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The purpose of this study was to compare the hemodynamics of the liver vasculature as determined by Doppler sonography for patients with Osler-Weber-Rendu disease and for healthy subjects.Real-time color Doppler sonography and pulsed Doppler sonography were used to study 10 patients with Osler-Weber-Rendu disease and 25 healthy subjects. Vessel diameter and flow velocity in the hepatic artery and its branches as well as in the portal vein and its segmental branches were determined. Flow patterns in the hepatic veins were analyzed. The clinical manifestations of the disease were correlated with the Doppler sonographic findings.The hepatic artery and its branches were dilated and tortuous, and flow velocity was greater in patients than in healthy subjects; hepatic artery velocities (mean +/- SD) were 153 +/- 65.2 cm/sec versus 64.9 +/- 11.4 cm/sec (p.005). The resistive index measured in the hepatic artery did not differ significantly between the two groups (p was not significant). No intrahepatic arteriovenous shunt sites were detected by color Doppler sonography. However, arterioportal shunts were observed in two patients, one of whom also had a portovenous shunt. A portovenous shunt and multiple shunts between hepatic veins were found in another patient. Portal venous flow was similar in both groups. No relationship between Doppler sonographic changes and clinical manifestations was found.The multiple microscopic arteriovenous shunts found in Osler-Weber-Rendu disease were not detected in our patients but resulted in dilatation and increased flow velocity in the hepatic artery. Flow velocity in the portal vein appeared to be undisturbed. Large intrahepatic shunts were easily outlined by Doppler sonography.

It has been suggested that CN (calcineurin, protein phosphatase-2B) regulates signal transduction, particularly in various secretory cells. In this study, we examined whether CN plays a role in stimulus-secretion coupling of gastric parietal cells.Localization of CN in gastric epithelial cells was examined immunohistochemically. The role of CN in the acid secretion pathway of gastric parietal cells was assessed by evaluating the effect of FK506, a specific inhibitor of CN, on gastric acid secretion in pylorus-ligated rats. In addition, the effect of FK506 on secretagogue (carbachol, tetragastrin and histamine)-stimulated acid secretion was investigated in lumen-perfused rats.CN was specifically expressed in gastric parietal cells and chief cells of the gastric mucosal epithelium immunohistochemically. FK506 dose-dependently inhibited gastric acid secretion in pylorus-ligated rats. In lumen-perfused rats, FK506 completely inhibited acid secretion prestimulated by carbachol and tetragastrin, agonists known to increase cytosolic Ca2+, but did not affect acid secretion prestimulated by histamine.Our findings demonstrate that FK506 has a potent antisecretory effect in parietal cells through inhibition of only Ca2+-mediated acid secretion pathways. As FK506 is known to specifically inhibit CN, which plays an important role in signal transduction in various secretory cells, protein dephosphorylation signalling might also be crucial for gastrin and M3 muscarine receptor-mediated stimulation of proton pump.

One of the important problems in experimentally induced small intestinal lesions is that there is no reproducible model of diffuse and stable mucosal lesion. In this paper, we studied in detail the effects of continuous perfusion of various concentrations of acetic acid on the rat small intestinal mucosa. In order to evaluate its applicability for screening of the preventive effect of drugs on gut damage, we also evaluated the efficacy of corticosteroid pretreatment in preventing acetic acid-induced mucosal lesion. Male Sprague-Dawley rats were fasted for 12 hr, and the small intestinal lumen was perfused with 1%, 1.5%, 2.5%, 3%, 3.75% (pH 2.4-2.6) acetic acid or saline (control) at 1 ml/min for 15 min. In separate experiments, the effect of preadministration of budesonide (0.5 or 0.75 mg/kg/day) and prednisone (0.75 mg/kg/day) on 1.5% acetic acid-induced mucosal damage was investigated. Macroscopic and microscopic lesions occurred diffusely in a concentration-dependent fashion. Histological findings revealed signs of transmural inflammation characterized by mucosal-submucosal edema, ulceration, and neutrophil infiltration. Mucosal-submucosal height had an inverse relation with the acetic acid concentrations perfused. Myeloperoxidase activity levels increased several-fold in the acetic acid-perfused groups. Corticosteroid pretreatment prevented microscopic damage and was associated with reduction of MPO activity levels in 1.5% acetic acid-perfused rats. We conclude that this simple and reproducible model could be applied for the screening of new drugs in the gastrointestinal tract in which large numbers of animals are taken into account.

