Síntesis, caracterización y evaluación de la actividad antimicrobiana contra E. coli de las nanopartículas de plata funcionalizadas acopladas al péptido VIHGW-alquino-G-NH2

Busca conjugar NPs metálicas con un péptido antibacteriano conteniendo el motivo ATCUN, el cual es una seria de 3 aminoácidos comenzando por el amino terminal en el cual el tercer aminoácido es la histidina (Libardo, Gorbatyuk, & Angeles-Boza, 2016), como una primera aproximación al desarrollo de vectores de transporte de proteínas basados en AgNPs. Estas NPs son consideradas debido a su comprobado poder bactericida. La conjugación entra las NPs y el péptido se lleva a cabo mediante una reacción click de Huisgen catalizada por Cu(I), la cual consiste en la reacción entre el grupo N3 y el grupo alquino catalizado por Cu(I) formando el anillo triazol. En este trabajo primero se funcionaliza las NPs con una molécula espaciadora SHPEG- N3), y finalmente se hace reaccionar el péptido VIHGW-alquino-G-NH2 con las NPs funcionalizadas en presencia de Cu(I) el cual actuará como catalizador. El producto obtenido luego de esta reacción fue caracterizado por medio de técnicas espectroscópicas como XPS, FTIR, DLS y microscópicas como TEM. Estas caracterizaciones estarán complementadas por un estudio de la actividad biocida hacia E. Coli usando la técnica de conteo en placas. Mediante espectroscopia FTIR se pudo demostrar que la reacción entre el péptido y las AgNPs-S-PEG-N3 se dio mediante la reacción click porque la señal del grupo N3 presente en las AgNPs-S-PEG-N3 desaparece una vez realizada la reacción con el péptido. Usando la técnica XPS se pudo determinar que la molécula espaciadora se unió a las AgNPs mediante el grupo funcional SH porque en el espectro de las AgNPs-S-PEG-N3 se observa que aparece un a 160 eV característico de los sulfuros metálicos. Por ultimo las pruebas antibacteriales nos dan como resultado que el producto final AgNPs-S-PEG-VIH presenta una mayor actividad antibacterial contra E. Coli que sus componentes individuales.Tesi

Expression of oxidative stress and antioxidant defense genes in the kidney of inbred mice after intestinal ischemia and reperfusion

PURPOSE: To determine the gene expressions profile related to the oxidative stress and the antioxidant response in the kidneys of mice subjected to intestinal ischemia and reperfusion. METHODS: Twelve inbred mice (C57BL/6) were randomly assigned to one of two groups: the control group (CG) underwent anesthesia and was observed for 120 min and the ischemia/reperfusion group (IRG), animals were anesthetized and subjected to laparotomy and ischemia for 60 minutes followed by 60 minutes of reperfusion. The expressions of 84 genes from the kidney were determined by the Reverse Transcription qualitative Polymerase Chain Reaction (RT-qPCR). All genes that were up regulated by more than threefold using the algorithm [2(ΔΔCt)] were considered statically significant (p<0.05). RESULTS: In the IRG group 29 (34.52%) of 84 genes, were up regulated by more than threefold. The genes that were differentially up regulated in the glutathione peroxidase cluster (10 genes): were Gpx2 and Gpx7. The genes that were up regulated in the peroxidase cluster (16 genes) were following: Duox1, Epx, Lpo, Mpo, Ptgs2, Rag2, Serpinb1b, Tmod1 and Tpo. The genes that up regulated in the reactive oxygen species cluster (16 genes): Il19, Il22, Nos2, Nox1, Noxa1, Noxo1, Recql4 and Sod2. The genes that were up regulated in the oxidative stress cluster (22 genes) were: Mpp4, Nudt15, Upc3 and Xpa. The genes that were up regulated in the oxygen carriers cluster (12 genes) were: Hbq1, Mb, Ngb, Slc38a1 and Xirp1. The peroxiredoxins genes (10) showed no consistent differential regulation. CONCLUSION: The genes related to oxidative stress and antioxidant defense showed increased expression in renal tissue trigged intestinal ischemia and reperfusion.São Paulo Federal University Postgraduate Program in Interdisciplinary Surgical SciencesMato Grosso do Sul Federal University Surgery DepartmentGrande Dourados Federal University Medical SchoolUFGD Medical SchoolUNIFESP Surgery Department Surgical Techniques and Experimental Surgery DivisionSão Paulo Federal University Postgraduate Program in Interdisciplinary Surgical SciencesUNIFESP, Surgery Department Surgical Techniques and Experimental Surgery DivisionSciEL

