Studies on cellular immunity against bile proteins in primary biliary cirrhosis by the leukocyte migration inhibition test (microdroplet method).

Cellular immunity against human bile proteins was investigated by the leukocyte migration inhibition test (LMIT) with 13 primary biliary cirrhosis (PBC) patients, 10 chronic aggressive hepatitis (CAH) patients and 21 healthy adults. Hepatic bile taken from patients operated on for lithiasis of the biliary tract was fractionated into five fractions with Sepharose 6B gel. A subtoxic dose of each fraction was determined in the healthy adults, and used as the antigen for LMIT. Out of the 5 fractions, only the third fraction led to an LMIT positive response in 8 out of 11 (73%) PBC patients and in 1 out of 10 (10%) CAH patients. The difference between PBC and CAH was significant (p less than 0.005). The remaining 3 PBC patients with LMIT negative responses were all under D-penicillamine treatment. Antibody to each fraction was prepared in rabbits. Using the antibodies after absorption with human serum, the localization of the antigens which were present in each fraction was investigated immunohistochemically using human liver sections. The antigen to the anti-first fraction antibody was detected specifically in the epithelial cells of the bile ducts and the ductules, and the antigen to the anti-third fraction antibody was detected specifically on the membrane of the bile canalicules. The third fraction was fractionated into three fractions by Sephadex G-200 gel. Only the first of the 3 fractions showed an LMIT positive response in 3 PBC patients, and its molecular weight was determined to be about 500,000. It is concluded that PBC patients develop cellular immunity against canalicular-antigen-containing fractions but not ductal-antigen-containing ones.</p

Activity of Rho-family GTPases during cell division as visualized with FRET-based probes

Rho-family GTPases regulate many cellular functions. To visualize the activity of Rho-family GTPases in living cells, we developed fluorescence resonance energy transfer (FRET)–based probes for Rac1 and Cdc42 previously (Itoh, R.E., K. Kurokawa, Y. Ohba, H. Yoshizaki, N. Mochizuki, and M. Matsuda. 2002. Mol. Cell. Biol. 22:6582–6591). Here, we added two types of probes for RhoA. One is to monitor the activity balance between guanine nucleotide exchange factors and GTPase-activating proteins, and another is to monitor the level of GTP-RhoA. Using these FRET probes, we imaged the activities of Rho-family GTPases during the cell division of HeLa cells. The activities of RhoA, Rac1, and Cdc42 were high at the plasma membrane in interphase, and decreased rapidly on entry into M phase. From after anaphase, the RhoA activity increased at the plasma membrane including cleavage furrow. Rac1 activity was suppressed at the spindle midzone and increased at the plasma membrane of polar sides after telophase. Cdc42 activity was suppressed at the plasma membrane and was high at the intracellular membrane compartments during cytokinesis. In conclusion, we could use the FRET-based probes to visualize the complex spatio-temporal regulation of Rho-family GTPases during cell division

Light- and Electron- microscopic and Immunohistochemical Studies of Human Rhabdomyosarcomas. Comparisons Among Primary Tumors, Heterotransplants in Nude Mice, and Cultured Cells from 13 Patients

Eighteen human rhabdomyosarcomas (RMS) were transplanted into the sub-cutaneous space on the back of nude mice. Thirteen of the tumors gave rise to transplantable tumors that were further examined morphologically and immuno-histochemically. The morphology of the transplanted tumors was similar to that of the primary tumors. Immunohistochemically, five primary tumors and six transplanted tumors were reacted with both desmin and myoglobin. However, in eleven cases cultured cells derived from the transplanted tumors, which showed elongated to strap-spindle-shaped cytoplasm resembling myotubules, reacted more intensely with both myoglobin and desmin. On ultrastructural examination, six primary tumors and seven transplanted tumors were found to have myofilaments or Z-bands. However, cultured cells showed myofilaments or Z-bands in their cytoplasm in all cases examined. We concluded that, on xeno-grafting, the histologic characteristics of the primary tumor are essentially con-served, and that tumor cells under culture conditions undergo an increased differentiation of skeletal muscle. These human RMS strains in nude mice and in cell lines will provide an excellent model system for investigating the biology of RMS and for further study of the molecular events underlying the genesis of this tumor

Biochemical and morphological study on hepatotoxicity of azathioprine in rat.

