Biomedical subjects

D G Scarpelli

Publications and source records attributed to D G Scarpelli.

At least 37 records · Page 2Linked to original sources

PubMed1990

Transdifferentiation of ductular cells into hepatocytes in regenerating hamster pancreas.

The transdifferentiation of regenerating pancreatic cells into hepatocytes in the Syrian golden hamster and Fischer rat is an example of the surprising plasticity of cells in the adult animal. While earlier experiments suggested that these might be derived from acinar cells, unequivocal evidence of this in both models has not been forthcoming. In this paper, we document that pancreatic ductular epithelium in the hamster is the cell of origin and that presumptive hepatocytes can be identified morphologically as early as the 3rd day after the induction of regeneration. The patterns of development of various organelles as characterized by their acquisition of liver-specific protein markers parallels those that have been reported by others during the differentiation of embryonic and postnatal hepatocytes. Colloidal gold-labeled antibody staining of carbamoyl phosphate synthetase, a liver-specific mitochondrial enzyme, and catalase, a peroxisomal enzyme, first appeared 60 hours and 4.5 days postregeneration, respectively. At these times, the morphologic features of the hepatocyte phenotype in the pancreas were ambiguous. Morphometric analysis showed that for both enzymes, the number of gold particles/organelle increased to a maximum by the 9th week. In contrast, urate oxidase, a liver-specific peroxisomal enzyme, was not identified by ultrastructural immunochemistry until the 14th day after regeneration and remained at low levels through the 9th week.

Animals↗

PubMed1989

DNA alkylation in the hamster induced by two pancreatic carcinogens.

N-Nitrosobis(2-oxopropyl)amine (BOP) and N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) alkylate DNA and other macromolecules in the liver, kidney, pancreas, and lungs when injected s.c. in the Syrian hamster. Two of the most abundant DNA adducts found were the N7-methylguanine and O6-methylguanine, which in the liver accounted for about 60% of total DNA alkylation. A third adduct which was invariably found in liver and kidneys, but could not always be detected in pancreas and lungs, was identified as N7-(2-hydroxypropyl)guanine. Quantitation of N7-methylguanine by its UV spectrum and radioactivity, following administration of single-labeled [1-14C]BOP or HPOP, showed that the specific activity of this adduct was one half that of the nitrosamine. This excludes participation of the gamma carbons of these nitrosamines in methylation reactions and indicates that intermediates in which scrambling of the alpha and gamma carbons is possible are not involved in yielding the ultimate methylating agent. Finally, a comparison of the alkylation levels caused by equivalent doses of BOP and HPOP showed that BOP targeted DNA and other cytoplasmic components of kidney, lungs, and pancreas more extensively than HPOP. Ratios of N7-methylguanine in BOP versus HPOP treated hamsters, at doses less than 40 mg/kg body weight, were 3.1 in the kidney, 7.0 in the pancreas, 3.9 in the lung, and only 3.5 in the liver. These ratios are in accordance with the greater carcinogenic potency of BOP compared to HPOP and also the different organotropic properties of the two carcinogens.

Alkylation↗

PubMed1989

Toxicology of the pancreas.

The pancreas of mammals is capable of biotransforming drugs and other chemicals and subject to toxic injury by the resultant reactive metabolites. In the case of experimental animals, especially rodents, numerous chemical toxicants have been identified for the exocrine pancreas and, to a lesser degree, the endocrine component. These include alcohol, alloxan, azaserine, dimethylbenzo[a]anthracene, ethionine, 4-hydroxyaminoquinoline-1-oxide, beta-oxidized derivatives of dipropylnitrosamine, oleic acid, and streptozotocin. Depending on the toxicant, the cellular effects span the gamut from acute injury and death, to hyperplasia, metaplasia, and malignant transformation. In contrast to the well-established toxic effects of chronic alcohol abuse on the human pancreas, its susceptibility to toxic injury by therapeutic drugs has become clinically apparent more recently. A drug should be considered as toxic only when it induces acute pancreatitis documented by clinical findings and laboratory tests; the condition improves when the drug is discontinued and worsens when the patient is challenged by readministration of the drug. At present, only 8 therapeutic drugs have fulfilled these criteria and are classified as toxic for pancreas. These are, azathioprine, estrogens, furosemide, methyldopa, pentamidine, procainamide, sulfonamides, and thiazide diuretics. Controlled exposure to cultured component cells of pancreas may be of potential value in identifying such drugs, documenting their toxicity, and allowing for biochemical and toxicologic investigation of the mechanisms involved.

