Heparin modulates the expression of genes encoding pro and anti-apoptotic proteins in endothelial cells exposed to intestinal ischemia and reperfusion in rats

PURPOSE: To investigate if expression of genes encoding pro and anti-apoptotic proteins in the rat enteric endothelial cells stimulated by intestinal ischemia followed by reperfusion (IR) can be modified by treatment with heparin (HP).METHODS: Eighteen adult Wistar rats were divided in three groups: sham group submitted to laparotomy only (SG), ischemia followed by reperfusion group (IRG); ischemia followed by reperfusion plus pretreatment with HP 100 mg.kg-1 (IRG+HP). Ischemia was performed by clamping of the superior mesenteric artery. After 60 min of ischemia, metal clamps were removed for reperfusion for 120 min. Gene expression of encoding pro (Casp1, Casp6, Casp3, Cflar, Fas and Pgl) and anti-apoptotic (Bcl2, Bcl2l1 and Naip2) proteins in rat enteric endothelial cells was evaluated by PCR microarray method.RESULTS: Compared to rat endothelial cells of SG, the expression of pro-apoptotic genes was up-regulated in IRG while anti-apoptotic genes were down-regulated. In contrast, the expression of anti-apoptotic genes in IRG+HP was up-regulated while pro-apoptotic genes was down-regulated compared to SG.CONCLUSION: The attenuation by heparin of intestinal ischemia-reperfusion previously demonstrated in rodents could be related with ability of this drug to stimulate and reduce gene expression of encoding anti and pro-apoptotic proteins, respectively.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Federal University of São Paulo Department of SurgeryUNIFESP Pharmacology DepartmentUNIFESP Gynecology DepartmentUniversidade Federal de São Paulo (UNIFESP) Department of BiochemistryUniversidade Federal de São Paulo (UNIFESP) Department of SurgeryUNIFESP, Department of SurgeryUNIFESP, Pharmacology DepartmentUNIFESP, Gynecology DepartmentUNIFESP, Department of BiochemistryUNIFESP, Department of SurgerySciEL

Ischemic preconditioning and the gene expression of enteric endothelial cell biology of rats submitted to intestinal ischemia and reperfusion

PURPOSE: To investigate the effects of ischemic preconditioning (IPC) on the expression of pro and anti-apoptotic genes in rat endothelial cells undergoing enteric ischemia (I) and reperfusion (R). METHODS: Thirty rats underwent clamping of the superior mesenteric vessels. Sham group (GS) laparotomy only; Ischemia (GI): intestinal ischemia (60 min); Ischemia and Reperfusion (GIR): ischemia (60 min) and reperfusion (120 min); Ischemia and intestinal ischemic preconditioning (GI + IPC) : 5 minutes of ischemia followed by 10 min of reperfusion before sustained ischemia (60 min) ischemia and reperfusion and IPC (GIR + IPC): 5 min ischemia followed by 10 min of reperfusion before sustained ischemia (60min) and reperfusion (120 min). Rat Endothelial Cell Biology (PCR array) to determine the expression of genes related to endothelial cell biology. RESULTS: Gene expression of pro-apoptotic markers (Casp1, Casp6, Cflar, Fas, and Pgl) was down regulated in GI+IPC and in GIR + IPC. In contrast, the expression of anti-apoptotic genes (Bcl2 and Naip2), was up-regulated in GI + IPC and in GIR + IPC. CONCLUSION: Ischemic preconditioning may protect against cell death caused by ischemia and reperfusion.UNIFESPUNIFESPSciEL

Evaluation of the Intensity and the Extension of Thermal Alterations Produced by High Frequency Surgery in the Uterine Cervix

Purpose: to evaluate the incidence of thermal damage to the specimens excised through large loop excision of the transformation zone (LLETZ) and to determine qualitatively and quantitatively the thermal injury to the ectocervical and endocervical epithelia as well as the influence of the menstrual phase on such process. Methods: we performed a prospective study of 100 patients with high-grade squamous intraepithelial lesions (HGSIL). Thermal damage was subdivided into three grades according to Messing et al¹. Results: thermal injury occurred in all the cases, however, through statistical analysis we found that in 91% of the cases it was insignificant, thus leading to a precise histological evaluation, hence to measure the thermal injury was unnecessary. The grade and extent of thermal damage in excised specimens using LLETZ had no relation to the menstrual phase. The extension of thermal tissue alteration in the endocervical epithelia was 271,6 mu while the extension in ectocervical epithelia was 254,8 mu, showing that the extension of thermal damage is significantly higher in endocervical epithelia. Of the one hundred patients, 80 were in menacme and 20 in menopause correlating the grade and extension of thermal damage with the menstrual state. Conclusion: there was no significant difference in both qualitative and quantitative evaluations. There is no need to measure the thermal damage.Objetivo: estudar a ocorrência de dano térmico tissular nos espécimes excisados por cirurgia de alta freqüência e avaliar qualitativa e quantitativamente o dano térmico sobre o epitélio ectocervical e o endocervical e a provável influência do estado menstrual sobre sua gênese. Método: estudo prospectivo de 100 pacientes com lesão cervical intra-epitelial de alto grau. O dano térmico encontrado foi subdividido em três graus, leve, moderado e grave, de acordo com os critérios de Messing et al.¹ Resultado: a análise estatística permitiu avaliar que o dano térmico ocorreu em todos os casos, porém em 91% das vezes ele foi insignificante, permitindo a correta avaliação histopatológica. A extensão da alteração térmica tecidual sobre o epitélio endocervical foi de 271,6 mi, ao passo que a extensão no epitélio ectocervical foi de 254,8 mi, mostrando desta forma que a extensão do dano térmico é significantemente maior no epitélio endocervical. Das 100 pacientes avaliadas, 80 estavam no menacme e 20 na menopausa. O grau e a extensão do dano térmico tissular não variaram com o estado menstrual. Conclusão: não se observou qualquer diferença significativa na avaliação tanto do ponto de vista qualitativo quanto quantitativo. Por outro lado, evidenciou-se que na prática cotidiana não há necessidade de se fazer a mensuração do dano térmico.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de GinecologiaUNIFESP, EPM, Depto. de GinecologiaSciEL

