Decrease of Serum Triglyceride in Normal Rats Fed with 2000 ppm Aluminum Diet for 67 Days: Feeding Young Rats Sucrose, Lactose, Milk, Casein or Soy-Protein Diets with Addition of Aluminum Chloride

Decrease of Serum Triglyceride in Normal Rats Fed with 2000ppm Aluminum Diet for 67 Days. I. Feeding Young Rats Sucrose.Lactose, Milk, Casein or Soy-Protein Diets with Addition ofAluminum Chloride. SUGAWARA, C, SUGAWARA, N., KJYOSAWA, H.,AND MIYAKE, H. (1988) Fundam Appl Toxicol 10, 607–615.Aluminum (Al) compounds are widely used in drugs and food additivesbut the toxicity of such compounds is not known in detail exceptin patients with renal insufficiency (J. W. Coburn and A. C.Alfrey, 1986, Kidney Int 29, Suppl. 18). In this experiment,toxicity of ingested Al was investigated in relation to nutritionalconditions in normal rats having no renal insufficiency. Sucrose,lactose, milk, casein and soy-protein diets were prepared. Asthe Al source, aluminum chloride (A1C1³) was added to thesediets at the level of 2000 µg/g (ppm). Male weanling Wistarrats were fed for 67 days without any Al effect on body weightgain. After a half-day starvation they were terminated. Thesignificance of difference resulting from Al treatment was statisticallytested between rats consuming diet with or without added Al.Serum Al concentrations did not exceed 20 ng/ml in any of thegroups. Tibia Al concentration doubled in rats consuming addedAl in every diet but lactose. Liver Al concentration increasedsignificantly in the Sucrose, Milk, and Casein groups comparedto each Control group consuming diet without addition of Al.No lactose effect on Al accumulation was observed. With Al treatment,anemia and hypophosphatemia were not observed, but a decreasein tibia weight was observed with every diet. Aluminum-dependent decreases in serum triglyceride (TG) concentrationwere also observed in all dietary groups, without any effecton serum cholesterol or phospholipid (P-lipid) concentrations.These results indicate that dietary factors affecting Al accumulationand toxicity were small. Effects of oral Al on nutritional metabolismshould be investigated even though there was no significantincrease in serum Al Concentrations.     

Low prevalence of hepatitis C virus infection in patients with auto-immune hepatitis type 1

Hepatitis C virus (HCV) antibodies were measured in 28 patients with auto-immune hepatitis type 1 using six different assay kits, three for C100–3 antibody and three for second generation HCV antibody, and two confirmatory tests to determine the prevalence of HCV infection in auto-immune hepatitis. These patients were confirmed to have human leucocyte antigen DR 4 or 2 which is susceptible to auto-immune hepatitis in Japanese. Of the 28 patients, four (14.3%) were positive for HCV antibody in all assays and reacted positively in at least one of the two confirmatory tests, indicating a true positive finding. Eight were positive for HCV antibody only by the Ortho ELISA kit and were negative in both confirmatory tests. The cut-off level for these results was low and became negative soon after the patients received corticosteroid treatment. Thus, these eight patients are presumed to be false-positive reactors. Hepatitis C virus RNA was detected in the serum of two of the four patients with HCV antibody and in none of 24 patients without HCV antibody. No significant difference was observed between the patients with and without HCV antibody in terms of clinical background, liver function tests and auto-antibodies. Our results showed that the prevalence of a past or present HCV infection in patients with auto-immune hepatitis in Japan is low; thus, auto-immune hepatitis is thought to be distinct from hepatitis type C. However, it is also suggested that HCV infection can potentially trigger auto-immune hepatitis.     

Development of hepatocellular carcinoma in a man with auto-immune chronic active hepatitis

A 57 year old man with auto-immune chronic active hepatitis, regularly treated with immunosuppressive therapy, had hepatocellular carcinoma (HCC) 10 years after diagnosis of the hepatitis. Assays of the hepatitis C virus antibodies against capsid and non-structural proteins revealed seronegativity in serial serum samples of this patient stored in the previous 10 years during follow up. The seronegative hepatitis C antibodies excluded hepatitis C virus as the cause of the HCC. The occurrence of HCC in this case suggests the necessity of surveillance for early detection of liver cancer in patients with auto-immune chronic active hepatitis undergoing long-term immunosuppressive therapy.     