Animals↗

PubMed1989

Carcinogenicity of N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine, N-nitrosobis(2-hydroxypropyl)amine and cis-N-nitroso-2,6-dimethylmorpholine administered continuously in the Syrian hamster, and the effect of dietary protein on N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine carcinogenesis.

The effect of continuous week-long administration of the three pancreatic carcinogens N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP), N-nitrosobis(2-hydroxypropyl)amine (BHP), and cis-N-nitroso-2,6-dimethylmorpholine (cis-NNDM), by a s.c. implanted osmotic pump, was examined in Syrian hamsters. HPOP at total doses of 220-250 mg/kg body weight induced ductal adenocarcinomas in the pancreas (41%), and cholangiomas (18%) and cholangiocarcinomas (18%) in the liver, 25 weeks following the initiation of treatment. Higher doses of HPOP resulted in severe hepatic injury and increased mortality (LD50 = 280 mg/kg). Cis-NNDM and BHP were less toxic than HPOP and induced pancreatic lesions at doses of 950 mg/kg. These data document that a week-long schedule of continuous administration of HPOP for the induction of pancreatic cancer compares favorably with those involving weekly injections. Application of this model to study the effect of dietary protein in HPOP-induced carcinogenicity showed that the number of cystic, intermediate and tubular complexes in the pancreas was significantly higher in animals fed a 20% as compared to an 8% protein diet 2 weeks prior to HPOP administration. Furthermore, the incidence of pancreatic adenocarcinomas and in situ carcinomas was only 13% in the hamsters fed the low-protein diet as compared to 46% in those fed the high-protein diet.

Adenocarcinoma↗

PubMed1989

Role of periductal and ductular epithelial cells of the adult rat pancreas in pancreatic hepatocyte lineage. A change in the differentiation commitment.

Development of pancreatic hepatocytes in adult rats maintained on copper deficient diet containing 0.6% trien (CuDT) has been reported recently. To elucidate the histogenesis of hepatocytes a sequential study was undertaken using morphologic, histochemical, immunochemical, in situ hybridization, and Northern blot analysis. Male F-344 rats weighing 80 to 90 g were fed CuDT for 8 weeks and returned to normal rat chow. Beginning from 4 weeks of copper depletion, there was a progressive loss of acinar cells and by 8 weeks more than 90% of the acinar tissue was lost. During this period, there was an increase in the number of adipocytes in the interstitium, and in the number of interstitial and ductular cells. Morphologic observations were confirmed by immunoblot and Northern blot analysis, in which the amount of pancreatic proteins and their mRNAs decreased between 5 and 8 weeks. During this period, a progressive increase in the level of albumin mRNA was observed. In situ hybridization, performed at 7 weeks of copper deficiency, showed localization of albumin mRNA over interstitial and ductular cells. Pancreatic hepatocytes were identified immediately after the rats were returned to a normal diet and gradually increased in number. The hepatocytes occupied almost 60% of the pancreatic volume by 8 weeks. During the early recovery phase, hepatocytes were identified in ductules as well as in the interstitium. Based on these studies, it is concluded that both the ductular cells and interstitial cells, which resemble oval cells of liver, are capable of transforming into pancreatic hepatocytes and these cells may be considered stem-cell equivalent.

Animals↗

PubMed1989

Histopathologic and flow cytometric analysis of adenomatous colonic polyps.