Gene expression profile of oxidative stress in the lung of inbred mice after intestinal ischemia/reperfusion injury

PURPOSE:To determine the gene expression profile associated with oxidative stress and antioxidant defense in the lung tissue of mice subjected to intestinal ischemia and reperfusion.METHODS:Twelve male, inbred mice (C57BL/6) were randomly assigned to one of two groups. The control group (CG) underwent anesthesia and laparotomy and was observed for 120 minutes; the ischemia/reperfusion group (IRG) was subjected to anesthesia, laparotomy, and ischemia of the small intestine for 60 minutes and to 60 minutes of reperfusion. A pool of six mice from each group was subjected to a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to analyze the oxidative stress and antioxidant defense genes. All genes that were up-regulated or down-regulated greater than three-fold, based on the algorithm [2Federal University of Grande Dourados Medical SchoolUFGD Medical SchoolFederal University of Mato Grosso do Sul Surgery DepartmentUniversitary Center Sao CamiloUNIFESP Surgery DepartmentUNIFESP, Surgery DepartmentSciEL

Ischemia and reperfusion of the soleus muscle of rats with pentoxifylline

BACKGROUND: Reperfusion of the skeletal muscle worsens existing lesions during ischemia, since the production of reactive oxygen species, associated with intense participation of neutrophils, increases the inflammatory reaction that induces tissue changes. OBJECTIVE: To evaluate the morphological and immunohistochemical changes of the skeletal (soleus) muscle of rats submitted to ischemia and reperfusion with pentoxifylline. METHODS: Sixty rats were submitted to ischemia of the pelvic limb for 6 hours induced by clamping the left common iliac artery. After ischemia, group A animals (n = 30) were observed for 4 hours and group B animals (n = 30) for 24 hours. Six animals constituted the sham group. Pentoxifylline was applied only in the reperfusion period A2 (n = 10) and B2 (n = 10), and in ischemia and reperfusion periods in A3 (n = 10) and B3 (n = 10). The soleus muscle was evaluated by histological (fiber disruption, leukocyte infiltrate, necrosis) and immunohistochemical (apoptosis through caspase-3 expression) analysis. The non-parametric tests Kruskal-Wallis and Mann-Whitney (p < 0.05) were applied. RESULTS: The changes were more intense in group B1, with fiber disruption mean scores of 2.16±0.14; neutrophilic infiltrate of 2.05±0.10; and caspase-3 expression in the perivascular area of 4.30±0.79; and less intense in group A3, with means of 0.76±0.16; 0.92±0.10; 0.67±0,15, respectively (p < 0.05). Caspase-3 was more expressive in group B1 in the perivascular area, with mean of 4.30±0.79 when compared with group B1 in the perinuclear area, with mean of 0.91±0.32 (p < 0.05) CONCLUSIONS: The lesions were more intense when observation time was longer after reperfusion, and pentoxifylline attenuated these lesions, above all when used in the beginning of ischemia and reperfusion phases.CONTEXTO: A reperfusão de músculo esquelético piora as lesões já presentes no período de isquemia, pois a produção de espécies reativas de oxigênio, associadas à intensa participação de neutrófilos, amplia a reação inflamatória que induz alterações teciduais. OBJETIVO: Avaliar as alterações morfológicas e imuno-histoquímicas de músculo esquelético (sóleo) de ratos submetidos a isquemia e reperfusão com pentoxifilina. MÉTODOS: Sessenta ratos foram submetidos a isquemia do membro pélvico, por 6 horas, pelo clampeamento da artéria ilíaca comum esquerda. Após isquemia, os animais do grupo A (n = 30) foram observados por 4 horas, e os do grupo B (n = 30), por 24 horas. Seis animais constituíram o grupo simulado. Administrou-se pentoxifilina apenas no período de reperfusão em A2 (n = 10) e B2 (n = 10) e nos períodos de isquemia e reperfusão em A3 (n = 10) e B3 (n = 10). O músculo sóleo foi avaliado por análise histológica (dissociação de fibras, infiltrado leucocitário, necrose) e imuno-histoquímica (apoptose pela expressão da caspase-3). Foram aplicados os testes não-paramétricos de Kruskal-Wallis e Mann-Whitney (p < 0,05). RESULTADOS: As alterações foram mais intensas no grupo B1, com médias de escore da dissociação de fibras musculares de 2,16 ± 0,14, infiltrado neutrofílico de 2,05 ± 0,10 e expressão da caspase-3 na área perivascular de 4,30 ± 0,79; e menos intensas no grupo A3, com respectivas médias de 0,76 ± 0,16, 0,92 ± 0,10 e 0,67 ± 0,15 (p < 0,05). A caspase-3 mostrou-se mais expressiva no grupo B1 na área perivascular, com média de 4,30 ± 0,79, em comparação com o grupo B1 na área perinuclear, com média de 0,91 ± 0,32 (p < 0,05). CONCLUSÕES: As lesões são mais intensas quando o tempo de observação é maior após a reperfusão, e a pentoxifilina atenua essas lesões, sobretudo quando usada no início das fases de isquemia e de reperfusão.SBACVUniversidade Federal de Mato Grosso do Sul Hospital UniversitárioUniversidade Federal de São Paulo (UNIFESP) Escola Paulista de MedicinaUFMSUNIFESP-EPM Departamento de PatologiaUFMS Departamento de Clínica CirúrgicaUFMS Hospital Universitário Comissão de Residência MédicaUNIFESP, EPM, Depto. de PatologiaSciEL

