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Biomedical subjects

M H Zile

Publications and source records attributed to M H Zile.

At least 37 records · Page 2Linked to original sources

Vitamin A homeostasis endangered by environmental pollutants.

Normal vitamin A function depends on adequate stores of the vitamin, a finely regulated supply of the vitamin to target tissues, and an ability of cells to generate functionally active forms of the vitamin. Both endogenous and exogenous factors can adversely affect vitamin A homeostasis. Polyhalogenated aromatic hydrocarbons are ubiquitous environmental pollutants and cause severe disturbances in vitamin A metabolism, manifested by an accelerated metabolism and breakdown of vitamin A and its metabolites and a depletion of vitamin A from the body; this sequence of events accounts for the vitamin A deficiency-like symptoms associated with PHAH intoxication. The mechanism(s) responsible for these events most likely includes altered activities of enzymes that are either directly or indirectly involved in critical vitamin A metabolic pathways. Human populations that continue to be exposed to environmental pollutants, may accumulate critical levels of polyhalogenated aromatic hydrocarbons and will be at risk for inadequate vitamin A function as well as for other health impairments that have been difficult to link to any specific causes. Therefore, it is important to seriously evaluate the similarities in physiological disturbances across species that have become apparent in studies with wildlife inhabiting polluted environments similar to ours; the relevance to human health is evident.

Animals↗

Production of a hybridoma cell line secreting retinoic acid-specific monoclonal antibody.

A stabilized hybridoma cell line secreting anti-retinoic acid monoclonal antibodies of subclass IgG1 with kappa chains was produced by fusing NS-1 myeloma cells with the spleen cells from BALB/c female mice immunized with all-trans-4-oxoretinoic acid-oxime-chicken IgG conjugate. The antibody titer of mice ascitic fluid ranged from 1/12,800 to 1/25,600, as determined by competitive indirect enzyme-linked immunosorbent assay (ELISA). 50% inhibition dosage of all-trans-retinoic acid at a 1/20,000 dilution of mice ascitic fluid was 6.6 ng/ml, as determined by ELISA. The anti-retinoic acid monoclonal antibody was generated in mice ascitic fluid and purified by protein G affinity chromatography. Cross-reactivity of the monoclonal antibody was determined at 0.1 microgram/ml concentration of retinoids and indicated high specificity to both all-trans-retinoic acid (86% inhibition) and 13-cis-retinoic acid (87% inhibition), and strong cross-reactivity with 4-oxoretinoic acid (77%) and 4-oxoretinoic acid oxime (109%). Specificity was confirmed by the horseradish peroxidase-linked immunostaining method and immunoradioassay. The affinity constant of the monoclonal antibody, K, was determined to be 3.6 X 10(9) l/mol. A calibration curve for retinoic acid using the monoclonal antibody to retinoic acid was developed; the detection limit for all-trans-retinoic acid is 1 ng/ml in the competitive indirect ELISA. The antibody counteracts the effect of retinoic acid on growth inhibition and differentiation in HL-60 cells.

Animals↗

Effect of retinoic acid and apo-RBP on serum retinol concentration in acute renal failure.

We recently demonstrated a rapid up-regulation of serum retinol-retinol binding protein-transthyretin concentration in rats with short-term acute renal failure. We examine the effect of retinoic acid and apo-retinol binding protein (apo-RBP) on the up-regulation of serum retinol in renal failure. Injection of retinoic acid (10 micrograms) into rats with acute renal failure or sham-operated rats increased circulatory retinoic acid concentration 29-fold within 2 h but did not influence serum retinol concentration in either group. Injection of a large dose of retinoic acid (100 micrograms) decreased serum retinol concentration in rats with acute renal failure (19%) and sham-operated rats (29%). These results suggest that changes in serum retinoic acid concentration within the near-physiological range have no effect on regulation of hepatic retinol release. Injection of a large dose of retinoic acid may depress serum retinol indirectly via a retinol sparing effect in target tissues. In rats with renal failure the serum retinol concentration, elevated 44-52% above that of sham-operated controls, was also increased to 70-164% above controls by the injection of 52-63 micrograms of apo-RBP. This suggests that circulatory apo-RBP can up-regulate serum retinol. Circulatory apo-RBP may be a positive physiological feedback signal from peripheral tissues for hepatic release of retinol.