We evaluated the histopathology, DNA content, and proliferative activity of colonic polyps independently. Paraffin-embedded specimens were used as source material. In each case, additional sections were cut at 3 microns and stained with hematoxylin-eosin and trichrome for histopathologic analysis. For DNA analysis and measurement of proliferative activity, the polyp parts were dissected and the nonpolypoid tissue was discarded. The study was limited to those specimens that were received in our department in the years 1972 and 1977. Of the 104 polyps that were submitted for flow cytometric analysis, 36 could not be analyzed owing to excessive debris or insufficient nuclei. DNA aneuploidy was identified in 32% of the cases, with a higher value noted in larger polyps and in severely dysplastic polyps, but these values were not statistically significant. Multiple adenomas from the same patient often showed different DNA histograms. When analyzed according to the percentage of cells in S phase, no significant difference was found in proliferative activity of polyps according to DNA content or size of the polyps. These results suggest that the diagnostic significance of aneuploidy and proliferative activity in polyps must be interpreted with caution.

Adenoma↗

PubMed1988

Almost total conversion of pancreas to liver in the adult rat: a reliable model to study transdifferentiation.

Study of transdifferentiation provides an excellent opportunity to investigate various factors and mechanisms involved in repression of activated genes and derepression of inactivated genes. Here we describe a highly reproducible in vivo model, in which hepatocytes are induced in the pancreas of adult rats that were maintained on copper-deficient diet containing a relatively non-toxic copper-chelating agent, triethylenetetramine tetrahydrochloride (0.6% w/w) for 7-9 weeks and then returned to normal rat chow. This dietary manipulation resulted in almost complete loss of pancreatic acinar cells at the end of copper-depletion regimen, and in the development of multiple foci of hepatocytes during recovery phase. In some animals, liver cells occupied more than 60% of pancreatic volume within 6-8 weeks of recovery. Northern blot analysis of total RNA obtained from the pancreas of these rats revealed the expression of albumin mRNA. Albumin was demonstrated in these pancreatic hepatocytes by immunofluorescence. The advantages of this model over the previously described models are: a) low mortality (10%), b) depletion of acinar cells, and c) development of multiple foci of hepatocytes in 100% of rats.

Animals↗

PubMed1988

Lack of expression of glutathione-S-transferase P, gamma-glutamyl transpeptidase, and alpha-fetoprotein messenger RNAs in liver tumors induced by peroxisome proliferators.

Many structurally unrelated nonmutagenic peroxisome proliferators induce altered areas, neoplastic nodules, and hepatocellular carcinomas in rats. Unlike the lesions induced by genotoxic hepatocarcinogens, these lesions do not stain positively for the phenotypic markers gamma-glutamyl transpeptidase (GGT) and glutathione-S-transferase P (GST-P). To ascertain whether the absence of immunocytochemically detectable GST-P and GGT proteins in peroxisome proliferator-induced neoplastic lesions is due to the absence of specific mRNAs, we analyzed the total RNA isolated from hepatocellular carcinomas induced by three different peroxisome proliferators (ciprofibrate, Wy-14643, and BR-931) and the genotoxic carcinogens, 2-acetylaminofluorene and aflatoxin B1 (AFB), for the presence of GST-P, GGT, and alpha-fetoprotein (AFP) mRNAs. Northern and dot blot analysis of total RNA isolated from liver tumors induced by three different peroxisome proliferators revealed no detectable GST-P, GGT, and AFP mRNAs. GST-P mRNA was also not detected in a transplantable hepatocellular carcinoma established from a liver tumor induced by ciprofibrate. In contrast, GST-P mRNA levels were high in primary liver tumors induced by both 2-acetylaminofluorene and AFB and the two transplantable hepatocellular carcinomas established from such tumors. By immunoblot method, GST-P protein was found to be abundant in both primary and transplantable liver tumors induced by genotoxic carcinogens but not in those derived from peroxisome proliferator treatment. The GGT and AFP mRNAs were also not found in all 18 liver tumors induced by peroxisome proliferators that were analyzed and also in the ciprofibrate-derived transplantable liver tumor. The expression of GGT and AFP genes in liver tumors induced by 2-acetylaminofluorene and AFB was variable. These studies with peroxisome proliferators show that the GST-P and GGT gene derepression is not essential for the hepatocarcinogenesis or successful tumor transplantation. Further characterization of the molecular basis for the differential expression, particularly of the GST-P gene in liver tumors, may help identification of the critical event(s) in hepatocarcinogenesis by genotoxic carcinogens and nongenotoxic peroxisome proliferators.

