Description of a new experimental model skin flap for studying skin viability in rats.

PURPOSE: To describe a new experimental flap for studying skin viability in rats. METHODS: Twenty male Wistar rats weighing between 250-300g were divided into two groups: group A - McFarlane technique, a 4 x 10cm flap was used (McFarlane); and in group B modified McFarlane technique, a 3 x 10cm flap was used. Seven days later, the animals were sacrificed and the area of necrosis was evaluated in both groups. RESULTS: Group A presented necrosis in 3% of the total area of the flap (CI: 0.01-0.05), Group B presented necrosis in 37% of the total area of the flap (CI: 0.29-0.46), (p<0.001). CONCLUSION: The modified McFarlane flap presented a larger area of necrosis and could be an adequate experimental model of skin flap viability.

Description of a new experimental model skin flap for studying skin viability in rats

To describe a new experimental flap for studying skin viability in rats. METHODS: Twenty male Wistar rats weighing between 250-300g were divided into two groups: group A - McFarlane technique, a 4 x 10cm flap was used (McFarlane); and in group B modified McFarlane technique, a 3 x 10cm flap was used. Seven days later, the animals were sacrificed and the area of necrosis was evaluated in both groups. RESULTS: Group A presented necrosis in 3% of the total area of the flap (CI: 0.01-0.05), Group B presented necrosis in 37% of the total area of the flap (CI: 0.29-0.46), (p<0.001). CONCLUSION: The modified McFarlane flap presented a larger area of necrosis and could be an adequate experimental model of skin flap viability.

Evaluation of the number of goblet cells in crypts of the colonic mucosa with and without fecal transit.

OBJECTIVE: To measure the thickness of the crypts and quantify the number of goblet cells of the colonic mucosa with and without intestinal transit, relating them to exclusion time. METHODS: Sixty Wistar rats were divided into three groups of 20 animals each according to the time of the final operation for the removal of the colon, in six, 12 or 18 weeks. In each group 15 animals underwent colonic exclusion by left colon proximal colostomy and distal mucous fistula, and five underwent only laparotomy (control). The colons with and without fecal stream were removed, processed and submitted to histological sections stained with hematoxylin-eosin. The height of the colonic crypts and the number of goblet cells were measured by computerized morphometry. We used the Student t test and Kruskal-Wallis test for comparison and analysis of variance, using a significance level of 5% (p <0.05). RESULTS: The height of the crypts decreased in segments without fecal stream (p =0.0001), reducing from six to 12 weeks of exclusion (p = 0.0003), stabilizing thereafter. The number of goblet cells in the crypts was smaller in segments without transit after 12 and 18 weeks (p = 0.0001), but increased as the time of exclusion progressed (p = 0.04) CONCLUSION: The exclusion of intestinal transit decreases the thickness of the colonic crypts and the number of goblet cells in the segments without transit. There is an increased number of goblet cells in the course of time exclusion.

Avaliação do número de células caliciformes nas criptas da mucosa colônica com e sem trânsito intestinal / Evaluation of the number of goblet cells in crypts of the colonic mucosa with and without fecal transit

OBJETIVO: Medir a espessura das criptas e quantificar o número de células caliciformes comparando a mucosa cólica com e sem trânsito intestinal, relacionando-as ao tempo de exclusão. MÉTODOS: Sessenta ratos Wistar, foram distribuídos em três grupos com 20 animais segundo a operação final para a retirada dos cólons, realizadas em seis, 12 ou 18 semanas. Em cada grupo, 15 animais foram submetidos à derivação do trânsito por colostomia proximal no cólon esquerdo e fístula mucosa distal e cinco apenas à laparotomia (controle). Os cólons com e sem trânsito fecal foram removidos, processados, submetidos a cortes histológicos corados pela hematoxilina-eosina. A altura das criptas colônicas e o número de células caliciformes foram mensurados por morfometria computadorizada. Foram utilizados os testes t de Student e Kruskal-Wallis para comparação e análise de variância, estabelecendo-se nível de significância de 5% (p<0,05). RESULTADOS: A altura das criptas diminui nos segmentos sem trânsito fecal (p=0,0001), reduzindo entre seis e 12 semanas de exclusão (p=0,0003), estabilizando-se após este período. O número de células caliciformes nas criptas é menor nos segmentos sem trânsito após 12 e 18 semanas (p=0,0001), porém aumenta com o decorrer do tempo de exclusão (p=0,04) CONCLUSÃO: A exclusão do trânsito intestinal diminui a espessura das criptas colônicas e o número de células caliciformes nos segmentos sem trânsito. Existe aumento do número de células caliciformes com o decorrer do tempo de exclusão.

OBJECTIVE: To measure the thickness of the crypts and quantify the number of goblet cells of the colonic mucosa with and without intestinal transit, relating them to exclusion time. METHODS: Sixty Wistar rats were divided into three groups of 20 animals each according to the time of the final operation for the removal of the colon, in six, 12 or 18 weeks. In each group 15 animals underwent colonic exclusion by left colon proximal colostomy and distal mucous fistula, and five underwent only laparotomy (control). The colons with and without fecal stream were removed, processed and submitted to histological sections stained with hematoxylin-eosin. The height of the colonic crypts and the number of goblet cells were measured by computerized morphometry. We used the Student t test and Kruskal-Wallis test for comparison and analysis of variance, using a significance level of 5% (p <0.05). RESULTS: The height of the crypts decreased in segments without fecal stream (p =0.0001), reducing from six to 12 weeks of exclusion (p = 0.0003), stabilizing thereafter. The number of goblet cells in the crypts was smaller in segments without transit after 12 and 18 weeks (p = 0.0001), but increased as the time of exclusion progressed (p = 0.04) CONCLUSION: The exclusion of intestinal transit decreases the thickness of the colonic crypts and the number of goblet cells in the segments without transit. There is an increased number of goblet cells in the course of time exclusion.

