A new systematization of histological analysis for the diagnosis of Hirschsprung's disease

Background: Hirschsprung's Disease (HD) is characterized by intestinal sub-occlusion and the absence of enteric ganglion cells. A rectal biopsy examination is performed to confirm the diagnosis. In a recent study, we demonstrated that the analysis of 60 sections of rectal mucosa and submucosa stained by H&E may ensure a 90% diagnostic accuracy. Although the need to analyze so many sections makes the process of reading the slides more time-consuming, this encouraged us to study their distribution in the healthy rectal submucosa, to simplify the diagnosis. Objectives: To develop a method that facilitates HD diagnosis by studying the distribution of ganglion cells in the submucosal plexus. Methods: Using the calretinin technique, we studied the distribution of plexuses in 60 fragments of rectal submucosa from 19 cadavers. After the study, the reading method created was used for diagnosis in 47 cases of suspected HD, using H&E staining. The accuracy was verified by comparing the results obtained with H&E to those obtained with the acetylcholinesterase technique, the golden standard in our laboratory. Results: The study of submucosal plexus distribution showed that just by examining the submucosal region every 20 µm, approximately, it is possible to locate a ganglionic plexus, and we have already been able to diagnose HD with 93% accuracy. Conclusion: The study of ganglion cell distribution enabled the creation of a simplified method for reading the slides. The method applied achieved good accuracy and it can be used as an alternative method in HD diagnosis

Estimación indirecta de emisiones de CO2 a partir de información satelital en áreas quemadas de ambientes insulares del delta del río Paraná (Argentina)

Fires in island environments in Delta del río Paraná (Argentina) have recurrence controlled by anthropogenic, climax and topographic factors. As a result, variations in the emissions of greenhouse gases are produced, added to local consequences (vegetable degradation, soil erosion, impacts on human health) or global consequences (air pollution, climate). Delta del río Paraná covers an area of approximately 17.500 km2, conforming a food plain characterized by pulses of flooding and droughts, associated to tidal and fluvial systems. This situation leads to a complex environmental structuring; dominate forest formations in the northern part and herbaceous in the distal portion in the Delta. In this last area, fire is applied recurrently as part of practice of handling livestock, without considering in most cases the interactions of other environmental variables (climate). During the period March/June 2008 there were a series of fires which affected mainly grassland and rushes in the Delta. The aim of this study was to estimate indirectly CO2 emissions that occurred during the period of burning of herbaceous plant communities in Delta del río Paraná. To do so, vegetations units were characterized from an unsupervised classification of the EVI index (Enhanced Vegetation Index), product MOD 13 Q1, in an annual series of compounds of 16 days of MODIS images (base year: 2007). Ten pieces of coverage coverage were identified; four of them corresponded to herbaceous vegetation. The burnt area was determined by applying the index BAI (Burnt Area Index) from bands from near infrared (2) and middle infrared (7) of the same compound. Estimates of CO2 emissions were made based on data from own herbaceous biomass, bibliographic and field data, using a parametric model that includes as variables of proportion of burnt biomass (0,6), the relation of C (Ct), total set in biomass (0,46) and the relation Ct/CO2 (3,67). Fires in Delta del Paraná produced CO2 emissions close to 20 Mg nC ha-1 for grasses and rushes prairies, while for the pre-Delta areas the emissions were estimate between 18 and 34 Mg C ha-1 for canutillares and cortaderales. Estimated emissions for the evaluated period are higher than 7,7 Tg C-CO2. These levels of CO2 emissions are comparable to the annually produced by an Argentinean city with 2.000.000 inhabitants.Fil: Sione, Walter Fabian. Universidad Autónoma de Entre Ríos; Argentina. Universidad Nacional de Luján; ArgentinaFil: Aceñolaza, Pablo Gilberto. Universidad Autónoma de Entre Ríos; Argentina. Provincia de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Universidad Autónoma de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción; ArgentinaFil: Zamboni, Lisandra Pamela. Universidad Autónoma de Entre Ríos; ArgentinaFil: Serafini, Maria Cristina. Universidad Nacional de Luján; ArgentinaFil: del Valle, Hector Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; ArgentinaFil: Gallardo Lancho, Juan. Consejo Superior de Investigaciones Científicas; Españ

Immediate expression of c-fos and c-jun mRNA in a model of intestinal autotransplantation and ischemia-reperfusion in situ