Animals↗

PubMed1988

Development of hepatocytes in the pancreas of hamsters treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Transdifferentiation is a process in which one differentiated cell type is converted to another. A unique example of transdifferentiation is the development of hepatocytes from pancreatic cells in adult hamsters and rats. In this communication we report the induction of pancreatic hepatocytes in hamsters that were given 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Two or 6 intraperitoneal (ip) injections of TCDD at a dose of 100 micrograms/kg body weight at 4-wk intervals induced pancreatic hepatocytes in 75% and 89% of the animals respectively. In animals given only two doses of TCDD each pancreas contained one to two hepatic foci, whereas when six injections were administered multiple hepatic foci were observed. By hematoxylin and eosin stain and by periodic acid Schiff stain, the pancreatic hepatocytes were morphologically identical to those in normal liver. Although the exact mechanism by which TCDD induces the transformation is not clear, it is conceivable that TCDD acting through receptor-mediated mechanisms is activating the repressed liver-specific genes in the pancreas.

Animals↗

PubMed1988

Uptake of N-nitrosobis(2-oxopropyl)amine by isolated rat and hamster hepatocytes: species differences and evidence for an active carrier-mediated transport process.

The rates of uptake of the carcinogen N-nitrosobis(2-oxopropyl)amine (BOP) by hepatocytes isolated from Fischer rats and Syrian hamsters were determined in order to investigate species differences in cellular transport of the carcinogen. Initial rates of uptake of [1-14C]BOP by hepatocytes were measured using a rapid centrifugation technique. At cell densities from 1.5 to 6 x 10(6) cells/ml, initial rates of uptake were as much as 4-fold more rapid in hamster hepatocytes than in those of the rat. The cell/medium distribution ratio for hamster hepatocytes reached a value of 9.0 after a 20-min incubation with an extracellular BOP concentration of 20 microM. Under the same conditions, the cell/medium distribution ratio for rat hepatocytes was only 2.4. These results indicated that BOP uptake proceeded against a concentration gradient and was more rapid in hamster hepatocytes. In both species, the rates of uptake were saturable with increasing concentration (2-685 microM) and displayed biphasic kinetics characteristic of high-affinity (Km less than 20 microM) and low-affinity (Km greater than 30 microM) processes for the uptake of BOP. Evidence for the involvement of an ATP-dependent active carrier-mediated transport process was obtained from experiments in which hepatocytes were preincubated with metabolic inhibitors. Significant inhibition of uptake was observed in the presence of KCN, carbonyl cyanide-3-chlorophenylhydrazone, antimycin A, oligomycin and other agents which interfere with electron transport or ATP generation. Based on the reduction in uptake rates, rat hepatocytes were more sensitive to the effects of these inhibitors. These results suggest that the entry of BOP into hepatocytes is under cellular regulation and that the more rapid rate of uptake in liver cells of the hamster may be one factor responsible for the observation that BOP is a more potent hepatotoxin and carcinogen in this species.

Animals↗

PubMed1988

Carcinogenicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the Syrian golden hamster.

The carcinogenic potential of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was examined in Syrian golden hamsters, at different dose levels. Twenty-one percent of the hamsters that received a total of 600 micrograms/kg body weight of TCDD either by the s.c. or i.p. route developed squamous cell carcinomas of the skin of facial region within 12-13 months from the beginning of the experiment. Neoplasms were not observed in any other organs. These studies suggest that TCDD may be a complete carcinogen in hamsters, the species most resistant to the toxic effect of this compound.

