Introducción: Se describe que las características de la vivienda son un factor importante para llevar a cabo adecuadamente el tratamiento de diálisis peritoneal, sin embargo, no se conoce la real relación entre las características de la vivienda y el desarrollo de la peritonitis. Métodos: Se realizó una cohorte retrospectiva que incluyó a todos los pacientes mayores de 18 años de edad que iniciaron el tratamiento de diálisis peritoneal dentro del periodo enero 2002 - diciembre 2011 en el Hospital Nacional Guillermo Almenara Irigoyen. Se describió los tiempos de seguimiento según la fecha de inicio de DP y la primera peritonitis u otros eventos (paso a hemodiálisis, trasplante, muerte o abandono). Las variables fueron evaluadas según su tipo, utilizando estadística descriptiva e inferencial. Resultados: El análisis incluyó a 218 pacientes con una media de edad de 54 años ± 16. El principal lugar donde se realiza el procedimiento de diálisis peritoneal es el dormitorio (77,3%), en su mayoría se encuentra limpio (54,3%), ordenado (71,3%) y despejado (61,8%). Los insumos para el procedimiento se almacenan bajo techo (95,7%) y el mismo paciente ejecuta el tratamiento en la mayoría de los casos (73,1%). La incidencia encontrada fue de 0,17 episodios/paciente-año. La media del tiempo de seguimiento fue de 975 días ± 750. De acuerdo al análisis realizado no se encontró resultados significativos que muestren la relación entre las variables de vivienda y el desarrollo de la primera peritonitis. Conclusión: No se encontró asociación entre las condiciones de vivienda y el desarrollo de peritonitis en pacientes en tratamiento de diálisis peritoneal. Es necesario evaluar y mejorar el programa de visitas domiciliarias / Background: The features of housing are an important risk factor to properly carry out the treatment of peritoneal dialysis. However, the actual situation is that it doesn’t know the real relationship between the characteristics of housing and the development of peritonitis. Methods: A retrospective cohort was made including all patients over 18 years who started PD between the period January 2002 and December 2011 at the Hospital National Guillermo Almenara Irigoyen. The follow-up times was described as the beginning date of use DP and the first peritonitis or other events (hemodialysis, transplant, death or abandonment). The variables were evaluated according to their type using descriptive and inferential statistic. Results: The analysis included 218 patients with a mean age of 54 years ± 16. The main place where the patients did the procedure of peritoneal dialysis is the bedroom (77,3%), most of it is clean (54,3%), organized (71,3%) and clear (61,8%). The inputs for the procedure are stored indoors (95,7%) and the person who executes the procedure is the patient in most cases (73,1%). The observed incidence was 0,17 episodes/patient-year. The mean follow-up time was 975 days ± 750. No significant results were found between the housing conditions and the development of the first peritonitis in patients who are treated by peritoneal dialysis. Conclusion: No association was found between the housing conditions and the development of the first peritonitis. It is necessary to evaluated and improve the home visit program in order to obtain better information of housing condition.

Um die intravaskuläre Hämolyse bei Pferden zu untersuchen, wurden die Konzentration des freien Hämoglobin (fr. Hb) bei gesunden Pferden, Pferden mit Kolik oder akuter Kolitis, Pferden mit Hyperbilirubinämie, equiner infektiöser Anämie (EIA) und Pferden mit equiner granulozytärer Ehrlichiose (Anaplasmose) gemessen. Der Tumor-Nekrose-Faktor (TNF-alpha wurde bei Pferden mit Kolik oder Kolitis bestimmt, um Korrelationen zwischen dem fr. Hb und systemischen Entzündungsreaktionen zu finden. Fr. Hb ist ein wichtiger Parameter für die Diagnose der intravaskulären Hämolyse. Die photometrische Methode nach KAHN kann auch für die Messung von fr. Hb in Pferdeplasma eingesetzt werden. Bei Kolikpferden mit Strangulation oder akuter Kolitis bzw. Peritonitis in Zusammenhang mit SIRS können auch hohe Werte von fr. Hb und TNF-alpha vorkommen. Wenn die Konzentrationen von TNF-alpha und fr. Hb stark ansteigen ist die Prognose für eine Heilung schlecht. Bei Pferden mit der inapparenten Form der EIA zeigte sich, dass die intravaskuläre Hämolyse nur bei 25 % der Patienten auftritt. Weiterhin konnte durch die Messung des fr. Hb nachgewiesen werden, dass die Erythrozytolyse bei Pferden mit Anaplasmose nicht auf eine intravaskuläre Hämolyse zurückzuführen ist. Für die Differentialdiagnostik ist das fr. Hb ein nützlicher Parameter bei der Klärung der Ursachen einer kontinuierlichen unkonjugierten Hyperbilirubinämie.