Prognostic impact of the lymph node metastatic ratio on 5-year survival of patients with rectal cancer not submitted to preoperative chemoradiation

Lymph node metastases are a major prognostic factor in colorectal cancer. Inadequate lymph node resection is related to shorter survival. The lymph nodes ratio (LNR) has been used as a prognostic factor in patients with colon cancer. Few studies have evaluated the impact of LNR on the 5-year survival of patients with rectal cancer. OBJECTIVE: To evaluate the impact of LNR on the survival of patients with rectal cancer not submitted to preoperative chemoradiotherapy. METHODS: Ninety patients with rectal cancer excluding colon tumors, synchronous tumors, hereditary colorectal cancer and those undergoing preoperative chemoradiation. The patients were divided into three groups according t < 0.05). RESULTS: The 5-year survival was related to the Dukes classification, TNM, number of metastatic lymph nodes and LNR. A difference was observed in 5-year survival between the different classes of LNR. Patients classified as LNR-0 had a survival rate of 85%, while classes LNR-1 and LNR-2, 73 and 19%, respectively (p=0.0001). CONCLUSIONS: The results showed that the LNR has an impact on 5-year survival of patients with rectal cancer not submitted to neoadjuvant therapy. (AU)

Metástases linfonodais representam um dos principais fatores prognósticos no câncer colorretal. A ressecção linfonodal inadequada relaciona-se à menor sobrevida. A proporção entre linfonodos metastáticos (PLM) vem sendo utilizada como fator prognóstico em doentes com câncer de cólon. Poucos estudos avaliaram o impacto da PLM na sobrevida de doentes com câncer retal. OBJETIVO: Avaliar o impacto da PLM na sobrevida de doentes com câncer de reto não submetidos à quimioradioterapia pré-operatória. MÉTODOS: Foram incluídos 90 doentes com adenocarcinoma retal excluindo-se tumores de cólon, tumores sincrônicos, câncer colorretal hereditário e aqueles submetidos a tratamento radioquimioterápico pré-operatório. Os doentes foram divididos em três grupos segundo a PLM: PLM-0, sem linfonodos comprometidos; PLM-1, 1 a 20% dos linfonodos comprometidos; e PLM-2, mais de 21% dos linfonodos comprometidos. A identificação do ponto de corte da amostra selecionada foi obtida a partir da curva de características de operação do receptor (curva ROC). A sobrevida foi avaliada pelo teste de Kaplan-Meier, a diferença entre os grupos pelo teste de Cox-Mantel e a correlação entre as variáveis pelo teste de Pearson, adotando-se um nível de significância de 5% (p<0,05). RESULTADOS: A sobrevida em cinco anos relacionou-se à classificação de Dukes, TNM, número de linfonodos metastáticos e PLM. Houve diferença na sobrevida ao compararem-se as diferentes classes de PLM. Doentes classificados como PLM-0 apresentaram sobrevida de 85%, enquanto os pertencentes às classes PLM-1 e PLM-2, de 73 e 19%, respectivamente (p=0,0001). CONCLUSÃO: Os resultados encontrados mostraram que a PLM tem impacto na sobrevida de doentes com câncer de reto não submetidos à neoadjuvância. (AU)

Hippocampal plasticity in rats submitted to a gastric restrictive procedure.

Bariatric surgery has been the most effective therapeutic intervention for morbidly obese patients. However, recent evidence has shown that this procedure may cause serious neurological complications such as Wernicke encephalopathy, depression, and memory impairment. With this in mind, we conducted an experimental study to investigate whether weight-reduction surgery would promote morphological changes in the hippocampal formation, a brain region linked to cognitive and emotional processes. To do so, the present study evaluated the hippocampal expression of parvalbumin interneurons in rats submitted to a gastric restrictive procedure (experimental phytobezoar). Our results demonstrated that rats with gastric-reduced capacity presented a significant increase in the expression of the parvalbumin interneurons in the hippocampal CA1 and CA3 subfields. These data are the first experimental evidence that restrictive bariatric surgery may alter hippocampal cytoarchitecture.

Reduction in oxidative stress levels in the colonic mucosa without fecal stream after the application of enemas containing aqueous Ilex paraguariensis extract.

PURPOSE: To evaluate the antioxidant effects of enemas containing aqueous extract of Ilex paraguariensis, comparing segments with and without fecal stream and correlating the segments with the duration of intervention. METHODS: Twenty-six Wistar rats were subjected to a diversion of the fecal stream in the left colon by a proximal colostomy and distal mucosal fistula. The rats were distributed randomly into two experimental groups of 13 animals each based on the time of sacrifice after surgical procedure (two or four weeks). Each group was then divided into two experimental subgroups that received either second daily enemas containing 0.9% saline solution or aqueous extract of Ilex paraguariensis at 0.2g/100g. Colitis was diagnosed by histopathological analysis and the detection of oxidative tissue damage by measuring the levels of malondialdehyde. The Mann-Whitney test was used to compare the tissue levels of malondialdehyde between colon segments with and without fecal stream in each experimental group, and the Kruskal-Wallis test was used to verify the variance between the levels of oxidative stress according the duration of the irrigation; both tests determined significance at 5% (p<0.05). RESULTS: The levels of malondialdehyde in the animals subjected to intervention in the colon with saline with and without fecal stream after two and four weeks of irrigation were 0.05±0.006 and 0.06±0.006, and 0.05± 0.03 and 0.08 ±0.02, respectively. The malondialdehyde levels in the animals irrigated with Ilex paraguariensis with and without fecal stream after two and four weeks of irrigation were 0.010±0.002 and 0.02±0.004, and 0.03±0.007 and 0.04±0.01, respectively. After two and four weeks of intervention, the levels of malondialdehyde were lower in the animals irrigated with Ilex paraguariensis regardless of the time of irrigation (p=0.0001 and p=0.002, respectively). CONCLUSION: The daily rectal application of enemas containing aqueous extract of Ilex paraguariensis decreases oxidative tissue damage in the colon without fecal stream regardless of the time of irrigation.

