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A K Roy

Publications and source records attributed to A K Roy.

At least 127 records · Page 7Linked to original sources
PubMed1988

Hydrocarbon-induced hyaline droplet nephropathy in male rats during senescence.

Male rats administered unleaded gasoline rapidly develop nephropathy characterized by accumulation of hyaline droplets in cells of the proximal convoluted tubules (PCT). This acute response is implicated in development of renal carcinoma in male rats exposed chronically to wholly volatilized gasoline. A major constituent of hyaline droplets is alpha 2 mu-globulin, a protein of hepatic origin for which the rate of synthesis declines during aging. Little information, however, is presently available on possible age-dependent susceptibility of male rats to hydrocarbon-induced nephropathy. In kidneys of untreated male Fischer 344 rats the number of constitutive hyaline droplets declined progressively with increasing age. Electrophoresis of renal cortical homogenates revealed a protein with Mr about 18 X 10(3), probably alpha 2 mu-globulin, in young (3.5 months old) male rats and total absence of this protein in aged (26 months old) males. RIA confirmed that constitutive levels of renal and hepatic alpha 2 mu-globulin in old rats were less than 1.5% of those in young adults. Unleaded gasoline (0.4 ml/kg/day, po, 5 days) caused accumulation of hyaline droplets in renal PCT of 3.5-month-old males accompanied by a marked increase (about twofold) in the renal content of alpha 2 mu-globulin, whereas the same treatment was without effect in 26-month-old rats. Finally, in the renal cortex of young rats the activities of the lysosomal proteases cathepsin B and D were increased following gasoline administration, presumably in response to protein accumulation. However, in 26-month-old rats cathepsin B activity was unaffected, while cathepsin D was increased by gasoline administration. Thus, we conclude that animal age is an important determinant in the development of hydrocarbon-induced nephropathy and only rats which produce large amounts of alpha 2 mu-globulin are susceptible to development of this pathology.

Aging↗
PubMed1988

Leupeptin-mediated alteration of renal phagolysosomes: similarity to hyaline droplet nephropathy of male rats exposed to unleaded gasoline.

alpha 2u-Globulin, a protein of hepatic origin found in the urine of male rats, is accumulated in the kidney cortex during exposure to unleaded gasoline and has been implicated in the development of fuel hydrocarbon-induced nephropathy and renal neoplasia. The principal morphological feature of gasoline-induced nephropathy is accumulation of hyaline droplets (enlarged secondary lysosomes or phagolysosomes) in epithelial cells of the proximal convoluted tubule S1 and S2 segments. Inhibition of cathepsin B (a major lysosomal peptidase) by treatment of male rats with leupeptin causes rapid accumulation of phagolysosomes and alpha 2u-globulin in the kidney very similar to gasoline exposure. Further, the renal cortical subcellular distribution of alpha 2u-globulin, determined with an electron microscopic immunochemical method, is almost totally confined to phagolysosomes following administration of either gasoline or leupeptin. These results, taken together, indicate that the mechanism of nephrotoxicity of gasoline involves inhibition of renal phagolysosomal proteolysis.

Alpha-Globulins↗
PubMed1988

Induction of theophylline toxicity and inhibition of clearance rates by ranitidine.

PURPOSE: Theophylline is metabolized by the hepatic microsomal oxidase system, as is ranitidine, although the latter has a much lower affinity for the system. The incidence of theophylline toxicity is rare when the two drugs are administered simultaneously. We observed the development of clinical and chemical toxicity in three elderly patients receiving oral theophylline therapy for chronic obstructive pulmonary disease after the administration of ranitidine therapy for peptic ulcer disease. PATIENTS AND METHODS: Three patients with chronic obstructive pulmonary disease who had been receiving theophylline for prolonged periods of time were studied. Two patients presented with complaints of epigastric pain, which was attributed to acid-peptic disease; the other patient presented with hematemesis, which was shown to be due to gastric and duodenal ulcers. Ranitidine was administered for treatment of acid-peptic disease. Theophylline clearance rates were determined before, during, and after ranitidine treatment. When symptoms of theophylline toxicity developed, the dose of theophylline was either stopped or reduced. Subsequently, patients were readministered their usual dose of theophylline. RESULTS: The effect of simultaneous administration of both drugs resulted in similar reductions in theophylline clearance rates. Serum theophylline levels returned to pretreatment values after cessation of ranitidine treatment. In one patient, rechallenge with ranitidine during steady-state theophylline treatment resulted in recurrence of clinical and chemical theophylline toxicity. CONCLUSION: These observations suggest that treatment with ranitidine may cause clinically apparent interactions with theophylline since both drugs are metabolized by the same cytochrome P-450 isozymes.