Acute Kidney Injury↗

Metabolism and secretion of retinol transport complex in acute renal failure.

Tissue uptake and distribution of retinol from circulatory vitamin A transport complex was studied in order to determine the origin of the increased serum retinol in rats with short-term acute renal failure. In rats with acute renal failure, serum retinol increased 37-70% within 2 h after surgery. After an injection of donor plasma containing 1.8 muCi of [3H]retinol in retinol transport complex, in rats with renal failure the ability to clear radioactivity was decreased 36% by 0.5 h and 57% by 2 h, as compared to sham-operated rats. The uptake and distribution of radioactivity by nonrenal tissues was similar in rats with acute renal failure and with intact kidneys. The lack of renal function did not alter hepatic cycling of [3H]retinol from the circulation and thus could not account for the increased serum retinol in renal failure. When hepatic release of retinol-retinol binding protein was blocked by colchicine, the up-regulation of serum retinol, normally observed in rats with acute renal failure, was abolished. Our studies provide strong evidence that kidney has an important role in maintaining serum retinol homeostasis by influencing the release of retinol-retinol binding protein from liver into circulation. Peripheral tissue uptake of circulatory retinol and hepatic cycling of nonutilized retinol are not directly influenced by the kidney.

Acute Kidney Injury↗

Upregulation of serum retinol in experimental acute renal failure.

Serum vitamin A homeostasis was studied in rats with nonfiltering kidneys prepared by ligation of renal arteries. Within 1-2 h of acute renal failure, the serum retinol level increased by 11-73% and was maintained for at least 4 h. More than 90% of the increase in serum retinol was associated with retinol in the retinol binding protein-transthyretin (RBP-TTR) complex. The activities of acyl-CoA:retinol acyltransferase and retinyl-palmitate hydrolase were not altered by short-term acute renal failure. Oral administration of 3H-labeled retinol 3 h before surgery resulted in 350% more tritium in the serum retinol-RBP-TTR complex of rats with acute renal failure as compared to sham-operated rats; this increase represented the fraction of retinol in RBP-TTR contributed by hepatic retinol from newly absorbed 3H-labeled retinol. Total retinol in the retinol-RBP-TTR complex was increased by only 60%. We conclude that short-term acute renal failure causes rapid upregulation of serum retinol-RBP-TTR; the extent of the increase depends on the magnitude of hepatic vitamin A stores, particularly the retinol pools. We hypothesize that kidney modulates the regulation of hepatic release of retinol-RBP from the pool of newly acquired retinol.

Acute Kidney Injury↗

Effect of tetrachlorodibenzo-p-dioxin (TCDD) on the glucuronidation of retinoic acid in the rat.

Administration of a single oral dose (10 micrograms/kg) of tetrachlorodibenzo-p-dioxin (TCDD) caused a 33% decrease in retinyl esters in the livers of male rats, but a 13-fold increase in retinyl esters in the kidney and a 3-fold increase in serum retinol. Liver and kidney microsomal uridine diphosphoglucuronosyltransferase (UDPGT) activity toward all-trans-retinoic acid was increased 3.7- and 2.6-fold, respectively, ten days following exposure to TCDD. Verification of the in vitro formation of [3H]retinoyl beta-glucuronide (RG) was by cochromatography with authenic RG on reversed phase high pressure liquid chromatography (HPLC), identification of retinoic acid as the hydrolysis product after beta-glucuronidase treatment, and the characterization of the all-trans-retinoyl glucuronide by negative fragment mass spectroscopy, fast atom bobardment. We conclude that increased retinoic acid glucuronidation may be a contributing factor to the hepatic depletion of vitamin A and the increased excretion of vitamin A metabolites following TCDD exposure.

Animals↗

Effect of hexachlorobiphenyl on vitamin A homeostasis in the rat.