Reduction in oxidative stress levels in the colonic mucosa without fecal stream after the application of enemas containing aqueous Ilex paraguariensis extract / Redução dos níveis de estresse oxidativo na mucosa cólica sem trânsito intestinal após aplicação de enemas contendo extrato aquoso de Ilex paraguariensis

PURPOSE: To evaluate the antioxidant effects of enemas containing aqueous extract of Ilex paraguariensis, comparing segments with and without fecal stream and correlating the segments with the duration of intervention. METHODS: Twenty-six Wistar rats were subjected to a diversion of the fecal stream in the left colon by a proximal colostomy and distal mucosal fistula. The rats were distributed randomly into two experimental groups of 13 animals each based on the time of sacrifice after surgical procedure (two or four weeks). Each group was then divided into two experimental subgroups that received either second daily enemas containing 0.9 percent saline solution or aqueous extract of Ilex paraguariensis at 0.2g/100g. Colitis was diagnosed by histopathological analysis and the detection of oxidative tissue damage by measuring the levels of malondialdehyde. The Mann-Whitney test was used to compare the tissue levels of malondialdehyde between colon segments with and without fecal stream in each experimental group, and the Kruskal-Wallis test was used to verify the variance between the levels of oxidative stress according the duration of the irrigation; both tests determined significance at 5 percent (p<0.05). RESULTS: The levels of malondialdehyde in the animals subjected to intervention in the colon with saline with and without fecal stream after two and four weeks of irrigation were 0.05±0.006 and 0.06±0.006, and 0.05± 0.03 and 0.08 ±0.02, respectively. The malondialdehyde levels in the animals irrigated with Ilex paraguariensis with and without fecal stream after two and four weeks of irrigation were 0.010±0.002 and 0.02±0.004, and 0.03±0.007 and 0.04±0.01, respectively. After two and four weeks of intervention, the levels of malondialdehyde were lower in the animals irrigated with Ilex paraguariensis regardless of the time of irrigation (p=0.0001 and p=0.002, respectively). CONCLUSION: The daily rectal application of enemas containing aqueous extract of Ilex paraguariensis decreases oxidative tissue damage in the colon without fecal stream regardless of the time of irrigation.

OBJETIVO: Avaliar os efeitos antioxidantes de clisteres contendo extrato aquoso de Ilex paraguariensis comparando segmentos com e sem trânsito fecal e correlacionando com o tempo de intervenção. MÉTODOS: Vinte e seis ratos Wistar machos foram submetidos à derivação do trânsito no cólon esquerdo pela confecção de colostomia proximal e fístula mucosa distal. Foram divididos de forma randomizada em dois grupos experimentais de 13 animais, de acordo com o sacrifício ser realizado após duas ou quatro semanas do procedimento cirúrgico. Cada grupo foi dividido em dois subgrupos experimentais segundo a aplicação diária ter sido realizada com enemas diários contendo solução fisiológica a 0,9 por cento ou extrato aquoso de Ilex paraguariensis na concentração de 0,2 g/100 ml. O diagnóstico de colite foi estabelecido por análise histológica e os níveis de estresse oxidativo pela dosagem tecidual de malondialdeído Para comparar os níveis de malondialdeído entre os cólons com e sem trânsito fecal em cada grupo experimental adotou-se o teste de Mann-Withney, e o teste de Kruskal-Wallis para verificar a variação dos níveis de estresse oxidativo segundo o tempo de irrigação, estabelecendo-se para ambos o nível de significância de 5 por cento (p<0,05). RESULTADOS: Os níveis de malondialdeído nos animais submetidos à intervenção com soro fisiológico nos cólons providos e desprovidos de trânsito fecal após duas e quatro semanas de irrigação foram de 0,05±0,006, 0,06±0,006 e 0,05±0,03, 0,08±0,02 respectivamente. Os níveis de malondialdeído nos animais irrigados com Ilex paraguariensis, nos cólons providos e desprovidos de trânsito, após duas e quatro semanas de irrigação foram de 0,010±0,002, 0,02±0,004 e 0,03±0,007, 0,04±0,01, respectivamente. Após duas e quatro semanas de intervenção os níveis de malondialdeído foram menores nos animais irrigados com Ilex paraguariensis, independente do tempo de irrigação (p=0,0001 e p=0,002, respectivamente). CONCLUSÃO: A aplicação diária de extrato aquoso de Ilex paraguariensis reduz os níveis de dano oxidativo tecidual nos segmentos cólicos sem trânsito fecal, independente do tempo de irrigação.

Cytometric evaluation of abdominal subcutaneous adipocytes after percutaneous CO2 infiltration.

OBJECTIVE: To evaluate the effects of carbon dioxide infusion to abdominal wall adipocytes. METHODS: Fifteen volunteers were subjected to sessions of CO2 infusion for three consecutive weeks (two sessions per week with intervals of two to three days between each). The volume of carbon dioxide infused per session, at points previously marked, was always calculated on the basis of surface area to be treated, with a fixed infused volume of 250 ml/100 cm² of treated surface. The infiltration points were marked respecting the limit of 2 cm equidistant between them. At each point 10 ml was injected per session, with a flow of 80 ml/min. Fragments were collected from subcutaneous tissue of the anterior abdominal wall before and after treatment. The number and histomorphological changes of adipocytes (mean diameter, perimeter, length, width and number of adipocytes per field of observation) were measured by computerized cytometry. The results were analyzed with paired Student t test, adopting a significance level of 5% (p <0.05). RESULTS: There was a significant reduction in the number of adipocytes in the abdominal wall, as well as the area, diameter, perimeter, length and width of the adipocytes, after the infusion of CO2 (p = 0.0001). CONCLUSION: The percutaneous infiltration of CO2 reduces the population of adipocytes of the anterior abdominal wall and modifies their morphology.

Avaliação citométrica dos adipócitos localizados no tecido subcutâneo da parede anterior do abdome após infiltração percutânea de CO2 / Cytometric evaluation of abdominal subcutaneous adipocytes after percutaneous CO2 infiltration

OBJETIVO: Avaliar os efeitos da infiltração de dióxido de carbono em adipócitos presentes na parede abdominal. MÉTODOS: Quinze voluntárias foram submetidas a sessões de infusão de CO2 durante três semanas consecutivas (duas sessões por semana com intervalos de dois a três dias entre cada sessão). O volume de gás carbônico infundido por sessão, em pontos previamente demarcados, foi sempre calculado com base na superfície da área a ser tratada, com volume infundido fixo de 250 mL/100cm² de superfície tratada. Os pontos de infiltração foram demarcados respeitando-se o limite eqüidistante 2cm entre eles. Em cada ponto se injetou 10mL, por sessão, com fluxo de 80mL/min. Foram colhidos fragmentos de tecido celular subcutâneo da parede abdominal anterior antes e após o tratamento. O número e as alterações histomorfológicas dos adipócitos (diâmetro médio, perímetro, comprimento, largura e número de adipócitos por campos de observação) foram mensurados por citometria computadorizada. Os resultados foram analisados com o teste t de Student pareado, adotando-se nível de significância de 5 por cento (p<0,05). RESULTADOS: Encontrou-se redução significativa no número de adipócitos da parede abdominal e na área, diâmetro, perímetro, comprimento e largura após o uso da hipercapnia (p=0,0001). CONCLUSÃO: A infiltração percutânea de CO2 reduz a população e modifica a morfologia dos adipócitos presentes na parede abdominal anterior.

