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The correlation between the cytochemistry (glycoprotein, glycogen, glucose-6-phosphatase, catalase, alkaline phosphatase) and the growth rate of the fast-growing Morris hepatoma 3924A and the slow-growing Morris hepatoma 9618A was studied by utracytochemical techniques. By the chromic acid-phosphotungstic acid technique, acid glycoprotein is stained in glycocalyx, Golgi saccules and vesicles, and secretory granules of the tumor cells of both hepatomas. However, the hepatoma 3924A cells contain thicker glycocalyx and more numerous glycoprotein-rich granules than hepatoma 9618A cells. Abundant alpha and beta glycogen particles are found in hepatoma 3924A. Moderate glucose-6-phosphatase activity is observed in the cisternae of endoplasmic reticulum and nuclear envelope of hepatoma 9618A, but it is totally absent in hepatoma 3924A. High catalase activity is present in numerous peroxisomes of hepatoma 9618A. Hepatoma 3924A contains only a few catalase-positive microperoxisomes. Weak to moderate alkaline phosphatase is present in the plasma membrane and nuclear envelope of hepatoma 9618A cells, while hepatoma 3924A shows no activity of the enzyme. All the cytochemical parameters except glycoprotein show an inverse relationship with the growth rate of the hepatomas. The higher intracellular glycoprotein content of hepatoma 3924A may be related to differences in cell coat secretion (composition and activity) from the slower-growing hepatoma 9618A
Anat Rec 1982 Feb
PMID:Correlation between growth rate and cytochemistry in Morris hepatomas. 627 86

Dairy cows were injected with 1 alpha-hydroxycholecalciferol (1 alpha-HCC) and, or, cloprostenol at 275 days of gestation. Blood samples were taken daily from 270 days of gestation until seven days after parturition and analysed for calcium, inorganic phosphate, magnesium and hydroxyproline. In all treated and control cows concentrations of calcium, inorganic phosphate and magnesium decreased around the time of parturition. Concentrations of hydroxyproline increased from the second to the fourth day after parturition. This increase was slightly smaller in cows injected with cloprostenol but was unaffected by 1 alpha-HCC. There was a greater indicence of retained placenta and endometritis in cows receiving cloprostenol. The injection of cloprostenol with 1 alpha-HCC at 275 days of gestation did not prevent milk fever.
Vet Rec 1981 Oct 17
PMID:Study of combined injections of 1 alpha-hydroxy-cholecaldiferol and cloprostenol in the prevention of parturient paresis. 703 60

The efficacy of 1 alpha-hydroxycholecalciferol (1 alpha-HCC) for the prevention of milk fever has been tested in a controlled field trial using a total of 601 cows on 18 farms with a history of a high incidence of milk fever. The trial protocol proposed two doses of 1 alpha-HCC with the first injection seven days before the predicted calving date and a second injection five to seven days later if the cow had not calved. Of 301 cows receiving either one or two injections of 1 alpha-HCC, 70 had milk fever, and of 300 cows that received a placebo, 102 had milk fever. The difference in incidence between the two groups was statistically significant (P = 0.005) and indicated an overall efficacy of 32 per cent for treatment with 1 alpha-HCC. An improved efficacy was demonstrated in 67 cows that received two injections of 1 alpha-HCC with the second injection administered either on the day of calving or one day previously (60 per cent efficacy). For 42 cows that received a single injection of 1 alpha-HCC between two and five days before calving an efficacy of 79 per cent was achieved.
Vet Rec 1981 Sep 26
PMID:Field trial to determine the efficacy of two doses of 1 alpha-hydroxycholecalciferol in the prevention of milk fever. 703 76

Varying doses of 1 alpha-hydroxycholecalciferol (1 alpha-HCC) (50, 150, 250 and 350 micrograms) in propylene glycol were injected intramuscularly into 30 dry adult Israeli Friesian cows. Fourteen of these animals received a second dose; four were given 250 or 350 micrograms 48 hours after the first dose and 10 were given 350 micrograms 72 hours after the first dose. Plasma calcium rose after 24 hours at all dose levels except 50 micrograms. A dose-dependent peak in plasma calcium was reached after three to four days, followed by a return to baseline five days (150 micrograms) and eight days (250 and 350 micrograms) post injection respectively. Repeating the injection 48 or 72 hours later increased the time span by three and four days respectively. The effect of plasma inorganic phosphate was double that on plasma calcium. Plasma magnesium declined slightly three days post injection. High calcium feeding in conjunction with one or two injections of 350 micrograms 1 alpha-HCC did not modify the response of plasma calcium. An injection of 350 micrograms of 1 alpha-HCC was given once to 40 parturient paresis-prone cows of the same breed and twice at 72-hour intervals to 37 such cows. Six of the animals received 5 mg of flumethasone together with the second injection and 13 received it 48 hours later. This was to induce parturition, which occurred within 24 to 48 hours. None of the cows injected earlier than 24 hours prepartum developed parturient paresis in comparison with 22 out of 60 control animals which did. The results suggest that 1 alpha-HCC is useful in the prevention of bovine parturient paresis.
Vet Rec 1980 Jun 21
PMID:Observations of the use of 1 alpha hydroxycholecalciferol in the prevention of bovine parturient paresis. 743 21

