Detection and differentiation of Entamoeba histolytica and Entamoeba dispar in clinical samples through PCR-denaturing gradient gel electrophoresis.

Amebiasis is one of the twenty major causes of disease in Mexico; however, the diagnosis is difficult due to limitations of conventional microscopy-based techniques. In this study, we analyzed stool samples using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to differentiate between Entamoeba histolytica (pathogenic) and E. dispar (non-pathogenic). The target for the PCR amplification was a small region (228 bp) of the adh112 gene selected to increase the sensitivity of the test. The study involved 62 stool samples that were collected from individuals with complaints of gastrointestinal discomfort. Of the 62 samples, 10 (16.1%) were positive for E. histolytica while 52 (83.9%) were negative. No sample was positive for E. dispar. These results were validated by nested PCR-RFLP (restriction fragment length polymorphism) and suggest that PCR-DGGE is a promising tool to differentiate among Entamoeba infections, contributing to determine the specific treatment for patients infected with E. histolytica, and therefore, avoiding unnecessary treatment of patients infected with the non-pathogenic E. dispar.

Evaluación del riesgo de fenómenos trombóticos tras el uso de factor VII activado / Risk assessment of thrombotic events after the use of activated factor VII

Introducción: El objetivo del estudio fue analizar la incidencia de efectos trombóticos relacionada con la administración de factor VII humano recombinante activo (rFVIIa) en el tratamiento de la hemorragia grave tras la cirugía cardiaca. Material y métodos: Estudio retrospectivo de casos-controles pareado, de 2 años de duración, que incluyó a 72 niños ingresados en cuidados intensivos y tratados con rFVIIa por una hemorragia grave, durante o tras la cirugía cardiaca. Utilizamos un grupo control de 63 pacientes, estadísticamente comparables en cuanto a sexo, peso, diagnóstico, riesgo quirúrgico según la clasificación RACHS-1 y las características quirúrgicas. Resultados: No existieron diferencias significativas en la incidencia de fenómenos trombóticos (20% en casos y 28% en controles, p = 0,540), ni en la mortalidad (16% en casos y 9,5% controles, p = 0,208). Conclusión: En nuestra serie, el tratamiento con rFVIIa ha demostrado ser útil en el control de la hemorragia incoercible en niños sometidos a cirugía cardiaca, y no parece aumentar el riesgo de fenómenos trombóticos ni la mortalidad en el periodo postoperatorio (AU)

Introduction: The objective of this study was to analyze the incidence of thrombotic complications related to recombinant human factor VII a (rFVIIa) therapy for severe postoperative bleeding in cardiac surgery. Material and methods: A retrospective matched case-control study was conducted over two years, including 72 children admitted to intensive care unit and treated with rFVIIa because of a severe bleeding during or after cardiac surgery. A control group of 63 patients was chosen, who were statistically comparable in sex, weight, diagnosis, surgical risk according RASCH-1 score, and surgical characteristics, was chosen. Results: There were no significant differences between cases and controls either in the rate of thrombosis (20% vs 28%, P=0.540), or in the mortality rate (16% vs 9.5%, P=0.208). Conclusions: In our study, the rFVIIa therapy was shown to be useful in controlling severe operative bleeding in pediatric cardiac surgery, but does not seem to increase the risk of thrombotic complications or mortality rate in the postoperative period (AU)

Pancreatitis aguda: nuestra experiencia / Acute pancreatitis: our experience

Introducción: La pancreatitis aguda es un cuadro de dolor abdominal poco frecuente en la infancia. Nuestro objetivo es describir las características de los pacientes ingresados entre1988 y 2008 en un hospital terciario con este diagnóstico. Métodos: Estudio observacional retrospectivo por revisión de historias clínicas. Se recogen datos epidemiológicos, etiológicos, clínicos, de laboratorio, radiológicos, de manejo terapéutico y evolución clínica. Resultados: Incluimos 27 pacientes (un 66% varones), con una media de edad de 7,2 años (rango: 6 meses-16 años). El52% presenta enfermedades crónicas relevantes. El mecanismo etiológico más frecuente fue el estructural (37%), aunque hubo otros agentes causales, como ingesta de fármacos, infecciones, shock e inmunodeficiencias. Un 26% de los casos fueron catalogados como idiopáticos. El síntoma guía en todos los pacientes estables fue el dolor abdominal, de localización y características variables, acompañado de vómitos (55%), fiebre(33%), anorexia (19%) y otros síntomas relacionados con el agente causal. En todos se observó un aumento de las enzimas pancreáticas. La prueba de imagen más realizada fue la ecografía (patológica en un 74% de los casos), y el aumento del tamaño de la glándula fue el hallazgo más común. En el 89%de los pacientes el tratamiento fue médico, con dieta absoluta durante una mediana de 48 horas. Dos pacientes precisaron la realización de una colangiopancreatografía retrógrada endoscópica para el tratamiento de una coledocolitiasis. La duración media del ingreso fue de 9 días, y el 40% de los casos evolucionó favorablemente. Tres pacientes evolucionaron hacia pancreatitis crónica y 6 fallecieron por causas extradigestivas. Conclusiones: La pancreatitis aguda es una entidad poco frecuente en la edad pediátrica, con una amplia variedad de factores etiológicos que condicionan un pronóstico muy heterogéneo(AU)

