequality in mortality between 160 regions of 15 European countries in the early 1990s. Soc Sci Med. 2000; 50(7-8):1047–58.Spinakis A, Anastasiou G, Panousis V, Spiliopoulos K, Palaiologou S, Yfantopoulos J. Expert Review and Proposals for Measurement of Health Inequalities in the European Union. European Commission. Technical report,Luxembourg: European Commission Directorate General for Health and Consumers; 2011. http://ec.europa.eu/health/social_determinants/docs/full_quantos_en.pdf .Staehr K. Economic transition in Estonia. Background, reforms and results In: Rindzeviciute E, editor. Contemporary Change in Estonia. Baltic and East European Studies. Sodertorns hogskola: Baltic and East European Studies: 2004. p. 437–67.Trnka L, Dankova D, Zitova J, Cimprichova L, Migliori GB, Clancy L, Zellweger J. Survey of BCG vaccination policy in Europe: 1994-96. Bull World Health Organ. 1998; 76(1):85–91.United Nations Inter–agency Group for Child Mortality Estimation. Levels & Trends in Child Mortality: Report 2013. New York: Technical report, United Nations Children’s Fund; 2013. Avaliable at www.who.int/maternal_child_adolescent/documents/levels_trends_child_mortality_2013.pdf Accessed 27 Oct 2016.Vågerö D. The east–west health divide in Europe: Growing and shifting eastwards. Eur Rev. 2010; 18(01):23–34.Vaupel JW, Zhang Z, van Raalte AA, Vaupel JW, Zhang Z, van Raalte AA. Life expectancy and disparity: an international comparison of life table data. BMJ Open. 2011; 1:e000128.Wickham H, Chang W. devtools: Tools to Make Developing R Packages Easier. R package version 1.11.1. 2016. https://CRAN.R-project.org/package=devtools .Wilcox R. Introduction to robust estimation and hypothesis testing, 3rd Edition.San Diego: Academic Press; 2012

Immunomodulation From Moderate Exercise Promotes Control of Experimental Cutaneous Leishmaniasis

Physical exercise has been described as an important tool in the prevention and treatment of numerous diseases as it promotes a range of responses and adaptations in several biological systems, including the immune system. Studies on the effect of exercise on the immune system could play a critical role in improving public health. Current literature suggests that moderate intensity exercise can modulate the Th1/Th2 dichotomy directing the immune system to a Th1 cellular immune response, which favors the resolution of infections caused by intracellular microorganisms. Leishmaniasis is a group of diseases presenting a wide spectrum of clinical manifestations that range from self-limiting lesions to visceral injuries whose severity can lead to death. The etiological agents responsible for this group of diseases are protozoa of the genus Leishmania. Infections by the parasite Leishmania major in mice (Balb/c) provide a prototype model for the polarization of CD4+ T cell responses of both Th1 (resistance) or Th2 (susceptibility), which determines the progression of infections. The aim of this study was to evaluate the effect of exercise on the development of L. major experimental infections by scanning the pattern of immune response caused by exercise. Groups of Balb/c mice infected with L. major were divided into groups that preformed a physical exercise of swimming three times a week or were sedentary along with treatment or not with the reference drug, meglumine antimoniate. Animals in groups submitted to physical exercise did not appear to develop lesions and presented a significantly lower parasite load independent of drug treatment. They also showed a positive delayed hypersensitivity response to a specific Leishmania antigen compared to control animals. The IFN-γ/IL-4 and IFN-γ/IL10 ratios in trained animals were clearly tilted to a Th1 response in lymph node cells. These data suggest that moderate intensity exercise is able to modulate the Th1 response that provides a protective effect against the development of leishmanial lesions

Delay of EGF-Stimulated EGFR Degradation in Myotonic Dystrophy Type 1 (DM1)