OBJECTIVE: To evaluate the effects of carbon dioxide infusion to abdominal wall adipocytes. METHODS: Fifteen volunteers were subjected to sessions of CO2 infusion for three consecutive weeks (two sessions per week with intervals of two to three days between each). The volume of carbon dioxide infused per session, at points previously marked, was always calculated on the basis of surface area to be treated, with a fixed infused volume of 250 ml/100cm² of treated surface. The infiltration points were marked respecting the limit of 2cm equidistant between them. At each point 10 ml was injected per session, with a flow of 80ml/min. Fragments were collected from subcutaneous tissue of the anterior abdominal wall before and after treatment. The number and histomorphological changes of adipocytes (mean diameter, perimeter, length, width and number of adipocytes per field of observation) were measured by computerized cytometry. The results were analyzed with paired Student t test, adopting a significance level of 5 percent (p <0.05). RESULTS: There was a significant reduction in the number of adipocytes in the abdominal wall, as well as the area, diameter, perimeter, length and width of the adipocytes, after the infusion of CO2 (p = 0.0001). CONCLUSION: The percutaneous infiltration of CO2 reduces the population of adipocytes of the anterior abdominal wall and modifies their morphology.

Morphofunctional malignancy grading is a valuable prognostic factor for colorectal cancer.

CONTEXT: Novel strategies are needed to identify more efficient biomarkers to accurately diagnose prognose and improve the treatment outcome of colorectal cancer. OBJECTIVES: To analyze the functional and morphological features of colorectal cancer to identify the neoplastic patterns that affect patient survival. METHODS: Forty-five patients with colorectal cancer were followed for a minimum of 3 years. Blood levels of carcinoembryonic antigen (CEA) were measured by chemiluminescence and immunohistochemical analysis of tissue expression followed by computer-assisted image processing. Tumors were assigned to three morphofunctional classes. The morphofunctional classification was based on combination between histological differentiation and cell polarization. The functional characterization was based on the CEA cell polarization. The tissue polarization of CEA was classified in well-polarized, moderately polarized or nonpolarized cells. Morphofunctional staging was defined by the association between morphofunctional class (polarization and histological differentiation) and TNM by score given to each one classification. RESULTS: There was an association between increased CEA tissue expression and loss of histological differentiation (P = 0.01) or loss of polarization capacity (P = 0.03). There was a progressive increase in tissue CEA quantities in accordance with the proposed morphofunctional grading system. Plasma levels of CEA were increased in advanced tumor stages. Blood levels of CEA were increased in advanced morphofunctional stages (P = 0.001). There was a relationship between survival outcome and morphofunctional staging (P = 0.005). CONCLUSION: Morphofunctional staging is a valuable prognostic factor for colorectal cancer and it correlates with plasma CEA levels.

Morphofunctional malignancy grading is a valuable prognostic factor for colorectal cancer / Graduação morfofuncional de malignidade como valioso fator prognóstico para o câncer colorretal

CONTEXT: Novel strategies are needed to identify more efficient biomarkers to accurately diagnose prognose and improve the treatment outcome of colorectal cancer. OBJECTIVES: To analyze the functional and morphological features of colorectal cancer to identify the neoplastic patterns that affect patient survival. METHODS: Forty-five patients with colorectal cancer were followed for a minimum of 3 years. Blood levels of carcinoembryonic antigen (CEA) were measured by chemiluminescence and immunohistochemical analysis of tissue expression followed by computer-assisted image processing. Tumors were assigned to three morphofunctional classes. The morphofunctional classification was based on combination between histological differentiation and cell polarization. The functional characterization was based on the CEA cell polarization. The tissue polarization of CEA was classified in well-polarized, moderately polarized or nonpolarized cells. Morphofunctional staging was defined by the association between morphofunctional class (polarization and histological differentiation) and TNM by score given to each one classification. RESULTS: There was an association between increased CEA tissue expression and loss of histological differentiation (P = 0.01) or loss of polarization capacity (P = 0.03). There was a progressive increase in tissue CEA quantities in accordance with the proposed morphofunctional grading system. Plasma levels of CEA were increased in advanced tumor stages. Blood levels of CEA were increased in advanced morphofunctional stages (P = 0.001). There was a relationship between survival outcome and morphofunctional staging (P = 0.005). CONCLUSION: Morphofunctional staging is a valuable prognostic factor for colorectal cancer and it correlates with plasma CEA levels.

CONTEXTO: Novas estratégias são necessárias à identificação de marcadores que promovam precisão no diagnóstico, no prognóstico e melhorem o tratamento de pacientes com câncer colorretal. OBJETIVO: Analisar características funcionais e morfológicas do câncer colorretal para identificar padrões de neoplasia que modificam a sobrevida. MÉTODOS: Quarenta e cinco pacientes com adenocarcinoma colorretal foram seguidos por no mínimo 3 anos. Níveis séricos de antígeno carcinoembrionário (CEA) foram mensurados por quimioluminescência e a análise imunoistoquímica da expressão tecidual do antígeno por meio de processamento de imagem assistida por computador. Tumores foram divididos em três classes morfofuncionais. A classificação morfofuncional foi baseada na combinação entre grau histológico e polarização do CEA celular. A polarização do CEA foi classificada em bem polarizada, moderadamente polarizada e não-polarizada. O estádio morfofuncional foi definido pela associação entre as classes morfofuncionais (polarização e grau histopatológico) e pontuação dada a cada uma das classificações. RESULTADOS: Houve associação entre aumento de expressão de CEA tecidual e perda do grau de diferenciação (P = 0.01) ou perda da capacidade de polarização (P = 0.03). Houve aumento progressivo dos níveis de proteínas teciduais em acordo com o sistema de classificação morfofuncional proposto. Níveis plasmáticos de CEA estavam aumentados com a progressão dos estádios tumorais (P = 0.001). Houve relação entre sobrevida e estádio morfofuncional (P = 0.005). CONCLUSÃO: Estádio morfofuncional é um valioso fator prognóstico para o câncer colorretal e se correlaciona com níveis séricos de CEA.