Introdução: Diálise peritoneal (DP) é a modalidade de diálise mais utilizada em pacientes pediátricos portadores de doença renal crônica (DRC) em todo o mundo. Peritonites e infecções de óstio e túnel do cateter de diálise peritoneal constituem as duas primeiras causas de morbidade e falha de tratamento em pacientes em programa de diálise peritoneal. Objetivo: Caracterizar o perfil dos episódios de peritonites e infecções de óstio e túnel nos pacientes em programa de diálise peritoneal crônica na Unidade de Nefrologia Pediátrica do Instituto da Criança do HC FMUSP. Métodos: Avaliou-se o período compreendido entre janeiro de 1994, quando foi iniciado o programa de diálise peritoneal crônica nessa unidade, e dezembro de 2005. Foram incluídos todos os pacientes que estiveram em programa de diálise peritoneal crônica no período, cujos prontuários estivessem acessíveis para análise. Realizou-se revisão dos prontuários quanto aos dados demográficos dos pacientes, peso e estatura ao início e ao final do programa de diálise, etiologia e evolução clinica de cada episódio infeccioso, e presença de fatores de risco para peritonites e infecções relacionadas ao cateter de DP. Realizou-se análise descritiva dos dados gerais, cálculo da incidência de episódios infecciosos no período estudado e freqüência de fatores de risco para infecção. Analisaram-se descritivamente os perfis de todos os episódios infecciosos. Resultados: Cinqüenta e três pacientes foram estudados, sendo 32 (60,4%) do sexo masculino, com mediana de idade de 9,5 (0,3 a 18,2) anos. A etiologia da DRC foi má-formação urológica em 32 (60,4%) pacientes, glomerulopatias em 15 (28,3%), tubulopatias em 4 (7,5%) e outras causas em 2 (3,8%). A mediana do tempo em diálise foi de 20,5 (3,9 a 87,1) meses. Houve 132 episódios infecciosos no período observado, em 42 (79%) pacientes, sendo 65 peritonites, 54 infecções de óstio e túnel e 13 infecções de óstio e túnel associadas a peritonite, num total de 1329,3 paciente-mês. Assim sendo, houve 0,49 infecções de óstio e túnel/ paciente-ano, 0,59 peritonites/ paciente-ano e 0,12 infecções de óstio e túnel associadas a peritonite/ paciente-ano. Os agentes mais encontrados nas infecções de óstio e túnel foram S. aureus, isolado em 23 culturas (43% das culturas de óstio positivas), Staphylococcus coagulase negativo em 7 (13%) e Pseudomonas aeruginosa em 7 (13%). Nas peritonites, os agentes mais encontrados foram S. aureus, isolado em 15 culturas (27% das culturas de líquido peritoneal positivas), Staphylococcus coagulase negativo em 13 (24%) e Acinetobacter baumanii em 4 (7%). Nos casos de peritonite associada a infecção de óstio e túnel, S. aureus foi isolado em 6 culturas (67% das culturas de secreção de óstio) e Staphylococcus coagulase negativo em 2 (22%). Staphylococcus sp, Pseudomonas aeruginosa e fungos foram os agentes mais relacionados a complicações como colonização do óstio, evolução para peritonite, necessidade de troca do cateter e perda funcional peritoneal. Devido ao reduzido número de pacientes que não apresentaram infecções, não foi possível identificar fatores de risco na casuística estudada. Conclusões: As taxas encontradas de peritonites e infecções de óstio e túnel são elevadas. Staphylococcus sp foram os agentes mais encontrados e um dos mais relacionados a complicações. Medidas profiláticas no sentido de diminuir a contaminação por esse agente podem constituir uma intervenção eficaz para a redução das taxas de infecção diálise-relacionadas. / Introduction: Peritoneal dialysis (PD) is one of the most frequently used modality of chronic dialysis in children presenting with end stage renal disease (ESRD) worldwide. Peritonitis and exit site and/or tunnel infections (ESI/TI) are the major causes of morbidity and treatment failure in patients undergoing peritoneal dialysis. Objective: To study the episodes of peritonitis and ESI/TI in patients undergoing maintenance peritoneal dialysis in the Pediatric Nephrology Unit of Instituto da Criança - HC FMUSP. Methods: All patients whose files were available for analysis were included. Data was collected from January 1994, at the beginning of the program of chronic peritoneal dialysis in this unit, until December 2005. Demographic data, weight and height at the start and at the end of the period of study, etiology and clinical outcome of each infectious episode and risk factors for peritonitis and catheter related infections, were analyzed. Descriptive analysis of demographic data was performed. Incidence rates of peritonitis and ESI/TI and frequency of possible risk factors rates were calculated. Results: Fifty three patients were studied, 32 (60.4%) male, with median age of 9.5 (0.3 to 18.2) years. Primary renal disease was urological malformation in 32 (60.4%) patients, glomerular diseases in 15 (28.3%), tubular diseases in 4 (7.5%) and other causes in 2 (3.8%). Median time on dialysis was 20.5 (3.9 a 87.1) months. In 1329.3 patient-month, 132 episodes of peritonitis and/or ESI/TI occurred in 42 (79%) patients, including 54 episodes of ESI/TI, 65 episodes of peritonitis and 13 episodes of peritonitis associated with ESI/TI, yielding an annualized rate of 0.49, 0.59 and 0.12 episodes/patientyear, respectively. The most frequent etiologic agents in the ESI/TI group were S. aureus, isolated in 23 exit site cultures (43% of positive cultures), Staphylococcus coagulase negative in 7 (13%) and Pseudomonas aeruginosa in 7 (13%). In the peritonitis group, the most frequent agents were S. aureus, isolated in 15 peritoneal fluid cultures (27% of positive cultures), Staphylococcus coagulase negative in 13 (24%) and Acinetobacter baumanii in 4 (7%). In the episodes of peritonitis associated with ESI/TI, S. aureus was isolated in 6 (67%) exit site cultures and Staphylococcus coagulase negative in 2 (22%). Complications such as exit site colonization, evolution to peritonitis, catheter removal and peritoneal failure were mostly associated with Staphylococcus sp, Pseudomonas aeruginosa and fungi infections. No risk factors could be identified due to the reduced number of patients not affected by infections. Conclusion: Peritonitis and ESI/TI rates in this population were high. Staphylococcus sp was the most frequently recovered agent. Infections caused by this agent presented a high rate of complications. Prophylactic measures to decrease contamination with this microorganism could be an efficient intervention to reduce dialysis associated infection rates.

