Multiplex real-time PCR monitoring of intestinal helminths in humans reveals widespread polyparasitism in Northern Samar, the Philippines

The global socioeconomic importance of helminth parasitic disease is underpinned by the considerable clinical impact on millions of people. While helminth polyparasitism is considered common in the Philippines, little has been done to survey its extent in endemic communities. High morphological similarity of eggs between related species complicates conventional microscopic diagnostic methods which are known to lack sensitivity, particularly in low intensity infections. Multiplex quantitative PCR diagnostic methods can provide rapid, simultaneous identification of multiple helminth species from a single stool sample. We describe a multiplex assay for the differentiation of Ascaris lumbricoides, Necator americanus, Ancylostoma, Taenia saginata and Taenia solium, building on our previously published findings for Schistosoma japonicum. Of 545 human faecal samples examined, 46.6% were positive for at least three different parasite species. High prevalences of S. japonicum (90.64%), A. lumbricoides (58.17%), T. saginata (42.57%) and A. duodenale (48.07%) were recorded. Neither T. solium nor N. americanus were found to be present. The utility of molecular diagnostic methods for monitoring helminth parasite prevalence provides new information on the extent of polyparasitism in the Philippines municipality of Palapag. These methods and findings have potential global implications for the monitoring of neglected tropical diseases and control measures.

Enteropathogenic Escherichia coli O125:H6 triggers attaching and effacing lesions on human intestinal biopsy specimens independently of Nck and TccP/TccP2

Typical enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) employ either Nck, TccP/TccP2, or Nck and TccP/TccP2 pathways to activate the neuronal Wiskott-Aldrich syndrome protein (N-WASP) and to trigger actin polymerization in cultured cells. This phenotype is used as a marker for the pathogenic potential of EPEC and EHEC strains. In this paper we report that EPEC O125:H6, which represents a large category of strains, lacks the ability to utilize either Nck or TccP/TccP2 and hence triggers actin polymerization in vitro only inefficiently. However, we show that infection of human intestinal biopsies with EPEC O125:H6 results in formation of typical attaching and effacing lesions. Expression of TccP in EPEC O125:H6, which harbors an EHEC O157-like Tir, resulted in efficient actin polymerization in vitro and enhanced colonization of human intestinal in vitro organ cultures with detectable N-WASP and electron-dense material at the site of bacterial adhesion. These results show the existence of a natural category of EPEC that colonizes the gut mucosa using Nck- and TccP-independent mechanisms. Importantly, the results highlight yet again the fact that conclusions made on the basis of in vitro cell culture models cannot be extrapolated wholesale to infection of mucosal surfaces and that the ability to induce actin polymerization on cultured cells should not be used as a definitive marker for EPEC and EHEC virulence.

Niveles de biológicos: eficacia e inmunogenicidad del tratamiento para enfermedad inflamatoria intestinal

Programa de doctorado: Avances en Medicina Interna. La fecha de publicación es la fecha de lectura

Pré-condicionamento isquêmico em diferentes tempos e seu efeito na translocação bacteriana induzida por isquemia e reperfusão intestinal em ratos

Para avaliar os efeitos de diferentes tempos de pré-condicionamento isquêmico (IPC) em translocação bacteriana intestinal (BT). MÉTODOS: Trinta ratos Wistar pesando 280 ± 27g foram divididos em cinco grupos. No grupo IV (n = 6), a laparotomia foi realizada e a artéria mesentérica superior foi obstruído por um microclampe atraumática durante 30 minutos. Nos quatro grupos de pré-condicionamento (n = 6 cada) antes dos 30 minutos de isquemia-reperfusão (I / R), os ratos foram submetidos a IPC para duas, cinco, dez e 15 minutos, seguido pelo mesmo momento da reperfusão. A fim de avaliar se o tempo de pré-condicionamento influenciaram o surgimento de translocação bacteriana, as amostras de nódulos linfáticos mesentéricos, fígado e baço foram colhidas em condições estéreis, 24 horas após os procedimentos para a quantificação de unidades formadoras de colónias de bactérias por grama de tecido (CFU / g). O sangue foi recolhido para a medição de citoquinas. RESULTADOS: No grupo I / R, o total de CFU / g em gânglios linfáticos mesentéricos, baço, fígado, bem como o soro de TNF-a, IL-1A e IL-6 foram significativamente mais elevados do que nos outros grupos (p <0,05). Pré-condicionamento por 15 minutos significativamente atenuada BT e citocinas séricas quando comparado a outros períodos de pré-condicionamento (p <0,05). CONCLUSÃO: Nossos dados sugerem que o pré-condicionamento como um fator chave para reduzir a translocação bacteriana intestinal em I / R. Numa escala de dois a 15 minutos, o melhor tempo de pré-condicionamento isquémico pela atenuação da translocação bacteriana foi de 15 minutos