Vitamin A status and turnover were examined in rats that had been exposed to chronic dietary treatment of 3,4,5,3',4',5'-hexachlorobiphenyl (HCB), 1 mg/kg diet. HCB caused hepatic depletion and renal accumulation of vitamin A, and a 1.7-fold increase in the serum retinol concentration. Intravenously administered [3H]retinol bound to retinol binding protein-transthyretin complex (RBP-TTR complex) was used to study the dynamics of circulatory retinol in these rats. In HCB-treated rats, the plasma turnover rate of retinol was increased compared to vitamin A-adequate untreated controls. HCB caused a 50% reduction of total radioactivity in liver, and, except for 0.5 h after the [3H]retinol-RBP-TTR dose, the specific activity of the hepatic retinyl ester pool was greater compared to control rats. The kidneys of HCB-treated rats accumulated radioactivity in the retinyl ester fraction. HCB also caused a 50% reduction in adrenal radioactivity compared with control rats. Urinary and fecal excretion of radioactivity was 3-fold higher in HCB-treated rats as compared to controls. Our findings demonstrate that chronic HCB feeding results in expansion of plasma vitamin A mass, in changes of liver and kidney retinol and retinyl ester pool dynamics and in an increased metabolism of vitamin A.

Animals↗

Alterations in vitamin A metabolism by polyhalogenated aromatic hydrocarbons.

Adequate stores and adequate tissue levels of vitamin A are maintained by a balance of tissue demands and dietary intake of the vitamin and are modified by many factors, including xenobiotics. It is well established that exposure to polyhalogenated aromatic hydrocarbons (PHAH) decreases hepatic content of vitamin A. Recent findings indicate that hepatic depletion of vitamin A is accompanied by an increase in serum and renal vitamin A content and enhanced excretion of vitamin A metabolites in urine and feces. Examination of tissue retinoid profiles reveals that PHAH exposure causes the generation of increased amounts of polar retinoids. It is very likely that PHAH affect enzymes crucial for regulation of vitamin A storage as well as enhance activities of specific enzymes in vitamin A metabolic pathway.

Animals↗

Effect of dietary retinoic acid on circulatory vitamin A homeostasis in polybrominated biphenyl-treated rats.

Retinoic acid (RA) supplementation is known to lower the amount of retinol in circulation. In contrast, the feeding of polyhalogenated aromatic hydrocarbons results in an elevated level of circulatory retinol. We investigated the effect of short-term dietary exposure to RA on the amount of serum retinol in female Sprague-Dawley rats fed either a basal diet (control rats) or the basal diet containing 100 mg of polybrominated biphenyls (PBBs)/kg diet (PBB-treated rats). After feeding of the above diets for 137 d, RA (12 mg/kg diet) was included in both the control and PBB-containing diets. The rats were fed the RA-containing diet for 3 d and then killed (d 140). Blood samples were obtained before and after RA treatment. Chronic PBB treatment of rats resulted in lower hepatic vitamin A and higher kidney vitamin A than in control rats. Serum retinol concentration was significantly higher in rats treated with PBB for 137 d than in controls; the subsequent treatment with RA lowered serum retinol to a level that was not different from that of control rats treated with RA. Our observations agree with earlier findings that 1) PBB treatment alters vitamin A homeostasis, and 2) dietary RA lowers the amount of circulatory retinol. An important new observation is that serum retinol homeostasis in PBB-treated rats appears to be regulated by a mechanism similar to that of normal rats. Polyhalogenated aromatic hydrocarbons may thus be useful tools to study the control mechanisms of vitamin A homeostasis.

Animals↗

Vitamin A metabolism in rats chronically treated with 3,3',4,4',5,5'-hexabromobiphenyl.