Introdução: Diálise peritoneal (DP) é a modalidade de diálise mais utilizada em pacientes pediátricos portadores de doença renal crônica (DRC) em todo o mundo. Peritonites e infecções de óstio e túnel do cateter de diálise peritoneal constituem as duas primeiras causas de morbidade e falha de tratamento em pacientes em programa de diálise peritoneal. Objetivo: Caracterizar o perfil dos episódios de peritonites e infecções de óstio e túnel nos pacientes em programa de diálise peritoneal crônica na Unidade de Nefrologia Pediátrica do Instituto da Criança do HC FMUSP. Métodos: Avaliou-se o período compreendido entre janeiro de 1994, quando foi iniciado o programa de diálise peritoneal crônica nessa unidade, e dezembro de 2005. Foram incluídos todos os pacientes que estiveram em programa de diálise peritoneal crônica no período, cujos prontuários estivessem acessíveis para análise. Realizou-se revisão dos prontuários quanto aos dados demográficos dos pacientes, peso e estatura ao início e ao final do programa de diálise, etiologia e evolução clinica de cada episódio infeccioso, e presença de fatores de risco para peritonites e infecções relacionadas ao cateter de DP. Realizou-se análise descritiva dos dados gerais, cálculo da incidência de episódios infecciosos no período estudado e freqüência de fatores de risco para infecção. Analisaram-se descritivamente os perfis de todos os episódios infecciosos. Resultados: Cinqüenta e três pacientes foram estudados, sendo 32 (60,4%) do sexo masculino, com mediana de idade de 9,5 (0,3 a 18,2) anos. A etiologia da DRC foi má-formação urológica em 32 (60,4%) pacientes, glomerulopatias em 15 (28,3%), tubulopatias em 4 (7,5%) e outras causas em 2 (3,8%). A mediana do tempo em diálise foi de 20,5 (3,9 a 87,1) meses. Houve 132 episódios infecciosos no período observado, em 42 (79%) pacientes, sendo 65 peritonites, 54 infecções de óstio e túnel e 13 infecções de óstio e túnel associadas a peritonite, num total de 1329,3 paciente-mês. Assim sendo, houve 0,49 infecções de óstio e túnel/ paciente-ano, 0,59 peritonites/ paciente-ano e 0,12 infecções de óstio e túnel associadas a peritonite/ paciente-ano. Os agentes mais encontrados nas infecções de óstio e túnel foram S. aureus, isolado em 23 culturas (43% das culturas de óstio positivas), Staphylococcus coagulase negativo em 7 (13%) e Pseudomonas aeruginosa em 7 (13%). Nas peritonites, os agentes mais encontrados foram S. aureus, isolado em 15 culturas (27% das culturas de líquido peritoneal positivas), Staphylococcus coagulase negativo em 13 (24%) e Acinetobacter baumanii em 4 (7%). Nos casos de peritonite associada a infecção de óstio e túnel, S. aureus foi isolado em 6 culturas (67% das culturas de secreção de óstio) e Staphylococcus coagulase negativo em 2 (22%). Staphylococcus sp, Pseudomonas aeruginosa e fungos foram os agentes mais relacionados a complicações como colonização do óstio, evolução para peritonite, necessidade de troca do cateter e perda funcional peritoneal. Devido ao reduzido número de pacientes que não apresentaram infecções, não foi possível identificar fatores de risco na casuística estudada. Conclusões: As taxas encontradas de peritonites e infecções de óstio e túnel são elevadas. Staphylococcus sp foram os agentes mais encontrados e um dos mais relacionados a complicações. Medidas profiláticas no sentido de diminuir a contaminação por esse agente podem constituir uma intervenção eficaz para a redução das taxas de infecção diálise-relacionadas. / Introduction: Peritoneal dialysis (PD) is one of the most frequently used modality of chronic dialysis in children presenting with end stage renal disease (ESRD) worldwide. Peritonitis and exit site and/or tunnel infections (ESI/TI) are the major causes of morbidity and treatment failure in patients undergoing peritoneal dialysis. Objective: To study the episodes of peritonitis and ESI/TI in patients undergoing maintenance peritoneal dialysis in the Pediatric Nephrology Unit of Instituto da Criança - HC FMUSP. Methods: All patients whose files were available for analysis were included. Data was collected from January 1994, at the beginning of the program of chronic peritoneal dialysis in this unit, until December 2005. Demographic data, weight and height at the start and at the end of the period of study, etiology and clinical outcome of each infectious episode and