Animals↗

PubMed1988

Comparative histopathology of the development of selected neoplasms of the liver, pancreas, and urinary bladder in rodents.

The valid extrapolation of carcinogenesis data from one species to another depends, in part, on strong similarities of the metabolic and cellular mechanisms involved in the carcinogenic process and similarities in the nature and behavior of the various lesions that appear during the development of neoplasia between the species involved. Although there are many biological differences between the various rodent species used in carcinogenesis research, there are more similarities, in keeping with the surprising unity of basic cellular and tissue organization and function that is evident throughout biological systems at every level of evolutionary development. An understanding of intraspecies similarities and differences, especially as these modify the morphologic responses of the host to carcinogenic chemicals, is of central importance if carcinogenesis data from one species are to be used to predict carcinogenic risk in another. In this manuscript the histopathology of the various lesions that appear during chemically induced cancer of the liver, pancreas, and bladder in several rodent species has been selected to compare and contrast similarities and differences that exist among them and among the spontaneous premalignant lesions and carcinomas of these organs in humans.

Animals↗

PubMed1987

Species specificity in the metabolism of N-nitrosobis(2-oxopropyl)amine and N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine to mutagens by isolated rat and hamster hepatocytes.

The metabolic activation of the carcinogens N-nitrosobis(2-oxopropyl)amine (BOP) and N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) by Fischer rat and Syrian hamster hepatocytes was investigated in order to determine the existence of species differences in the induction of cell mutation. The conversion of BOP and HPOP into forms mutagenic to V79 cells was studied by using the hepatocyte-mediated mutagenicity assay. Mutations at the hypoxanthine:guanine phosphoribosyltransferase locus and the Na-K-ATPase locus were scored by the induction of 6-thioguanine resistance (TGr) or ouabain resistance (Ouar), respectively. Hepatocytes of both species were capable of converting BOP and HPOP to mutagens for V79 cells in a dose-dependent manner. Metabolism of BOP by rat hepatocytes resulted in higher mutation frequencies than that by hamster hepatocytes. At a BOP concentration of 240 microM, rat hepatocyte metabolism yielded 90.7 TGr mutants and 19.5 Ouar mutants per 10(5) V79 cells. At the same concentration, hamster hepatocyte metabolism of BOP yielded 54.1 TGr mutants and 13.0 Ouar mutants per 10(5) V79 cells. These results did not correlate with the known carcinogenic potency of BOP in the hamster as compared to the rat. Hamster hepatocytes carried out the catabolism of BOP to CO2 at faster rates than rat hepatocytes; therefore, the species difference in mutagenic activation was not due to a defect in BOP uptake or metabolism by hamster hepatocytes. In contrast, metabolism of HPOP by hamster hepatocytes resulted in significantly higher mutation frequencies than that by rat hepatocytes. At an HPOP concentration of 240 microM, hamster hepatocyte metabolism yielded 83.5 TGr mutants per 10(5) V79 cells; rat hepatocyte metabolism yielded only 19.8 TGr mutants per 10(5) V79 cells. This species difference in mutagenic activation correlated well with the known potency of HPOP as a carcinogen for the hamster as compared to the rat. Since hamster pancreatic cells and subcellular fractions are known to have very limited capacity to perform the metabolic activation of HPOP, the results of this study imply that liver metabolism plays an important role in the conversion of HPOP to an agent(s) which subsequently affects the hamster pancreas. The mutagenic potency of BOP versus HPOP was compared after metabolism by hepatocytes from both species. Following their metabolism by hamster hepatocytes, the two compounds were nearly equivalent in mutagenic potency. After metabolism by rat hepatocytes, BOP was significantly more potent mutagen than HPOP.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1987

Tannic acid-induced nucleolar changes in hepatocytes transplanted into syngeneic or xenogeneic host and in hepatocytes maintained in primary culture.

