Vasorelaxant Activity of AP39, a Mitochondria-Targeted H2S Donor, on Mouse Mesenteric Artery Rings In Vitro

This is the final version. Available from MDPI via the DOI in this record. The data presented in this study are available on request from the corresponding author.Mitochondria-targeted hydrogen sulfide (H2S) donor compounds, such as compound AP39, supply H2S into the mitochondrial environment and have shown several beneficial in vitro and in vivo effects in cardiovascular conditions such as diabetes and hypertension. However, the study of their direct vascular effects has not been addressed to date. Thus, the objective of the present study was to analyze the effects and describe the mechanisms of action of AP39 on the in vitro vascular reactivity of mouse mesenteric artery. Protein and gene expressions of the H2S-producing enzymes (CBS, CSE, and 3MPST) were respectively analyzed by Western blot and qualitative RT-PCR, as well the in vitro production of H2S by mesenteric artery homogenates. Gene expression of CSE and 3MPST in the vessels has been evidenced by RT-PCR experiments, whereas the protein expression of all the three enzymes was demonstrated by Western blotting experiments. Nonselective inhibition of H2S-producing enzymes by AOAA abolished H2S production, whereas it was partially inhibited by PAG (a CSE selective inhibitor). Vasorelaxation promoted by AP39 and its H2S-releasing moiety (ADT-OH) were significantly reduced after endothelium removal, specifically dependent on NO-cGMP signaling and SKCa channel opening. Endogenous H2S seems to participate in the mechanism of action of AP39, and glibenclamide-induced KATP blockade did not affect the vasorelaxant response. Considering the results of the present study and the previously demonstrated antioxidant and bioenergetic effects of AP39, we conclude that mitochondria-targeted H2S donors may offer a new promising perspective in cardiovascular disease therapeutics.The Royal SocietySao Paulo Research Foundation (FAPESP)Sao Paulo Research Foundation (FAPESP)Brazilian National Council for Scientific and Technological Development (CNPq)Brazilian National Council for Scientific and Technological Development (CNPq)Coordination for the Improvement of Higher Education Personnel (CAPES

Formyl Peptide Receptor as a Novel Therapeutic Target for Anxiety-Related Disorders

Formyl peptide receptors (FPR) belong to a family of sensors of the immune system that detect microbe-associated molecules and inform various cellular and sensorial mechanisms to the presence of pathogens in the host. Here we demonstrate that Fpr2/3-deficient mice show a distinct profile of behaviour characterised by reduced anxiety in the marble burying and light-dark box paradigms, increased exploratory behaviour in an open-field, together with superior performance on a novel object recognition test. Pharmacological blockade with a formyl peptide receptor antagonist, Boc2, in wild type mice reproduced most of the behavioural changes observed in the Fpr2/3(-/-) mice, including a significant improvement in novel object discrimination and reduced anxiety in a light/dark shuttle test. These effects were associated with reduced FPR signalling in the gut as shown by the significant reduction in the levels of p-p38. Collectively, these findings suggest that homeostatic FPR signalling exerts a modulatory effect on anxiety-like behaviours. These findings thus suggest that therapies targeting FPRs may be a novel approach to ameliorate behavioural abnormalities present in neuropsychiatric disorders at the cognitive-emotional interface

Co-Administration of a Plasmid DNA Encoding IL-15 Improves Long-Term Protection of a Genetic Vaccine against Trypanosoma cruzi

Background: Immunization of mice with the Trypanosoma cruzi trans-sialidase (TS) gene using plasmid DNA, adenoviral vector, and CpG-adjuvanted protein delivery has proven highly immunogenic and provides protection against acute lethal challenge. However, long-term protection induced by TS DNA vaccines has not been reported. the goal of the present work was to test whether the co-administration of a plasmid encoding IL-15 (pIL-15) could improve the duration of protection achieved through genetic vaccination with plasmid encoding TS (pTS) alone.Methodology: We immunized BALB/c mice with pTS in the presence or absence of pIL-15 and studied immune responses [with TS-specific IFN-gamma ELISPOT, serum IgG ELISAs, intracellular cytokine staining (IFN-gamma, TNF-alpha, and IL-2), tetramer staining, and CFSE dilution assays] and protection against lethal systemic challenge at 1 to 6 months post vaccination. Mice receiving pTS alone developed robust TS-specific IFN-gamma responses and survived a lethal challenge given within the first 3 months following immunization. the addition of pIL-15 to pTS vaccination did not significantly alter T cell responses or protection during this early post-vaccination period. However, mice vaccinated with both pTS and pIL-15 challenged 6 months post-vaccination were significantly more protected against lethal T. cruzi challenges than mice vaccinated with pTS alone (P6 months post immunization. Also, these TS-specific T cells were better able to expand after in vitro restimulation.Conclusion: Addition of pIL-15 during genetic vaccination greatly improved long-term T cell survival, memory T cell expansion, and long-term protection against the important human parasite, T. cruzi.National Institutes of HealthFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Millennium Institute for Gene TherapySt Louis Univ, Dept Internal Med, St Louis, MO 63103 USAUniversidade Federal de São Paulo, Ctr Terapia Celular & Mol, Escola Paulista Med, São Paulo, BrazilSt Louis Univ, Dept Mol Microbiol, St Louis, MO 63103 USAUniv Fed Minas Gerais, Inst Ciencias Biol, Dept Microbiol, Belo Horizonte, MG, BrazilUniversidade Federal de São Paulo, Ctr Terapia Celular & Mol, Escola Paulista Med, São Paulo, BrazilNational Institutes of Health: RO1 AI040196CNPq: 420067/2005-1Web of Scienc

