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Biomedical subjects

A C Ross

Publications and source records attributed to A C Ross.

At least 55 records · Page 3Linked to original sources
PubMed1995

Regulation of thyroid-stimulating hormone beta-subunit and growth hormone messenger ribonucleic acid levels in the rat: effect of vitamin A status.

T3 inhibits transcription of the rat TSH beta gene, and two T3 response elements have been identified that bind T3 receptors and that share sequence homology with the consensus sequence that is also recognized by retinoic acid receptors (RARs). We, therefore, asked whether RA was a regulator of TSH beta gene expression in vivo. Using RNase protection analysis, we found that vitamin A deficiency led to a 2-fold increase in rat pituitary TSH beta messenger RNA (mRNA) levels, which returned to normal 18 h after treatment with RA (20 micrograms/rat). Vitamin A deficiency had no effect on TSH beta mRNA levels in hypothyroid rats. Coadministration of RA and T3 (10 micrograms/100g body wt) to either vitamin A-deficient or vitamin A-deficient, hypothyroid animals caused decreases in TSH beta mRNA content that were indistinguishable from those seen with T3 alone. Surprisingly, vitamin A deficiency had no significant effect on GH mRNA levels in euthyroid or hypothyroid rats. Furthermore, treatment of vitamin A-deficient, hypothyroid animals with RA for either 18 or 72 h had no effect on GH mRNA levels, whereas T3 caused 11-fold and 18-fold increases in GH mRNA, respectively, at these times. We also used transient transfection to test for direct, retinoid receptor-mediated regulation of TSH beta gene transcription by RA. A plasmid TSH beta luciferase, containing 0.8 kilobases of rat TSH beta gene 5'-flanking sequences, exon 1, and 150 base pairs of intron 1, was transfected into CV-1 cells. Cotransfection with RAR alpha and retinoid X receptor-beta induced TSH beta expression by 3.5-fold, and treatment with RA suppressed this induction by 46%. These results show that vitamin A levels play a significant role in regulating the expression of the TSH beta gene, but not the GH gene, in vivo and suggest that RA may suppress TSH beta gene transcription directly by an RAR-retinoid X receptor heterodimer-mediated mechanism.

Animals↗
PubMed1995

Cell-type specific interactions between retinoic acid and thyroid hormone in the regulation of expression of the gene encoding ornithine aminotransferase.

The purposes of this study were to determine whether expression of the gene encoding ornithine aminotransferase (OAT) in the rat liver and kidney is regulated by retinoic acid (RA) and to characterize further the role of thyroid hormone in regulating the expression of this gene. The level of OAT messenger RNA (mRNA) was reduced 70% in the liver of animals fed a vitamin A-deficient diet relative to that in animals fed a vitamin A-sufficient diet. RA, administered at a dose of 20 micrograms/rat to A-deficient rats for 1 or 3 days, restored OAT mRNA to near the level observed in animals fed the A-sufficient diet. Retinol was also effective in this regard. T3, when injected alone at a dose of 10 micrograms/100 g BW, had no effect on the level of OAT mRNA in the liver. However, when injected concurrently with RA, T3 blocked the ability of RA to induce OAT mRNA in the liver of rats fed the vitamin A-deficient diet. Animals made both vitamin A deficient and hypothyroid responded to RA in a manner similar to vitamin A-deficient animals. The vitamin A-deficient, hypothyroid rats responded somewhat differently to T3, however. T3 was unable to block the induction of OAT mRNA in the liver of vitamin A-deficient, hypothyroid rats when injected concurrently with RA for 1 day, but did block the induction of OAT mRNA by RA when these two hormones were injected concurrently for 3 days. These data indicate that RA and T3 exert opposing effects on the level of OAT mRNA in the liver. The effects of RA and T3 on OAT mRNA were markedly different in the kidney. Neither vitamin A deficiency nor RA had any apparent affect on the level of OAT mRNA in the kidney. T3, however, increased the level of OAT mRNA in the kidney of vitamin A-deficient rats. In the kidney of vitamin A-deficient, hypothyroid rats, T3 was unable to increase OAT mRNA when injected for 1 day, but did increase this mRNA when injected for 3 days. Together, these data indicate cell-type specific effects of both RA and T3 on the OAT gene.

Animals↗
PubMed1995

Hepatic uptake and metabolism of chylomicron retinyl esters: probable role of plasma membrane/endosomal retinyl ester hydrolases.

