The development of biological and non-biological artificial liver support (ALS) therapy is reviewed. Of the various currently available nonbiological ALS, the combination of plasma exchange and hemodiafltration using high performance membranes, we originally developed, is the most effective because it can sustain patients with the severest liver failure that may be comparable to an anhepatic state. This ALS has already been adopted by 80% of major hospitals and is currently accepted as the standard method of liver support in Japan Several biological ALS systems have been developed, and some of them are clinically available. However, they must overcome major obstacles such as low biocompatibility, low durability, and compensation of only narrow liver function. Recently, a cell line which overcome most of these defects has been generated. Experimentally it can support 90% hepatectomized rats. Such cells may eventually facilitate biological liver support.

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Although periductal fibrosis is a well-known risk factor for progressive hepatic fibrosis in biliary atresia, the fibrogenic process is not well understood. Histological examination was used to determine the cell type responsible for periductal fibrosis in biliary atresia. Hepatic specimens were obtained by radical operation of 20 patients with biliary atresia, ranging in age from 1 to 4 months. Alpha smooth muscle actin and CD34 antibodies were used as probes in immunohistochemical labeling experiments. In all cases, cytoplasm stained positive for alpha smooth muscle actin. The number of cells in the CD34 positive peribiliary capillary plexus was greater in BA patients, and the capillaries had more frequent connections with periductal mesenchymal cells. Electron microscopy revealed that spindle-shaped mesenchymal cells, regarded as periductal mesenchymal cells, surrounded the interlobular bile duct in 16 cases, and were occasionally located in spaces between the capillaries and bile duct epithelia. These results suggest that periductal mesenchymal cells are derived from pericytes of the peribiliary capillary plexus in biliary atresia.

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The aim of this study was to investigate the pathophysiology of fecal incontinence in patients with diabetes mellitus. Methods. Two groups of diabetic patients were studied. Group A consisted of 7 subjects (48 to 76 years, mean age 53.5 years) with fecal incontinence and Group B consisted of 9 subjects (38 to 79 years, mean age 62.1 years) without fecal incontinence. A third group of 10 healthy volunteers (Group C) were included as control subjects. All subjects underwent anorectal manometry and rectal sensitivity tests, and pudendal nerve terminal motor latency ( PNTML) . Results. Anal resting pressure was significantly lower in Group A (38 cm H₂O) compared to Group B (60 cm H₂O) and Group C (59 cm H₂O) . Maximum squeeze pressure was significantly lower in Group A (98 cm H₂O) and Group B (160 cm H₂O) compared to Group C (240 cm H₂O) . Initial sensation volume was significantly higher in Group A (80 cm H₂O) compared to Group C (30 cm H²O) . Pudendal nerve terminal motor latency was significantly higher in Group A (3.15 msec) compared to Group B (2.45 msec) and Group C (2.03 msec) . There was no significant difference in the length of the high-pressure zone, urgency, or maximum tolerable volume between the three groups. Conclusion. Our results demonstrate that autonomic neuropathy causes internal sphincter or initial rectal sensation damage, and that somatic neuropathy, such as pudendal neuropathy, may play an important role in fecal incontinence in diabetic patients.

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Myocardial oxygen consumption (MVO₂) may be disproportionately high relative to contractile function in post-ischemic, reperfused myocardium. This study investigated the mechanism of dissociation between MVO₂ and contractile function in post-ischemic, reperfused myocardium using isolated rat hearts. Mitochondrial dysfunction secondary to increased calcium uptake has been implicated as an important mediator of reperfusion injury in the heart. In post-ischemic, isovolumic, antegrade-perfused rat hearts, MVO₂ rate and contractile function were studied in relation to mitochondrial function. Left ventricular pressure, coronary blood flow, and MVO₂ were determined. Mitochondrial respiration and membrane potential were measured by polarography and flow cytometry. To examine the role of mitochondrial calcium uptake in ischemic reperfusion injury, isolated rat hearts perfused with ruthenium red (an inhibitor of calcium uptake) were compared to control perfused hearts. Hearts were subjected to 60 minutes of no-flow ischemia, followed by 60 minutes of reperfusion. At 15 minutes after the onset of reperfusion, there was poor recovery of left ventricular pressure to 54% of the control level, but MVO₂ increased to 149% of the control. Addition of 2.5μM ruthenium red to the perfusate resulted in a decrease in MVO₂. The oxygen consumption rate in state 3 mitochondria decreased similarly following reperfusion both in control and in ruthenium red hearts. The mitochondrial membrane potential was reduced to 89% (logarithmic scale) after 15 minutes of reperfusion. This data suggests that the dissociation between MVO₂ and contractile function following early reperfusion is partly caused by the repair of intracellular damage resulting from reduction of the mitochondrial membrane potential.

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Ryanodine receptors (RyRs) are calcium (Ca²⁺) release channel proteins with Ca²⁺-induced Ca²⁺ release properties. To analyze the structurefunction relationships in RyRs, ten serial deletion clones of type 1 RyR (RyR 1) were. constructed. Each was made by excising a cDNA cassette (dCs) encoding approximately 500 amino acids from the full length cDNA which encodes 5, 037 amino acids. All of these deletion clones transiently expressed RyR 1 as green fluorescence (GFP) fusion proteins in cultured Chinese hamster ovary (CHO) cells, and in addition showed the same cytoplasmic localization as the full length RyR 1. An increase of intracellular Ca²⁺ concentration in response to 4-chloro-3-ethylphenol (4-CEP), a RyR stimulating agent, was only observed in the dCs8 clone while the other dCs clones did not respond to 4-CEP (up to 500μM) . These results suggest that the amino acids between ³²²⁶Glu and ³⁷²⁴Ala of RyR, which were lacking in the dCs8 clone, do not have a critical role in the functional formation of the RyRI/Ca²⁺ release channel protein.