Translocation of 99mTc labelled bacteria after intestinal ischemia and reperfusion

Ischemia and reperfusion of the small intestine disrupts gut barrier, causes bacterial translocation and activates inflammatory responses. An experimental study was planned to evaluate if 99mTc labelled Escherichia coli translocates to mesenteric lymph nodes, liver, spleen, lung and serum of rats submitted to mesenteric ischemia/reperfusion. Additionally, it was observed if the time of reperfusion influences the level of translocation. METHODS: Forty male Wistar rats underwent 45 minutes of gut ischemia by occlusion of the superior mesenteric artery. The translocation of labelled bacteria to different organs and portal serum was determined in rats reperfused for 30 minutes, 24 hours, sham(S) and controls(C), using radioactivity count and colony forming units/g (CFU). RESULTS: All the organs from rats observed for 24 hours after reperfusion had higher levels of radioactivity and positive cultures (CFU) than did the organs of rats reperfused for 30 minutes, C and S, except in the spleen (p<0,01). CONCLUSION: The results of this study indicated that intestinal ischemia/reperfusion led to bacterial translocation, mostly after 24 hours of reperfusion

Intestinal obstruction by giant Meckel's diverticulum. Case report

Most cases of Meckel’s diverticulum (MD) are asymptomatic and discovered by chance. Management of MD is controversial. The authors describe an exceptional case of intestinal obstruction caused by a giant MD in a patient who had previously undergone appendectomy. A review of the contradictory literature on this subject leads to the conclusion that careful consideration of clinical and morphological data (patient's age, ASA score, the surgical procedure to be performed, morphology and position of the MD, any fibrotic bands) is required before deciding whether or not to resect an asymptomatic MD.

Emergency emicolectomy for intestinal primary Aspergillosis in acute myeloid leukemia

ntestinal aspergillosis is an infection with a very high death rate especially in leukemic patients. Here we describe a case of a 46 years old woman with acute myeloid leukemia (LAM M5) who developed intestinal primary aspergillosis. This patient was diagnosed with LAM M5 through bone marrow aspiration and bone biopsy in March 2004. Symptoms of the disease were slight persistent fever, weight loss, asthenia, anemia, thrombocytopenia,and leukocytosis with high number of blasts in peripheral blood. After induction chemotherapy with ICE (Ifosfamide, Carboplatin, Etoposide), she developed neutropenia and high fever without apparent infective foci. She was treated with empiric antibiotic therapy, nevertheless she developed an intense diarrhea and ileo-cecal distention. Diagnostic exams didn’t show signs of a focal lesion. Despite the change in antibiotic treatment and the transfusions of granulocytes and blood cells, the patient developed extremely critical conditions with persistence of neutropenia and abdominal distention. A surgical treatment was decided at the time. We treated the patient with a two steps surgical procedure. The first step was a right abdominal ileostomy followed by improvement of general conditions and then the second step a right colectomy. The histological morphology confirmed necrotizing colitis with Aspergillus ife. At that time , treatment with voriconazole was started. The general conditions of the patient improved rapidly and we were able to treat the patient with other medical anti-leukemic therapies. The patient is now cured and in healthy state. We obtained a good clinical result as only in other few cases described in literature.