Previous studies have indicated the presence of both neutral and acid, bile salt-independent retinyl ester hydrolases associated with plasma membrane and endosome fractions of rat liver homogenates. In the present studies, chylomicrons containing tritium-labeled retinyl esters were injected intravenously into rats in order to study the initial metabolism of retinyl esters during and after uptake into the liver. At various times after chylomicron injection, plasma was obtained and the liver was homogenized and subjected to analytical subcellular fractionation. Labeled retinyl esters were rapidly cleared from plasma (half-time approximately 10 min) and appeared in the liver. Within the liver, label first appeared in plasma membrane/endosomal fractions that were also enriched in both neutral and acid, bile salt-independent retinyl ester hydrolase activities. At no time were the labeled esters significantly associated with fractions enriched in lysosomes. Rather, it appeared that the labeled esters were hydrolyzed and/or transferred to fractions enriched in endoplasmic reticulum. These studies demonstrate the co-localization of newly delivered retinyl esters and bile salt-independent retinyl ester hydrolase enzyme activities and thus, suggest a probable role for these enzymes in the initial hepatic metabolism of chylomicron retinyl esters. This conclusion was further supported by the observation that plasma membrane/endosomal fractions were active in catalyzing the hydrolysis of chylomicron remnant retinyl esters in vitro.

Animals↗
PubMed1994

The function of vitamin A in cellular growth and differentiation, and its roles during pregnancy and lactation.

Recent advances in the molecular biology of the retinoids have provided a mechanistic explanation for the observations, first made several decades ago, that vitamin A profoundly influences the differentiation of tissues throughout the body. A central concept has recently emerged, namely that retinoids seldom exist "free" in solution but, rather, are nearly always associated with specific retinoid-binding proteins. In plasma, these include RBP and the chylomicron whereas, in cells two distinct classes of retinoid-binding proteins exist: the cellular (cytoplasmic) proteins (CRBPs and CRABPs) and the nuclear receptors proteins (RARs and RXRs). Whereas the cellular retinoid-binding proteins serve as buffers and as chaperones during metabolism (Ross, 1993b), the nuclear receptors are now recognized to be the direct mediators of retinoid actions on the genome. Both the cytoplasmic and nuclear classes of retinoid-binding proteins are expressed early in development and are proposed to control the concentration of retinoic acid and the transcription of retinoid-responsive genes, respectively. Given the profound effects of retinoic deficiency or excess on the developing fetus, it is not surprising that mechanisms have evolved to control the placental transfer of vitamin A. Transfer is nearly uniform over a rather wide range of maternal dietary vitamin A intake. The importance of RBP in transporting retinol to tissues is suggested by the observations that the visceral yolk sac and the liver of the fetus transcribe and translate RBP. In comparison to pregnancy, vitamin A transport during lactation is much more responsive to variations in maternal vitamin A intake. The young of mothers with good vitamin A nutriture may thus accumulate significant retinol reserves during the suckling period. Conversely, young nursed by mothers with poor vitamin A status and low intake during lactation may fail to develop adequate stores and be vulnerable to vitamin A deficiency if the post-weaning diet is also poor in vitamin A. In populations with low vitamin A status, the lactation period provides an excellent window of opportunity for supplementing mothers and, indirectly, their offspring, with vitamin A to replenish the mother's vitamin A reserves and assure that the infant's growth and development are not limited by an inadequate quantity of this essential nutrient.

Cell Differentiation↗
PubMed1994

Antibody response against tetanus toxoid is enhanced by lipopolysaccharide or tumor necrosis factor-alpha in vitamin A-sufficient and deficient rats.

Reduced antibody response to tetanus toxoid (TT) was previously demonstrated during vitamin A deficiency but the response to bacterial lipopolysaccharide (LPS) was normal. We addressed whether anti-TT IgG responses are enhanced in vitamin A-sufficient and deficient rats by immunization with LPS plus TT. Antibody responses in vitamin A-sufficient and deficient rats increased significantly after coimmunization (LPS + TT) compared with the response of rats immunized with TT alone. In additional studies, recombinant tumor necrosis factor-alpha (TNF-alpha) also significantly increased the anti-TT IgG concentrations. Because pretreatment with anti-TNF before coimmunization or immunization with TT alone markedly reduced the anti-TT IgG responses, we infer TNF to be a mediator of both the adjuvanticity of LPS and the unstimulated response to TT. In conclusion, vitamin A-deficient rats can be stimulated to respond to TT by coimmunization with LPS or by treatment with TNF.

Adjuvants, Immunologic↗
PubMed1994

The antibody response of vitamin A-deficient rats to pneumococcal polysaccharide is enhanced through coimmunization with lipopolysaccharide.