Molecular Characterization and Phylogenetic Study of Coxsackievirus A24v Causing Outbreaks of Acute Hemorrhagic Conjunctivitis (AHC) in Brazil

Coxsackievirus A24 variant (CA24v) is the most prevalent viral pathogen associated with acute hemorrhagic conjunctivitis (AHC) outbreaks. Sixteen years after its first outbreak in Brazil, this agent reemerged in 2003 in Brazil, spread to nearly all states and caused outbreaks until 2005. In 2009, a new outbreak occurred in the northeast region of the country. In this study, we performed a viral isolation in cell culture and characterized clinical samples collected from patients presenting symptoms during the outbreak of 2005 in Vitória, Espírito Santo State (ES) and the outbreak of 2009 in Recife, Pernambuco State (PE). We also performed a phylogenetic analysis of worldwide strains and all meaningful Brazilian isolates since 2003.Sterile cotton swabs were used to collect eye discharges, and all 210 clinical samples were used to inoculate cell cultures. Cytopathic effects in HEp-2 cells were seen in 58 of 180 (32%) samples from Vitória and 3 of 30 (10%) samples from Recife. Phylogenetic analysis based on a fragment of the VP1 and 3C gene revealed that the CA24v causing outbreaks in Brazil during the years 2003, 2004 and 2005 evolved from Asian isolates that had caused the South Korean outbreak of AHC during the summer of 2002. However, the 2009 outbreak of AHC in Pernambuco was originated from the reintroduction of a new CA24v strain that was circulating during 2007 in Asia, where CA24v outbreaks has been continuously reported since 1970.This study is the first phylogenetic analysis of AHC outbreaks caused by CA24v in Brazil. The results showed that Asian strains of CA24v were responsible for the outbreaks since 1987 and were independently introduced to Brazil in 2003 and 2009. Phylogenetic analysis of complete VP1 gene is a useful tool for studying the epidemiology of enteroviruses associated with outbreaks

Delivering medical abortion at scale: a study of the retail market for medical abortion in Madhya Pradesh, India.

BACKGROUND: Medical abortion (mifepristone and misoprostol) has the potential to contribute to reduced maternal mortality but little is known about the provision or quality of advice for medical abortion through the private retail sector. We examined the availability of medical abortion and the practices of pharmacists in India, where abortion has been legal since 1972. METHODS: We interviewed 591 pharmacists in 60 local markets in city, town and rural areas of Madhya Pradesh. One month later, we returned to 359 pharmacists with undercover patients who presented themselves unannounced as genuine customers seeking a medical abortion. RESULTS: Medical abortion was offered to undercover patients by 256 (71.3%) pharmacists and 24 different brands were identified. Two thirds (68.5%) of pharmacists stated that abortion was illegal in India. Only 106 (38.5%) pharmacists asked clients the timing of the last menstrual period and 38 (13.8%) requested to see a doctor's prescription - a legal requirement in India. Only 59 (21.5%) pharmacists correctly advised patients on the gestational limit for medical abortion, 97 (35.3%) provided correct information on how many and when to take the tablets in a combination pack, and 78 (28.4%) gave accurate advice on where to seek care in case of complications. Advice on post-abortion family planning was almost nonexistent. CONCLUSIONS: The retail market for medical abortion is extensive, but the quality of advice given to patients is poor. Although the contribution of medical abortion to women's health in India is poorly understood, there is an urgent need to improve the practices of pharmacists selling medical abortion

TSPO ligand residence time influences human glioblastoma multiforme cell death/life balance

Abstract Ligands addressed to the mitochondrial Translocator Protein (TSPO) have been suggested as cell death/life and steroidogenesis modulators. Thus, TSPO ligands have been proposed as drug candidates in several diseases; nevertheless, a correlation between their binding affinity and in vitro efficacy has not been demonstrated yet, questioning the specificity of the observed effects. Since drug-target residence time is an emerging parameter able to influence drug pharmacological features, herein, the interaction between TSPO and irDE-MPIGA, a covalent TSPO ligand, was investigated in order to explore TSPO control on death/life processes in a standardized glioblastoma cell setting. After 90 min irDE-MPIGA cell treatment, 25 nM ligand concentration saturated irreversibly all TSPO binding sites; after 24 h, TSPO de-novo synthesis occurred and about 40 % TSPO binding sites resulted covalently bound to irDE-MPIGA. During cell culture treatments, several dynamic events were observed: (a) early apoptotic markers appeared, such as mitochondrial membrane potential collapse (at 3 h) and externalization of phosphatidylserine (at 6 h); (b) cell viability was reduced (at 6 h), without cell cycle arrest. After digitonin-permeabilized cell suspension treatment, a modulation of mitochondrial permeability transition pore was evidenced. Similar effects were elicited by the reversible TSPO ligand PIGA only when applied at micromolar dose. Interestingly, after 6 h, irDE-MPIGA cell exposure restored cell survival parameters. These results highlighted the ligand-target residence time and the cellular setting are crucial parameters that should be taken into account to understand the drug binding affinity and efficacy correlation and, above all, to translate efficiently cellular drug responses from bench to bedside