Aged↗
PubMed1988

Rapid postexposure decay of alpha 2u-globulin and hyaline droplets in the kidneys of gasoline-treated male rats.

Unleaded gasoline induces nephropathy, characterized by rapid accumulation of hyaline (protein resorption) droplets in epithelial cells of the renal proximal convoluted tubules, only in male rats. The hepatic synthesis of the male rat-specific protein alpha 2u-globulin, a constituent of renal hyaline droplets, is unaltered by gasoline treatment (Olson et al., 1987). Renal alpha 2u-globulin content increased to 210% of control within 18 h of a single oral dose of gasoline (2.0 ml/kg); maximal levels (320% of control) were attained following gasoline administration for 3 d. Increases in renal alpha 2u-globulin caused by gasoline were accompanied by concurrent proliferation of hyaline droplets. However, within 3 d of terminating gasoline administration renal alpha 2u-globulin content decreased to the same level as that in unexposed rats, although renal hyaline droplet number returned to pretreatment levels somewhat more slowly. The conjoint effect of postexposure recovery and estradiol (an inhibitor of hepatic alpha 2u-globulin synthesis) administration was also determined in male rats. On postexposure d 3, 6, and 9, estradiol treatment (1 mg/kg, sc, 4 d, starting on d 9 of gasoline treatment) decreased renal alpha 2u-globulin content to 75%, 59%, and 48%, respectively, of that in rats allowed to recover from gasoline with no hormone treatment. Hepatic alpha 2u-globulin content in estradiol-treated rats was decreased by 74%, 97%, and 96% at the same intervals. Estradiol treatment during recovery from gasoline also appeared to increase the removal of accumulated hyaline droplets from the renal cortex. Thus, accumulation of alpha 2u-globulin-containing hyaline droplets after subacute exposure of male rats to gasoline is rapidly reversible, dependent on continuous exposure to gasoline and maintenance of the normal rate of hepatic alpha 2u-globulin synthesis. These results emphasize the dynamic state of renal cortical hyaline droplets and suggest strongly that gasoline hydrocarbons cause hyaline droplet accumulation by prolonging the half-time of degradation of alpha 2u-globulin.

Alpha-Globulins↗
PubMed1988

Metabolism of K-region derivatives of 1-nitropyrene by rat liver in vitro.

The metabolism of K-region derivatives of 1-nitropyrene was studied in order to provide insight into factors that may contribute to their mutagenic activities and to obtain information on unknown metabolites of 1-nitropyrene. Using 9000 g supernatant from livers of Aroclor-treated rats, 1-nitro-pyrene-4,5-diol, a mutagenic metabolite of 1-nitropyrene, was metabolized to 1-aminopyrene-4,5-diol, a mixture of 1-nitropyrene-4,5,9,10-tetraols, 1-amino-4,5-pyrenedione and 1-nitro-4,5-pyrenedione. 1-Nitro-5H-phenanthro[4,5-bcd]pyran-5-one, a highly mutagenic lactone, was not detected. The metabolism of 1-nitro-4,5-pyrenedione yielded only 1-amino-4,5-pyrenedione; the lactone was not observed. No metabolites were detected when the lactone was incubated under conditions identical to those employed for 1-nitro-4,5-diol and 1-nitro-4,5-pyrenedione. Upon re-examination of the metabolism of 1-nitropyrene, we were able to detect 1-nitropyrene-4,5,9,10-tetraol, 1-amino-4,5-pyrenedione and 1-nitro-4,5-pyrenedione as minor metabolites in addition to the major metabolites reported previously. The results of this study, combined with the mutagenicity data for the K-region derivatives of 1-nitropyrene, suggest that nitroreduction of 1-nitropyrene-4,5-diol and 1-nitro-4,5-pyrenedione to the corresponding hydroxylamines is an important pathway for their metabolic activation in Salmonella typhimurium and their metabolic activation in Salmonella typhimurium and possibly in mammalian systems.

