Role of the Intramural Vascular Network of the Extrahepatic Bile Duct for the Blood Circulation in the Recipient Extrahepatic Bile Duct Used for Duct-to-Duct-Biliary-Anastomosis in Living Donor Liver Transplantation.

A duct-to-duct-biliary-anastomosis is the preferred biliary reconstruction technique in liver transplantation; biliary complications remain the major concerns for the technique. We examined the significance of the intramural vascular network of the extrahepatic bile duct (EBD) and its relevant vessels. We microscopically examined the axial sections of the EBD with 5 mm intervals of 10 formalin-fixed deceased livers. The luminal-areas of the 3 and 9 o'clock arteries correlated significantly and positively with the distance from the bifurcation of the right and left hepatic ducts (the 3 o'clock artery, r = 0.42, p < 0.001; the 9 o'clock artery, r = 0.39, p < 0.001); the ratios of the numbers of the intramural vessels to the areas of the corresponding sections of the EBD significantly correlated positively with the distance from the bifurcation of the right and left hepatic ducts (total vessels, r = 0.78, p < 0.001; arterioles, r = 0.52, p < 0.001; venules, r = 0.45, p < 0.001). This study demonstrated that there is a significant locoregional distributional heterogeneity of the intramural vessels among the EBD. The hepatic arteries neighboring the EBD primarily supply the blood flow to the EBD; thus, when the broader isolation of the EBD from the neighboring arteries is necessary, this locoregional distributional heterogeneity of the intramural vessels may render the EBD likely to suffer ischemia of the anastomotic site.

New bufadienolides extracted from Rhinella marina inhibit Na,K-ATPase and induce apoptosis by activating caspases 3 and 9 in human breast and ovarian cancer cells.

Bufadienolide compounds have been used for growth inhibition and apoptosis induction in tumor cells. Those families of cardiotonic steroids can bind the Na,K-ATPase, causing its inhibition. The use of bufadienolides is widely described in the literature as an anticancer function. The aim of this study was to evaluate the effects of bufadienolides and alkaloid isolated from venom samples from R. marina on tumor cells. We performed cytotoxicity assay in MDA-MB-231 and TOV-21G cells and evaluated the activity of Caspases (3 and 9), Na, K-ATPase, PMCA and SERCA. Four compounds were extrated from the venom of R. marina. The compound 1 showed higher cytotoxicity in MDA-MB-231cells. Compound 1 also showed activation of Caspase 3 and 9. This compound caused an inhibition of the activity and expression of Na, K-ATPase, and also showed activation of both caspase-9 and caspase-3 in MDA-MB-231 cells. We also observed that Compound 1 had a direct effect on some ATPases, such as Na, K-ATPase, PMCA and SERCA. Compound 1 was able to inhibit the activity of the purified Na, K-ATPase enzyme from the concentration of 5 µM. It also caused inhibition of PMCA at all concentrations tested (1 nM-30 µM). However, the compound 1 led to an increase of the activity of purified SERCA between the concentrations of 7.5-30 µM. Thus, we present a Na, K-ATPase and PMCA inhibitor, which may lead to the activation of caspases 3 and 9, causing the cells to enter into apoptosis. Our study suggests that compound 1 may be an interesting molecule as an anticancer agent.

Tylophorine Abrogates G2/M Arrest Induced by Doxorubicine and Promotes Increased Apoptosis in T47D Breast Cancer Cells

Background: The effects of tylophorine, a natural alkaloid found in Tylophora indica, administered as a single compound or in combination with doxorubicin on cell cycling and apoptosis were assessed in T47D breast cancer cells, selected as a model system for breast cancer. Methods: Cell cycle distribution and apoptosis were examined by flow cytometry. Caspase 3 and 9 expression was determined by immunocytochemistry.Result: We found that tylophorine did not significantly influence the cell cycle distribution of T47D cells. However, the alkaloid did prevent accumulation of cells in the G2/M phase. In addition, tylophorine increased the number of apoptotic cells. Expression of proapoptotic proteins (caspases 3 and 9) was up-regulated upon administration of tyloporine alone or in combination with doxorubicin. Conclusions: Tylophorine alone or in combination with doxorubicin induced apoptosis in T47D breast cancer cells through modulation of the cell cycle and affecting the expression of caspases 3 and 9.

