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Biomedical subjects

M M Compton

Publications and source records attributed to M M Compton.

At least 19 recordsLinked to original sources
PubMed2004

Responses of broilers to dietary zinc concentrations and sources in relation to environmental implications.

1. Since high concentrations of zinc are associated with reduced crop yields, environmental concerns are emerging regarding zinc accumulation in areas where poultry production is prevalent. This study investigates growth performance and zinc utilisation during the life cycle of broilers when diets were supplemented with various concentrations of zinc from two different sources. 2. A total of 740 Cobb 500 1-d-old male broiler chicks was randomly distributed into 88 battery cages. Excreta were collected over a 48-h period on d 10, 17, 24, 31, 38, and 45 to measure zinc excretion on a dry matter basis. 3. During the 45-d experimental period, broilers were given a maize-soybean meal basal diet (30 mg/kg zinc) supplemented with 0, 20, 40, and 80 mg/kg of zinc from zinc sulphate, Availa Zn amino acid complex, or an equal combination of zinc sulphate and Availa Zn amino acid complex. 4. Progressive additions of zinc to the basal diet significantly increased body weight gain of broilers, but did not affect feed conversion or mortality. Optimum body weight gain was achieved at 80 mg/kg supplemental zinc, which exceeds the NRC recommendations of 40 mg/kg. 5. During each excreta collection period, increased supplemental zinc concentrations significantly increased zinc excretion. However, the zinc source did not influence zinc utilisation. 6. As compared with zinc sulphate supplementation, the cumulative zinc excretion data indicate that adding zinc to the basal diet in the form of Availa Zn amino acid complex decreased zinc excretion.

Animal Feed↗
PubMed2003

The expression of calbindin in chicks that are divergently selected for low or high incidence of tibial dyschondroplasia.

Three experiments were conducted with broiler chicks that were divergently selected for low or high incidence of tibial dyschondroplasia (LTD and HTD, respectively) to determine if the expression of intestinal calbindin-28 kD mRNA and protein differed between the 2 strains. In addition, levels of intestinal vitamin D receptor mRNA and plasma thyroid hormone concentrations were also examined. In experiment 1, LTD and HTD chicks were fed a corn-soybean meal diet that was adequate in all nutrients except cholecalciferol (D3), which was titrated to 5 or 40 microg/kg diet in a completely randomized 2 x 2 factorial arrangement. At 4 and 8 d of age, HTD chicks fed 5 microg of D3/kg of diet had a lower (P < 0.05) expression level of calbindin-28 kD mRNA than the LTD chicks fed the same diet. At 4 and 8 d of age, HTD chicks fed 5 microg of D3 had the lowest intestinal expression of calbindin-28 kD protein. Expression of vitamin D receptor mRNA did not differ for broiler strains at either level of D3 supplementation. In experiment 2, there was no significant difference in the expression of calbindin-28 kD mRNA or vitamin D receptor mRNA between day-of-hatch LTD, HTD, and commercial broiler chicks. Experiment 3 was similar in design to the first experiment except that the birds were fed for 18 d. Calbindin-28 kD and vitamin D receptor mRNA expression levels at 18 d were similar to those observed in experiment 1. Plasma triiodothyronine and free-triiodothyronine concentrations were greater for LTD chicks, regardless of dietary D3 supplementation levels. These results suggest that divergent selection of broilers for LTD or HTD alters the physiological response to nutritionally inadequate levels of dietary D3.

Animals↗
PubMed2001

TNFalpha induces and insulin inhibits caspase 3-dependent adipocyte apoptosis.

Regulation of fat cell number by apoptosis is proposed to be part of a normal physiological cycle in adipose growth and development. To investigate this process, cultured rat adipocytes were treated with various concentrations of tumor necrosis factor alpha (TNFalpha) and/or insulin to determine the roles of these factors in adipocyte apoptosis. The cells were analyzed by flow cytometry using a TUNEL assay. TNFalpha increased adipocyte apoptosis in a dose-dependent fashion. TNFalpha-mediated apoptosis was detectable within 6 h of treatment and continued to increase with time. Decreasing media insulin concentration from 8.5 to 0.85 nM resulted in increased adipocyte apoptosis, whereas high doses of insulin protected adipocytes from TNFalpha-induced apoptosis. TNFalpha-activated apoptosis was accompanied by an increase in caspase 3 activity and could be inhibited by a caspase 3-specific inhibitor. These data suggest that adipose tissue cell number is regulated, in part, by an apoptotic signaling pathway that involves TNFalpha, insulin, and caspase 3.

Adipocytes↗
PubMed2001

The analysis of cThy28 expression in avian lymphocytes.