risk factors for peritonitis and catheter related infections, were analyzed. Descriptive analysis of demographic data was performed. Incidence rates of peritonitis and ESI/TI and frequency of possible risk factors rates were calculated. Results: Fifty three patients were studied, 32 (60.4%) male, with median age of 9.5 (0.3 to 18.2) years. Primary renal disease was urological malformation in 32 (60.4%) patients, glomerular diseases in 15 (28.3%), tubular diseases in 4 (7.5%) and other causes in 2 (3.8%). Median time on dialysis was 20.5 (3.9 a 87.1) months. In 1329.3 patient-month, 132 episodes of peritonitis and/or ESI/TI occurred in 42 (79%) patients, including 54 episodes of ESI/TI, 65 episodes of peritonitis and 13 episodes of peritonitis associated with ESI/TI, yielding an annualized rate of 0.49, 0.59 and 0.12 episodes/patientyear, respectively. The most frequent etiologic agents in the ESI/TI group were S. aureus, isolated in 23 exit site cultures (43% of positive cultures), Staphylococcus coagulase negative in 7 (13%) and Pseudomonas aeruginosa in 7 (13%). In the peritonitis group, the most frequent agents were S. aureus, isolated in 15 peritoneal fluid cultures (27% of positive cultures), Staphylococcus coagulase negative in 13 (24%) and Acinetobacter baumanii in 4 (7%). In the episodes of peritonitis associated with ESI/TI, S. aureus was isolated in 6 (67%) exit site cultures and Staphylococcus coagulase negative in 2 (22%). Complications such as exit site colonization, evolution to peritonitis, catheter removal and peritoneal failure were mostly associated with Staphylococcus sp, Pseudomonas aeruginosa and fungi infections. No risk factors could be identified due to the reduced number of patients not affected by infections. Conclusion: Peritonitis and ESI/TI rates in this population were high. Staphylococcus sp was the most frequently recovered agent. Infections caused by this agent presented a high rate of complications. Prophylactic measures to decrease contamination with this microorganism could be an efficient intervention to reduce dialysis associated infection rates.

Orientador: Pasqual Barretti / Resumo: Peritonite por bacilos Gram-negativos não fermentadores (BGNNF) é complicação importante da diálise peritoneal (DP), com curso clínico grave e elevada taxa de falência do método. Fatores associados à virulência, resistência antimicrobiana, formação de biofilme, entre outros, têm sido relatados, mas o limitado conjunto de evidências não permite concluir sobre os fatores responsáveis pelo pior curso clínico dessas infecções. O objetivo deste trabalho foi avaliar a influência das características microbiológicas, das condições clínicas do paciente e do tratamento na evolução de peritonites por BGNNF, ocorridas num único centro, em período de 18 anos. A sensibilidade in vitro aos antimicrobianos, produção de biofilme, além da análise do perfil clonal das bactérias pela técnica de eletroforese em gel de campo pulsado foram realizadas em todos os isolados bacterianos. Foram pesquisados genotipicamente, em isolados de Pseudomonas aeruginosa, a presença de marcadores de virulência (alginato, exoenzima S, fosfolipases C, exotoxina A, protesase alcalina, elastase e ramnolipídeos). Associações entre as características microbiológicas do paciente e tratamento com a taxa de resolução da peritonite foram estudadas. A espécie mais frequente foi Pseudomonas aeruginosa (45,59%), seguida por isolados do complexo Acinetobacter baumannii (17,65%). O estudo dos fatores de virulência da Pseudomonas aeruginosa revelou a presença de fatores de virulência em 100% dos casos, exceto exoenzima S (58,33%)... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Peritonitis due to non-fermentative Gram-negative bacilli (NFGNB) is a serious complication of peritoneal dialysis (PD), with a severe clinical course and high technique failure rate. Factors as bacterial virulence, antimicrobial resistance, biofilm formation, among others, have been reported, but the limited amount of evidence does not allow to conclude on the factors responsible for the worst clinical course of these infections. The objective of this study was to evaluate the influence of the microbiological characteristics, patients conditions, and treatment on evolution of peritonitis episodes at a single center in an 18 - year period. In vitro susceptibility, biofilm