Effect of maternal diabetes and hypercholesterolemia on fetal liver of albino Wistar rats.

OBJECTIVE: The aim of this study was to predict the development of hepatic lesions and impairment of function during the development of fetuses (13-, 15-, 17-, and 19-d-old embryos) of diabetic and hypercholesterolemic mothers. METHODS: Eighty virgin and fertile male rats (one male/three females) of Wistar strain with an average body weight of 150 to 180 g were used. Mating was carried out, and pregnancy was determined by examining sperm in vaginal smears. Pregnant rats were arranged into three groups; control, diabetic (single intraperitoneal injection [i.p.] of 60 mg streptozotocin/kg) and hypercholesterolemic groups (fed on a diet containing 3% cholesterol for 6 wk before conception and throughout gestation) (n = 20). Pregnant rats were sacrificed and 13-, 15-, 17-, and 19-d-old embryos and livers were incised and subjected to histological and transmission electronic microscopical (TEM) investigations, assessments of alkaline phosphatase (Al-Pase) isoenzymes electrophoresis, DNA fragmentation, and comet assay. Flow cytometric analysis of apoptosis and caspases 3 and 9 in the livers of mother rats and their 19-d-old fetuses was determined. RESULTS: Histologic findings of diabetic and hypercholesterolemic mothers revealed apparent damage of hepatocytes, accumulation of lipid-laden cells, and vascular steatosis, while the 13-, 15-, 17- or 19-d-old fetuses of either diabetic or hypercholesterolemic mothers revealed disorganized hepatic architecture and massive cell damage. TEM of diseased mothers and their fetuses possessed increased incidence of pyknotic hepatocytes with massive vesicuolation of rough endoplasmic reticulum and degeneration of mitochondria. Al-Pase isoenzymes were altered and genomic DNA of both double and single helical structures were markedly damaged, especially in fetuses of maternally diabetic and hypercholesterolemic mothers. Flow cytometry revealed an increase in apoptosis and caspases 3 and 9 in diabetic and hypercholesterolemic mothers and their 19-d-old fetuses. CONCLUSION: These results suggested that maternal diabetes and hypercholesterolemia predicted early hepatitis and increased apoptosis in mothers and their fetuses as a result of oxidative stress and elevated apoptic markers caspases 3 and 9.

The Study of the Cytotoxic Mechanism of Cisplatin in HeLa Cells / 대한산부인과학회잡지

Cis-diamminedichloroplatinum II (cisplatin) has been reported to induce cell death. However, the mechanism by which cisplatin is induced the apoptosis of cancer cells is still unclear. To evaluate the mechanistic insights of apoptosis by cisplatin, we tested the activities of apoptotic signaling pathway in HeLa cells. Apoptotic death of HeLa cells by cisplatin was confirmed by ladder-pattern fragmentation of genomic DNA. Cisplatin induced the activation of caspase-3 and 9 proteases in a time dependent manner. The caspase-3 protease activation and the cleavage of poly (ADP-ribose) polymerase (PARP) and procaspase-3 was demonstrated. We also showed that the expression of Bcl-2 and Bcl-xL was markedly decreased by the addition of cisplatin in HeLa cells. Moreover, expression of Fas and FasL proteins was increased by cisplatin. These data suggest that cisplatin triggers the activation of apoptotic signaling pathway in human cervical cancer, HeLa cells, via affects the expression of Fas, FasL and Bcl-2 families as well as triggers the activation of caspase-3 and -9 proteases.