Pré-condicionamento isquêmico em diferentes tempos e seu efeito na translocação bacteriana induzida por isquemia e reperfusão intestinal em ratos

Para avaliar os efeitos de diferentes tempos de pré-condicionamento isquêmico (IPC) em translocação bacteriana intestinal (BT). MÉTODOS: Trinta ratos Wistar pesando 280 ± 27g foram divididos em cinco grupos. No grupo IV (n = 6), a laparotomia foi realizada e a artéria mesentérica superior foi obstruído por um microclampe atraumática durante 30 minutos. Nos quatro grupos de pré-condicionamento (n = 6 cada) antes dos 30 minutos de isquemia-reperfusão (I / R), os ratos foram submetidos a IPC para duas, cinco, dez e 15 minutos, seguido pelo mesmo momento da reperfusão. A fim de avaliar se o tempo de pré-condicionamento influenciaram o surgimento de translocação bacteriana, as amostras de nódulos linfáticos mesentéricos, fígado e baço foram colhidas em condições estéreis, 24 horas após os procedimentos para a quantificação de unidades formadoras de colónias de bactérias por grama de tecido (CFU / g). O sangue foi recolhido para a medição de citoquinas. RESULTADOS: No grupo I / R, o total de CFU / g em gânglios linfáticos mesentéricos, baço, fígado, bem como o soro de TNF-a, IL-1A e IL-6 foram significativamente mais elevados do que nos outros grupos (p <0,05). Pré-condicionamento por 15 minutos significativamente atenuada BT e citocinas séricas quando comparado a outros períodos de pré-condicionamento (p <0,05). CONCLUSÃO: Nossos dados sugerem que o pré-condicionamento como um fator chave para reduzir a translocação bacteriana intestinal em I / R. Numa escala de dois a 15 minutos, o melhor tempo de pré-condicionamento isquémico pela atenuação da translocação bacteriana foi de 15 minutos

Caraterização in vitro dos efeitos tóxicos da patulina na integridade do epitélio intestinal e potenciais efeitos protetores da cisteína

A mucosa intestinal é a primeira barreira biológica encontrada pelas micotoxinas presentes nos alimentos, sendo a patulina, uma micotoxina produzida por fungos do género Penicillium spp., uma preocupação particular atendendo a que a exposição humana a esta micotoxina pode conduzir a efeitos imunológicos, neurológicos e gastrointestinais. Considerando estes efeitos para a saúde, o presente estudo tem como objetivos a avaliação do efeito tóxico da exposição intestinal a patulina, bem como a determinação do potencial efeito protetor da coadministração de patulina e cisteína na membrana intestinal, utilizando para o efeito células Caco-2. A integridade da membrana intestinal foi determinada pela medição da resistência elétrica transepitelial (TEER). Os resultados evidenciaram um decréscimo acentuado nos valores de TEER após 24 horas de exposição celular a 95 μM de patulina. Para as concentrações mais reduzidas verificou-se uma redução máxima inferior a 25% após 24 horas de exposição. A coadministração de patulina (95 μM) e cisteína (40 μM) revelou um decréscimo nos valores de TEER. O tratamento com cisteína em concentrações superiores ( 400 μM) revelou efeito protetor da membrana intestinal, tendo em conta os valores de TEER. Estes resultados contribuem para uma avaliação do risco mais precisa associada à exposição a contaminantes alimentares.