Animals↗
PubMed1988

Aflatoxin contamination of some common drug plants.

This paper deals with the natural occurrence of aflatoxins in some common drug plants collected from storehouses in Bihar, India. Of 15 samples analyzed, 14 were aflatoxin positive. The highest level of aflatoxin contamination was detected in the seeds of Piper nigrum (1.20 micrograms/g), followed by the level detected in the seeds of Mucuna prurita (1.16 micrograms/g), and the lowest level was detected in the bark of Acacia catechu (0.09 micrograms/g). Of 158 isolates of Aspergillus flavus obtained from as many samples of drug plants, 49 were found to be toxigenic. The amount of aflatoxin B1 elaborated by the toxigenic isolates was in the range of 0.86 to 5.24 micrograms/ml of culture filtrate.

Aflatoxins↗
PubMed1987

Cloning and expression of the rat liver cDNA for peroxisomal enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase in lambda GT11. Transcriptional regulation of enzyme activity by Wy-14643 in primary cultures of rat hepatocytes.

Proliferation of rat liver peroxisomes by the hypolipidemic drug Wy-14643 is associated with a concomitant induction of peroxisomal enzymes involved in the beta-oxidation of fatty acids. In order to explore the molecular mechanism of this induction process we have cloned the cDNA for the peroxisomal bifunctional enzyme enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase (ECH) in the lambda gt11 expression vector. The library was screened with the monospecific rabbit antiserum to ECH. Hybrid-selected-mRNA translation established that the immunoreactive clones contain the cDNA sequences of the ECH bifunctional enzyme. The cloned cDNA was used to define the early events associated with enzyme induction in primary cultures of rat hepatocytes. Dot-blot hybridization of the total hepatocyte RNA with the ECH cDNA probe showed that the ECH mRNA begins to rise at about 10-15 h following incubation with Wy-14643. At 24 h and 48 h of incubation the stimulation of the ECH mRNA over the vehicle-treated control reached 26-fold and 47-fold respectively. Run-off experiments in the isolated nuclei of hepatocytes showed no increase in the transcription rate of the ECH gene at 5 h after drug treatment and a 2-fold and 11-fold increase at 10 h and 20 h of drug treatment. From these results we conclude that the increase in ECH activity by Wy-14643 is due to an enhancement of the rate of transcription of the ECH gene. However, the relatively long lag period of about 10-15 h after exposure of hepatocytes to Wy-14643 suggests that the induction of the ECH mRNA may involve an indirect effect of the drug on the transcription of this gene.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1987

Cytoplasmic androgen binding protein of rat liver: molecular characterization after photoaffinity labeling and functional correlation with the age-dependent synthesis of alpha 2u-globulin.