During the course of a screening procedure designed to isolate gene products that mediate avian lymphocyte apoptosis, a 1070 bp cDNA encoding a 242 amino acid protein, cThy28 (GenBank accession number U34350), was isolated. Based on the deduced amino acid sequence, cThy28 is a conserved protein that shares greater than 90% amino acid similarity with several putative mammalian homologues. A structural analysis of the protein suggests that is a nuclear-localized, phosphoprotein with potential glycosylation and myristolation sites. Compared to other non-lymphoid tissues, the cThy28 protein and its transcript are present in immune organs at elevated levels. When the expression of cThy28 was analyzed in cultured bursal lymphocytes undergoing apoptosis, the cellular levels of this protein and its transcript declined and underwent degradative processes as a function of the apoptotic process. Treatment of lymphocytes with phorbol esters inhibited apoptosis and protected the cellular levels of the cThy28 protein and its transcript from undergoing this degradative process; however, analysis of transcripts of a "housekeeping" gene, GAPDH (glyceraldehyde phosphate dehydrogenase), revealed a similar response to this apoptosis treatment regimen. Thus, the apoptotic-mediated alterations in cThy28 expression and its role in lymphocyte function remain undefined; however, the potential task of this gene in apoptosis may be related to the putative functions of the cThy28 mammalian homologues identified in this report.

Amino Acid Sequence↗
PubMed2000

Induction of apoptosis by all-trans-retinoic acid and C2-ceramide treatment in rat stromal-vascular cultures.

Apoptosis of preadipocytes and adipocytes contributes to the balance of adipose tissue mass by reducing adipocyte number. To address this phenomenon, we treated cultured rat S-V cells with all-trans-retinoic acid (RA) (10 microM) or C2-ceramide (50 microM) during adipogenesis. Gel electrophoresis of DNA from treated cells cultured in serum-free medium showed that 10 microM RA or 50 microM ceramide induced a distinct laddering pattern of DNA fragments. Cellular caspase 3 activity, another marker of apoptosis, was increased by RA (10 microM) (P < 0.05), but not by 50 microm C2-ceramide. RT-PCR results showed that RA (10 microM) decreased the expression of Bcl-2 mRNA. These results suggest that fat cell loss by apoptosis can be regulated, in part, by RA (10 microM) which increases caspase 3 activity and decreases Bcl-2 expression in rat S-V cells. C2-ceramide apparently works through a different cellular mechanism to induce apoptosis.

Adipocytes↗
PubMed2000

Immunization of chickens with quail and turkey perivitelline membrane proteins: production of antibodies and their effects on fertility.

The cross-reactivity of antibodies developed against zona pellucida proteins and their subsequent deleterious effect on fertility in heterologous species is well documented. However, similar investigations have not been undertaken in avian species. In Experiment 1, White Leghorn hens were immunized with intact germinal discs (GD) of quail and chickens. Chicken GD proteins did not elicit an immune response in chicken hens, whereas quail GD proteins were highly immunogenic. Anti-quail GD antibodies did not bind with chicken inner perivitelline membrane (IPM) proteins as determined by immunoblot analysis. To examine the fertility of immunized hens, artificial insemination was performed at weekly intervals for 4 wk following the booster immunization. No significant differences were detected in fertility or hatchability of immunized hens when compared with unimmunized control hens. In Experiment 2, White Leghorn hens were immunized with intact turkey GD, solubilized turkey perivitelline membrane (PM) modified with dinitrophenol (DNP), and solubilized chicken IPM modified with DNP. High titers of antibodies were detected against the turkey GD and the DNP-modified turkey PM proteins. A weak immune response was observed in hens immunized with modified chicken IPM proteins. The fertility and hatchability of eggs laid by immunized hens, however, were not significantly different from those of unimmunized hens. Antibodies from immunized hens were further analyzed using an in vitro assay that assesses sperm penetration of intact IPM. Sperm penetration of intact IPM was inhibited to the same extent when IPM was preincubated with preimmune as well as anti-PM immunoglobulins. Collectively, these results suggested that the antibodies developed in these hens did not cross-react with the chicken IPM proteins involved in sperm-egg interaction and thus did not influence the fertility.

Acrosome Reaction↗
PubMed1999

Multiparametric assessment of bursal lymphocyte apoptosis.

When bursal lymphocytes are placed in cell culture, they undergo an apoptotic form of cell death that can be inhibited by phorbol esters and protein synthesis inhibitors. The goal of the current study was to evaluate the time course of this process and the inhibition of this process using several different assays to detect apoptosis: (1) terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) of lymphocyte DNA strand breaks with dUTP-FITC; (2) propidium iodide (PI) staining of lymphocyte chromatin; (3) chloromethyl-x-rosamine (CMX-Ros) binding to lymphocyte mitochondria; (4) merocyanine-540 (MC-540) binding to the lymphocyte plasma membrane; (5) flow cytometric analysis of light scatter from lymphocytes; (6) analysis of genomic DNA from lymphocytes by agarose gel electrophoresis; and (7) cellular caspase-3 activity of lymphocytes. When bursal lymphocyte apoptosis was analyzed as a function of time, or inhibited by phorbol esters or cycloheximide, all of these assays corroborated the apoptotic process. However, treatment of lymphocytes with a cytotoxic level of the proteinase inhibitor, n-ethylmaleimide (NEM) resulted in a putative, necrotic form of cell death that revealed discrepancies among the various assays in the detection of apoptotic cells. Specifically, the CMX-Ros and MC-540 assays erroneously detected the necrotic cells as being apoptotic cells following NEM treatment. These findings indicate the need for additional assays and appropriate treatment controls to verify the apoptotic process when using the CMX-Ros and MC-540 assays.