Translocation of 99mTc labelled bacteria after intestinal ischemia and reperfusion

Ischemia and reperfusion of the small intestine disrupts gut barrier, causes bacterial translocation and activates inflammatory responses. An experimental study was planned to evaluate if 99mTc labelled Escherichia coli translocates to mesenteric lymph nodes, liver, spleen, lung and serum of rats submitted to mesenteric ischemia/reperfusion. Additionally, it was observed if the time of reperfusion influences the level of translocation. METHODS: Forty male Wistar rats underwent 45 minutes of gut ischemia by occlusion of the superior mesenteric artery. The translocation of labelled bacteria to different organs and portal serum was determined in rats reperfused for 30 minutes, 24 hours, sham(S) and controls(C), using radioactivity count and colony forming units/g (CFU). RESULTS: All the organs from rats observed for 24 hours after reperfusion had higher levels of radioactivity and positive cultures (CFU) than did the organs of rats reperfused for 30 minutes, C and S, except in the spleen (p<0,01). CONCLUSION: The results of this study indicated that intestinal ischemia/reperfusion led to bacterial translocation, mostly after 24 hours of reperfusion

Estudio Comparativo de Parasitismo Intestinal, realizado en los alumnos de Primero a Tercer Grado de los Centros Escolares Cantón Ojo de Agua y colonia El Cocal del Municipio y departamento de Usulután en el periodo de junio a agosto de 2014

El parasitismo intestinal, representa un problema de salud pública de mucha importancia a nivel mundial, sobre todo en zonas rurales y urbanas de los países en desarrollo, donde un gran número de habitantes viven en hacinamiento con graves problemas sanitarios y ausencia de hábitos higiénicos. Los parásitos en los niños son causa importante de trastornos en el desarrollo físico y mental de los mismos, el mecanismo y vía de contagio varía, la mayoría de los parásitos se adquieren al ingerir agua o alimentos contaminados con sus quistes o huevecillos. No obstante; todas las personas a cualquier edad pueden ser portadores de parásitos. El Objetivo del presente estudio fue comparar el porcentaje de alumnos de primero a tercer grado que presentan parasitismo intestinal en los Centros Escolares Cantón Ojo de Agua y Colonia El Cocal del Municipio y Departamento de Usulután en el periodo de junio a agosto de 2014. Metodología es una investigación de tipo prospectivo, transversal, comparativa, descriptiva y de laboratorio; la investigación incluyó a todos los alumnos de los grados antes mencionados en ambos Centros Escolares, se aplicaron criterios de inclusión y exclusión para establecer la población, además se administró una cédula de entrevista a los responsables de los niños para recopilar la información necesaria acerca de las condiciones de vida, hábitos higiénicos y alimenticios de los niños incluidos en el estudio. Se realizó el examen general de heces para poder observar la presencia de parásitos intestinales. Se utilizó una hoja de reporte de examen general de heces lo cual permitió realizar la tabulación de datos. Resultados: De un total de 83 niños estudiados en ambos Centros Escolares 44 (53.1%) resultaron negativos, 39 (46.9%) resultaron positivos para algún parasito; en el Centro Escolar Cantón Ojo de Agua ubicado en la zona rural 51 alumnos se sometieron al estudio de estos 26 (50.9%) resultaron con parasitismo intestinal, mientras que en el Centro Escolar Colonia Cocal se analizaron 32 muestras de heces de las cuales 13 (40.6%) resultaron con parásitos. De los 39 casos positivos de ambos Centros Escolares, 22 (56.4%) resultaron con un solo tipo de parásito y 17 (43.6%) con multiparasitismo. Los parásitos encontrados con mayor frecuencia fueron: Endolimax nana (51.3%) y Entamoeba histolytica (48.7%), el parasito encontrado con menor frecuencia fue: Iodamoeba butschlii (7.7%). Conclusión: Estadísticamente el porcentaje de parasitismo intestinal de los alumnos del Centro Escolar Cantón Ojo de Agua fue igual que el de los alumnos del Centro Escolar Colonia El Cocal.