The liver of the mature male rat contains a moderate affinity (Kd = 10(-8)M), low-capacity, cytoplasmic androgen binding protein (CAB) whose appearance during puberty and disappearance during senescence correlate with the androgen-dependent synthesis of alpha 2u-globulin. Molecular properties of CAB were examined by photoaffinity labeling with tritiated methyltrienolone (R-1881), a synthetic androgen, and by its localization within the hepatocytes which are competent to produce alpha 2u-globulin. Photoaffinity labeling of the liver cytosol derived from postpubertal male rats, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography, showed a predominant androgen binding band corresponding to Mr 31,000. This 31-kilodalton (kDa) binding component was conspicuously absent in the liver of androgen-insensitive prepubertal and senescent male rats and in adult male rats treated with estradiol-17 beta. In addition, unlike the cytoplasmic extract, the nuclear lysate of the male rat hepatocytes did not contain the 31-kDa androgen binder. Disappearance of the 31-kDa androgen binding band from the cytosolic fraction of androgen-insensitive animals was associated with a concomitant appearance of a minor androgen binding component of apparent Mr 29,000. The livers of postpubertal male rats normally contain two subpopulations of hepatocytes, only one of which is highly active (competent) in alpha 2u-globulin synthesis. Separation of these two subpopulations through a fluorescence-activated cell sorter followed by whole cell labeling showed more than a 2-fold higher uptake of R-1881 by the competent cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Affinity Labels↗
PubMed1987

Molecular cloning and characterization of cDNA for androgen-repressible rat liver protein, SMP-2.

SMP-2 is a rat liver protein whose synthesis is influenced by both androgens and aging. The steady-state level of its mRNA is repressed by the androgen. Compared to the adult male, SMP-2 mRNA is found in higher amounts in the prepubertal and senescent male rat livers which show relative androgen insensitivity. A cDNA library in the plasmid pBR322 was constructed from the female rat liver which contains a high level of SMP-2 mRNAs. Recombinant plasmids were screened by differential colony hybridization to 32P-labeled single-stranded cDNAs from adult female and adult male hepatic poly(A)+ RNAs. From a total of 3500 recombinant clones, 11 highly female specific clones were identified. From these female specific colonies the SMP-2 cDNA-containing plasmid (pSP11) was identified by its ability to select an mRNA species whose translation product is immunochemically and electrophoretically indistinguishable from SMP-2. This insert represents a 571-base pair portion of the SMP-2 cDNA. Rescreening of the library at a high colony density using the 32P-labeled cDNA insert of pSP11 identified several positive clones with larger inserts. Hybrid-selected mRNA translation again confirmed these clones to carry SMP-2 cDNA sequences. The plasmid pSP4a containing a 1040-base pair cDNA insert of SMP-2 was characterized by DNA sequence analysis. The size of the cDNA insert of pSP4a is close to the estimated size of the SMP-2 mRNA. The cDNA sequence provides an open reading frame of 282 amino acid residues. A comparison of the translated amino acid sequence with the protein sequences of NBRF-PIR, PSQNEW, and LOSALA data bases did not establish any sequence homology with known proteins. Northern blot analysis using the 32P-labeled cDNA insert of pSP4a confirms the androgenic repression of the SMP-2 mRNA.

Amino Acid Sequence↗
PubMed1987

Androgenic regulation of hepatic gene expression.

Androgen-dependent synthesis of alpha 2u globulin in the rat liver has been used in our laboratory as a model for studying the effect of sex hormones on hepatic gene expression. alpha 2u Globulin is a group of low molecular weight (Mr approximately 18,000) male specific urinary proteins synthesized and secreted by hepatocytes. In the male rat hepatic synthesis of alpha 2u globulin begins at puberty (approximately 40 days), reaches a peak level (approximately 20 mg/day) at about 75 days and declines during old age. Androgens can induce alpha 2u globulin in ovariectomized female rats in vivo and in the liver perfusion system in vitro. However, both prepubertal and senescent (greater than 800 days) male rats not only do not produce alpha 2u globulin but are also refractory to androgen administration. alpha 2u Globulin is coded by a multigene family comprising about 20-30 gene copies per haploid genome. All of these gene copies seem to express translationally active mRNAs giving rise to individual isoforms of alpha 2u globulin. Appearance and disappearance of the cytoplasmic androgen-binding protein (CAB) correlates with the androgen responsiveness of hepatocytes. Photoaffinity labeling of the hepatic cytosol shows that the biologically active binding protein, found in the cytosol of the mature male rat liver, has a molecular weight of 31 kDa. A molecular transition of the 31-kDa CAB to a biologically inactive 29-kDa form may be the basis of hepatic androgen insensitivity during prepuberty and senescence.

