Surgical Treatment for Biliary Carcinoma Arising After Pancreatoduodenectomy

The clinicopathological features and surgical treatment of biliary carcinoma around the major hepatic duct confluence arising after pancreatoduodenectomy (PD) due to initial bile duct carcinoma are described in three patients. Occurrence of biliary carcinoma more than 12 years after initial surgery and a histological finding of cholangiocellular carcinoma mixed with hepatocellular carcinoma suggested metachronous incidence of biliary carcinoma after PD. Extended right hemihepatectomy with complete removal of the residual extrahepatic bile duct and segmental, resection of the jejunal loop were carried out safely without operative death or severe postoperative complications. Two patients died of tumor recurrence 6 months after surgery, and the remaining patient is currently living a normal life without evidence of recurrence 17 months after surgery. These surgical procedures are a therapeutic option in patients with biliary carcinoma around the major hepatic duct confluence arising after PD

Analytical Studies on the Colorimetric Estimation of the Activity of Succinic Dehydrogenase System with Neotetrazolium Chloride

Despite the excellence in bringing about reactions, there are still many difficulties and discrepancies in the use of neotetrazolium chloride for measuring the activity of succinic dehydrogenase system of tissue homogenate. Therefore, with the purpose to find out a measuring method simpler and more accurate, the authors performed some experimets, first on the enzymic reactions of tissue homogenate and then after stopping these reactions by using various agents, tested the ease with which reduction products are extracted with various solvents, and finally compared the absorption spectra of these liquid extracts with those of the products reduced chemically by sodium hydrosulfite. As the results, the extraction of the reduction products with acetone-ether (mixed in equal proprtion v/v) after stopping the reaction with 10 per cent formalin solution was found to be the simplest and best method, and moreover, the absorption spectra of extracts of the products obtained by enzymatical or chemical reduction proved to he uniform and the both of which showed the maximum absorption at the wave length of 520 mμ. In addition, the concentrations of sodium succinate and neotetrazolium chloride, the content and kinds of tissues, pH values and concentration of phosphate buffer solutions, the duration of reaction, aerobic or anaerobic conditions, various factors either enhancing or in hibitory to reaction, were all regulated absolutely or relatively; and by drawing absorption curves of the reduction products, these enzymatic reactions were analyzed and scrutinyzed Then the authors established a method which is general, simple and accurate in measuring the enzymatic activities of a small quantity of fresh tissues in contrast with those of hist chemically stained specimens

Influence of simulated microgravity on the activation of the small GTPase Rho involved in cytoskeletal formation – molecular cloning and sequencing of bovine leukemia-associated guanine nucleotide exchange factor

BACKGROUND: The irregular formation of cytoskeletal fibers in spaceflown experimental cells has been observed, but the disorganization process of fibers is still poorly understood. It is well known that the activation of the small GTPase Rho leads to actin stress fibers assembly. This study was performed to evaluate the effect of simulated microgravity on the activation of Rho that is involved in actin fiber remodeling in cells. RESULTS: Clinorotation influences actin fiber remodeling and its related signaling pathways that involve the small GTPase Rho. Actin stress fiber remodeling was significantly inhibited to a greater extent in cells cultured under clinorotation than in static cultured cells. From the gene and protein expression analyses, we found that the expression level of leukemia-associated Rho guanine nucleotide exchange factor (LARG), which activates Rho, was downregulated under clinorotation. Moreover, we identified the full-length LARG cDNA. The amount of GTP-bound RhoA, that is, the active form of RhoA, decreased under this condition. CONCLUSION: The activation of the small GTPase Rho was influenced by simulated microgravity generated by a three-dimensional (3D) clinostat. Furthermore, the full-length cDNA of bovine LARG, a member of the Rho guanine nucleotide exchange factor (GEF) family, was identified, and its gene expression was observed to be downregulated under clinorotation. This downregulation subsequently resulted in the repression of RhoA activation. These results indicated that the disorganization of the actin fibers was caused by the inhibition of Rho activation by 3D clinorotation

A simple and high-sensitivity method for analysis of ubiquitination and polyubiquitination based on wheat cell-free protein synthesis

<p>Abstract</p> <p>Background</p> <p>Ubiquitination is mediated by the sequential action of at least three enzymes: the E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme) and E3 (ubiquitin ligase) proteins. Polyubiquitination of target proteins is also implicated in several critical cellular processes. Although Arabidopsis genome research has estimated more than 1,300 proteins involved in ubiquitination, little is known about the biochemical functions of these proteins. Here we demonstrate a novel, simple and high-sensitive method for <it>in vitro </it>analysis of ubiquitination and polyubiquitination based on wheat cell-free protein synthesis and luminescent detection.</p> <p>Results</p> <p>Using wheat cell-free synthesis, 11 E3 proteins from Arabidopsis full-length cDNA templates were produced. These proteins were analyzed either in the translation mixture or purified recombinant protein from the translation mixture. In our luminescent method using FLAG- or His-tagged and biotinylated ubiquitins, the polyubiquitin chain on AtUBC22, UPL5 and UPL7 (HECT) and CIP8 (RING) was detected. Also, binding of ubiquitin to these proteins was detected using biotinylated ubiquitin and FLAG-tagged recombinant protein. Furthermore, screening of the RING 6 subgroup demonstrated that At1g55530 was capable of polyubiquitin chain formation like CIP8. Interestingly, these ubiquitinations were carried out without the addition of exogenous E1 and/or E2 proteins, indicating that these enzymes were endogenous to the wheat cell-free system. The amount of polyubiquitinated proteins in the crude translation reaction mixture was unaffected by treatment with MG132, suggesting that our system does not contain 26S proteasome-dependent protein degradation activity.</p> <p>Conclusion</p> <p>In this study, we developed a simple wheat cell-free based luminescence method that could be a powerful tool for comprehensive ubiquitination analysis.</p

The Relationship between Inferior Vena Cava Distensibility and Arterial Blood Pressure

The purpose of this study was to clarify the relationship between the inferior vena cava distensibility and blood pressure. Six Japanese healthy males volunteered to participate in this study. We measured the cross-sectional area of inferior vena cava (CSAivc) and blood pressure at rest and during passive leg raising in supine position. We calculated the change rates of each parameter, based on the value at rest in supine position. We observed a negative correlation between the change rate of CSAivc and systolic blood pressure (P<0.05). These results suggest that the inferior vena cava distensibility affects partially systolic blood pressure