Chronic dietary administration of 3,3',4,4',5,5'-hexabromobiphenyl (HBB), 1 mg/kg diet, caused a decrease in retinol (20-fold) and retinyl esters (23-fold) in the livers of female rats, but resulted in a 6.4-fold increase in retinol and 7.4-fold increase in retinyl esters in the kidneys. Liver acyl-CoA:retinol acyltransferase and retinyl palmitate hydrolase activities were reduced while serum concentration of retinol was unaffected by HBB feeding. Metabolism of a physiological dose of [11-3H]retinyl acetate (10 micrograms), was examined in rats fed either vitamin A-adequate diet, or marginal amounts of vitamin A, or vitamin A-adequate diet containing HBB. A 13-fold greater amount of the administered vitamin A was found in kidneys of HBB-treated rats. In rats fed adequate or low amounts of vitamin A, kidney radioactivity was primarily in the retinol fraction, while in HBB-fed rats the radioactivity was associated mostly with retinyl esters. Fecal and urinary excretion of radioactivity was greatly increased in HBB-treated rats. Chronic HBB feeding results in a loss of ability of liver to store vitamin A, and severely alters the uptake and metabolism of vitamin A in the kidneys. We conclude that HBB causes major disturbances in the regulation of vitamin A metabolism.

Acyltransferases↗

Induction of differentiation of human promyelocytic leukemia cell line HL-60 by retinoyl glucuronide, a biologically active metabolite of vitamin A.

We examined the differentiation activity of retinoyl beta-D-glucuronide, a biologically active physiological metabolite of vitamin A, using the human promyelocytic leukemic cell line HL-60, which can be induced to differentiate with retinoic acid. Retinoyl beta-D-glucuronide (1 microM) inhibited HL-60 cell proliferation by 55-75%, inhibited tritiated thymidine incorporation into DNA by 63-80%, and induced 38-50% of the cells to differentiate into mature granulocytes. The potency of growth inhibition and induction of differentiation by retinoyl beta-D-glucuronide was similar to that of all-trans-retinoic acid. The continuous presence of either retinoyl beta-D-glucuronide or all-trans-retinoic acid was not required to obtain maximum growth arrest and differentiation: a 1-hr exposure of HL-60 cells to the retinoids gave the same response (measured after a total incubation time of 48 hr) as a 24-hr or 48-hr continuous treatment. Retinoyl beta-D-glucuronide (0.1-0.2 mM) was 50% less cytotoxic to HL-60 cells than all-trans-retinoic acid at an equimolar concentration. Retinoyl beta-D-glucuronide was not significantly metabolized to other retinoids; retinoic acid was not formed during incubation. We conclude that retinoyl beta-D-glucuronide can arrest HL-60 cell proliferation and induce their differentiation into mature granulocytes; it may act by itself or by being hydrolyzed to retinoic acid, which could be immediately utilized and metabolized. The therapeutic use of this retinoid as an antineoplastic agent is suggested.

Cell Differentiation↗

Quantitation of biological retinoids by high-pressure liquid chromatography: primary internal standardization using tritiated retinoids.

A single method is described for quantitation of 14 retinoids found in biological material. The method consists of reversed-phase HPLC, internal standardization, and carrier extraction procedures with three synthetic retinoids. Primary standardization of HPLC uv detector is achieved using tritiated all-trans-retinoic acid, all-trans-retinol, all-trans-retinyl palmitate, and all-trans-retinyl acetate. Extraction methods are standardized by correlating the uv absorbance of retinoids at 340 nm with radioactivity of tritiated retinoids of known specific activity. Quantitation of 10 pg of tritiated or 5 ng of nonradioactive retinoid per 0.1 g sample in a polarity range from 4-oxo-retinoic acid to retinyl stearate can be achieved in a single, 50-min chromatographic run. A single HPLC pump, a C18 reversed-phase analytical column, a multistep three-solvent gradient, and inexpensive solvents based on methanol, water, and chloroform comprise this cost-effective chromatographic system. Our primary standardization method allows investigators employing different procedures to compare results between laboratories by standardizing the HPLC uv detector with commercially available tritiated retinoids. With this method we were able to quantitate nanomolar amounts of endogenous retinoic acids and retinyl esters, that "HPLC uv only" conditions usually would not detect in the circulation and liver of rats under physiological conditions.