production, and clonal profile analysis of bacteria by pulsed-field gel electrophoresis (PFGE) were performed in all isolates. The presence of virulence markers (alginate, exoenzyme S, phospholipases C, exotoxin A, alkaline protease, elastase, and ramnolipids) was genotyped in bacterial isolates of Pseudomonas aeruginosa. From the data referring to the patient and causal agent, associations between the microbiological, patient characteristics, and treatment on the resolution rate of peritonitis were analyzed. The most frequent species was Pseudomonas aeruginosa (45.59%), followed by Acinetobacter baumannii complex (17.65%). The study of the virulence factors of Pseudomonas aeruginosa revealed the presence of virulence factors in 100% of the cases, except for exonzyme S (58.33%) and hemolytic phospholipase C ... (Complete abstract click electronic access below) / Mestre

Introduction: Globally, one in four persons infected with the human immunodeficiency virus (HIV) who are living with the acquired immunodeficiency syndrome (AIDS) will die of tuberculosis (TB). The estimated number of HIV-infected children who die of tuberculosis, especially extrapulmonary TB (EPTB), in Haiti, is only loosely based on facts or investigation. This study proposes to describe demographics of children with EPTB in a pediatric TB hospital in Haiti. The objectives are two-fold. The first objective is to describe the population of children discharged from Grace Children's Hospital with a confirmed diagnosis of tuberculosis overall, and broken down by whether or not the child had an extrapulmonary manifestation of the TB disease. Specifically, we describe the demographic characteristics and the prevalence of HIV and other co-morbidities of the children, in-hospital mortality, and the diagnostic tools used to determine TB infection including the sputum test, and th e documentation of family members also infected. As part of the descriptive process, by examining those with only pulmonary TB (PTB) and EPTB separately, we investigate if they appear to be different sub-populations based on demographic characteristics and clinical measures. The second objective of this work is to determine if there is a positive association between HIV infection and the EPTB manifestation in children with a confirmed diagnosis of TB, both crudely and after adjusting for demographic variables and co-morbidities. Methods: A cross-sectional study design was used to review medical charts of clinically diagnosed pediatric TB cases for a five-year period (January 1, 1999 -- December 31, 2003). This included 492 pulmonary TB and 210 extrapulmonary TB cases. Variables measured included clinical measures and demographic characteristics. Results: Data for 615 hospitalized, clinically diagnosed pediatric TB cases were reviewed. There were 315 (51.4%) males and 298 (48.6%) females with a mean age of 5.40 years (range 0.17 - 14 years), with 214 (37.9%) of the patients aged 0-2 years. Percent males were 47.8% and 57.9% in PTB and EPTB groups respectively (p<0.05). One hundred and seven (17.4%) of patients were HIV positive. Three hundred eighty-eight (63%) of the patients had one or more additional co-morbidities: [anemia 299 (48.6%), intestinal parasites 93 (15.1%), malaria 58 (9.4%) and gastroenteritis 19 (3.1%)]. Nearly 85% of the children were undernourished. Eighty-three child patients (13.5%) died in the hospital. Children with EPTB were much more likely to be over the age of two (74% vs. 56% in PTB group), resulting in a highly significant Chi-square stati stic. The overall difference in mean age, however, was only borderline significant with children with EPTB being slightly older [p=0.059] and age was only weakly associated with TB group. They were much less likely to be HIV positive (8.6% vs. 22%, p<0.01). Children in the EPTB group were somewhat less likely to die in the hospital (10.0% vs. 15.4%, p=0.066). The OR was greater than 4 for HIV and was greater than 2 for poor nutrition status [p<0.01 for each]. Conclusion: There was no association in this model between EPTB and mortality. The apparent univariate association between EPTB and reduced mortality can be explained by lower prevalence of HIV and poor nutrition status in this sub-sample. This study has implications for hospital-based pediatric TB diagnosis and epidemiology in resource-poor countries.