Introduction: Acute pancreatitis is an unusual disorder in childhood that commonly onsets with sudden abdominal pain. Our goal is to describe the features of the patients diagnosed of acute pancreatitis between 1988 and 2008 who were admitted to our hospital. Methods: Retrospective observational study by review of medical records. We collected data referred to epidemiology, aetiology, clinical symptoms, analytical (amylase, lipase) and radiological findings, medical treatment administered and outcome. Results: We included 27 patients (66% males), with a mean age of 7.2 years (range: 6 months-16 years). 52% of them suffered from chronic diseases. The most common cause was a structural anomaly (37%). Other aetiologies included: drugs, infections, shock and immunodeficiencies. 26% of cases were catalogued as idiopatic. The presenting symptom in non-critical patients was abdominal pain, in different areas and with diverse features, associated with vomiting (55%), fever (33%),anorexia (19%) and other symptoms related to the a etiological agent. Pancreatic enzyme levels were elevated in all patients. The most widely used radiological test was ecography (abnormalin 74%), being the most common image the enlargement of the gland. 89% of patients were managed conservatively with complete bowel rest during a median of 48 hours. ERCP was indicated in 2 patients to treat coledocolitiasis. Median hospital stay was 9 days, with favourable outcome in 40% of patients. Recurrence was described in 3 patients and 6 died because of non-digestive causes. Conclusions: Acute pancreatitis is an unusual disease in childhood with a great variety of aetiological factors that determine different prognosis(AU)

[Risk assessment of thrombotic events after the use of activated factor VII]. / Evaluación del riesgo de fenómenos trombóticos tras el uso de factor vii activado.

INTRODUCTION: The objective of this study was to analyze the incidence of thrombotic complications related to recombinant human factor viia (rFVIIa) therapy for severe postoperative bleeding in cardiac surgery. MATERIAL AND METHODS: A retrospective matched case-control study was conducted over two years, including 72 children admitted to intensive care unit and treated with rFVIIa because of a severe bleeding during or after cardiac surgery. A control group of 63 patients was chosen, who were statistically comparable in sex, weight, diagnosis, surgical risk according RASCH-1 score, and surgical characteristics, was chosen. RESULTS: There were no significant differences between cases and controls either in the rate of thrombosis (20% vs 28%, P=.540), or in the mortality rate (16% vs 9.5%, P=.208). CONCLUSIONS: In our study, the rFVIIa therapy was shown to be useful in controlling severe operative bleeding in pediatric cardiac surgery, but does not seem to increase the risk of thrombotic complications or mortality rate in the postoperative period.

Pericarditis recurrente secundaria a un quiste pericárdico / Recurring secondary pericarditis due to pericardial cyst

Los procesos expansivos intratorácicos son infrecuentes en la infancia, y se localizan habitualmente en el mediastino. La escasa expresión clínica inicial condiciona un diagnóstico tardío y dificulta el tratamiento efectivo. Se clasifican según la localización topográfica (anterior, media o posterior), lo que permite establecer una orientación diagnóstica basada en las estructuras de cada compartimento. La patología tumoral es la más frecuente, aunque no hay que descartar la etiología infecciosa o malformativa (quística-vascular) y la patología tiroidea opericárdica, como el caso que motiva esta publicación. La radiografía de tórax es la técnica diagnóstica inicial, aunque generalmente se requiere la realización de una tomografía computarizada (mediastino anterior y medio) o una resonancia magnética (mediastino posterior), que localizan y delimitan la lesión e informan de su consistencia y contenido. La ecografía es coste efectiva, no invasiva, está libre de radiación y permite evaluar el timo o el pericardio. Las masas pericárdicas son infrecuentes, generalmente congénitas, aunque pueden tener un origen inflamatorio; su tratamiento es conservador, y está indicada la cirugía si el tamaño es grande o la sintomatología persistente(AU)

The intrathoracic expansive mass are infrequent during infancy, being the common localization the mediastinic, the scarce clinical expression conditions to a late diagnosis obstructing the effective treatment. They are classified according to topographic localization (front, middle or back) allowing a diagnosis orientation based on the structures of each compartment. The tumoral pathology is the most frequent, not being despised the infectious etiology, malformative (cystic or vascular), the thyroid pathology or pericardial, as the case, which this articleis based on. The thorax x-ray is the initial diagnosis technique, although generally a cat scan is needed of the (front mediastinum or middle) or a magnetic resonance (back mediastinum)that localizes and defines the lesion, informing of the consistency and context. The ecography is cheap not invasive, radiation free and allows evaluating the thymus and pericardium. The pericardial masses are infrequent, generally congenital, although they might have an inflammatory origin; its treatment is conservative, surgery is indicated if the size is big or the group of symptoms is persistent(AU)

Clonidine decreases intraoperative bleeding in middle ear microsurgery.