The effect of human recombinant tumor necrosis factor alpha (h rec TNF-alpha) on the growth of Morris hepatoma 5123 implanted in the skeletal muscles of the thigh of Buffalo rats was investigated. The cytokine was repeatedly given in an intratumor administration (i.t.) in dose of 1.5 x 10(4) U once a day in regimens of four or eight days. Comparative groups consisted of animals which were given saline i.t. Control groups included healthy rats subjected to local cytokine effect. The experiments revealed an inhibitory effects of the preparation on the growth of tumors. Biometric parameters of the tumors induced indicated that the inhibition of Morris hepatoma was most effective after the eighth dose of h rec TNF-alpha. The administration of fourfold dose resulted in an initial loss of body mass increase. However, when injected eight times, the factor produced a relative tolerance reflected in minor reduction of actual body mass. The estimation of survival time in rats injected i.t. with h rec TNF-alpha, compared to those given saline, revealed statistically significant differences at the eighth repeated dose.
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PMID:Evaluation of biometric parameters of Morris hepatoma after application of human recombinant tumor necrosis factor. 761 82

The effect of the human recombinant tumor necrosis factor alpha (h rec TNF-alpha) on the transplantable Morris hepatoma 5123 was studied in Buffalo rats. The cytokine was repeatedly administered intratumorly (i.t.) in a dose of 1.5 x 10(4) U once a day in a cycle of four and eight days. The control groups consisted of animals given saline i.t. The experiments revealed an inhibitory effect of the h rec TNF-alpha upon the growth of neoplastic tumors. The biometric parameters of the tumors indicated that the inhibition of the Morris hepatoma was most effective after eight repeated doses of TNF. After injections of TNF-alpha, the tumors presented extensive hemorrhagic necrosis, the regressive alterations being found mainly in the central and intermediate tumor zones. In the early phase of the tumor growth, neoplastic tissue necrosis prevailed, as well as hemorrhages within the necrotic masses, necrosis of the blood vessel walls and thrombi in their lumina. In the later period, numerous fibres of the fibrous tissue, richly vascularized, occurred in the peripheral and intermediate zones. Clusters of eosinophilic granulocytes and macrophages with apoptotic bodies in the cytoplasm were seen on the border of the necrotic foci.
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PMID:Inhibitory effect of the human recombinant tumor necrosis factor on the growth of the Morris hepatoma in rats. 771 25

Four monoclonal antibodies were raised against crude gap junction fractions of rat liver to clarify the distribution of gap junctions during animal development and to analyze gap junction expression in vivo and the polarity of hepatocytes in vitro. Among the monoclonal antibodies obtained, HAM8 antibody recognized the 27-kDa rat liver gap junction protein connexin 32. This antibody recognized gap junctions at the contiguous faces of hepatocytes, and the antigen was also observed in exocrine pancreas and salivary gland but not in kidney, heart, esophagus, or thymus. HAM8 did not react with amphibian or fish liver, heart, esophagus, stomach, or intestine as assessed via the immunofluorescence method on frozen sections. A few hepatocytes and many hemopoietic cells were seen in rat fetal liver at 15 days of gestation. HAM8 antigen was expressed on some hepatocytes but not on any hemopoietic cells. As the fetus grew, the number of hepatocytes in the liver increased gradually, together with the amount of HAM8 antigen. The distribution of HAM8 antigen at 25 days after birth was similar to that in adult liver. When the expression of HAM8 antigen was examined in primary cultured hepatocytes using the immunofluorescence method, the antigen was observed clearly between the hepatocytes. However, most of the HAM8 antigen on the free surface of hepatocytes disappeared within 4 hr. HAM8 antigen was not expressed on AH-7974 rat hepatoma cells when they formed small islets in the rat peritoneal cavity or within the liver. When HAM8 IgG antibody was injected intravenously, the HAM8 signal was expressed in the liver.
Anat Rec 1993 Mar
PMID:Immunohistochemical analysis of rat liver using a monoclonal antibody (HAM8) against gap junction. 838 23

