High-performance liquid chromatographic separation and quantification of the four biliverdin dimethyl ester isomers of the IX series.
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Biomedical subjects
Publications and source records attributed to B H Ruebner.
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The spontaneous hepatocellular neoplasms of C3H (MTV-ve) male mice were compared with the hepatic tumors induced in these animals by diethylnitrosamine (DEN) and dieldrin. No morphologic differences could be detected by light or electron microscopy between the spontaneous and induced lesions. However, the animals given diethylnitrosamine or dieldrin developed the lesions earlier, in greater numbers and of larger size. The earliest change was the development of foci composed of clear cells. Later nodules appeared which were composed of clear or basophilic cells. These lesions were followed by and presumably progressed to nodules of trabecular hepatocellular carcinomas. It is postulated that in this series, the first morphological step in the neoplastic transformation is the appearance of unusually clear hepatocytes. Ultrastructurally, the clear cells had increased glycogen and lipid droplets and a decrease in smooth endoplasmic reticulum. The basophilic cells seen later resembled the clear cells except for having a greatly increased rough endoplasmic reticulum. Trabecular hepatocellular carcinomas differed from benign nodules in the greater secretory activity of the rough endoplasmic reticulum, in the development of basement membranes at the vascular pole and of microvilli along the lateral cell membranes. The stepwise progression of normal hepatocytes to hepatocellular carcinoma is discussed on the basis of these sequential light microscopic and ultrastructural observations.
1. Bilirubin-IXalpha monoglucuronide was the predominant bilirubin in biles and meconiums of newborn humans and rhesus monkeys. Rhesus-monkey baby biles contained slightly more diglucuronide than did human baby biles. 2. Bilrubin-IXalpha glucoside, bilirubin-IXalpha xyloside and bilirubin-IXbeta were also constituents of human and rhesus-monkey baby biles and meconiums. Bilirubin-IXalpha glucuronide glucoside was present in human and rhesus-monkey baby biles but not in meconiums. The identity of the bilirubins was confirmed by u.v.-visible and mass spectroscopy of the azodipyrroles obtained by treating the bilirubins with diazotized ethyl anthranilate. The resulting azodipyrroles were identical with the corresponding azodipyrroles obtained from human adult biles and also from reduced isomers of biliverdin. 3. Bilirubin-IXbeta was present in much higher proportions in the extracts of meconiums than in the extracts of biles from the same babies. 4. Oxidation of bilirubins to biliverdins occurs in utero to a small but undetermined extent. The resulting green pigments were present in meconiums collected from the lower small and large intestines of newborn babies and rhesus monkeys. 5. Butanol extracted most of the bilirubins present in biles. This modified method proved to be quick and easy. Little hydrolysis of bilirubins took place during extraction or separation by t.l.c.
A young male with Buerger's disease who had previously required a left leg amputation died in renal failure and sepsis. Postmortem examination revealed an obliterative lesion of the celiac artery, which resulted in hepatic, splenic, and pancreatic infarctions. Celiac artery involvement represents an unusual manifestation of thromboangiitis obliterans.
Closed needle liver biopsy routinely provides 60-100 mg tissue from rhesus monkeys (Macaca mulatta). No mortality or morbidity was observed in over 400 needle biopsies during a two-year period in which some monkeys were biopsied over 30 times. Biopsy tissue was used as enzyme source in low-volume in vitro systems for measurement of hepatic monooxygenase activity.
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Hepatopathy associated with glucocorticoid therapy or with naturally occurring hyperadrenocorticism was detected retrospectively in hepatic biopsy specimens from 22 of 60 dogs, by histologic review. The hepatopathy was characterized by centrilobular vacuolization, perivacuolar glycogen accumulation within hepatocytes, and focal centrilobular necrosis. The predominant clinical findings were hepatomegaly, increased serum enzyme activity associated with hepatic disease, and increased bromsulphalein retention. The hepatopathy appeared to be reversible.