Quantification by computerized morphometry of tissue levels of sulfomucins and sialomucins in diversion colitis in rats / Quantificação por morfometria computadorizada dos níveis teciduais de sulfomucinas e sialomucinas na colite de exclusão em ratos

PURPOSE: To quantify the intensity of sulfomucin and sialomucin expression in the colon mucosa, by means of computer-assisted image processing, comparing segments with and without fecal stream and correlating with the duration of fecal transit exclusion. METHODS: Forty-five Wistar rats were subjected to diversion of the fecal stream in the left colon by means of constructing a proximal colostomy and distal mucosal fistula. They were distributed randomly into three experimental groups of 15 animals, of which 10 were subjected to colon diversion (experimental subgroup) and five were only subjected to laparotomy, without colon diversion (control subgroup). The three experimental groups were formed according to the sacrifice date, which was to be performed six weeks after the surgical procedure (Group A), 12 weeks (Group B) and 18 weeks (Group C). The sulfomucin and sialomucin expression in the colon mucosa was evaluated using the histochemical technique of high iron diamine-alcian blue (HID-AB). The tissue expression was quantified for each animal, in the segments with and without fecal stream, at a location where there were four complete contiguous crypts in two random fields, with the aid of the computer-assisted image analysis software. The final value was taken to be the mean reading from the two fields selected, in the segments with and without fecal stream. To compare the expressions of the two mucin subtypes in the segments with and without fecal stream, the paired Student t test was used. To analyze variance according to duration of exclusion, ANOVA with the Newman-Keuls post-test was used, setting the significance level at 5 percent (p<0.05). RESULTS: There were significant reductions in tissue sulfomucin and sialomucin content in the colon without fecal stream, independent of the duration of exclusion considered. There was increased tissue sulfomucin content and decreased tissue sialomucin in the segments without fecal stream, ...

OBJETIVO: Quantificar, por meio de processamento de imagem assistida por computador, a intensidade de expressão de sulfomucinas e sialomucinas na mucosa cólica comparando segmentos com e sem trânsito e relacionando-a ao tempo de exclusão de trânsito fecal. MÉTODOS: Quarenta e cinco ratos Wistar machos foram submetidos à derivação do trânsito no cólon esquerdo pela confecção de colostomia proximal e fístula mucosa distal. Foram divididos de forma randomizada em três grupos experimentais de 15 animais, nos quais 10 foram submetidos à derivação do trânsito cólico (subgrupo experimental) e cinco somente a laparotomia exploradora sem desvio do trânsito fecal (subgrupo controle). Os três grupos experimentais foram formados segundo o sacrifício ter sido realizado em seis (grupo A), 12 (Grupo B) e 18 semanas (Grupo C). A avaliação da expressão de sulfomucinas e sialomucinas na mucosa cólica foi realizada pela técnica histoquímica da diamina de ferro alto alcian-blue (HID-AB). A quantificação da expressão tecidual foi determinada, para cada animal, nos segmentos com e sem trânsito, em local onde existiam quatro criptas contíguas e íntegras em dois campos aleatórios com auxílio de programa de análise de imagem assistida por computador. Adotou-se como valor final a média das leituras dos dois campos selecionados, nos segmentos providos e desprovidos de trânsito fecal. Na comparação entre a expressão dos dois subtipos de mucinas nos segmentos com e sem trânsito fecal utilizou-se o teste t de Student pareado. Para análise de variância segundo o tempo de exclusão utilizou-se o teste de ANOVA com o pós-teste de Newmann-Keuls, estabelecendo-se nível de significância de 5 por cento (p<0,05). RESULTADOS: Houve redução significante no conteúdo tecidual de sulfomucinas e sialomucinas no cólon desprovido de trânsito fecal, independente do tempo de exclusão considerado. Houve aumento no conteúdo tecidual de sulfomucinas e diminuição de sialomucinas nos segmentos ...

Quantification by computerized morphometry of tissue levels of sulfomucins and sialomucins in diversion colitis in rats.

PURPOSE: To quantify the intensity of sulfomucin and sialomucin expression in the colon mucosa, by means of computer-assisted image processing, comparing segments with and without fecal stream and correlating with the duration of fecal transit exclusion. METHODS: Forty-five Wistar rats were subjected to diversion of the fecal stream in the left colon by means of constructing a proximal colostomy and distal mucosal fistula. They were distributed randomly into three experimental groups of 15 animals, of which 10 were subjected to colon diversion (experimental subgroup) and five were only subjected to laparotomy, without colon diversion (control subgroup). The three experimental groups were formed according to the sacrifice date, which was to be performed six weeks after the surgical procedure (Group A), 12 weeks (Group B) and 18 weeks (Group C). The sulfomucin and sialomucin expression in the colon mucosa was evaluated using the histochemical technique of high iron diamine-alcian blue (HID-AB). The tissue expression was quantified for each animal, in the segments with and without fecal stream, at a location where there were four complete contiguous crypts in two random fields, with the aid of the computer-assisted image analysis software. The final value was taken to be the mean reading from the two fields selected, in the segments with and without fecal stream. To compare the expressions of the two mucin subtypes in the segments with and without fecal stream, the paired Student t test was used. To analyze variance according to duration of exclusion, ANOVA with the Newman-Keuls post-test was used, setting the significance level at 5% (p<0.05). RESULTS: There were significant reductions in tissue sulfomucin and sialomucin content in the colon without fecal stream, independent of the duration of exclusion considered. There was increased tissue sulfomucin content and decreased tissue sialomucin in the segments without fecal stream, with increasing duration of exclusion. CONCLUSIONS: Diversion of the fecal transit decreased the tissue sulfomucin and sialomucin content in the segments without fecal stream. Notwithstanding the reduction in the levels of both subtypes of acid mucin in the segments without fecal stream, there was increased tissue sulfomucin content and decreased tissue sialomucin with increasing duration of intestinal diversion.