LEC rats spontaneously develop hepatocellular carcinoma with cholangiofibrosis after chronic hepatitis, but the mechanism of development of the hepatic injury is not clear. To investigate the role of hepatic stellate cells in induction or suppression of hepatic fibrosis, we morphologically examined the liver of LEC rats. Accumulation of copper was analyzed by the Danscher-Timm's sulfide-silver method. Histopathological changes were evaluated by hematoxylin and eosin staining, and by Masson's trichrome method. Activated stellate cells were identified by immunostaining method for alpha-smooth muscle actin. Cytological alterations of the stellate cells were investigated by transmission electron microscopy. To evaluate the lipid content in the stellate cells, we analyzed the area of lipid droplets of the cells by morphometric analysis. Also for evaluation of the changes in the number of stellate cells, the numbers of nucleated stellate cells and parenchymal cells were counted and statistically analyzed. Hepatic parenchymal cells showed excessive accumulation of copper at 5 weeks of age. Submassive necrosis was observed at 19 weeks of age. The liver of LEC rats 1.5 years of age showed cholangiofibrosis and subcellular injury of hepatic parenchymal cells. However, no diffuse hepatic fibrosis was observed in the liver, and hepatic stellate cells around the regions of cholangiofibrosis were negative for alpha-smooth muscle actin. The area of lipid droplets of a stellate cell in the liver of LEC rats was 1.6 to 1.8 times as large as that of normal Wistar rats. The hepatic stellate cells did not participate in the accumulation of collagen fibers around themselves when the cells contained a large amount of vitamin A-lipid droplets, even though the development of hepatic lesions was in progress. Our present data are consistent with our previous hypothesis that there is an antagonistic relationship between the storage of vitamin A and the production of collagen in stellate cells.
Anat Rec 2000 04 01
PMID:Storage of lipid droplets in and production of extracellular matrix by hepatic stellate cells (vitamin A-storing cells) in Long-Evans cinnamon-like colored (LEC) rats. 1073 52

Metastasis, a major factor contributing to poor prognosis of cancer patients, is caused by a complex series of events that involve many genes. To investigate this process, we analyzed by differential display three cell lines that had been established from a murine colon adenocarcinoma (colon 26), NL4, NL17, and NL22, each of which possessed a different potential for metastasis in mice. We report here the identification of a novel gene, ream (reduced expression associated with metastasis), which showed significantly lower expression in NL17 and NL22 with a high potential for metastasis than in NL4 without a metastatic potential. The human counterpart of murine ream expressed two sizes of transcript, 4.4 kb and 1.8 kb, both encoding the same 367-amino acid peptide, which appeared to contain four membrane-spanning regions. The cDNA showed no significant homology to any known genes in the public database. Human REAM was found to lie within an 800-kb segment of 8p21.3-22, where we had previously identified a commonly deleted region in colorectal and hepatocellular carcinomas. Its expression was reduced in more than half of the human colorectal cancers we examined, particularly in advanced stages with liver metastasis. Furthermore, we identified somatic mutations of this gene in a colorectal cancer, a hepatocellular carcinoma, and a nonsmall lung cancer among 111 human tumors of various stages examined.
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PMID:Isolation of a novel gene on 8p21.3-22 whose expression is reduced significantly in human colorectal cancers with liver metastasis. 1091 88

Aim of the study was to investigate the genotoxic effects of methyl isothiocyanate (MITC), a compound widely distributed in the environment as a constituent of certain vegetables, a soil fumigant and breakdown product of carbamate pesticides. MITC caused only marginal mutation induction in reversion assays with Salmonella strains TA100 and TA98 and, the maximum effect (<2-fold increase over the background rate) was seen at 100microg/ml. In differential DNA-repair assays with E. coli (strains 343/763 uvrB/recA and 343/765 uvr(+)/rec(+)), a pronounced dose-response effect (induction of repairable DNA-damage) was seen at low concentrations (>/=4microg/ml). In both bacterial assays, addition of activation mix (rat liver S-9) led to a reduction of the genotoxic effects. In micronucleus assay and in single cell gel electrophoresis assay with human hepatoma cells (HepG2), clear cut positive results were obtained at exposure concentrations of <5microg/ml. On the contrary, only marginal effects were seen in differential DNA-repair host-mediated assays where E. coli indicator cells were recovered from different inner organs of mice that had been treated orally with a high dose (90mg/kg bw) of the test compound. Further in vitro experiments showed that MITC is inactivated by body fluids (saliva, gastric juice) and that its DNA-damaging properties are attenuated by non-enzymatic protein binding. Since exposure of HepG2 cells to MITC led to formation of thiobarbituric acid reactive substances, it is likely that its DNA-damaging effects involve lipid peroxidation processes. Overall, our findings show that MITC induces only marginal effects at extremely high (almost lethal) doses in inner organs in vivo, but it causes DNA-damage at low concentrations in vitro.
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PMID:Genotoxic effects of methyl isothiocyanate. 1115 66


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