In this study we have investigated the effect of a single dose of tannic acid, administered s.c., on the nucleolar ultrastructure of hepatocytes transplanted into a syngeneic or xenogeneic host in order to evaluate the validity of our hepatocyte transplantation system as an in vivo alternative to the use of whole animals to test for species and strain differences to the effects of hepatotoxins. Within 4-6 h following tannic acid injection, rat hepatocytes transplanted into the anterior chamber of eye and inguinal fat pads of rat and athymic nude mouse, showed changes of nucleolar components, with separation of ribonucleoprotein containing granules into discrete dark zones. These dark areas were surrounded by light areas consisting of granular and fibrillar components of the nucleolus. These changes were identical to tannic acid-induced nucleolar alterations in the homotopic liver. Hamster and rat hepatocytes xenotransplanted into athymic nude mice also displayed prominent nucleolar alterations in response to tannic acid. The similarity and extent of nucleolar alterations observed in transplanted hepatocytes and the in situ homotopic liver cells attest to the usefulness of the hepatocyte transplantation system for the evaluation of species differences in biological response to toxic/carcinogenic effects of xenobiotics.

Animals↗

PubMed1987

Atypical acinar cell lesions of the pancreas in mice induced by 4-hydroxyaminoquinoline-1-oxide.

The carcinogenic effect of 4-hydroxyaminoquinoline-1-oxide (4-HAQO) on exocrine pancreas of mice after a single intravenous (i.v.) injection was investigated. At a dose of 24 mg/kg body weight, 100% of the mice killed at the end of 40 weeks had developed atypical acinar cell foci (AAF) in the pancreas. Each pancreas contained an average of 18.3 +/- 2.7 AAF. Cells in AAF were arranged as acini and their cytoplasm contained eosinophilic zymogen granules. The nuclei were large, round to oval, basally located and contained 1-2 prominent nucleoli. Cells in AAF showed a markedly increased mitotic activity with an index of 4 +/- 0.7/1000 cells as compared to a value of 0.25 +/- 0.25 in the acinar cell population of control pancreas. Autoradiographic studies showed a labeling index of 23 +/- 3 in AAF as compared to only 1.5 +/- 0.29 in the pancreatic acinar cells of controls. These findings indicate that 4-HAQO induces pancreatic lesions in mice which are quite similar to the acidophilic foci observed in rat pancreas and may serve as another useful model for studies of pancreatic carcinogenesis.

4-Hydroxyaminoquinoline-1-oxide↗

PubMed1987

Structural relationships of pancreatic nitrosamine carcinogens.

N.m.r. spectroscopy demonstrates that N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) exists as a mixture of four isomers, A, B, C and D, the equilibrium ratios of which are 57:8:16:19, respectively, at 25 degrees C. Two of these isomers, A and B, are rotomers of the open chain conformer, while the other two, C and D, are rotomers of the ring tautomer of HPOP and are derived from A and B, respectively, via an intramolecular cyclization reaction. A syn orientation of the nitroso and carbonyl groups favors an open chain configuration (isomer A), while an anti orientation favors cyclization of the molecule (isomer D). Two forms of HPOP (I and II) which are mixtures of isomers A and C, and D and B, respectively were separated chromatographically. These two forms interconvert to each other. The first rate order constants for the interconversion reactions were determined to be 4.7 X 10(-3) and 12.8 X 10(-3)/min, respectively. During these reactions isomers A and D interconvert via the intermediate formation of isomer C. This suggests that rotomerization of C and D is thermodynamically more favorable than rotomerization of their open-chain tautomers A and B, and suggests an intramolecular interaction between the carbonyl and nitroso groups. Isomers A and D are formed during the metabolism of N-nitrosobis(2-oxopropyl)amine (BOP) and cis N-nitroso-2,6-dimethylmorpholine (NNDM), respectively, by hamster liver microsomes and NADH or NADPH. The stereo-specificity of reduction of BOP and the hydroxylation of cis NNDM results in the formation of two slowly interconvertible isomers of HPOP. This, in combination with a possible different metabolic fate of the cyclic and open tautomers of this compound, may have a significant impact on the mechanism of activation of pancreatropic nitrosamines which share HPOP as a common metabolite.