Manometria anorretal no divertículo de reto / Anorectal manometry in rectal diverticulum

Divertículo localizado no reto é um achado excepcional, estimando-se que existam pouco mais de 50 casos publicados. A doença apresenta aspectos controversos, quanto a ser de origem congênita ou adquirida. Recentemente, distúrbios defecação vêm sendo relacionado à maior possibilidade do desenvolvimento da doença. Contudo, até a presente data, as alterações manométricas em portadores de divertículo do reto ainda não foram estudadas. OBJETIVO: O objetivo do presente estudo é demonstrar os resultados de estudo eletromanométrico anorretal, realizado em dois doentes portadores divertículo do reto. CASUÍSTICA E MÉTODO: Um homem e uma mulher, com 56 e 58 anos, respectivamente, foram submetidos à colonoscopia, enema opaco, ultrassonografia endorretal e ressonância magnética da pelve, para confirmação e documentação diagnóstica de divertículo localizado no reto. Os enfermos foram submetidos à eletromanometria anorretal com cateter de oito canais sob perfusão de água a 0,3 ml/min/canal, através de sistema de infusão capilar pneumático e hidráulico. RESULTADOS: O resultado dos exames em ambos os doentes mostrou perfil pressórico esfincteriano normal, tanto em repouso, como em contração voluntária máxima, não se encontrando assimetrias esfincterianas. O reflexo reto-anal inibitório encontrava-se presente e dentro de valores normais, assim como a sensibilidade e complacência retal. A análise pelo vetor volume não mostrou alterações significativas concluindo-se por estudo manométrico ano-retal normal. CONCLUSÃO: O estudo manométrico anorretal não demonstrou existência de distúrbios pressóricos nos esfíncteres anorretais reforçando a possibilidade de que o divertículo de reto possa ter origem congênita, desenvolvendo-se em pontos onde exista maior fraqueza da parede retal.

Diverticulum located in the rectum is an exceptional find with fewer than 50 cases published. The etiology of the diverticulum of the rectum still has controversial aspects regarding its origin is congenital or acquired. Recently, alterations of the evacuative dynamic had been associated with the development of the disease. Nevertheless, until the present date, the manometric alterations in patients with diverticulum of the rectum were not studied. OBJECTIVE: The aim of the present study is to demonstrate the results of electromanometric study carried out in a patient with isolated diverticulum of the rectum. CASE REPORT: A man and a woman, 56 and 58 years-old, respectively, were submitted to colonoscopy, barium enema, endorectal ultrasound and magnetic resonance of the pelvis to diagnose and document the presence of diverticulum located in the rectum. The patients were submitted to manometry using catheter with eight-channel perfusion under water at 0.3 ml/min/channel, through pneumatic and hydraulic capillary infusion system. The anorectal manometry showed normally sphincter pressure profile both in rest and squeezes pressure state, and did not find sphincter asymmetries. The anorectal inhibition reflex was present with normal values and the sensibility and rectal complaisance were between the normal limits. The vector volume analysis did not show any significant alterations and it was concluded by normal manometric anorectal study. CONCLUSION: The anorectal manometry study did not demonstrate the existence of pressure disturbances in anorectal sphincter, enhancing the possibility that the rectal diverticulum can be congenital, developing at points where there is major weakness of the rectal wall.

Is peritoneal reflection the best anatomical repair landmark in experimental colorectal surgery on rats?

PURPOSE: To validate Peyer's patch as an anatomical repair landmark for colorectal surgery in rats and to measure the collagen content in segments of the colon containing or not containing Peyer's patch. METHODS: The distance between Peyer's patch and the peritoneal reflection was measured in forty-five Wistar rats. The colon and rectum were resected for quantification of collagen content by means of computer-assisted image analysis in regions of the colon with and without Peyer's patch. RESULTS: There was great variation in the distance between Peyer's patch and the peritoneal reflection when the male and female rats were considered as a single group (p=0.04). Comparison between the genders showed that the distance between the patch and the peritoneal reflection was greater in female than in male rats (p=0.001). The colonic segment containing Peyer's patch was observed to have lower tissue collagen content than the segment in which this structure was not present (p=0.02). CONCLUSION: Peyer's patch can be indicated as an anatomical repair landmark, and there is a need to study the healing of colorectal anastomoses in rats based on differing quantities of tissue collagen existing in the colonic wall with or without this structure.

Is peritoneal reflection the best anatomical repair landmark in experimental colorectal surgery on rats? / A reflexão peritoneal é o melhor reparo anatômico na cirurgia experimental colorretal em ratos?

PURPOSE: To validate Peyer's patch as an anatomical repair landmark for colorectal surgery in rats and to measure the collagen content in segments of the colon containing or not containing Peyer's patch. METHODS: The distance between Peyer's patch and the peritoneal reflection was measured in forty-five Wistar rats. The colon and rectum were resected for quantification of collagen content by means of computer-assisted image analysis in regions of the colon with and without Peyer's patch. RESULTS: There was great variation in the distance between Peyer's patch and the peritoneal reflection when the male and female rats were considered as a single group (p=0.04). Comparison between the genders showed that the distance between the patch and the peritoneal reflection was greater in female than in male rats (p=0.001). The colonic segment containing Peyer's patch was observed to have lower tissue collagen content than the segment in which this structure was not present (p=0.02). CONCLUSION: Peyer's patch can be indicated as an anatomical repair landmark, and there is a need to study the healing of colorectal anastomoses in rats based on differing quantities of tissue collagen existing in the colonic wall with or without this structure.