In order to study the cytoprotective function of colonic heat shock proteins (HSPs) in vivo, the effect of specific preinduction of HSP60 by thyrotropin-releasing hormone (TRH) administration on the development of acetic acid-induced colonic mucosal lesion was investigated. Expression of 60-kDa, 72-kDa, and 90-kDa heat shock proteins (HSP60, HSP72, and HSP90, respectively) in rat colonic mucosa was investigated by western blot and immunohistochemical analyses before and after TRH administration. Following pretreatment with or without TRH administration, the rats received intrarectal infusion of 5% acetic acid. The colonic mucosal damage was macroscopically evaluated 24 hr after the intrarectal infusion of acetic acid. Expression of HSP60 was significantly increased by TRH administration in the colonic mucosa, whereas HSP72 and HSP90 did not increase. Immunohistochemical study also showed a significant increase in HSP60 in colonic mucosal cells, especially at the surface of the colonic mucosa after TRH administration. No histopathologic alteration was observed in the colonic mucosa after TRH administration. The colonic mucosal damage caused by intrarectal infusion of 5% acetic acid was not prevented by preinduction of HSP60. We demonstrated that specific preinduction of HSP60 by TRH administration did not show cytoprotective function in the colonic mucosa, although this protein plays a crucial role for cytoprotection in the pancreatic acinar cells. Our results indicate that the role of HSP60 may be different in each organ with respect to cytoprotection.

A controlled trial was conducted to compare the efficacy of interferon (IFN) between two groups of patients with type C liver. Thirty-five patients were randomly assigned to group A (17 patients) or group B (18 patients). The former received 3 megaunits (MU) of human lymphoblastoid IFN six days per week for two weeks, followed by three days per week for 50 weeks; the latter group received 6 MU six days per week for two weeks followed by three days per week for 24 weeks. The percentages of biological sustained responders (B-SR) and virological sustained responders (V-SR) were 29.4 and 23.5%, respectively, in group B, and 17.6% for both in group A. The therapeutic effects were not different between two groups. HCV genotype 2 accounted for significantly higher percentage of B-SR and V-SR (both 57.1%, respectively). These findings indicate that IFN is effective in type C cirrhosis with genotype 2.

To provide a genetic survey of hereditary ataxia, we performed PCR screening of SCA1, SCA2, MJD1 (SCA 3), SCA6, DRPLA, with 71 patients in 61 families living in Akita prefecture (1,205,571 population in 1997) in Japan. Of 71 patients in 61 families, 18 MJD1, 14 SCA6, 5 DRPLA, 1 SCA1 and 1 SCA2 patients were detected. Eighty percent of autosomal dominant inherited spinocerebellar degeneration (AD-SCD) including 7 spoladic patients genetically diagnosed as AD-SCD was MJD1 (45.7%) and SCA6 (34.3%). These suggest the prevalence rate of hereditary ataxias in Akita prefecture; 1.5 and 1.2/100,000 of MJD1 and SCA6, respectively. Only one patient of SCA1 was detected, which was frequently reported in Hokkaido and Tohoku area in Japan.

It is well documented that gastric mucosa can increase its resistance to mucosal damage caused by aspirin during repeated long-term administration of aspirin. However, the underlying mechanism of this adaptation is not well established. In the present study, we investigated the effect of long-term (chronic) administration of aspirin on expression of heat shock proteins (HSPs), which are known as endogenous cytoprotectants, in rat gastric mucosa. Rats were administered aspirin (100 mg/kg) daily for up to 20 days. After various periods of aspirin administration, a high dose of aspirin (250 mg/kg) was administered, and the mucosal damage was assessed. Expression of heat shock proteins (HSPs) in gastric mucosa was evaluated by Western blot. Intracellular localization of each HSP was studied immunohistochemically. Prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) levels were also investigated. Long-term aspirin administration resulted in development of resistance to aspirin-induced mucosal damage, and the increase of HSP72 expression correlated with mucosal resistance to aspirin. No significant increase was observed in HSP60 and HSP90 levels. Immunohistochemical study showed an increase of HSP72 in the cytoplasm of mucosal surface cells. The PGE2 level was suppressed and no change in the level of LTB4 was observed. It is possible that HSP72 could play important roles in gastric mucosal adaptation when the PGE2 level is suppressed by NSAIDs.