Alpha-Globulins↗
PubMed1987

Accumulation of alpha 2u-globulin in the renal proximal tubules of male rats exposed to unleaded gasoline.

Saturated branched-chain aliphatic hydrocarbons, found in motor fuels, induce nephrotoxicity in male rats. Treatment of male rats with unleaded gasoline (0.04-2.0 ml/kg body wt, po) for 9 days increased markedly the number and size of hyaline (protein resorption) droplets in epithelial cells of the renal proximal convoluted tubules (PCT) and enhanced cellular exfoliation at high dose levels. No other treatment-related pathological effects were observed in the glomeruli, distal tubules, or medulla. The renal content of alpha 2u-globulin, a major urinary protein of male rats, was increased maximally by about 4.4-fold after gasoline administration (1.0 ml/kg, po, 9 days); no further increase was observed at higher doses. Immunoperoxidase staining of kidney tissue sections for alpha 2u-globulin revealed large accumulations of antigen localized in many of the PCT epithelial cells which contained hyaline droplets. The hepatic content of alpha 2u-globulin and its mRNA were not altered by gasoline administration. These data show, for the first time, that alpha 2u-globulin is accumulated in the kidneys of gasoline-intoxicated male rats and sequestered specifically in some of the hyaline droplets characteristic of gasoline-induced nephropathy. A hydrocarbon-induced defect in the renal lysosomal degradation of low-molecular-weight urinary proteins, rather than increased synthesis of these proteins, appears to cause hyaline droplet accumulation.

Administration, Oral↗
PubMed1987

Sex-independent synthesis of alpha 2u-globulin and its messenger ribonucleic acid in the rat preputial gland: biochemical and immunocytochemical analyses.

alpha 2u-Globulin is the principal urinary protein of the mature male rat. The major urinary source of this protein is the liver where it is synthesized and secreted by hepatocytes under hormonal regulation. High levels of alpha 2u-globulin and its messenger RNA (mRNA) are also present in the preputial gland of both male and female rats, and neither castration nor ovariectomy significantly alters the preputial concentration of this protein and its mRNA. Per unit mass of RNA and protein, the preputial gland as compared to liver contains about 3-fold higher level of alpha 2u-globulin mRNA and about 300-fold higher level of alpha 2u-globulin. Despite a 3-fold (300%) difference in the content of alpha 2u-globulin mRNA, nuclear run-off experiments show only a 30% higher rate of alpha 2u-globulin gene transcription in the preputial gland than in the liver. Immunocytochemical analyses reveal that the liver possesses two alpha 2u-globulin cell populations, one showing higher immunoreactivity than the other. In contrast, the preputial gland contains only one type of alpha 2u-globulin containing acinar cells, and a large amount of alpha 2u-globulin accumulates in the ductal lumen. From these results we conclude that the 300% higher level of alpha 2u-globulin mRNA in the preputial gland is not due to a corresponding difference in the rate of transcription of alpha 2u-globulin gene. Such a difference may represent tissue-specific regulation at a posttranscriptional level of mRNA metabolism. Furthermore, the huge difference in the alpha 2u-globulin content of the preputial gland and the liver is primarily due to the cellular and ductal accumulation of this protein in the preputial gland vs. its rapid secretion by the liver.

Alpha-Globulins↗
PubMed1987

Rapid androgenic stimulation of alpha 2u-globulin synthesis in the perfused rat liver.