Animals↗

Influence of dietary retinyl acetate on normal rat mammary gland development and on the enhancement of 7,12-dimethylbenz[a]anthracene-induced rat mammary tumorigenesis by high levels of dietary fat.

The effect of high levels of dietary fat and retinyl acetate (ROA) on 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary tumor development and growth was examined. Female Sprague-Dawley rats, 51-53 days of age, were treated ig with 5 mg DMBA. At 55-57 days of age, the animals were divided into the following dietary treatment groups: A) 4.5% fat [control fat (CF)]; B) CF + 1.0 mmol ROA/kg diet (CF + ROA); C) 20.0% fat [high fat (HF)]; and D) HF + ROA. HF diets significantly increased mammary tumor multiplicity, with or without ROA, but did not significantly influence mammary tumor growth. ROA treatment reduced mammary tumor multiplicity regardless of the level of dietary fat and inhibited mammary tumor growth in the presence of normal levels of dietary fat. High levels of dietary fat did not significantly influence normal mammary gland growth and development. ROA significantly decreased normal mammary gland growth and development regardless of the level of dietary fat. Blood retinoids in rats fed ROA were primarily in the form of retinyl esters, i.e., retinyl linoleate, retinyl palmitate-oleate, and retinyl stearate. Free retinol levels in blood were not significantly influenced by ROA feeding. Blood retinyl ester levels were lower in rats fed the HF + ROA diet as compared to rats fed the CF + ROA diet.

9,10-Dimethyl-1,2-benzanthracene↗

Effect of moderate vitamin A supplementation and lack of dietary vitamin A on the development of mammary tumors in female rats treated with low carcinogenic dose levels of 7,12-dimethylbenz(a)anthracene.

We examined the effect of moderately increased and of marginal continued dietary supplementation of vitamin A (retinyl acetate) and the effect of lack of dietary vitamin A on the initiation and promotion stages of mammary tumorigenesis in female Sprague-Dawley rats treated with a single low (0.5 mg/100 g body weight) or very low (0.1 mg/100 g body weight) dose of i.v.-administered 7,12-dimethylbenz(a)anthracene. The number of mammary tumors was significantly (P less than 0.05) reduced if prior to and during initiation with 7,12-dimethylbenz(a)anthracene the rats were fed a moderately increased (30 micrograms/day) or marginal (3 micrograms/day) amount of vitamin A, compared to rats fed an adequate (10 micrograms/day) amount of vitamin A. The number of mammary tumors was also significantly (P less than 0.05) reduced when a moderately increased or marginal amount of vitamin A was provided during the tumor promotion phase. In addition, the number of mammary tumors was significantly (P less than 0.05) reduced by the lack of dietary vitamin A during both the initiation and promotion stages of this tumorigenic process, when compared to vitamin A adequate, ad libitum-fed rats, but not when compared to vitamin A adequate, food-restricted controls. The reduction in numbers of mammary tumors observed in these studies was reflected primarily in significant (P less than 0.05) decreases in mammary fibroadenomas; the number of mammary carcinomas was often reduced, but due to a low frequency of the carcinomatous lesions, this reduction did not reach the 5% level of statistical probability. Plasma and liver vitamin A levels were determined during both the initiation and promotion stages. As the dietary supplementation of vitamin A increased from 0 to 30 micrograms/day, there was an increase in mean liver and plasma vitamin A levels. No consistent correlation between plasma and liver vitamin A levels and the occurrence of mammary tumors was observed, except with the moderately increased (30 micrograms/day) intake of vitamin A, that resulted in a small, but statistically significant (P less than 0.05) increase of serum retinol at initiation; this may account for the observed reduction in mammary tumors. These results provide evidence that moderate alterations in vitamin A consumption can modulate low-dose chemically induced mammary gland tumorigenesis. Most importantly, suppression of mammary gland tumorigenesis can be achieved by moderately increased, frequent, and regular consumption of vitamin A; prolonged consumption of vitamin A-deficient diets or diets marginal in vitamin A does not enhance the risk of mammary tumor development.