Animals↗

PubMed1987

Species differences in the metabolism of N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine.

Hamsters and rats metabolize [1-14C]N-nitroso(2-hydroxypropyl)(2-oxopropyl)amine (HPOP) and N-nitrosobis(2-oxopropyl)amine (BOP) to yield N-nitrosobis(2-hydroxypropyl)-amine (BHP), glucuronic acid conjugates of HPOP and BHP, the sulfate ester of HPOP and 14C-labeled urea, all of which are excreted, and 14CO2 which is both incorporated in the urea cycle, and exhaled. The extent of metabolism and the ratios of these metabolites does not vary significantly with age or sex of the animal, however, marked species differences are evident in the metabolite composition of urine 6 h following administration of HPOP. Hamsters sulfate HPOP several times more rapidly, and reduce it to BHP more efficiently than rats. In contrast, the rat excretes more unchanged HPOP and its glucuronic acid conjugate than the hamster. Since sulfation and glucuronidation of HPOP may be involved in its activation and detoxication, these reactions were examined in detail in order to elucidate the reason(s) for their distinctively different contributions to its metabolism in rats and hamsters. Conjugation of HPOP with glucuronic acid and sulfate occurs in the livers of both rats and hamsters and is catalyzed by microsomal glucuronyl transferases and cytosolic sulfotransferases, respectively. The levels of glucuronyl transferase activity for conjugating phenolic compounds are comparable in the livers of two species; however, glucuronidation of HPOP is catalyzed by an isozyme the activity of which is three times greater in rat than in the hamster. In contrast to glucuronidation, sulfation of HPOP is catalyzed approximately 10 times faster by hamster than rat liver cytosol. Although rat liver can catalyze sulfation of phenolic compounds very effectively, it has low activity in sulfating aliphatic alcohols and beta-hydroxynitrosamines. Since both aliphatic alcohols and HPOP are sulfated by hamster liver cytosolic preparations and since these reactions are not significantly affected by the classic phenol sulfotransferase inhibitors, it appears that beta-hydroxynitrosamines may be sulfated by the aliphatic (hydroxysteroid) sulfotransferase isozymes. The failure of the rat to extensively sulfate HPOP in vivo may be attributed to the high Km of rat hydroxysteroid sulfotransferases for this compound. Of the four isomers of HPOP, only isomer A, in which the nitroso group is syn to the free keto group, is sulfated in vitro to an appreciable extent. The other three isomers either are not sulfated, or become unstable and decompose when they undergo such a reaction.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1987

Prognostic implications of proliferative activity and DNA aneuploidy in colonic adenocarcinomas.

Paraffin-embedded surgical specimens from 120 patients who underwent resections for primary untreated colonic adenocarcinoma were examined for proliferative activity, DNA aneuploidy, DNA index, and the proportion of aneuploid cells by flow cytometry. The results were correlated with survival times and clinical characteristics of the patients. The presence of metastases, both distant and restricted to local lymph nodes, was found to be a more potent adverse prognostic indicator than any DNA flow cytometry-derived parameter. Additional analyses were performed following stratification of patients into two groups on the basis of presence or absence of metastases. Analysis of 60 patients without metastatic involvement revealed a significant correlation between high proliferative activity, defined as more than 20% of cells in S-phase, and DNA aneuploidy. In fact, high proliferative activity was shown to be a more powerful adverse prognostic indicator in relation to survival than DNA aneuploidy in these cases after multivariate analysis. These results suggest that differences in proliferative activity may be an important biologic factor operative in the variable prognosis seen in colonic adenocarcinoma. In addition, they re-emphasize the importance of stratifying patients into groups based on metastatic involvement when evaluating other possible prognostic features in this disease.

Adenocarcinoma↗