OBJETIVO: Validar a placa de Peyer como reparo anatômico para a cirurgia colorretal em ratos e mensurar a quantidade de colágeno em segmentos da parede cólica que contém ou não a placa de Peyer. MÉTODOS: Foi aferida a distância entre a placa de Peyer e a reflexão peritoneal em 45 ratos Wistar. O cólon e o reto foram ressecados, para a quantificação do colágeno, por meio de análise de imagem assistida por computador, em regiões do cólon que continham ou não a placa de Peyer. RESULTADOS: Existe grande variação entre a distância da placa de Peyer e a reflexão peritoneal quando se consideraram os animais de ambos os gêneros como grupo único (p= 0.04), sendo a distância entre a placa e a reflexão peritoneal maior entre as fêmeas (p=0.001). Constatou-se que o segmento cólico que contém a placa de Peyer apresenta conteúdo menor de colágeno quando comparado ao segmento onde a estrutura não estava presente (p=0.02). CONCLUSÃO: A placa de Peyer pode ser indicada como reparo anatômico e no estudo da cicatrização de anastomoses colorretais em ratos, baseado nas diferentes quantidades de colágeno tecidual existente na parede cólica que contém ou não esta estrutura.

Avaliação da expressão tecidual do gene de reparo MLH1 e dos níveis de dano oxidativo ao DNA em doentes com câncer colorretal / Evaluation of expression of mismatch repair gene MLH1 and levels of oxidative DNA damage in normal and neoplastic tissues of patients with colorectal cancer

O dano oxidativo ao DNA provocado por radicais livres de oxigênio representa um dos principais mecanismos responsáveis pelas etapas iniciais da carcinogênese colorretal. O estresse oxidativo ocasiona erros de pareamento de bases possibilitando o aparecimento de mutações em genes controladores do ciclo celular. As células possuem um sistema de defesa representado pelos genes de reparo do DNA que corrigindo os erros de pareamento impedem o desenvolvimento de mutações. Poucos estudos avaliaram a relação entre dano oxidativo ao DNA e a expressão tecidual do gene de reparo MLH1. OBJETIVO: O objetivo do presente estudo foi avaliar os níveis de estresse oxidativo ao DNA e a expressão tecidual do gene de reparo MLH1 nas células da mucosa cólica normal e neoplásica de doentes com câncer colorretal. MATERIAL E MÉTODO: Foram estudados 44 doentes com diagnóstico de adenocarcinoma colorretal. Foram excluídos os doentes com câncer colorretal hereditário, portadores de câncer relacionado às doenças inflamatórias intestinais e os submetidos à radioquimioterapia neoadjuvante. Para a avaliação dos níveis de dano oxidativo ao DNA utilizou-se a técnica da eletroforese alcalina em gel de célula isolada (ensaio do cometa) avaliando 100 células obtidas dos tecidos normal e neoplásico. Para a avaliação da expressão do gene MLH1 utilizou-se a técnica de reação de polimerase em cadeia em tempo real (RT-PCR) com primer especificamente desenhados para amplificação do gene. A comparação dos resultados encontrados para os níveis de estresse oxidativo ao DNA, e expressão do gene MLH1 nos tecidos normais e neoplásicos foi feito pelo teste t de Student, adotando-se nível de significância de 5 por cento (p<0,05). RESULTADOS: Os níveis de dano oxidativo ao DNA no tecido neoplásico foram significativamente mais elevados quando comparados ao tecido normal (p=0,0001). A expressão tecidual do gene MLH1 no tecido neoplásico foi significativamente menor quando comparado ao tecido...

The oxidative DNA damage caused by oxygen free radicals is one of the most important mechanisms responsible for the initial steps of colorectal carcinogenesis. The oxidative stress can cause errors in the pairing of nitrogenous bases that form the DNA, allowing mutations in controlling genes of the cell cycle. The cells have a defense system represented by the DNA mismatch repair genes that correct the errors of matching prevent the development of DNA mutations. Few studies have evaluated the relationship between oxidative DNA damage and the tissue expression of mismatch repair genes. AIM: The aim of the present study was evaluate the levels of oxidative DNA and the tissue expression of MLH1 mismatch repair gene in the cells of normal and neoplastic colonic mucosa of patients with colorectal cancer. MATERIAL AND METHODS: Were studied 44 patients with diagnosis of colorectal adenocarcinoma. Were excluded patients with hereditary colorectal cancer, with colorectal cancer associate with inflammatory bowel diseases and those undergoing neoadjuvant radioquimiotherapy. To evaluate the levels of oxidative DNA damage was used the single cell gel electrophoresis (comet assay) evaluating 100 cells obtained from normal and neoplastic tissues. For the evaluation of the tissue expression of MLH1 gene was employed the technique of polymerase chain reaction in real time (RT-PCR) with primer specifically designed for MLH1 gene. The comparison among the levels of DNA oxidative stress and expression of MLH1 mismatch repair gene in normal and neoplastic tissues was done by Student t test adopting a significance level of 5 percent (p< 0.05). RESULTS: The levels of oxidative DNA damage in tumor tissue were significantly higher when compared to the level of the normal tissue (p = 0.0001). The tissue expression of MLH1 mismatch repair gene in tumor tissue was significantly lower when compared to normal tissue (p=0.02). CONCLUSION: The mismatch repair gene MLH1...

Proporção de linfonodos metastáticos como variável independente de prognóstico no câncer colorretal / Metastatic lymph node ratio as an independent prognostic variable in colorectal cancer

No câncer colorretal, o comprometimento linfonodal é um dos fatores prognósticos mais importantes. Objetivo: Determinar o valor prognóstico independentemente da relação entre linfonodos comprometidos e examinados em doentes com câncer colorretal. MÉTODO: Foram estudados 113 doentes (62 mulheres) portadores de câncer do cólon e reto superior. Quinze pertenciam ao estádio I da classificação TNM, 44 ao estádio II, 42 ao estádio III e 24 ao estádio IV. O índice de linfonodos comprometidos foi determinado pela relação entre número total de linfonodos comprometidos e examinados. Os doentes foram divididos segundo a proporção de linfonodos comprometidos em três grupos: ILC-0: sem comprometimento linfonodal; ILC-1: com comprometimento de até 20 por cento dos linfonodos examinados; e ILC-2: com comprometimento neoplásico em 21 por cento ou mais dos linfonodos examinados. As relações entre proporção de linfonodos comprometidos, número de linfonodos ressecados e número de linfonodos comprometidos foram determinadas. Excluíram-se doentes menores de 18 anos, submetidos à neoadjuvância e aqueles onde foram ressecados menos de 12 linfonodos. As variáveis anátomo-clínicas foram analisadas por estatística descritiva. A correlação entre as variáveis foi avaliada pelo teste de Spearman. A sobrevida foi determinada pelos testes de Kaplan-Meier, Log-rank e a análise multivariada pelo modelo de Cox estabelecendo nível de significância de 5 por cento. RESULTADOS: Houve diferença significante na sobrevida de cinco anos em doentes classificados nos diferentes grupos de ILC (p=0,009). Houve relação entre o número total de linfonodos comprometidos e o ILC (p=0,00001, com rs=0,977). Doentes do grupo ILC-0 apresentaram sobrevida global em cinco anos maior que 80 por cento, enquanto os pertencentes aos grupos ILC-1 e ILC-2 a sobrevida global foi menor que 60 por cento e 40 por cento, respectivamente. O comprometimento linfonodal isolado apresentava-se relacionado...