Characterization of in vitro effects of patulin on intestinal epithelial and immune cells

The intestinal mucosa is the first biological barrier encountered by natural toxins, and could possibly be exposed to high amounts of dietary mycotoxins. Patulin (PAT), a mycotoxin produced by Penicillium spp. during fruit spoilage, is one of the best known enteropathogenic mycotoxins able to alter functions of the intestine (Maresca et al., 2008). This study evaluated the effects of PAT on barrier function of the gut mucosa utilizing the intestinal epithelial cell model Caco-2, and scrutinized immunomodulatory effects using human peripheral blood mononuclear cells (PBMC) and human blood monocyte-derived dendritic cells (moDCs) as test systems. PAT exposure reduced Caco-2 cell viability at concentrations above 12 mM. As expected, the integrity of a polarized Caco-2 monolayer was affected by PAT exposure, as demonstrated by a decrease in TER values, becoming more pronounced at 50 mM. No effects were detected on the expression levels of the tight junction proteins occludin, claudin-1 and claudin-3 at 50 mM. However, the expression of zonula occludens-1 (ZO-1) and myosin light chain 2 (MLC2) declined. Also, levels of phospho-MLC2 (p-MLC2) increased after 24 h of exposure to 50 mM of PAT. T cell proliferation was highly sensitive to PAT with major effects for concentrations above 10 nM of PAT. The same conditions did not affect the maturation of moDC. PAT causes a reduction in Caco-2 barrier function mainly by perturbation of ZO-1 levels and the phosphorylation of MLC. Low doses of PAT strongly inhibited T cell proliferation induced by a polyclonal activator, but had no effect on the maturation of moDC. These results provide new information that strengthens the concept that the epithelium and immune cells of the intestinal mucosa are important targets for the toxic effects of food contaminants like mycotoxins

Trois expositions environnementales, un trouble : perturbations endocrine, du métabolisme monocarboné, et du microbiote intestinal maternel pendant la gestation déclenchent des phénotypes de type autistique chez le rat de type sauvage

Les Troubles du Spectre Autistique (TSA) sont caractérisés par deux principaux symptômes : des difficultés de communication sociale et des comportements stéréotypés et intérêts restreints. Les TSA touchent 5 fois plus les garçons que les filles et une augmentation de la prévalence exponentielle et continue a été observée aux États-Unis ces dernières décennies. Cette augmentation ne peut s’expliquer par les facteurs génétiques à eux seuls qui ne représentent que 5 à 15% des cas de TSA. Il est donc indispensable d’identifier de potentiels facteurs de risque environnementaux des TSA. Le but de ce travail est d’étudier différents facteurs environnementaux potentiellement modifiables dans le développement de phénotypes autistiques dans différents modèles précliniques des TSA. Les objectifs spécifiques sont : (i) caractériser les effets neurocomportementaux provoqués par une exposition périnatale simultanée à 5 perturbateurs endocriniens parmi les plus prévalent dans notre environnement quotidien (DEHP, DBP, DiNP, BDE-47, BDE-99) à de faibles doses pertinentes pour l’exposition humaine, (ii) identifier les effets neurocomportementaux associés à une altération périconceptionnelle du microbiote maternelle (iii) déterminer les effets neurocomportementaux associés à une altération périconceptionnelle du métabolisme monocarboné. Les résultats présentés dans cette thèse démontrent le potentiel de chacun de ces facteurs environnementaux d’altérer le développement cérébral fœtal. Chaque condition expérimentale a provoqué l’apparition de traits autistiques chez les rats, avec des spécificités comportementales pour chaque exposition développementale. Des déficits d’interactions sociales ont été observés dans chaque situation expérimentale, associés soit à de l’anxiété, de l’hyperactivité, des altérations d’intégration sensorimotrice, et/ou des stéréotypies. Cela nous force à considérer les TSA comme une pathologie aux multiples facettes où l’hétérogénéité des tableaux cliniques est représentative de l’hétérogénéité des causes possibles. La multitude des interactions environnementales courantes possibles avec l’épigénome pourrait être à la base de la grande diversité observée dans la sévérité des symptômes et / ou des comorbidités des TSA. Ce travail ouvre des perspectives futures de prévention ciblée des TSA fondées sur de potentielles modifications de l’environnement comme la réduction de l’exposition aux perturbateurs endocriniens, ou des supplémentations en donneurs monocarbonés (e.g. acide folique) et/ou probiotiques.