OBJECTIVE: Intestinal ischemia-reperfusion injury occurs in several clinical conditions and after intestinal transplantation. The aim of the present study was to investigate the phenomena of apoptosis and cell proliferation in a previously described intestinal ischemia-reperfusion injury autograft model using immunohistochemical markers. The molecular mechanisms involved in ischemia-reperfusion injury repair were also investigated by measuring the expression of the early activation genes c-fos and c-jun, which induce apoptosis and cell proliferation. MATERIALS AND METHODS: Thirty adult male Wistar rats were subjected to surgery for a previously described ischemia-reperfusion model that preserved the small intestine, the cecum and the ascending colon. Following reperfusion, the cecum was harvested at different time points as a representative segment of the intestine. The rats were allocated to the following four subgroups according to the reperfusion time: subgroup 1: 5 min; subgroup 2: 15 min; subgroup 3: 30 min; and subgroup 4: 60 min. A control group of cecum samples was also collected. The expression of c-fos, c-jun and immunohistochemical markers of cell proliferation and apoptosis (Ki67 and TUNEL, respectively) was studied. RESULTS: The expression of both c-fos and c-jun in the cecum was increased beginning at 5 min after ischemia-reperfusion compared with the control. The expression of c-fos began to increase at 5 min, peaked at 30 min, and exhibited a declining tendency at 60 min after reperfusion. A progressive increase in c-jun expression was observed. Immunohistochemical analyses confirmed these observations. CONCLUSION: The early activation of the c-fos and c-jun genes occurred after intestinal ischemia-reperfusion injury, and these genes can act together to trigger cell proliferation and apoptosis

Land Cover Map of the Fluvial/Littoral Parana River Complex (FLPRC) Using Geomatic Tools

El Complejo Fluvio/Litoral del río Paraná (CFLRP) es un humedal que posee características de sistemas ambientales complejos y heterogéneos, las cuales están dadas por la acción conjunta de procesos hidrológicos, geomorfológicos, biológicos y de origen antrópico asociados con el uso del suelo y la gestión del territorio. En el caso de los procesos de origen antrópico, se ha observado que las principales actividades económicas en el CFLRP se han centrado en los últimos a˜nos en la ganadería de isla y la forestación de salicáceas, si bien se pueden nombrar otras actividades como la urbanización, la pesca, la apicultura, el transporte fluvial, etc. Estas actividades han amenazado al CFLRP, ante lo cual se han realizado algunos esfuerzos de conservación que han conducido a la declaración de una parte de la región como área natural protegida. En este escenario, la cartografía de la cobertura y especialmente las cubiertas vegetales son un insumo básico para los planes de gestión territorial. Los datos de teledetección constituyen una base importante para la cartografía y seguimiento de cubiertas y permiten lograr mapas precisos y a escalas adecuadas. Teniendo lo anterior en cuenta, el presente trabajo se propuso generar un mapa de cobertura de suelo para el CFLRP mediante el uso de datos del sensor Landsat 8 OLI que sirva como propuesta metodológica para la generación de cartografía que puedan emplear los responsables de la gestión del territorio. Se descargó la escena 227-82 del servidor EarthExplorer (USGS, NASA, 2013). Como ventana temporal se utilizaron fechas representativas contrastantes en la región: primavera (período de aguas bajas) y oto˜no (período de aguas altas). De esta forma se incluyó la variabilidad temporal propia del área de estudio. Con el software e-Cognition (Trimble, 2000) se segmentó un compuesto multitemporal de 12 bandas (primavera y oto˜no). Se usaron como criterios un factor de escala de 100, de color de 0.9 y de forma de 0.1, a partir de lo cual