Although the antibody response of vitamin A-deficient rats immunized with pneumococcal polysaccharide (SSS-III) was barely detectable, coimmunization with SSS-III plus 2 or 20 micrograms of Pseudomonas aeruginosa lipopolysaccharide (LPS) resulted in an anti-SSS-III response that was equal to or greater than that of vitamin A-sufficient rats immunized only with SSS-III. Doses of 2 and 20 micrograms of LPS, but not 0.1 and 0.5 micrograms, produced significant adjuvanticity, and LPS functioned as an adjuvant when given concomitantly with SSS-III (day 0) or up to 3 days later. Thus, vitamin A deficiency per se does not preclude the anti-SSS-III response, and LPS or factors released in response to LPS, even after initiation of the response to SSS-III, may substantially enhance antibody production to this thymus-independent, type 2 antigen.

Animals↗
PubMed1994

Effect of vitamin A deficiency and retinoic acid repletion on intestinal and hepatic apolipoprotein A-I mRNA levels of adult rats.

Apolipoprotein A-I (apoA-I) gene expression is known to be regulated by nutritional and hormonal factors. Experiments were conducted to determine the effects of vitamin A deficiency and retinoic acid repletion on the in vivo expression of apoA-I in rat intestine and liver. The relative abundance of apoA-I mRNA (apoA-I/beta-actin ratio) in the intestine did not differ significantly between vitamin A-deficient and -sufficient rats. However, the relative abundance of hepatic apoA-I mRNA of vitamin A-deficient rats was 2.2- to 6-times that of sufficient rats. Even marginal vitamin A status resulted in a significant increase in hepatic apoA-I mRNA expression. Treatment of vitamin A-deficient rats with a single dose of retinoic acid (20 micrograms, 20 h before tissues were collected) reduced the hepatic apoA-I mRNA/beta-actin ratio by about 40%, while further reduction (about 60-65%) was observed after two treatments with retinoic acid. By nuclear run-on assay, the increase in hepatic apoA-I mRNA in vitamin A-deficient rats was attributable to increased transcription of the apoA-I gene. However, immunoblot analysis showed no apparent differences in apoA-I protein in either liver homogenates or plasma of vitamin A-deficient and -sufficient rats. These data indicate that apoA-I gene expression in vivo is sensitive to retinoid status and suggest that there is additional regulation of post-transcriptional events.

Animals↗
PubMed1993

Cellular metabolism and activation of retinoids: roles of cellular retinoid-binding proteins.

Vitamin A and related molecules--the retinoids--play essential roles in vision, growth, reproduction, and cellular differentiation beginning in early development. Retinoic acid, a key regulatory retinoid formed intracellularly through the oxidation of retinol, functions as ligand for a family of nuclear retinoic acid receptors that regulate transcription from target genes. Metabolic events are critical to maintaining physiological concentrations of retinoic acid. In the cytoplasm, retinol, retinaldehyde, and retinoic acid are associated with retinoid-binding proteins, most of which are approximately 15-kDa proteins belonging to the fatty acid-binding protein/cellular retinol-binding protein family. The ligand binding properties and molecular features of these proteins have been well characterized. Recent experiments have highlighted the importance of the cellular retinoid-binding proteins in controlling the concentration of free retinoids and in directing protein-bound retinoids to key enzymes responsible for their metabolism. For example, the cellular retinol-binding protein, CRBP, has been implicated in retinol uptake, retinol esterification, mobilization of retinyl esters, and the initial oxidation of retinol to retinaldehyde. The ligands bound to other retinoid-binding proteins have also been shown to be available for enzymatic transformation. The new knowledge of the various ways these cytoplasmic proteins buffer the concentration of ligand, control their distribution, and determine their metabolism by specific enzymes is contributing to an improved understanding of the physiological control of retinoid action.

Animals↗
PubMed1993

Regulation of hepatic lecithin: retinol acyltransferase activity by retinoic acid.

The activity of the hepatic enzyme lecithin: retinol acyltransferase (LRAT), thought to catalyze the esterification of retinol for storage, was previously shown to vary directly with the vitamin A (retinol) status of the rat [Randolph, R. K., and Ross, A. C. (1991) J. Biol. Chem. 266, 16453-16457]. The present studies were designed to determine whether liver LRAT activity is regulated in vivo by retinoic acid, a principal active metabolite of retinol. LRAT activity was negligible in the livers of vitamin A-deficient rats. Following treatment with a single 2-micrograms dose of retinoic acid, LRAT activity increased significantly while, following treatment with a single 20-micrograms dose, liver LRAT activity equalled that of vitamin A-sufficient adult rats. Retinoic acid was more effective than an equimolar quantity of retinol in restoring LRAT activity in the vitamin A-deficient rat liver. The increase in hepatic LRAT activity after administration of retinoic acid occurred rapidly, reaching a maximum within 12-16 h but declining again after 48-72 h. Studies were conducted in vivo to gain insight into the level of regulation of LRAT by retinoic acid. The increase in LRAT activity by retinoic acid in the vitamin A-deficient rat was blocked completely by both actinomycin D and cycloheximide. The ability of liver to esterify retinol in vivo was correlated with the in vitro activity of LRAT after retinoic acid induction. We conclude that retinoic acid, an important end product of retinol metabolism, regulates a key aspect of hepatic retinol metabolism through its regulatory activity on liver LRAT.