BACKGROUND: The antihypertensive drug clonidine is a centrally acting alpha2 agonist useful as a premedicant because of its sedative, anxiolytic, and analgesic properties. We examined the effect of clonidine given as an oral preanesthetic medication in producing a bloodless surgical field in patients undergoing middle ear microsurgery. We also evaluated whether the administration of clonidine would alter the reflex cardiovascular response to laryngoscopy and endotracheal intubation, anesthetic requirement, postoperative pain intensity and consumption of analgesics, and pre- and postoperative sedation and anxiety. METHODS: A prospective, randomized, double-blind clinical trial was performed in 40 patients scheduled for elective middle ear surgery under general anesthesia. Twenty-one patients received clonidine (300 microg p.o.) 90 min prior to arrival at the operating theater and 19 received placebo (control group). The hemodynamic endpoint of the anesthetic management was maintenance of hypotension for producing a bloodless surgical field. The desired control of the cardiovascular system was attained with isoflurane (inspired concentration increments of 0.25 vol% up to a maximum of 1.5 vol%)+/-fentanyl (bolus of 1 microg. kg-1)+/-urapidil (bolus of 0.3 mg. kg-1) as needed. Intraoperative bleeding was assessed on a four-point scale from 0=no bleeding to 3=abundant bleeding. RESULTS: There was less bleeding in the clonidine group (mean+/-SEM) than in the control group (0.75+/-0.3 vs 1.1+/-0.4, P<0.05). Patients given clonidine required a mean inspired isoflurane concentration of 0.63+/-0.1 vol% as compared with 1.01+/-0.2 vol% in controls (P<0.05). Fentanyl requirements were also significantly lower (57.10 vs 79.42 microg. kg-1, P<0.05). Four clonidine-treated patients required urapidil to achieve satisfactory hypotension as compared with 11 controls (P<0.05). Clonidine attenuated the associated cardiovascular response following laryngoscopy and intubation, and was more effective than placebo in achieving a satisfactory preoperative sedation and decreasing intensity of postoperative pain. Preoperative anxiety and incidence of adverse events was similar in both groups. CONCLUSION: Premedication with clonidine reduced bleeding in middle ear microsurgery, attenuated hyperdynamic response to tracheal intubation, and reduced isoflurane, fentanyl, and urapidil requirements for controlled hypotension.

Entamoeba histolytica : a novel cysteine protease and an adhesin form the 112 kDa surface protein.

Here, we present evidence that a cysteine protease (EhCP112) and a protein with an adherence domain (EhADH112) form the Entamoeba histolytica 112 kDa adhesin. Immunoelectron microscopy and immunofluorescence assays using monoclonal antibodies (mAbAdh) revealed that, during phagocytosis, the adhesin is translocated from the plasma membrane to phagocytic vacuoles. mAbAdh inhibited 54% adherence, 41% phagocytosis, and 35% and 62% destruction of MDCK cell monolayers by live trophozoites and their extracts respectively. We cloned a 3587 bp DNA fragment (Eh112 ) with two open reading frames (ORFs) separated by a 188 bp non-coding region. The ORF at the 5' end (Ehcp112 ) encodes a protein with a cysteine protease active site, a transmembranal segment and an RGD motif. The second ORF (Ehadh112 ) encodes a protein recognized by mAbAdh with three putative transmembranal segments and four glycosylation sites. Northern blot, primer extension and Southern blot experiments revealed that Ehcp112 and Ehadh112 are two adjacent genes in DNA. Ehcp112 and Ehadh112 genes were expressed in bacteria. The recombinant peptides presented protease activity and inhibited adherence and phagocytosis, respectively, and both were recognized by mAbAdh. The EhCP112 and EhADH112 peptides could be joined by covalent or strong electrostatic forces, which are not broken during phagocytosis.

[The values of high-density lipoproteins and the risk of developing atherosclerosis]. / Valores de lipoproteínas de alta densidad y riesgo de desarrollar ateroesclerosis.

A serum search was performed at Centro municipality, Tabasco, Mexico. 393 samples were obtained with the aim of determining high density lipoproteins (HDL) values. A Framingham search was also done. 56.7 per cent of females showed values above 65 mg/dl; 15.7 per cent, between 45 and 65; and 27.6 per cent less than 45 mg/dl, with minor, medium and high risks for atherosclerosis, respectively. 56.9 per cent of males showed values above 55 mg/dl; 17.6 per cent between 35 and 55, and 25.5 per cent less than 35, with similar risk levels. In the Framingham search, 67.9 per cent of females showed 4.5 points and 32.1 per cent more than 4.6 for minor and major risk levels, respectively. 70.3 per cent of males obtained less than 5 points and 29.7 more than 5.1, for same risk.