Value of the hygroscopic dilator for visualization of the endocervical canal in conization by loop electrosurgical excision procedure

OBJECTIVE: to evaluate the effectiveness of the hygroscopic dilator in the colposcopic examination of the endocervical canal in patients with high-grade lesion in the cytopathology and unsatisfactory colposcopy. METHODS: prospective study, including 62 patients with unsatisfactory colposcopic examination and cytology compatible with high-grade intraepithelial lesion. The patients were submitted to dilation of the endocervical canal by means of a hygroscopic dilator. After dilation, the new colposcopic findings were recorded, and then conization was made through loop electrosurgical excision procedure. The incidence of neoplasic involvement of surgical margins was compared between patients with examinations modified toward satisfactory results and those that remained with unsatisfactory colposcopy. In order to compare the incidence of involved margins and the incidence of residual disease, two retrospective control-groups were used: the GinSat group (n = 35): patients with unsatisfactory colposcopy; GSat group (n = 38): patients with satisfactory colposcopy and endocervical atypy. RESULTS: 80.6% of the cases presented satisfactory colposcopic vision after dilation. 80.4% of those presented disease-free resection margins. The incidence of disease-free resection margins in patients with persistent unsatisfactory colposcopy after dilation was 36.3%. Affected surgical margins occurred in 28% of the group that had undergone dilation, 28.5% of the cases in GinSat group, and 31.5% in the Gsat group. Follow-up showed the incidence of residual disease in 7.5% of the patients under dilation, 28.5% in the GinSat group and 28.9% in the GSat group. CONCLUSION: the use of hygroscopic dilation improved visualization of lesions of difficult access to the colposcopic examination, thus permitting reduction in the percentage of residual neoplasic disease in patients with unsatisfactory colposcopy treated with loop electrosurgical excision procedure.OBJETIVO: avaliar a eficácia do dilatador higroscópico para auxiliar o exame colposcópico do canal endocervical, em pacientes com lesão de alto grau e colposcopia insatisfatória. MÉTODOS: estudo prospectivo no qual foram incluídas 62 pacientes com exames colposcópicos insatisfatórios e citologia sugestiva de lesão de alto grau. Todas foram submetidas à dilatação do canal por meio de dilatador higroscópico. Após a dilatação os novos achados colposcópicos foram registrados e procedeu-se à conização por cirurgia de alta freqüência (CAF). Comparamos a incidência de comprometimento neoplásico das margens cirúrgicas operatórias entre as pacientes que modificaram o exame para satisfatório e aquelas que persistiram insatisfatórios. Para comparação entre a incidência de margens comprometidas e a incidência de doença residual, foram utilizados dois grupos controle retrospectivos: grupo GinSat (n=35): pacientes com colposcopias insatisfatórias; grupo GSat (n=38): pacientes com colposcopias satisfatórias e atipia de localização endocervical. RESULTADOS: em 80,6% dos casos a visão colposcópica após a dilatação foi satisfatória. Destes, 80,4% apresentaram margens livres de doença. A incidência de margens livres nas pacientes com colposcopias insatisfatórias persistentes após a dilatação foi de 36,3%. O comprometimento das margens cirúrgicas ocorreu em 28,0% no grupo submetido à dilatação, em 28,5% dos casos no grupo GinSat e em 31,5% no grupo GSat. No acompanhamento verificou-se a incidência de 7,5% de doença residual nas pacientes submetidas à dilatação. No grupo GinSat a incidência de doença residual foi de 28,6% e no grupo GSat foi de 28,9%. CONCLUSÃO: o uso do dilatador higroscópico facilitou a visualização de lesões de difícil acesso ao exame colposcópico, permitindo diminuir a porcentagem de doença residual em pacientes com colposcopias insatisfatórias submetidas à conização por cirurgia de alta freqüência.Faculdade de Medicina de ItajubáUniversidade Federal de São Paulo (UNIFESP) Departamento de GinecologiaUNIFESP, Depto. de GinecologiaSciEL

Study of Heparin in Intestinal Ischemia and Reperfusion in Rats: Morphologic and Functional Evaluation

To study whether treatment with heparin (HEP) attenuates intestinal dysfunction caused by ischemia (I) and reperfusion (R), rats were treated with HEP (100 U/kg intravenously) or saline solution (SS) before I (60 min), which was produced by occlusion of the superior mesenteric artery, and R (120 min). After I or I/R, we mounted 2-cm jejunal segment in an organ bath to study neurogenic contractions stimulated by electrical pulses or KCl, using a digital recording system. Thin jejunal slices were stained with hematoxylin and eosin for optical microscopy. Compared with the sham group, jejunal contractions were similar in the I + HEP and the I/R + HEP groups, but reduced in the I + SS and the I/R + SS groups. the jejunal enteric nerves were damaged in the I + SS and the I/R + SS, but not in the I + HEP and the I/R + HEP cohorts. These results suggested that HEP attenuated intestinal dysfunction caused by I and I/R.Universidade Federal de São Paulo, Escola Paulista Med, Dept Surg, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biochem, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morphol, BR-04023900 São Paulo, BrazilFed Univ Great Dourados, Sch Med, Dourados, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Surg, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biochem, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morphol, BR-04023900 São Paulo, BrazilWeb of Scienc