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We examined the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in hepatocellular carcinoma (HCC) to elucidate the relationship between angiogenesis and lung metastasis. Liver samples obtained from 40 autopsy cases of primary HCC patients were evaluated (20 patients with lung metastasis) . Immunohistochemical studies were performed using anti-CD34 and anti-VEGF antibodies. The proportion of cases with high MVD was significantly greater in the lung metastasis positive group, compared to the negative group. VEGF expression was not associated with lung metastasis. Clinicopathological features did not correlate with MVD. We conclude that MVD in HCC is an independent risk factor for lung metastasis.

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We devised and tested a simple and practical measuring system for assessing drug-induced extrapyramidal signs (EPS) in schizophrenic patients. We first assessed the ability of the system to clinically measure the degree of EPS, and then investigated differences between conventional and atypical antipsychotic agents. Schizophrenic patients (28 males, 22 females) were compared with age-matched normal controls. A 14-item battery of measurements was analyzed in terms of established EPS scale scores and psychiatric symptoms. Measurements concentrated on the face, upper and lower limbs. Patients were impaired in 10 out of 14 quantitative subtests concerning movements when compared with normal controls. Facial characteristics that differed between patients and normal controls included oral agility and Myerson's sign; for the upper limbs included various fine, dominant-handed tasks; and for the lower limbs included gait. We then compared 16 patients receiving conventional agents and 18 patients receiving atypical agents, using the same system. Improvements in oral agility and fine finger movements were observed for patients on newer atypical agents. Our measurement system can reveal motor dysfunction and difficulties that cause problems in daily life for schizophrenic patients, and therefore provides a clinically useful assessment of drug-induced EPS. The most sensitive subtests for detecting drug-induced side effects were those that dealt with oral agility and rapid, repetitive finger movements.

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We examined the effects of a potent cholecystokinin (CCK) receptor antagonist, L-364, 718 and those of a trypsin inhibitor, camostat mesilate (camostat) after induction of acute pancreatitis with caerulein. Seven intraperitoneal caerulein injections (15μg/kg) were repeated in rats at hourly intervals. The rats were then divided into four treatment groups: oral administration of water plus intraperitoneal injection of a vehicle, polyethylene glycol and glycerin (PEG) ; oral administration of water plus intraperitoneal injection of L-364, 718 in PEG at a dose of 2mg/kg; oral administration of camostat 400 mg/kg a day plus intraperitoneal injection of PEG, and oral administration of camostat plus intraperitoneal injection of L-364, 718. Camostat or water alone was administered once a day beginning 1 h after the last caerulein injection. L-364, 718 or PEG alone was injected every 6h beginning 18h after the last caerulein injection. All observations were made on day 4. The rate of DNA synthesis was determined to evaluate pancreatic regeneration. L-364, 718 inhibited regeneration whether or not camostat was given, while camostat given alone increased plasma CCK, promoted pancreatic DNA synthesis, and decreased the histologic severity of the pancreatitis. Histologic damage in the pancreas was milder in rats receiving both camostat and L-364, 718 compared to those receiving only L-364, 718. Therefore, we conclude that CCK participates in the regeneration of acinar cells after acute pancreatitis in rats, together with other factors.

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In order to analyze the regulation of the ubiquitination of p21, an in vitro or in vivo model must first be developed. We describe the development of an in vitro ubiquitination system for p21. The substrate p21 was extracted from the lysate of ML-1 cells that had been treated with 15 Gy of γ-irradiation, using Protein G sepharose beads that were cross-linked with anti-p21 antibody. After binding of p21 to the beads, rabbit reticulocyte lysate containing El, ML-1 cell lysate containing E2/E3, and methylated ubiquitin, were added. Ubiquitinated p21 was detected by Western blot analysis using anti-p21 or anti-ubiquitin antibodies. Ubiquitinated p21 was also obtained using K48R ubiquitin, confirming the validity of this system. The ubiquitination activity in the ML-1 cell lysate was heat sensitive, suggesting protein composition. This in vitro system will provide a useful tool for studying the regulation of the ubiquitination of p21.

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We report a rare case of serrated adenoma of the appendix synchronously associated with adenocarcinoma of the sigmoid colon. A 69-year-old Japanese man admitted to our hospital for a routine physical checkup and colorectal endoscope was found to have a flat-elevated tumor measuring 23×8×15 mm in size in the sigmoid colon. He was healthy until this admission, with no abdominal symptoms in the previous couple of months. A surgical sigmoidectomy with routine additional appendectomy was performed. Histopathological examination of the resected tumor revealed an intra-mucosal adenocarcinoma and tubular adenoma with severe dysplasia in the sigmoid colon. The appendix, grossly unremarkable, harbored a serrated adenoma with no evidence of invasion or malignant transformation. Serrated adenoma of the appendix is extremely rare, and only one case report could be found on MEDLINE.

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