International multicenter observational study on assessment of ventilatory management during general anaesthesia for robotic surgery and its effects on postoperative pulmonary complication (AVATaR) : study protocol and statistical analysis plan

Introduction: Robotic-assisted surgery (RAS) has emerged as an alternative minimally invasive surgical option. Despite its growing applicability, the frequent need for pneumoperitoneum and Trendelenburg position could significantly affect respiratory mechanics during RAS. AVATaR is an international multicenter observational study aiming to assess the incidence of postoperative pulmonary complications (PPC), to characterise current practices of mechanical ventilation (MV) and to evaluate a possible association between ventilatory parameters and PPC in patients undergoing RAS. Methods and analysis: AVATaR is an observational study of surgical patients undergoing MV for general anaesthesia for RAS. The primary outcome is the incidence of PPC during the first five postoperative days. Secondary outcomes include practice of MV, effect of surgical positioning on MV, effect of MV on clinical outcome and intraoperative complications. Ethics and dissemination: This study was approved by the Institutional Review Board of the Hospital Israelita Albert Einstein. The study results will be published in peer-reviewed journals and disseminated at international conferences. Trial registration number: NCT02989415; Pre-results

I Diretriz brasileira de cardio-oncologia pediátrica da Sociedade Brasileira de Cardiologia

Sociedade Brasileira de Oncologia PediátricaUniversidade Federal de São Paulo (UNIFESP) Instituto de Oncologia Pediátrica GRAACCUniversidade Federal de São Paulo (UNIFESP)Universidade de São Paulo Faculdade de Medicina Instituto do Coração do Hospital das ClínicasUniversidade Federal do Rio Grande do Sul Hospital de Clínicas de Porto AlegreInstituto Materno-Infantil de PernambucoHospital de Base de BrasíliaUniversidade de Pernambuco Hospital Universitário Oswaldo CruzHospital A.C. CamargoHospital do CoraçãoSociedade Brasileira de Cardiologia Departamento de Cardiopatias Congênitas e Cardiologia PediátricaInstituto Nacional de CâncerHospital Pequeno PríncipeSanta Casa de Misericórdia de São PauloInstituto do Câncer do Estado de São PauloUniversidade Federal de São Paulo (UNIFESP) Departamento de PatologiaHospital Infantil Joana de GusmãoUNIFESP, Instituto de Oncologia Pediátrica GRAACCUNIFESP, Depto. de PatologiaSciEL