We analyzed muscle area in CT and muscle pathology in a patient with isolated ACTH deficiency who started with the difficulty of elevation of both arms. Cortisol treatment resulted in full recovery from severe muscle atrophy and contracture of major joints. Change of volume of major muscles in arm, thigh and calf was followed. Major muscles were identified in CT and the area of each muscle was calculated with computer assistance. The increase of total muscle area in sequential 3 times in CT was up to 74% after prednisolone treatment. This indicates that the deficiency of cortisol resulted in 42% reduction of muscle volume. This also suggests that reduction of muscle volume induces the limitation of range of motion of shoulder joint. ATPase of muscle biopsy revealed the influence on fiber type proportion; type 1 : type 2A : type 2B = 29.6 : 6.0 : 64.4% and 35.7 : 17.6 : 46.7% in pre-treatment and post-treatment of cortisol, respectively. Mean diameters of muscle fibers in type 1, type 2A and type 2B was 41.8, 41.8, 39.1 microns and 46.2, 44.0, 37.2 microns in pre-treatment and post-treatment of cortisol, respectively. These suggest that deficiency of glucocorticoid introduces the reduction of the activity of the motor neurons innervating type 1 and type 2A muscle fibers.

Bowel dysfunction such as irritable bowel syndrome caused by stress is well described. Previous reports suggest that stress is known to cause the release of endogenous substances such as catecholamine, beta-endorphine, 5-hydroxytryptamine, corticotropin-releasing factor, and thyrotropin-releasing hormone (TRH). However, the role played by these neurohormonal mediators in bowel dysfunction under stress conditions is not well known. We investigated the influence of water-immersion stress or TRH administration on the expression of 60-kDa, 72-kDa, and 90-kDa heat-shock proteins (HSP60, HSP72, and HSP90, respectively) in rat small intestinal mucosa by Western blot and immunohistochemical analyses. The cytoprotective function of preinduced HSPs on experimentally induced mucosal damage also was studied. In order to investigate the influence of preinduction of HSP60 on small intestinal damage, the small intestinal lumen was perfused with 1.5% acetic acid 1 ml/min for 15 min with or without pretreatment with water-immersion stress or TRH administration. Expression of HSP60 was significantly increased by water-immersion stress or TRH administration in the small intestinal mucosa, whereas HSP72 and HSP90 did not increase. Interestingly, expression of this protein showed the biphasic peak pattern after water-immersion stress or TRH administration. Each peak was observed 3-6 hr and 21-24 hr after the initiation of water-immersion stress or TRH administration. Immunohistochemical study also showed a significant increment of HSP60 in both the cytoplasm and nuclei of the small intestinal mucosal cells. No histopathologic alteration was observed in rat small intestinal mucosa after each treatment. Small intestinal damage caused by 1.5% acetic acid perfusion was not influenced by preinduction of HSP60. We demonstrated that water-immersion stress or TRH administration specifically induced HSP60, although preinduction of this protein did not show a cytoprotective function in the small intestinal mucosa.

Class II antigens are strongly expressed on the inflamed colonic epithelium in inflammatory bowel disease. However, the mechanism of this epithelial class II antigen induction is not fully understood. Increased activities of interferon (IFN)gamma, tumor necrosis factor (TNF)alpha, and interleukin (IL)2 have been shown in the inflamed mucosa of inflammatory bowel disease. Therefore, we studied whether these cytokines could induce class II antigens on the human colonic epithelium. By an organ culture technique, 284 normal colonic biopsy specimens obtained from 49 individuals were cultured in media containing different concentrations of cytokines with/without anti-IFNgamma R antibody. Colonic epithelial class II antigens were identified by the indirect immunoperoxidase staining method with anti-human leukocyte antigen (HLA)-DR, DP, and DQ monoclonal antibodies. IFNgamma, TNFalpha, and IL2 induced epithelial class II antigens in 16 of 16 cases (100%), 2 of 16 cases (12.5%), and 6 of 17 cases (35%), respectively. Epithelial class II antigen expression in response to TNFalpha was induced via IFNgamma but not via IL2. This is the first demonstration that: (i) the induction of class II antigens on the colonic epithelium in response to TNFalpha is mediated via IFNgamma, and (ii) that IL2 induces class II antigens.