Immunological responses against an autologous human hepatocellular carcinoma cell line

We performed a detailed analysis of immune responses in a hepatocellular carcinoma (HCC) cell line and effector cells obtained from a patient with HCC. We examined the cytotoxic activity of natural killer (NK) cells, lymphokine-activated killer (LAK) cells and cytotoxic T lymphocytes (CTL) against an autologous tumour cell line (SUHC-1) to investigate the immune mechanism of human lymphocytes against HCC cells. Cytotoxic T lymphocytes were induced by co-culturing of peripheral blood lymphocytes (PBL) and SUHC-1 cells, mixed lymphocyte and tumour cell culture (MLTC). The susceptibility of SUHC-1 to NK and LAK cells was similar to that of other allogeneic cell lines, such as K562, PLC/PRF/5 and Mahlavu. Effector cells induced in the primary MLTC had high cytotoxic acitivity but were not specific for SUHC-1. Cytotoxic T lymphocytes with specific activity against SUHC-1 were induced after PBL were stimulated five times at 7–10 day intervals with SUHC-1 and low-dose recombinant interleukin-2 (rIL-2), suggesting that as the culture progressed, broadly reactive effector cells disappeared and specific effector cells survived. The specific effector cells were identified as CD3⁺/CD4⁺ and CD⁺/CD8⁺ T-lymphocyte subsets. The recognition mechanisms of CD3⁺/CD4⁺ CTL remain unresolved because the cytotoxicities were not inhibited by anti-CD4 and anti-major histocompatibility complex (MHC) class II monoclonal antibodies (MoAb). Treatment of cells with anti-CD3, anti-CD8 and anti-MHC class I MoAb partially inhibited lysis. These results demonstrated that the T-cell receptor (TCR)/CD3 complex appeared to be involved in SUHC-1 specific antigen recognition and antigen recognition of CD3⁺/CD8⁺ CTL was MHC class I restricted.     

Sarcomatous Renal Tumor in A Child: A Case Report

A patient, a 7-yr 4-mo-old boy, with sarcomatous renal tumorwas presented. At the age of 3 yr and 1 mo, he had a right nephrectomyfor a renal tumor, group I, which was histologically evaluatedas an abortive subtype of the nephroblastic type of nephroblastoma.For 24 mo after surgery, the outlook for the patient seemedto be favorable. When he was 5 yr and 2 mo old, he complainedof pain and swelling of the left shoulder. By X-ray examination,an osteolytic mass in the left scapula was discovered. Afterbiopsy, it was histologically diagnosed as a metastasis of thesarcomatous renal tumor. During the next 29 mo, multiple bonemetastases appeared in the ribs, vertebrae, skull and long bonesand slowly increased in size. ⁶⁰Co irradiation of a total of 12,015 rads to four differentregions and various chemotherapy regimens consisting of actinomycinD (AMD), AMD+vincristine (VCR), VCR+adriamycin (ADR)+cyclophosphamide(CYD), vinblastine+6-mercaptopurine and VCR+AMD+CYD, were givenfor the 53 mo from surgery to death, but they were not effectiveagainst this tumor. The clinical course of this case may show the natural historyof sarcomatous renal tumor.     

Trimethadione tolerance test for the quantitative assessment of liver function in rats

The quantitative estimation of the hepatic functional volume in rats was attempted using the serum dimethadione (DMO)/trimethadione (TMO) ratio in a single blood sampling after oral administration of TMO, which we call the TMO tolerance test, in order to develop a means of pre-operatively assessing hepatic resectability. Serum DMO/TMO ratios correlated well with the total amount of the hepatic microsomal TMO-N-demethylase activity (enzyme activity) and with the remnant liver weight after 37% and 68% partial hepatectomy. These ratios increased after partial hepatectomy in parallel with the changes in the enzyme activity and the remnant liver weight. The results suggest that the quantitative functional changes of the remnant liver after hepatectomy in both CCl4-treated and untreated rats can be estimated pre-operatively by the TMO tolerance test.     

Ultrastructural studies of hepatocyte cytoskeleton in experimental cholestasis by quick-freezing and deep-etching method

The ultrastructural association between the cytoskeleton and other organelles was studied by the quick-freezing and deep-etching method in rats treated with alpha-naphthylisothiocyanate (ANIT), or phalloidin, and in rats with obstructive jaundice. Cytoplasmic filaments were classified by measuring their diameters, and actin filaments were identified by specific decoration with myosin subfragment 1 (S1). S1-positive actin filaments and S1-negative intermediate filaments (12-14 nm in diameter) were observed to form a three-dimensional network around bile canaliculi, and were more numerous than in controls, not only in phalloidin-treated rats and rats with obstructive jaundice, but also in ANIT-administered rats. In all cholestatic rats, vesicular structures were also more numerous than in controls in the pericanalicular regions, and were closely associated with the microfilaments and the intermediate filaments. Filaments of a new type were localized between the lamellae of rough-surfaced endoplasmic reticulum and mitochondria, and between the lamellae of Golgi sacs and vesicles. Other thin filaments were also observed within the network of actin filaments. These filaments were 4-6 nm in diameter on replica membranes and were never decorated with S1. They were also directly connected with the canalicular membranes. Cytoskeletal components associated with membrane-bound organelles, including these new filaments, were suggested to be involved in the localization and migration of organelles.