Acyltransferases↗
PubMed1993

Vitamin A as a hormone: recent advances in understanding the actions of retinol, retinoic acid, and beta carotene.

Within the past few years, much has been learned about the metabolism and actions of vitamin A and the carotenoids. This article reviews the biochemical and cellular events in retinoid metabolism that lead to production of retinoic acid, an active metabolite of vitamin A. Retinoic acid functions in a hormone-like manner to regulate the expression of a number of genes. Beta carotene is now under study as an anticancer agent and for its possible beneficial effects in a number of chronic diseases. Current recommendations for carotene intake exceed the usual daily intake nearly fourfold.

Animals↗
PubMed1993

Quantitative analysis of immunoglobulin G subclasses in the rat.

Quantitative ELISAs have been developed for the four immunoglobulin G (IgG) subclasses (IgG1, IgG2a, IgG2b and IgG2c) of the rat. These assays were shown to have excellent sensitivity, reproducibility and adequate specificity for studies of natural IgG and antigen-elicited IgG responses. The sensitivity (working range for each isotype assay) was: IgG1, 1.6 to 200 ng/ml; IgG2a, 8 to 200 ng/ml; IgG2b, 1.6 to 200 ng/ml; and IgG2c, 1.6 to 1000 ng/ml. The isotype sum and the direct assay of total plasma IgG agreed closely. The utility of these assays was demonstrated in a study of the development of total IgG and of the specific IgG response following immunization with tetanus toxoid (TT). For total IgG, the predominant isotype was IgG2b (64% of total IgG) followed by IgG2a (29%), IgG1 (4%) and IgG2c (3%). In contrast, the anti-TT response was mainly of the IgG1 subclass (57% of total anti-TT). These quantitative assays should prove useful for investigating the response to experimental vaccines and the influence of cytokines on class switching in vivo.

Animals↗
PubMed1993

Dietary vitamin A restriction produces marginal vitamin A status in young rats.

A model of marginal vitamin A deficiency was developed in young rats by limiting the vitamin A intake of dams and of offspring postweaning. Prior to and throughout pregnancy and lactation, female rats were fed diets in which the vitamin A concentration was either low marginal [0.18 retinol equivalent (RE)/g diet], high marginal (0.4 RE/g) or sufficient (4 RE/g). Vitamin A restriction had no effect on dams' reproduction or litter sizes, but total retinol in liver was depleted by the end of lactation. Pups fed all diets grew steadily from birth through 35 d. The milk curd total retinol concentration of 9-d-old pups' stomachs was significantly different among the low marginal, high marginal and vitamin A-sufficient groups. By 35 d, plasma retinol concentrations of pups in the low marginal group were less than half of those in the high marginal and vitamin A-sufficient groups. The liver total retinol concentrations and lecithin retinol:acyltransferase activity of 35-d-old pups in the low and high marginal groups were much lower than those of vitamin A-sufficient pups. When pups from dams fed low marginal diet were weaned onto vitamin A-free diet, frank vitamin A deficiency was evident by 33 d of age as judged by physical signs and biochemical alterations in vitamin A status. Thus, these dietary protocols are useful in inducing early onset of either marginal or frank vitamin A deficiency.

Animals↗
PubMed1993

Overview of retinoid metabolism.

Substantial and exciting progress has been made in understanding the cytoplasmic metabolism and the nuclear functions of vitamin A. Keys to this understanding have come from studies of the plasma and cytoplasmic proteins that bind retinol and retinoic acid, thereby regulating retinoid concentration and specifying the enzymatic reactions through which retinol is absorbed, stored as esterified retinol or oxidized to retinoic acid. In the nucleus, the hormone-like activity of retinoic acid is transduced by a family of retinoic acid receptors that regulate a large number of retinoic acid-responsive genes. The coordination of cytoplasmic retinoid metabolism and the nuclear effects of retinoic acid or other retinoid metabolites is now recognized as critical to embryonic development and the control of cellular differentiation throughout life.