Comparison of nonstructural glycoprotein NSP4 gene sequences from 22 rotavirus strains originating from six host species and of 14 different combinations of G and P types revealed the presence of three distinct NSP4 alleles, represented by strains Wa, KUN and AU-1. Genetic distances between any of these alleles (18.0%) were significantly greater than those within each allele (5.5%) and phylogenetic analysis suggested that divergence into three distinct alleles had occurred at about the same time during evolution. While amino acid variation among strains was minimal in the amino-terminal two-thirds of the protein (aa 1-130), variability increased toward the carboxy terminus of the enterotoxic peptide region (aa 114-135) and was greatest between residues 135 and 141. Comparison of the amino acid sequences corresponding to the enterotoxic peptide region between strains isolated from asymptomatic neonates and those from children with diarrhoea failed to identify any conserved changes that correlated with the capacity of the virus to cause disease. Amino acids were relatively conserved in the domains important for viral morphogenesis.

We previously reported that water-immersion stress specifically induced the synthesis of a 60-kDa heat-shock protein (HSP60, chaperonin homolog) in pancreatic cells and preinduction of HSP60 completely prevented development of cerulein-induced pancreatitis in the rat in an HSP60 quantitatively dependent manner. In order to study the cytoprotective function of a 72-kDa heat-shock protein (HSP72, stress-inducible hsp70), the effect of specific preinduction of HSP72 by hyperthermia on cerulein-induced pancreatitis was investigated and compared with the effect of preinduction of HSP60 in this study. Expression of HSP60 and HSP72 in the pancreas was investigated by immunoblot before and after water immersion or hyperthermia. Following pretreatment with water-immersion stress or hyperthermia, the rats were injected with cerulein (40 micrograms/kg, intraperitoneally). The pancreas wet weight and serum amylase concentration were measured before and after cerulein injection. Hyperthermia (42.5 degrees C, 20 min) specifically induced HSP72 in the pancreas. The synthesis of HSP60 was specifically induced by water-immersion stress in the pancreas. Cerulein-induced pancreatitis was clearly prevented by specific preinduction of HSP60 by water-immersion stress. However, preinduction of HSP72 by hyperthermia had no preventive effect on cerulein-induced pancreatitis. Our findings suggest that HSP60 and HSP72 have distinct functions in the pancreas, and their induction mechanisms are also different in vivo. These results could be important for understanding the mechanism of "adaptive cytoprotection" in the pancreas mediated by heat-shock proteins.

In order to study the cytoprotective function of heat shock proteins (HSPs) in vivo, the effect of preinduction of HSPs by hyperthermia on acetic acid-induced colitis was investigated. Expression of 60-kDa, 72-kDa, and 90-kDa heat shock proteins (HSP60, HSP72, and HSP90, respectively) in rat colonic mucosa was investigated by Western blot analysis and immunohistochemical study before and after hyperthermia. Following pretreatment with or without hyperthermia, the rats received intrarectal infusion of various doses of acetic acid. The colonic mucosal damage was evaluated by macroscopic and microscopic assessments 24 hr after the intrarectal infusion of acetic acid. Expression of HSPs was significantly increased by hyperthermia in rat colonic mucosa. Immunohistochemical study also showed the increments of HSPs in the colonic mucosal cells after hyperthermia. Acetic acid-induced colitis was dramatically prevented by pretreatment with hyperthermia when HSP72 and HSP90 were preinduced. On the other hand, induction of HSP60 did not correlate with mucosal protection. Our findings suggest that HSP72 and HSP90 may have cytoprotective function against acetic acid-induced mucosal damage. These results may be important for understanding the mechanism of "adaptive cytoprotection" mediated by HSPs.