se obtuvo un vectorial de polígonos de clases. Por otra parte, se realizó una clasificación no supervisada con el clasificador KMEAN (Técnica de Análisis de Auto-Organización Iterativa de Datos), empleando como parámetros 60 clases y 10 iteraciones. El mapa raster resultante se integró al vectorial producto de la segmentación a fin de obtener estadísticas (moda y diversidad) sobre el comportamiento de los píxeles en cada polígono. Finalmente, en la interpretación de los resultados, se le asignó la clase al polígono a partir de la moda e información de terreno. Se discriminaron 11 clases de cobertura, algunas de las cuales se asociaron con las características fisonómicas de la vegetación (fisonomías arbóreas, arbustivas, herbáceas de gran porte y herbáceas de baja altura). Otras de las clases de cobertura identificadas fueron agua, suelo, áreas urbanas, entre otras.Fluvio/Littoral Parana River complex (FLPRC) is a major wetland that has the characteristics of complex and heterogeneous environmental systems, which are given by the joint action of hydrological, geomorphological, biological and anthropogenic processes associated to land use practices. In the case of anthropogenic processes, it has been observed that the main economic activities have been focused, in recent years, on livestock farming and Salicaceae forestation, and other activities such as fishing, beekeeping, river transport, and recently urbanization. These activities have threatened the FLPRC, where some conservation efforts have been done, and led to the creation of some protected areas. In this context, land cover cartography is a basic input for land management plans. Remote sensing data are an important base for mapping and monitoring of land cover and allow achieving accurate maps. With this in mind, the present work was proposed to generate a map of land cover for CFLRP by using data from Landsat 8 OLI sensor, which serve as methodological proposal for generating cartography that can be used by those responsible of land management. 227-82 scene images of EarthExplorer server (USGS, NASA, 2013) were downloaded. Two temporal windows representing contrasting dates were used: spring (low water period) and autumn (high water period). Thus, the temporal variability of the study area was included. With e-Cognition software (Trimble, 2000), a segmentation of a multi-temporal product of 12 bands (spring and fall) was done. The result was a feature class. Furthermore, an unsupervised classification with KMEAN classifier using as parameters 60 classes and 10 iterations was performed. The resulting raster map was joined to the segmentation product to obtain statistics on the behavior of the pixels in each feature class. Finally, in the interpretation of the results, a class assignment was made to the feature class based on mode and field data. 11 land cover classes were discriminated, some of which were associated with the physiognomic characteristics of the vegetation (tree, shrubs, grass physiognomies). Other land cover classes were water, land, urban areas, among others.Fil: Aceñolaza, Pablo Gilberto. Provincia de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Universidad Autónoma de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción; ArgentinaFil: Zamboni, Lisandra Pamela. Universidad Autónoma de Entre Rí­os. Facultad de Ciencia y Tecnología. Centro Regional de Geomática; ArgentinaFil: Tentor, Fernando Raul. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Kalesnik, Fabio Alberto. Universidad Nacional de Luján; ArgentinaFil: Rodriguez, Estela Elizabeth. Provincia de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Universidad Autónoma de Entre Ríos. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Centro de Investigaciones Científicas y Transferencia de Tecnología a la Producción; ArgentinaFil: Sione, Walter Fabian. No especifica;Fil: Serafini, Maria Cristina. No especifica