<p>Nitric oxide (NO) is an endogenous gaseous signaling molecule produced by three NO synthase isoforms (NOS1, 2, 3) and important in host defense. The induction of NOS2 during bacterial sepsis is critical for pathogen clearance but its sustained activation has long been associated with increased mortality secondary to multiple organ dysfunction syndrome (MODS). High levels of NO produced by NOS2 incite intrinsic cellular dysfunction, in part by damaging macromolecules through nitration and/or nitrosylation. These include mitochondrial DNA (mtDNA) and enzymes of key mitochondrial pathways required for maintenance of normal O2 utilization and energy homeostasis. However, animal studies and clinical trials inhibiting NOS2 have demonstrated pronounced organ dysfunction and increased mortality in response to live bacterial infections, confirming that NOS2 confers pro-survival benefits. Of particular interest here, the constitutive NOS1 and NOS3 have been linked to the up-regulation of nuclear genes involved in mitochondrial biogenesis but no comparable role has been described for NOS2. <italic> Therefore, I hypothesized that NOS2 is indispensible for host protection but must be tightly regulated to ensure NO levels are high enough to activate mitochondrial and other pro-survival genes, but below the threshold for cellular damage.</italic></p><p>This hypothesis was explored with two major Aims. The <italic>first Aim</italic> was to define the role of NOS2 in the activation of mitochondrial biogenesis in the heart of <italic>E. coli</italic>-treated mice. The <italic>second</italic> was to investigate the ability of NOS2 to be transcriptionally regulated by an enzyme previously shown to induce mitochondrial biogenesis, heme oxygenase-1 (HO-1). This hypothesis was tested using an <italic>in vivo</italic> model of sublethal heat-killed <italic>E. coli</italic> (<italic>HkEC</italic>) peritonitis in C57B/L6 (Wt), NOS2-/-, and TLR4-/- mice. Additionally, <italic>in vitro</italic> systems of mouse AML-12 or Hepa 1-6 cells pretreated with HO-1 activators or <italic>Hmox1</italic> shRNA prior to inflammatory challenge with lipopolysaccharide (LPS) +/- tumor necrosis factor-&alpha; (TNF-&alpha;). For the first Aim, Wt, NOS2-/-, and TLR4-/- mice were treated with (<italic>HkEC</italic> and cardiac tissue analyzed for mitochondrial function, expression of nuclear and mitochondrial proteins needed for mitochondrial biogenesis, and histological expression of NOS2 and TLR4 relative to changes in mitochondrial mass. For the second Aim, Wt mice were pretreated with hemin or carbon monoxide (CO) to activate HO-1 prior to <italic>HkEC</italic>-peritonitis. Liver tissue in these animals was evaluated at four hours for HO-1 induction, <italic>Nos2</italic> mRNA expression, cytokine profiles, and nuclear factor (NF)-&kappa;B activation. Liver cell lines were pretreated with hemin, CO-releasing molecule (CORM), or bilirubin one hour before LPS exposure and the <italic>Nos2</italic> transcriptional response evaluated at two and 24 hours. The MTT assay was used to confirm that <italic>in vitro</italic> treatments were not lethal. </p><p>These studies demonstrated that <italic>HkEC</italic> induced mtDNA damage in the heart that was repaired in Wt mice but not in NOS2-deficient mice. In KO mice, sustained mtDNA damage was associated with the reduced expression of nuclear (NRF-1, PGC-1&alpha;) and mitochondrial (Tfam, Pol-&gamma;) proteins needed for mitochondrial biogenesis. The findings thus supported that NOS2 is required for mitochondrial biogenesis in the heart during Gram-negative challenge. Evaluation of the relationship between HO-1 and NOS2 in murine liver was more complex; HO-1 activation in <italic>HkEC</italic>-treated Wt mice attenuated 4-hour <italic>Nos2</italic> gene transcription. In liver cell lines, hemin, CORM, and bilirubin were unable to suppress <italic>Nos2</italic> expression at the time of maximal induction (2 hours). <italic>Nos2</italic> was, however, suppressed by 24 hours, suggesting that the regulatory impact of HO-1 induction was not engaged early enough to reduce <italic>Nos2</italic> transcription at 2 hours. It is concluded that NOS2 induction in bacterial sepsis optimizes the expression of the mitochondrial biogenesis transcriptional program, which subsequently can also be regulated by HO-1/CO in murine liver. This provides a potential new mechanism by which immune suppression and mitochondrial repair can occur in tandem during the acute inflammatory response.</p> / Dissertation

Made available in DSpace on 2016-12-23T13:56:06Z (GMT). No. of bitstreams: 1 Eliana Ferrraz de Brito.pdf: 1819028 bytes, checksum: f09d6294a2538b396426079d487e84b5 (MD5) Previous issue date: 2010-09-03 / Introdução. A acupuntura e a eletroacupuntura têm efeitos antiinflamatórios, demonstrados em modelos experimentais e em algumas observações em humanos, mas muito pouco se conhece sobre o seu impacto na evolução de infecções. Há uma observação sobre efeitos da acupuntura nas fases iniciais da sepse induzida por ligadura e punção do ceco em ratos, mostrando redução das bactérias nas primeiras 6 horas. Objetivos. Avaliar os efeitos da eletroacupuntura na evolução de uma peritonite polimicrobiana por inoculação intraperitoneal de fezes em camundongos. Material e Métodos. Camundongos suíços, machos, 6-8 semanas de idade, foram anestesiados com ketamina e xilazina e inoculados por via intraperitoneal com 4&#956;l/g de peso corporal de uma suspensão de fezes diluídas a 1:9 (peso/vol em salina). Imediatamente após, um grupo recebia