We reported a 66-year-old male patient with advanced gastric cancer accompanied by multiple liver metastasis, who responded to combination chemotherapy using UFT, CDDP and etoposide. The patient was administered three courses of 600 mg/body UFT po daily, 50 mg/body CDDP iv and 50 mg/body etoposide iv on days 1 and 8 every 4 weeks. As a result, both the primary and metastatic tumors decreased remarkably in size. Adverse reactions were leukocytopenia (Grade 2), thrombocytopenia (Grade 1) and nausea (Grade 1). he is alive 16 months after the beginning of therapy in a condition of partial response (PR). This combination therapy seemed to be effective for advanced gastric cancer.

Mammalian chaperonin homolog (HSP60) was purified from porcine livers cytosol using a tandem ATP-Sepharose column and Mono Q column chromatography. A partial amino acid sequence (96 amino acid residues) of this protein was determined and coincided with those of human HSP60 with 96.9% homology, which was deduced from the nucleotide sequence of the cDNA. The sequence of the NH2 termini of the purified protein (5 amino acid residues) coincided with the signal sequence of HSP60. These facts led to the identification of the 60-kDa liver protein with the chaperonin homolog. Dihydrofolate reductase was able to form a stable complex with the liver chaperonin homolog. The liver chaperonin homolog was detected by at least five spots around pI = 5.6 on two-dimensional gel electrophoresis. Immunoblotting studies using an antibody against chaperonin homolog showed that the chaperonin homolog was localized in the cytosol, mitochondrial, and nuclear fractions of porcine liver. The chaperonin homolog was localized both in the mitochondria and cytoplasm of rat kidneys at the electron microscopic level. The chaperonin homolog in the cytosol, but not in the other subcellular fractions, was cross-reacted with an antibody against the synthetic peptide corresponding to the signal peptide of HSP60 as well as the purified chaperonin homolog on immunoblotting. These results suggested that the functional chaperonin homolog in the cytosol may be transported into the mitochondria and the protein may be processed to mitochondrial HSP60 in the organella.

The morbidity rate for retinopathy of unknown origin was significantly higher (P0.001) in the chronic hepatitis C group, 27 (31.8%) of 85 cases, than in the control group, 6 (6.0%) of 100 cases. Among hepatitis C virus (HCV)-associated retinopathy patients, 6 cases (22.2%) were aware of ocular subjective symptoms, retinopathy recurred in 8 (29.6%) cases, binocular retinopathy occurred in 14 (51.9%) cases, and retinopathy related to liver dysfunction, in 13 (81.3%) of 16 tested cases. HCV-associated retinopathy involved hemorrhage at the posterior pole retina in 21 (77.8%), cotton-wool patches in 9 (33.3%), and hemorrhage at the peripheral retina in 7 (25.9%) cases. Sequelae occurred in only one case. Retinopathy worsened or recurred in all 7 cases treated with interferon. The risk factors for HCV-associated retinopathy were mild thrombocytopenia (P0.001), long-term hepatitis illness (P0.005), advanced age (P0.02), concurrence with liver cirrhosis (P0.02), history of systemic hypertension (P0.05), and female gender (P0.05).

Color Doppler sonography is frequently used for the diagnosis of abdominal abnormalities, especially in cases of liver and vascular disease. However, color Doppler findings of splenic diseases have been reported only rarely. We describe a case of a splenic hemangioma in which color Doppler sonography showed blood flow within the tumor. Many color echoes appeared in the tumor immediately after it was compressed by the probe, suggesting that flow in the vessels in the hemangioma was obliterated by the probe.