Analysis of the reversibility of biliary cirrhosis in young rats submitted to biliary obstruction followed by desobstruction

In this letter to the editor of Revista de Medicina, we present a summary of our study entitled “Analysis of the reversibility of biliary cirrhosis in young rats submitted to biliary obstruction followed by desobstruction”, presented at the XXXV COMU in 2016 - 1st place of Oswaldo Cruz award, surgical area.Biliary atresia and other liver conditions are relevant in pediatric clinic, due to its progression into biliary cirrhosis and eventually, necessity for liver transplant. It is known that the period during which biliary obstruction persists is determining for the development of cirrhosis and its reversibility after a biliary drainage procedure. However, there are no time or histological markers of biliary cirrhosis reversibility. An animal model of biliary obstruction and desobstruction was employed and after histologic and molecular analysis, we concluded that, considering the high mortality rate and the improvement in histologic and molecular changes after biliary drainage in most groups, cirrhosis and its histological and molecular changes occur early after biliary obstruction, are severe and potentially fatal, but can be reversed or at least delayed after biliary drainage

Optimizing the pharmacological component of integrated balance therapy

Drug treatment is an important option for the treatment of peripheral vestibular diseases. AIM: To identify the drug component associated with optimal integrated balance therapy (IBT) for Ménière s disease or other peripheral vestibular disorders. MATERIALS AND METHODS: Analysis of a series of patients with Ménière s disease patients or patients with other peripheral vestibular disorders that received IBT involving either no medication or betahistine, cinnarizine, clonazepam, flunarizine or Ginkgo biloba during 120 days. RESULTS: In Ménière s disease, significant differences were observed for all drug therapies (60 days) versus no medication; betahistine was significantly more effective than all other drugs at 60 and 120 days. For non-Ménière s disorders, significant differences were observed among betahistine, cinnarizine, clonazepam and flunarizine and no medication after 60 days; all drug therapies were significantly more effective than no medication after 120 days; betahistine, cinnarizine or clonazepam were equally effective and betahistine was more effective than flunarizine and EGb 761. All treatment options were well tolerated. CONCLUSIONS: Drug therapies were more effective than no medication in the IBT for patients with Ménière s disease or other peripheral vestibular disorders. Betahistine was the most effective medication for patients with Ménière s disease and was as effective as cinnarizine and clonazepam for other peripheral vestibular disorders.A farmacoterapia é opção importante no tratamento das vestibulopatias periféricas. OBJETIVO: Identificar a medicação que otimiza a terapia integrada da vertigem (TIV) na doença de Ménière e em outras vestibulopatias periféricas. MATERIAL E MÉTODO: Estudo de casos em que pacientes com doença de Ménière ou outras vestibulopatias periféricas receberam TIV com betaistina, cinarizina, clonazepam, flunarizina, Ginkgo biloba ou sem medicação durante 120 dias. RESULTADOS: Na doença de Ménière, TIV com qualquer um dos medicamentos foi mais eficaz do que TIV sem medicação, após 60 dias; a betaistina foi mais efetiva que todas as outras drogas, após 60 e 120 dias. Nas outras vestibulopatias periféricas, diferenças significantes foram observadas entre TIV com betaistina, cinarizina, clonazepam ou flunarizina e TIV sem medicação após 60 dias e todas as drogas foram mais efetivas que TIV sem medicação após 120 dias; betaistina, cinarizina ou clonazepam foram igualmente efetivos e betaistina foi mais efetiva que flunarizina e Ginkgo biloba. Os tratamentos foram bem tolerados. CONCLUSÕES: TIV incluindo medicação é mais efetiva que sem medicação na doença de Ménière ou em outras vestibulopatias periféricas. Betaistina foi o medicamento mais efetivo na doença de Ménière e tão eficaz quanto cinarizina ou clonazepam em outras vestibulopatias periféricas.Universidade Federal de São Paulo (UNIFESP)Universidade Bandeirante de São Paulo Programa de Mestrado em Ciências do Movimento CorporalUNIFESP-EPMUNIBANUNIFESP, EPMSciEL

Sepsis and cirrhosis in pediatrics: revisiting two-hit bdl-clp rat model and its pathological and immunological reliability

Anai

Effects of ischemic preconditioning in a pig model of large-for-size liver transplantation

OBJECTIVE: In most cases of pediatric liver transplantation, the clinical scenario of large-for-size transplants can lead to hepatic dysfunction and a decreased blood supply to the liver graft. The objective of the present experimental investigation was to evaluate the effects of ischemic preconditioning on this clinical entity. METHODS: Eighteen pigs were divided into three groups and underwent liver transplantation: a control group, in which the weights of the donors were similar to those of the recipients, a large-for-size group, and a large-for-size + ischemic preconditioning group. Blood samples were collected from the recipients to evaluate the pH and the sodium, potassium, aspartate aminotransferase and alanine aminotransferase levels. In addition, hepatic tissue was sampled from the recipients for histological evaluation, immunohistochemical analyses to detect hepatocyte apoptosis and proliferation and molecular analyses to evaluate the gene expression of Bax (pro-apoptotic), Bcl-XL (anti-apoptotic), c-Fos and c-Jun (immediate-early genes), ischemia-reperfusion-related inflammatory cytokines (IL-1, TNF-alpha and IL-6, which is also a stimulator of hepatocyte regeneration), intracellular adhesion molecule, endothelial nitric oxide synthase (a mediator of the protective effect of ischemic preconditioning) and TGF-beta (a pro-fibrogenic cytokine). RESULTS: All animals developed acidosis. At 1 hour and 3 hours after reperfusion, the animals in the large-for-size and large-for-size + ischemic preconditioning groups had decreased serum levels of Na and increased serum levels of K and aspartate aminotransferase compared with the control group. The molecular analysis revealed higher expression of the Bax, TNF-alpha, I-CAM and TGF-beta genes in the large-for-size group compared with the control and large-for-size + ischemic preconditioning groups. Ischemic preconditioning was responsible for an increase in c-Fos, IL-1, IL-6 and e-NOS gene expression. CONCLUSION: Ischemia-reperfusion injury in this model of large-for-size liver transplantation could be partially attenuated by ischemic preconditioning