9,10-Dimethyl-1,2-benzanthracene↗

Metabolism of all-trans-retinoic acid and all-trans-retinyl acetate. Demonstration of common physiological metabolites in rat small intestinal mucosa and circulation.

The kinetics and metabolism of physiological doses of all-trans-retinoic acid were examined in blood and small intestinal mucosa of vitamin A-depleted rats. A major portion of intrajugularly injected retinoic acid is rapidly (within 2 min) sequestered by tissues; subsequently 13-cis-retinoic acid and polar metabolites are released into circulation. All-trans-retinoic acid appears in small intestinal epithelium within 2 min after dosing and is the major radioactive compound there for at least 2 h. Retinoyl glucuronide and 13-cis-retinoic acid are early metabolites of all-trans-retinoic acid in the small intestine of bile duct-cannulated rats. Retinoyl glucuronide, the major metabolite of retinoic acid intestinal epithelium, in contrast to other polar metabolites, was not detected in circulation. An examination of [3H]retinyl acetate metabolites under steady state conditions in vitamin A-repleted rats demonstrates the occurrence of all-trans-retinoic acid and 13-cis-retinoic acid in circulation and in intestinal epithelium, in a pattern similar to that found after injection of retinoic acid into vitamin A-depleted rats. Our data establish that all-trans-retinoic acid, 13-cis-retinoic acid, and retinoyl glucuronide are physiological metabolites of vitamin A in target tissues, and therefore are important candidates as mediators of the biological effect of the vitamin.

Animals↗

Acute polybrominated biphenyl toxicosis alters vitamin A homeostasis and enhances degradation of vitamin A.

A single oral dose of HBB (3,3',4,4',5,5',-hexabromobiphenyl), a congener of the polybrominated biphenyls (PBBs), and a dioxin-type cytochrome P-450 enzyme inducer rapidly and dramatically altered the steady-state metabolism of vitamin A and caused an abnormal twofold enhancement in the metabolic output of degraded vitamin A in urine and feces of rats. The effects are most likely explained by an increased metabolism of vitamin A in kidney and deregulation of vitamin A metabolism in liver; this may lead to an increased dietary vitamin A requirement.

Animals↗

Comparison of fetal and adult retinol and retinoic acid binding proteins in bovine serum and pigment epithelium.

Cellular retinol binding protein (CRBP) and cellular retinoic acid binding protein (CRABP) were assayed in bovine eye pigment epithelium (PE) cytosol from both early fetal and young adult animals. Both fetal and adult PE contain CRBP but only fetal PE contains CRABP. Blood retinol binding protein (RBP) was also assayed in serum from fetal and adult animals. Both fetal and adult serum contain RBP. A novel albumin adsorption technique (Affi-Gel Blue) combined with refinements of the sucrose gradient centrifugation technique provide a better assay method for retinoid receptors than polyacrylamide disc gel electrophoresis. Our findings support the hypothesis that retinoid receptors may cause or be the consequence of differentiation in animal tissues.

Adsorption↗

Analysis of retinol and dideuterated retinol in rat plasma by gas chromatography combined mass spectrometry.

An isotope dilution method for determining the total body stores of vitamin A using a hydrocarbon derivative of rat plasma retinol, anhydroretinol (ANR) or dideuteroanhydroretinol (2H2ANR) has been evaluated by gas chromatography combined mass spectrometry (GC/MS). Rats with negligible vitamin A stores were dosed with retinyl acetate (ROA) or ROA and 11,12dideuteroretinyl acetate (2H2ROA). The 2H2ROA dose was allowed to equilibrate with total body stores for seven days and the deuterium/hydrogen ratio (D/H ratio) of rat plasma retinol fraction was determined. The results indicate that after the equilibration period the plasma D/H ratio is 73 to 108 percent of the loading dose D/H ratio. This study extends earlier reports that exogenous vitamin A storage is lower than the expected fifty percent figure in rats with low initial total body stores of vitamin A [1, 7, 8, 15, 17]. This work supports the concept that the estimation of total body stores of vitamin A by an isotope dilution method is useful for populations with negligible stores (i.e. liver stores) of vitamin A.

Animals↗