as agulhas nos pontos E-36 e VB-30, bilateralmente, permanecendo sem qualquer manipulação; o outro grupo recebeu as agulhas nos mesmos pontos, com estimulação elétrica (2 Hz) durante 30 minutos. Em um experimento se avaliou o número de neutrófilos exsudados na cavidade peritoneal 24h após a indução da peritonite e em outro experimento os animais foram acompanhados por 14 dias, quando foram eutanasiados para contagem e medida dos abscessos intraperitoneais. Resultados. Houve aumento significativo do número de neutrófilos na cavidade peritoneal e redução do número e tamanho dos abscessos intraperitoneais nos animais que receberam a eletroacupuntura. Conclusão. A eletroacupuntura aumentou a resistência dos camundongos à peritonite polimicrobiana induzida por uma suspensão de fezes. Este dado foi indicado pela maior exsudação de neutrófilos nas primeiras 24 horas e pelo menor número e tamanho dos abscessos intraperitoneais avaliados 14 dias depois da indução da peritonite. Uma explicação possível para o resultado observado seria o efeito da EA liberando mediadores do simpático e endorfinas que potencializariam a imunidade inata agindo em receptores &#945; adrenérgicos de macrófagos peritoneais e potencializando a ação de células citotóxicas naturais / Introduction. Acupuncture and electroacupuncture have anti-inflammatory effects as demonstrated in experimental models and some observations in humans, but little is known about their impact on the evolution of infections. There is one observation on effects of acupuncture in the early stages of sepsis induced by cecal ligation and puncture in rats, showing a reduction of bacteria in the first six hours. Objectives. To study the effects of electroacupuncture on the evolution of a polymicrobial peritonitis by intraperitoneal inoculation of feces in mice. Material and Methods. Male Swiss mice, 6-8 weeks old, were anesthetized with ketamine and xylazine and inoculated by intra-peritoneal route with 4&#956;l / g body weight of a suspension of feces diluted to 1:9 (weight / vol in saline). Immediately after, a group received the needles in points E36 and VB-30, bilaterally, remaining without any manipulation, and other group received the needles on the same points with electrical stimulation (2 Hz) for 30 minutes. In one experiment the neutrophils in the peritoneal exudates were counted 24 h after induction of peritonitis and in another experiment the animals were followed up for 14 days when they were euthanized for counting and measurement of intraperitoneal abscesses. Results. Significant increase in the number of neutrophils in the peritoneal cavity and reduction in number and size of intraperitoneal abscesses were observed in the animals receiving electroacupuncture. Conclusion. Electroacupuncture increased the resistance of mice to polymicrobial peritonitis induced by a suspension of feces. This conclusion is supported by the higher exudation of neutrophils in the first 24 hours and the lower number and size of intra-peritoneal abscesses evaluated 14 days after peritonitis induction. A possible explanation for the observed result would be the effect of EA releasing sympathetic mediators and endorphins enhancing the innate immunity by acting on &#945;-adrenergic receptors of peritoneal macrophages and stimulating the activity of natural killer cells

Submitted by Ana Cláudia Moro Lima dos Santos (anna.moro@hotmail.com) on 2018-05-07T20:15:18Z No. of bitstreams: 1 Dissertaçãofinal.pdf: 1989003 bytes, checksum: c02493762a88023038e4fc08eea12f1c (MD5) / Approved for entry into archive by Sulamita Selma C Colnago null (sulamita@btu.unesp.br) on 2018-05-09T16:37:26Z (GMT) No. of bitstreams: 1 santos_acml_me_bot.pdf: 1989003 bytes, checksum: c02493762a88023038e4fc08eea12f1c (MD5) / Made available in DSpace on 2018-05-09T16:37:26Z (GMT). No. of bitstreams: 1 santos_acml_me_bot.pdf: 1989003 bytes, checksum: c02493762a88023038e4fc08eea12f1c (MD5) Previous issue date: 2018-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Peritonite por bacilos Gram-negativos não fermentadores (BGNNF) é complicação importante da diálise peritoneal (DP), com curso clínico grave e elevada taxa de falência do método. Fatores associados à virulência, resistência antimicrobiana, formação de biofilme, entre outros, têm sido relatados, mas o limitado conjunto de evidências não permite concluir sobre os fatores responsáveis pelo pior curso clínico dessas infecções. O objetivo deste trabalho foi avaliar a influência das características microbiológicas, das condições clínicas do paciente e do tratamento na evolução de peritonites por BGNNF, ocorridas num único centro, em período de 18 anos. A sensibilidade in vitro aos antimicrobianos, produção de biofilme, além da análise do perfil clonal das bactérias pela técnica de eletroforese em gel de campo pulsado foram realizadas em todos os isolados bacterianos. Foram pesquisados genotipicamente, em isolados de Pseudomonas aeruginosa, a presença de marcadores de virulência (alginato, exoenzima S, fosfolipases C, exotoxina A, protesase alcalina, elastase e ramnolipídeos). Associações entre as características microbiológicas do paciente e tratamento com a taxa de resolução da peritonite foram estudadas. A espécie mais frequente foi Pseudomonas aeruginosa (45,59%), seguida por isolados do complexo Acinetobacter baumannii (17,65%). O estudo dos fatores de virulência da Pseudomonas aeruginosa revelou a presença de fatores de virulência em 100% dos casos, exceto exoenzima S (58,33%) e fosfolipase C não hemolítica (87,5%). Houve elevada proporção de BGNNF