Concepts of Health in Nursing Education in Undergraduate Schools of Brazil and Portugal

This article represents an extract of my PhD research regarding the concepts of health present in undergraduate schools of nursing of Brazil and Portugal. The objective of this research was to comprehend how the concepts of health are explicit in the documents of undergraduate courses and how they are expressed in the discourses of the nursing professors and students. For this, thematic content analysis was used to analyze the professional education of the nurses and the concepts of health present in the nursing courses. The presentation of the results was performed using the Collective Subject Discourse technique. The results showed that in the state of Santa Catarina in Brazil, the concepts of health follow the National Curriculum Guidelines and there is an emphasis directed towards reflection on the Brazilian National Health System (SUS); while in the city of Porto, in Portugal, these concepts follow the Study Plans of the Nursing courses and emphasize nursing care both for the healthy and the sick person. The data also reveal a concern in the construction of the concept of people-centered healthcare.Este artículo representa una parte de la investigación que realicé durante el doctorado sobre las concepciones de salud presentes en escuelas de graduación en enfermería de Brasil y Portugal. El objetivo de esta investigación fue comprender como las concepciones sobre la salud están explícitas en los documentos de los cursos de graduación y de qué forma se expresan en los discursos de los profesores y estudiantes de enfermería. Para esto, utilizamos el análisis de contenido temático sobre la educación profesional de los enfermeros y sobre las concepciones de la salud presentes en los cursos de enfermería. La presentación de los resultados fue realizada a través del Discurso del Sujeto Colectivo. Los resultados apuntaron que, en el estado de Santa Catarina, en Brasil, las concepciones de salud siguen las Directrices Curriculares Nacionales y existe un énfasis dirigido a reflexionar sobre el Sistema Único de Salud; en cuanto en la ciudad de Porto, en Portugal, esas concepciones siguen los Planos de Estudios de los cursos de Enfermería y se enfatiza el cuidado de Enfermería tanto a la persona saludable como a la enferma. Los datos revelan, también, una preocupación en la construcción del concepto de salud centrado en las personas.Este artigo representa um recorte de pesquisa de doutorado a respeito das concepções de saúde, presentes em escolas de graduação em enfermagem do Brasil e de Portugal. O objetivo foi compreender como as concepções de saúde estão explícitas nos documentos dos cursos de graduação e de que forma se expressam nos discursos dos professores e estudantes de enfermagem. Para tanto, utilizou-se análise de conteúdo temática sobre a educação profissional dos enfermeiros e sobre as concepções de saúde presentes nos cursos de enfermagem. A apresentação dos resultados foi realizada através do Discurso do Sujeito Coletivo. Os resultados apontaram que, no Estado de Santa Catarina, no Brasil, as concepções de saúde seguem as Diretrizes Curriculares Nacionais e há ênfase dirigida à reflexão sobre o Sistema Único de Saúde; enquanto na cidade do Porto, em Portugal, tais concepções seguem os Planos de Estudos dos Cursos de Enfermagem e se enfatiza o cuidado de Enfermagem tanto à pessoa saudável quanto à doente. Os dados revelam, ainda, preocupação na construção do conceito de saúde centrado na pessoa

Vineyard microclimate and yield under different plastic covers.

The use of plastic cover in vineyards minimizes effects of adverse weather conditions. The northwest of São Paulo State is one of the largest grape producing regions in Brazil; however, few studies investigate the effects of different plastic covers on vineyards in this region. This study compared the effect of black shading screen (BSS) and braided polypropylene film (BPF) on BRS Morena vineyard microclimate, grown on an overhead trellis system in the northwestern São Paulo. The experiments were carried out during three growing seasons (2012 ? 2014). BSS allowed superior incoming solar radiation (SR) transmissivity, resulting in higher net radiation (Rn), and higher ratio between photosynthetically active (PAR) and SR. No differences were observed between the average air temperatures (T) and relative humidity (RH) of covered environments (BPF and BSS) and outside condition (automatic weather station ? AWS), due to high air circulation, despite wind speed (WS) reduction caused by plastic covers. BPF provided better conditions for vineyard growth with higher fruit yield than vineyard under BSS regarding the number of shoots with bunches per plant, bunch and stem weights, longitudinal diameter of berries, quantity of fertile buds per shoot, and yield per shoot and per plant. BPF covers also influenced leaf size and growth speed of plants in vineyards. Keywords Black shading screen . Braided polypropylene film . BRS Morena . Leaf wetness duration . Yiel