Sprague-Dawley rats given azathioprine in the diet for 3 to 4 weeks developed severe liver damage. Elevations of serum alkaline phosphatase and gamma-glutamyl transpeptidase activities were associated with increased hepatic glucose 6-phosphate dehydrogenase levels and decreased liver glucose 6-phosphatase activities, i.e., conditions which were commonly observed in various hepatotoxin-induced liver injuries. Light and electron microscopic observations revealed centrolobular necrosis with large scars and the proliferation of the mitochondria and rough endoplasmic reticulum. This model could be used to study the mechanisms of azathioprine-induced liver damage and its prevention.</p

Reactivity of IgE in fish-allergic patients to fish muscle collagen

ABSTRACTBackground: In addition to parvalbumin, the well- known major allergen in fish, collagen was recently identified as a new allergen in the muscle of bigeye tuna and in the skin of several species of fish. The aim of the present study was to evaluate fish muscle colla- gens for their reactivity with IgE in fish-allergic patients and antigenic cross-reactivity.Methods: Collagen was purified from the white muscle of five species of fish (Japanese eel, alfonsin, mackerel, skipjack and bigeye tuna) by acid extraction and salt precipitation, whereas parvalbumin was purified from bigeye tuna by gel filtration and reverse- phase HPLC. The IgE reactivities to collagen and parvalbumin were examined by ELISA, whereas antigenic cross-reactivity among fish muscle collagens was investigated by ELISA inhibition experiments.Results: When 15 sera from fish-allergic patients were subjected to ELISA using bigeye tuna collagen and parvalbumin, 10 sera reacted only to parvalbumin, two reacted only to collagen, two reacted to both collagen and parvalbumin and one reacted to neither collagen nor parvalbumin. The sera containing specific IgE to bigeye tuna collagen also reacted to collagens from the other four species of fish. In the ELISA inhibition experiments, bigeye tuna collagen inhibited the binding of IgE not only to bigeye tuna collagen, but also to that from the other four species of fish, suggesting cross-reactivity among the collagens from five species of fish.Conclusions: These results demonstrate that some Japanese fish-allergic patients have specific IgE to fish muscle collagen and that fish muscle collagen is a cross-reactive allergen among various species of fish

Long-term survival in a case of hepatocellular carcinoma.

A patient with an unresectable hepatocellular carcinoma (HCC) who survived without active treatment 3 years and 8 months after histological diagnosis is described. The size of the liver, which was already quite huge at the time of diagnosis, changed little during the entire clinical observation. However, 2 months before death, his condition deteriorated rapidly following gastrointestinal bleeding due to the direct invasion of the stomach by HCC. A critical reason for the unusually long-term survival of the patient may stem from the facts that a well-differentiated and bile-producing HCC was extent in most encapsulated-tumor tissues and that liver cirrhosis was not present.</p

Inclusive dileptonic rare B decays with an extra generation of vector-like quarks

We investigate the leading effects of extending the Standard Model of electroweak interactions by an extra iso-singlet up- and down- type quark pair on various distributions and total branching ratio of the inclusive B-> X_s l^+ l^- (l =e,\mu) rare B decays. The presence of the extra vector-like down quark $D$ results in the non-unitarity of the extended quark mixing matrix V, which in turn leads to b-> s FCNC at the tree level proportional to (V^\dagger V)_{sb}. On the other hand, the effective penguin and box vertex functions are sensitive to the mass of the extra iso-singlet up quark m_U. The experimental upper bound on BR(B-> X_s \mu^+ \mu^-) is used to constrain the parameters of the model. It is shown that the shapes of the differential branching ratio and forward-backward asymmetry distribution are very sensitive to the value of the model parameters. We also calculate the CP aymmetry distribution of the dileptonic decay in the vector-like quark model. It is shown that, for a typical choice of the model parameters, asymmetries up to around 10% can be achieved for certain values of the dilepton invariant mass.Comment: 15 pages, 2 figure

Clinical and histological features of sporadic non-A, non-B hepatitis.

The incidence of hepatitis A (HA), hepatitis B (HB), and non-A, non-B hepatitis (NANBH) was 27%, 30% and 43% among 73 patients with sporadic hepatitis. Epidemiological data (geographical distribution, seasonal variation, age, sex, and occupation) were not distinguishing of the type of hepatitis. Neither intrafamilial infection nor previous contact with viral hepatitis patients could be demonstrated in the NANBH cases. Fever and jaundice were less frequent in NANBH than in HA. Maximum levels of SGPT, serum bilirubin, ZTT, and gamma-globulin were significantly lower in NANBH than in HA and HB. Ten of 29 NANBH patients (35%) presented abnormal SGPT activities for more than 6 months, and four (14%) more than 12 months. In the ten patients with prolonged courses, jaundice was more frequent and maximum levels of SGPT were higher than in patients with transient courses. Histopathologic findings were not markedly different from those of HA and HB. Bile duct damage, fatty deposition, and giant multi-nucleated cells were recognized in 6, 12, and 2 NANBH patients, respectively. There were no characteristic ultrastructural changes in NANBH.</p