resistentes aos antimicrobianos testados, em particular à amicacina (36,73%) e à ciprofloxacina (44,9%), sendo que a sensibilidade aos betalactâmicos esteve acima de 70%. Observou-se elevada proporção de isolados produtores de biofilme (73,08%). Os resultados da tipagem por PFGE revelaram um perfil policlonal para a maioria dos isolados, entretanto para isolados do complexo Acinetobacter baumannii a análise revelou um cluster, entre 2000-2008, com perfil de multiresistência aos antimicrobianos, sugerindo fonte hospitalar. A evolução dos episódios mostrou reduzida taxa de cura (35,29%). A sensibilidade à amicacina e cefepime, se associaram de modo independente à maior chance de cura, enquanto a presença concomitante de infecção do óstio de saída do cateter de DP foi preditor independente de não resolução do episódio. Não se observaram associações entre fatores de virulência, produção de biofilme e características do paciente e tratamento com o desfecho dos episódios. Em conclusão, peritonites em DP, por BGNNF, são infecções com reduzida taxa de cura; a resistência bacteriana é fator associado à menor chance de resolução e peritonite por bactérias do gênero Acinetobacter spp. podem representar infecção grave, potencialmente de origem hospitalar, o que deve fazer redobrar os cuidados quanto ao seu manejo clínico. / Peritonitis due to non-fermentative Gram-negative bacilli (NFGNB) is a serious complication of peritoneal dialysis (PD), with a severe clinical course and high technique failure rate. Factors as bacterial virulence, antimicrobial resistance, biofilm formation, among others, have been reported, but the limited amount of evidence does not allow to conclude on the factors responsible for the worst clinical course of these infections. The objective of this study was to evaluate the influence of the microbiological characteristics, patients conditions, and treatment on evolution of peritonitis episodes at a single center in an 18 - year period. In vitro susceptibility, biofilm production, and clonal profile analysis of bacteria by pulsed-field gel electrophoresis (PFGE) were performed in all isolates. The presence of virulence markers (alginate, exoenzyme S, phospholipases C, exotoxin A, alkaline protease, elastase, and ramnolipids) was genotyped in bacterial isolates of Pseudomonas aeruginosa. From the data referring to the patient and causal agent, associations between the microbiological, patient characteristics, and treatment on the resolution rate of peritonitis were analyzed. The most frequent species was Pseudomonas aeruginosa (45.59%), followed by Acinetobacter baumannii complex (17.65%). The study of the virulence factors of Pseudomonas aeruginosa revealed the presence of virulence factors in 100% of the cases, except for exonzyme S (58.33%) and hemolytic phospholipase C (87.5%). There was a high proportion of antimicrobial resistant, in particular to amikacin (36.73%) and ciprofloxacin (44.9%), with sensitivity to betalactam above 70%. A high (73.08%) proportion of biofilm producing isolates was observed. The results of the PFGE typing revealed a polyclonal profile for most of the isolates; however, for the Acinetobacter baumannii complex species the analysis revealed a cluster at interval from 2000 to 2008, with antimicrobial multi resistance profile, suggest that peritonitis by this agent had a hospital source. The evolution of the episodes showed a reduced resolution rate (35.29%). The susceptibility to amikacin and to cefepime were independently associated with a higher odds of resolution, while the concomitant presence of PD catheter exit site infection was an independent predictor of non-resolution. In conclusion, peritonitis due to NFGNB in PD are infections with reduced resolution rate; bacterial resistance is an independent predictor of lower odds of resolution. Peritonitis by Acinetobacter spp. can represent serious / 64736017.2.0000.5411

Sepsis-induced organ failure is associated with high morbidity and mortality rates. The onset of an exaggerated host response to microbial invasion and/or trauma, is believed to be the primary cause of excessive inflammation and the subsequent tissue hypoperfusion observed in patients with severe sepsis. In our mouse model of sepsis induced by cecal ligation and perforation (CLP), symptoms indicative of the disease, including diarrhea, increased ventilation and persistent hypothermia, are present at six hours after the surgery (T6). In the untreated CLP mice, mortality occurs starting at T15. As induced hypothermia has shown to exert immunomodulatory effects, this study is aimed at assessing its potential in attenuating inflammation and improving survival in experimental sepsis. Our data has shown that deep hypothermia initiated at T6, by means of cold chamber-induced cooling, prolongs survival. Plasma cytokine quantification by enzyme-linked immunosorbent assays (ELISA) also reveals that induced deep hypothermia reduces tumour necrosis factor(TNF)-α and interleukin (IL)-6 production in untreated CLP mice. In contrast, induced moderate hypothermia does not have such effect. Antibiotic (cefotaxime) and saline resuscitation initiated immediately following CLP ensures survival. However, when these supportive treatments are initiated at T6, >50% mortality is observed in the CLP mice with or without induced hypothermia. In summary, this preliminary study provides proof for a downregulated inflammatory response mediated by external cooling. However, to achieve a survival benefit, treatment strategies in addition to cooling and antibiotics may be required.