In patients with colorectal cancer, the compromising of the lymph node is one of the most important prognostic factors. Aim: The aim of this study is to determine the prognostic independent value of relation between compromised and examined lymph nodes, (LNR) in patients with colorectal cancer. METHOD: One hundred and thirteen 113 patients (62 women) with colon ad upper rectum cancer were studied. Fifteen patients were staged to the stadium I of the TNM classification, 44 to II, 42 to III and 24 to the stadium IV. The lymph node ratio rate was determined by the relation between total number of compromised and examined lymph nodes. The patients were divided in three groups according to the proportion of compromised lymph nodes: LNR-0: when there was no lymph node involvement; LNR-1: when there was compromising of up to 20 percent of the examined lymph nodes and LNR-2: when there was compromising in 21 percent or more of the examined nodes. The relations between lymph node ratio, number of removed lymph nodes and number of compromising lymph nodes by cancer were determined. Patients younger than 18 years old who were submitted to neoadjuvant quimioradiation protocol were excluded as well as the ones who had less than 12 lymph nodes removed. The clinical and histopathological variables were analyzed through descriptive statistic. The correlation between the variables was valued by Spearman correlation test. The five years survival was determined by the tests of Kaplan-Meier, Log-rank and the multivariate analysis of the prognostic variables by the Cox model establishing level of significance of 5 percent (p=0.05). RESULTS: There was significant difference in the five years overall survival in patients classified in the different groups of LNR (p=0.009). Patients of the group LNR-0 presented five years overall survival bigger than 80 percent, while in those classified to the groups LNR-1 and LNR-2, the five years overall survival was less...

Influência da irrigação de soluções nutricionais no colo excluso de trânsito intestinal: estudo experimental em ratos / Influence of irrigation of nutritional solutions in the colon excluded of fecal stream: experimental study in rats

A colite por exclusão é descrita como processo inflamatório que ocorre nos segmentos colorretais desprovidos do trânsito fecal. A deficiência dos ácidos graxos de cadeia curta vem sendo considerada como principal fator causal. OBJETIVO: O objetivo do presente estudo foi avaliar, em modelo experimental de colite de exclusão, a importância da irrigação do segmento desprovido de trânsito com soluções nutricionais na prevenção e tratamento do processo inflamatório. MÉTODO: Foram utilizados trinta ratos Wistar, machos, com peso inicial variando entre 350 e 500 gramas, submetidos à derivação do trânsito intestinal através da realização de colostomia proximal e fístula mucosa distal. Os animais foram divididos em três grupos de 10 animais segundo a irrigação do segmento excluso de trânsito ter sido realizada, empregando-se: Grupo SF: solução fisiológica a 0,9 por cento; Grupo GH: solução de glicose a 50 por cento; e Grupo AG: solução de ácidos graxos de cadeia curta. Em todos os animais, a irrigação do colo excluso foi realizada em intervalos de quatro dias sendo sacrificados sempre no 21º pós-operatório. Os fragmentos removidos dos segmentos intestinais foram corados pelas técnicas da hematoxilina-eosina e tricrômio de Masson. As variáveis histológicas estudadas foram: espessura da túnica mucosa, congestão vascular; infiltrado inflamatório e a deposição de colágeno. Os resultados encontrados foram submetidos a estudo estatístico considerando nível de significância de 5 por cento (p< 0,05). RESULTADOS: Verificou-se que no grupo onde se irrigou o cólon excluso com solução de ácidos graxos de cadeia curta houve menor congestão vascular, menor infiltrado inflamatório e menor deposição de colágeno quando comparado aos demais grupos experimentais. CONCLUSÃO: Os resultados do presente trabalho mostram que a irrigação de segmentos desprovidos de trânsito fecal com ácidos graxos de cadeia curta, encontra-se relacionada à melhora no processo inflamatório...

Diversion colitis is described as an inflammatory process that occurs in the colorectal segments excluded of the fecal stream. The deficiency of the short-chain fatty acids inside of intestinal lumen was considered the main etiologic factor. AIM: The aim of the present study was valued, in experimental model of diversion colitis, the importance of the irrigation of the segment without fecal stream with nutritional solutions in prevention and treatment of the colic mucosa inflammation. METHOD: There were used thirty Wistar males rats, with initial weight varying between 350 and 500 grammas, submitted to the derivation of the fecal stream through proximal colostomy and distal mucous fistula. The animals were divided in three groups with 10 animals according to the irrigation of the excluded colic segment had been carried out with: SF Group: 0.9 percent physiologic solution; GH Group: hypertonic glucose solution of 50 percent and AG Group: solution of short-chain fatty acids. In all the animals, the irrigation of the excluded colon was carried each four days and the animals were sacrificed ever in 21st post-operative day. The removed fragments of the intestinal segments were colored by the techniques of hematoxylin-eosin and Masson trichromium. The histological variables studied were: thickness of the mucous layer, vascular congestion; inflammatory infiltrate and the level of tissue collagen. The considered results were subjected to statistical study considering signification level of 5 percent (p <0.05). RESULTS: The obtained results showed that in the group where the excluded colon was irrigated with solution of short-chain fatty acids there was less vascular congestion, less inflammatory infiltrated and less collagen deposition when compared to another experimental groups. CONCLUSION: The results of the present study indicate that in the group receiving the short-chain fatty acid solution there was a decrease in the inflammatory infiltrate...