We report the case of a 13-year-old boy who was known to have Fanconi's anemia for five years. For treatment of this condition he was given androgens and corticosteroids. Two months before his death, severe varicella developed complicated by pneumonia, jaundice, and prolonged fever; all of which resolved during a five-week hospitalization. Three weeks later he died of Clostridium septicum sepsis caused by necrotizing enterocolitis. At autopsy he was found to have multiple hepatocellular neoplasms. A striking feature of the neoplasms was cholestasis. The liver also showed peliosis hepatis. The association of the use of certain androgenic steroids with hepatic neoplasms histologically resembling hepatocarcinomas, but characterized by lack of metastases and apparent reversibility, suggests the desirability of a new nomenclature for these hepatocellular lesions.
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Six young adult male rhesus monkeys were given diethylnitrosamine ip for 3-5 years. Liver biospies were done monthly. After 6 months, biopsy specimens showed individual hepatocytes and small foci of hepatocytes that were intensely positive for glycogen. During the second and later years, larger foci of such cells developed. In sections stained with hematoxylin and eosin, the glycogen-containing hepatocytes generally appeared unusually clear. Some hepatocytes, however, had eosinophilic or basophilic cytoplasm. Nuclear enlargement and atypic developed, particularly outside the foci. The hepatocytes within most foci were uniform in their histochemical features: glycogen was elevated, glucose-6-phosphatase was decreased, and ATPase activity was present not only along the bile canalicular surface but also along the enire cell membrane. After 3-5 years, neoplastic nodules and hepatocarcinomas developed in 5 of 6 animals. Two nodules and particularly the heptocarcinomas differed from the foci in one of more histochemical parameters. The findings suggested that the glycogen-containing, histochemically altered cells of the foci in one or more histochemical parameters. The findings suggested that the glycogen-containing, histochemically altered cells of the foci may be the first step in the development of neoplasia; further steps toward malignancy appeared to be frequently associated with additional alterations, such as loss of sinusoidal ATPase and re-formation of glucose-6-phosphatase.
Meconium of human infants and rhesus monkey infants (Macaca mulatta) contained only about 0.10 of the amount of bilirubin in gallbladder bile of the same individuals. Ninety-nine percent of the bilirubin in adult bile was conjugated. The proportion of conjugated bilirubin in infant bile and meconium was only slightly lower. Adult bile contained more bilirubin disconjugates than monoconjugates, whereas only 20% of the bilirubin in infant bile and meconium was in the form if disconjugates. The predominant azopigment in adult bile was azopigment delta (a glucuronide). Infant bile contained less azopigment delta, more azopigment alpha (azodipyrrole), and a so far unidentified conjugated azopigment (azopigment beta). Azopigment beta was also found in meconium but adult gallbladder bile contained only trace amounts of this pigment.
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Rhesus monkeys were tube-fed 100 calories per kg of a liquid diet based on casein in which 41% of the calories were derived from grain alcohol. The alcohol intake was 5.8 g per kg per day. Control diets contained isocaloric amounts of glucose. The protein content of the diet was 15% and fat supplied 21% of the calories. After 28 days the animals which had been fed ethanol developed hepatic fatty change and serum L.D.H. levels were elevated. The most striking electron microscopic changes in the alcohol animals were mitochondrial swelling, focal cytoplasmic degradation, and dilatation of the rough endoplasmic reticulum. In the monkeys which had received ethanol the metabolism of alcohol increased from 17.4 mg per 100 ml per hour to 26.6 mg and antipyrene half-life decreased from 61.0 minutes to 49.9 minutes. The carbohydrate animals showed no significant change in alcohol metabolism or antipyrene half life. The ethanol animals lost weight significantly while the carbohydrate animals gained significantly. The metabolic effects of alcohol thus were not reproduced by glucose. Administration of phenobarbital at 30 mg per kg for 5 days increased alcohol metabolism from 16.5 mg per hour to 22.5 mg per hour and shortened antipyrene half life from 76.5 minutes to 33.6 minutes. Alcohol and phenobarbital both induced enhanced drug metabolism but alcohol was a more powerful inducer of its own metabolism than phenobarbital. Phenobarbital on the other hand was a better inducer of antipyrene metabolism than alcohol.
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