Funding Information: This research was supported by the Isabel Gemio Foundation (P18–13) and was also partially supported by the “Fondo Europeo de Desarrollo Regional” (FEDER) from the European Union. E.A.-C. was supported by a pre-doctoral fellowship of Valhondo Calaff Foundation. S.C.-C. and E.U.-C. were supported by FPU fellowships (FPU19/04435 and FPU16/00684, respectively) from the Ministerio de Ciencia, Innovación y Universidades, Spain. M.P.-B. and A.G.-B. received fellowships from the “Plan Propio de Iniciación a la Investigación, Desarrollo Tecnológico e Innovación (Universidad de Extremadura). M.N.-S. was supported by the “Ramon y Cajal” Program (RYC-2016–20883), and P.G.-S., was funded by “Juan de la Cierva Incorporación” Program (IJC2019–039229-I), Spain. S.M.S.Y.-D. was supported by the Isabel Gemio Foundation and CIBERNED (CB06/05/0041). J.M.F received research support from the Isabel Gemio Foundation and the “Instituto de Salud Carlos” III, CIBERNED (CB06/05/0041). Publisher Copyright: © 2022 by the authors.Myotonic dystrophy type 1 (DM1) is an autosomal dominant disease caused by a CTG repeat expansion in the 3′ untranslated region of the dystrophia myotonica protein kinase gene. AKT dephosphorylation and autophagy are associated with DM1. Autophagy has been widely studied in DM1, although the endocytic pathway has not. AKT has a critical role in endocytosis, and its phosphorylation is mediated by the activation of tyrosine kinase receptors, such as epidermal growth factor receptor (EGFR). EGF-activated EGFR triggers the internalization and degradation of ligand–receptor complexes that serve as a PI3K/AKT signaling platform. Here, we used primary fibroblasts from healthy subjects and DM1 patients. DM1-derived fibroblasts showed increased autophagy flux, with enlarged endosomes and lysosomes. Thereafter, cells were stimulated with a high concentration of EGF to promote EGFR internalization and degradation. Interestingly, EGF binding to EGFR was reduced in DM1 cells and EGFR internalization was also slowed during the early steps of endocytosis. However, EGF-activated EGFR enhanced AKT and ERK1/2 phosphorylation levels in the DM1-derived fibroblasts. Therefore, there was a delay in EGF-stimulated EGFR endocytosis in DM1 cells; this alteration might be due to the decrease in the binding of EGF to EGFR, and not to a decrease in AKT phosphorylation.publishersversionpublishe

The MD Anderson prostate cancer patient-derived xenograft series (MDA PCa PDX) captures the molecular landscape of prostate cancer and facilitates marker-driven therapy development

BACKGROUND: Advances in prostate cancer (PCa) lag behind other tumor types partly due to the paucity of models reflecting key milestones in PCa progression. OBJECTIVE: To develop clinically relevant PCa models. DESIGN: Since 1996 we have generated clinically annotated patient-derived xenografts (PDXs) (the MDA PCa PDX series) linked to specific phenotypes reflecting all aspects of clinical PCa. RESULTS: We studied two cell line-derived xenografts and the first 80 PDXs derived from 47 human PCa donors. Of these, 47 PDXs derived from 22 donors are working models and can be expanded either as cell lines (MDA PCa 2a and 2b) or PDXs. The histopathologic, genomic, and molecular characteristics (AR, ERG, and PTEN loss) maintain fidelity with the human tumor and correlate with published findings. PDX growth response to mouse castration and targeted therapy illustrate their clinical utility. Comparative genomic hybridization and sequencing show significant differences in oncogenic pathways in pairs of PDXs derived from different areas of the same tumor. We also identified a recurrent focal deletion in an area that includes the SPOPL gene in PDXs derived from 7 human donors out of 28 studied (25%). SPOPL is a SPOP paralog, and SPOP mutations define a molecular subclass of PCa. SPOPL deletions are found in 7% of TCGA PCas, which suggests that our cohort is a reliable platform for targeted drug development. CONCLUSIONS: The MDA PCa PDX series is a dynamic resource that captures the molecular landscape of PCas progressing under novel treatments and enables optimization of PCa-specific, marker-driven therapy