Hepatic synthesis of alpha 2u-globulin and its mRNA in the male rat is dependent on androgen, glucocorticoid, T4, insulin, and GH. Some of these hormones may act directly on the liver, while others may influence alpha 2u-globulin synthesis through indirect physiological changes. In the present study the specific role of androgen in the synthesis of alpha 2u-globulin was examined in an in vitro liver perfusion system. The addition of 5 alpha-dihydrotestosterone to the medium perfused through livers from castrated rats resulted in a rapid increase (approximately 10-fold over the vehicle control within 120 min) in the circulating level of alpha 2u globulin. Labeling with [35S]-methionine showed that the androgen-mediated increase in the circulating level of alpha 2u-globulin is due to release of the newly synthesized protein. Quantification of alpha 2u-globulin mRNA in the perfused livers with and without androgen supplementation indicated that the increased mRNA level can only partially account for the elevation of the circulating level of this protein. From these results it is concluded that androgen can act directly on the liver to stimulate alpha 2u-globulin synthesis, and the hormone may influence more than one regulatory step.

Alpha-Globulins↗
PubMed1987

Independent regulatory influence of growth hormone on the hepatic synthesis of alpha 2u-globulin.

Administration of GH through sc injections to hypophysectomized male rats induces the hepatic mRNA for alpha 2u-globulin (a male-specific urinary protein) from an undetectable level to 43.4% of the normal male level. The same treatment administered to hypophysectomized-gonadectomized rats and androgen-insensitive Tfm rats induces alpha 2u-globulin mRNA to a level of only 5-10% of that in the normal male. However, none of these types of animals shows an appreciable response when GH is administered continuously through osmotic minipumps. Perfusion of the livers derived from hypophysectomized male rats with the blood from hypothyroid rabbits (also deficient in GH) was used to examine in vitro effects of GH on alpha 2u-globulin synthesis. Supplementation of the perfusion medium with GH and T4 failed to induce alpha 2u-globulin within the perfusion period of 120 min. These results show that GH can influence alpha 2u-globulin synthesis independent of the androgen and that the mode of administration of GH plays an important role in its biological response.

Alpha-Globulins↗
PubMed1986

Partial reversal of alpha 2u globulin gene expression by thyroxine in the liver of diabetic rats.

Synthesis of alpha 2u globulin and its mRNA has been used as an index to monitor the effect of thyroxine on specific gene expression in the liver of hypoinsulinemic male rats. Administration of a physiological dose of thyroxine can partially reverse (to approximately 30% of the normal control) the marked reduction (more than 90%) in the hepatic levels of alpha 2u globulin and its mRNA during streptozotocin-induced diabetes. Estimation of newly synthesized alpha 2u globulin RNA transcripts from the native chromatin of isolated liver nuclei by "nuclear runoff experiments" showed that thyroxine can elevate the rate of transcription of alpha 2u globulin gene in the diabetic rat. Hypoinsulinemic diabetes is also found to be associated with an approximately 35% reduction in the thyroid hormone receptor level as compared to the normal control. The stimulatory effect of thyroxine on the synthesis of alpha 2u globulin and its mRNA was also evident in spontaneous diabetic Wistar "BB" rats. From these studies it can be concluded that severe hypoinsulinemia can cause a decrease in thyroid hormone action at the level of specific gene expression.

Alpha-Globulins↗
PubMed1986

Synthesis of K-region derivatives of the carcinogen 1-nitropyrene.

The synthesis of potential metabolites of 1-nitropyrene, resulting from oxidation at the K-regions, is described. Reaction of 1-nitropyrene with OsO4 gave the cis-4,5- and 9,10-dihydrodiols. These were separated and oxidized with activated MnO2 to give the corresponding 4,5- and 9,10-diones; further oxidation to the highly mutagenic lactones, 1- and 3-nitro-5H-phenanthro[4,5-bcd]pyran-5-one, was observed in these reactions. Reduction of the 4,5-dione with KBH4 gave the trans-4,5-dihydrodiol, which was identical to one of the metabolites of 1-nitropyrene. Reduction of the 9,10-dione gave an unstable trans-9,10-dihydrodiol, which was characterized as its diacetate. The u.v. spectra, n.m.r. spectra and h.p.l.c. retention times of these compounds are presented.

Carcinogens↗