Animals↗
PubMed1993

Vitamin A status and immunoglobulin G subclasses in rats immunized with tetanus toxoid.

Vitamin A deficiency severely compromises the magnitude of the primary and secondary antibody response to tetanus toxoid (TT) but does not impair the development of immunologic memory. To further characterize this immunodeficiency in antibody production, we have quantified the immunoglobulin G (IgG) subclasses (IgG1, IgG2a, IgG2b, and IgG2c) during the primary and secondary response to TT in normal, vitamin A-deficient, and retinol-repleted rats. In the primary response in normal rats, anti-TT IgG1 and IgG2b predominated. In vitamin A-deficient rats the production of anti-TT IgG2b was severely impaired, with little change in either IgG1 or IgG2a. In the secondary response vitamin A-deficient rats produced low levels of all anti-TT IgG subclasses. However, when vitamin A-deficient rats were repleted with retinol 2 days before reimmunization their secondary anti-TT IgG response was normal both in magnitude and IgG subclass distribution. This result implies that although vitamin A deficiency during the primary antibody response impaired anti-TT IgG2b production, it did not inhibit Ig heavy chain recombination or the differentiation of lymphocytes that formed memory B cells for each subclass; furthermore, these cells were activated in the secondary response after vitamin A status was improved. Thus, these experiments further support the concept that memory cell formation remains normal during vitamin A deficiency despite low levels of antibody production.

Animals↗
PubMed1993

Infections complicating joint replacement and other orthopedic conditions.

The principles underlying the diagnosis and management of infection in the orthopedic patient are well established. Despite this, the patient with an infected implant still presents the orthopedic surgeon with a challenging problem. In this review, I consider the recent developments in the diagnosis and management of the infected arthroplasty, infected tibial nonunions, spinal infections, and disease transmission by bone allografts.

Bacterial Infections↗
PubMed1993

Radiation inactivation analysis of acyl-CoA:retinol acyltransferase and lecithin:retinol acyltransferase in rat liver.

Microsomes from liver and several other tissues esterify retinol through both fatty acyl-CoA-dependent and -independent reactions. Two activities, acyl-CoA:retinol acyltransferase (ARAT) and lecithin:retinol acyltransferase (LRAT) activities, have been characterized enzymatically but neither has yet been purified and characterized biochemically. We have used the method of radiation inactivation to determine the target sizes of ARAT and LRAT in intact microsomal membranes from rat liver. After exposure of frozen liver microsomes to ionizing radiation, the activity of ARAT decayed exponentially yielding a target size of 73 +/- 18 kDa (mean +/- SD, n = 6). The activity of LRAT was assayed both by monitoring the esterification of retinol bound to the cellular retinol-binding protein (CRBP) and of solvent-dispersed retinol. With both assays a single exponential was observed with radiation doses of 9 to 150 Mrads. The slopes obtained with both LRAT assays were similar, yielding target sizes of 52 +/- 10 kDa (n = 10) for the LRAT assay with CRBP-retinol and 56 +/- 7 kDa (n = 6) for the LRAT assay with dispersed retinol. These target sizes did not differ from each other but were significantly smaller than that of ARAT. These data provide the first physical evidence of the independent entities which catalyze the ARAT and LRAT reactions of liver microsomes.

Acyltransferases↗
PubMed1992

Characterization of a bile salt-dependent cholesteryl ester hydrolase activity secreted from HepG2 cells.

HepG2 cells and medium were assayed for cholesteryl ester hydrolase (CEH) activity in the presence and absence of sodium cholate. Although bile salt-dependent CEH activity was measured in the medium at 6 to 96 h (up to 4500 pmol/h per mg cell protein), there was very little activity detected in the corresponding cell homogenates (less than 70 pmol/h per mg cell protein). Activity in the medium was expressed only in the presence of trihydroxy bile salts and was maximal at 40 mM cholate and pH 7.5. Incubation of HepG2 cells with brefeldin A resulted in an 80 to 90% inhibition of secretion of the bile salt-dependent CEH activity, while only inhibiting total protein secretion by 42%. Bile salt-dependent CEH activity could also be detected in rat liver perfusates. Although there was measurable activity in all of 14 livers analyzed (47 +/- 10 and 53 +/- 17 nmol/h per g liver per h perfusion during two 5-min collections after 15 and 30 min of perfusion, respectively), it did not correlate with the activity found in corresponding liver homogenates, as only four livers had detectable bile salt-dependent CEH activity. These results provide evidence for the secretion of a bile salt-dependent CEH activity, from both a hepatic cell line and the intact liver, that has similar properties to the enzyme previously isolated from rat liver homogenates and rat pancreas.

Animals↗