A new autoantibody to 210kDa microtubule associated protein (MAP) obtained from rabbit liver excrude was detected in the serum with various liver diseases and SLE. As there were no reports concerning autoantibodies to MAPs in animals and human, it may say that anti-210kDa MAP autoantibody we detected was a new cytoskeleton antibody. Using western blotting method, the anti-210kDa MAP autoantibody was frequently found in the patients with alcoholic liver diseases (52.5%), PBC (55.6%), autoimmune hepatitis (83.3%), SLE (71.4%) but rarely in the patients with viral liver diseases (26.4%) and none in normal controls at a serum dilution of 1:10. In addition, at a serum dilution of 1:100, the anti-210kDa MAP autoantibody was found in the patients with alcoholic liver diseases (22.5%), PBC (44.4%), autoimmune hepatitis (66.7%), SLE (71.4%), viral liver diseases (17.0%) and none in normal controls. It was confirmed that the anti-210kDa MAP autoantibody was frequently detected in cases of alcoholic liver diseases, PBC, autoimmune hepatitis and SLE than in those of viral liver diseases and normal control.

Twelve patients with unresectable squamous cell carcinoma of the esophagus were treated with a combination chemotherapy regimen consisting of cis-diamminedichloroplatinum (CDDP), bleomycin (BLM) or peplomycin (PEP), and 5-fluorouracil (5-FU). Ten of them received radiation therapy additionally. CDDP was administered once every 4 weeks at a dose of 50 mg/m2. Methylprednisolone of 250 mg was given intravenously 4 times at the same day with infusion of CDDP. BLM or PEP was administered intravenously at a dose of 20 mg/m2 every 2 weeks and 5-FU was administered at a dose of 330 mg/m2 on days 1-5, 15-19, and afterwards every 4 weeks. All patients received at least two courses of chemotherapy. All of them were evaluable. Complete and partial responses were obtained in one and eight cases, respectively. Responsive rate was 75.0%. The median duration of response was 17.0 weeks. The median duration of survival was 44.0 weeks in all patients, 46.1 weeks in responders and 17.9 weeks in non-responders. Nausea, vomiting, leucopenia, fever, nephrotoxicity and radiation esophagitis were observed as side effects but most of them were mild and well tolerated. In conclusion, this regimen was considered to be very useful as the chemotherapy for primary esophageal carcinoma.

We have studied the protein composition of the pectoralis superficialis muscle of genetically dystrophic (New Hampshire line 413) and normal control (line 412) chickens by one- and two-dimensional gel electrophoresis. A protein, referred to hereafter as the 30 kDa abnormal protein, was specifically detected in the affected muscle. It was purified to homogeneity, and its molecular properties were studied. It is a monomer with a molecular mass of approximately 30 kDa and an isoelectric point of about pI 8.4. We have screened by Western blotting a variety of muscles from line 412 and line 413 chickens for the presence of the 30 kDa protein. While the pattern of total protein is very similar in all cases, the 30 kDa protein was not detected in the pectoralis superficialis muscle of line 412 chickens. However, the immunoreactive bands were detected in the sartorius muscle and the tensor fasciae latae muscle from dystrophic and normal chickens. Interestingly, the immunoreactive bands of normal skeletal muscles are smaller in molecular weight than those of dystrophic skeletal muscles. To determine the early time sequence of the appearance of the abnormal protein, we studied muscles from embryos and post-hatched chickens at various ages. The abnormal protein was detected in dystrophic muscles as early as 15 days ex ovo and occurred throughout development up to six months ex ovo. Although the implication of the dystrophy-associated appearance of the 30 kDa protein in the affected muscle is not clear at present, it would be of particular interest to elucidate the biochemical functions of the 30 kDa protein in the affected muscle (pectoralis superficialis muscle) of genetically dystrophic chicken.

The clinical significance of serum sialyl SSEA-1 antigen was evaluated using the sera of 1261 patients with malignant tumors and 717 patients with non-malignant diseases measured with Otsuka Assay Laboratories' RIA Kits. The results indicate that the antigen was frequently elevated in the sera from the patients with various adenocarcinomas, including lung (45%), pancreas (64%) and ovary (57%). The false positive incidence of antigens in the sera from the patients with non-malignant disorders was as low as 4.9%. No correlation was observed with other tumor markers examined in this study, and the diagnostic efficiency increased significantly by the combined determination of sialyl SSEA-1 antigen level with other markers. The serial determination of the serum sialyl SSEA-1 antigen level has a clinical utility also in monitoring the patients with adenocarcinoma receiving surgical operation and/or chemotherapy as indicated by the results of the longitudinal observation of the patients.