Animals↗
PubMed1999

Leptin mRNA expression and serum leptin concentrations as influenced by age, weight, and estradiol in pigs.

Two experiments (EXP) were conducted to determine the roles of age, weight and estradiol (E) treatment on serum leptin concentrations and leptin gene expression. In EXP I, jugular blood samples were collected from gilts at 42 to 49 (n = 8), 105 to 112 (n = 8) and 140 to 154 (n = 8) d of age. Serum leptin concentrations increased (P < 0.05) with age and averaged 0.66, 2.7, and 3.0 ng/ml (pooled SE 0.21) for the 42- to 49-, 105- to 112-, and 140- to 154-d-old gilts, respectively. In EXP II, RNase protection assays were used to assess leptin mRNA in adipose tissue of ovariectomized gilts at 90 (n = 12), 150 (n = 11) or 210 (n = 12) d of age. Six pigs from each age group received estradiol (E) osmotic pump implants and the remaining animals received vehicle control implants (C; Day 0). On Day 7, back fat and blood samples were collected. Estradiol treatment resulted in greater (P < 0.05) serum E levels in E (9 +/- 1 pg/ml) than C (3 +/- 1 pg/ml) pigs. Serum leptin concentrations were not affected by age, nor E treatment. Leptin mRNA expression was not increased by age in C pigs nor by F in 90- and 150-d-old pigs. However, by 210 d of age, leptin mRNA expression was 2.5-fold greater (P < 0.01) in E-treated pigs compared to C animals. Serum insulin concentrations were similar between treatments for 210-d-old pigs. However, insulin concentrations were greater (P < 0.05) in E than C pigs at 90 d and greater in C than E animals at 150 d. Plasma glucose and serum insulin-like growth factor-I concentrations were not influenced by treatment. These results demonstrate that serum leptin concentrations increased with age and E-induced leptin mRNA expression is age- and weight-dependent.

Adipose Tissue↗
PubMed1998

Down-regulation of CCAAT/enhancer binding proteins alpha, beta and delta in adipose tissue by intracerebroventricular leptin in rats.

In our previous report, intracerebroventricular (i.c.v.) administration of leptin caused fat depletion by an induced adipocyte apoptosis in addition to influencing lipid metabolism. To uncover the biochemical mechanisms that mediate this response, the present study was designed to determine whether CCAAT/enhancer binding proteins (C/EBP)alpha, -beta and -delta play a role in the leptin-induced fat depletion. Expressions of C/EBPalpha, -beta and -delta in epididymal fat tissues were examined by Western immunoblot and in situ immunocytochemical analysis after 5 days of i.c.v. treatment. Young and old rats (3 and 8 months old) were treated with or without 5 micrograms/day leptin. The expression of C/EBPalpha, -beta and -delta was decreased by i.c.v. leptin treatment in young rats as compared with controls (P<0.05). However, leptin did not influence the expression of C/EBPalpha, -beta and -delta in adipose tissues of 8-month-old rats. The basal level of expression of C/EBPbeta was greater in 8-month-old rats than in 3-month-old rats, (P<0.05) whereas the basal expression of C/EBPalpha and -delta was not different between age groups. These results were confirmed by in situ immunocytochemical analysis. The present study suggests that leptin-induced down-regulation of C/EBPalpha, -beta and -delta might influence adipocyte differentiation and growth in a number of ways.

Adipose Tissue↗
PubMed1998

Leptin regulation of peroxisome proliferator-activated receptor-gamma, tumor necrosis factor, and uncoupling protein-2 expression in adipose tissues.

It has previously been reported that intracerebroventricular (i.c.v.) administration of leptin induced adipose tissue apoptosis in addition to influencing lipid metabolism. The objective of the present study was to determine if the expressions of peroxisome proliferator-activated receptor-gamma (PPAR gamma), uncoupling protein-2 (UCP2), and tumor necrosis factor (TNF alpha) were influenced by in vivo leptin treatment. Expression of PPAR gamma, UCP2, and TNF alpha in epididymal fat tissue was examined by Western immunoblot and in situ immunocytochemical analysis after 5 days of i.c.v. leptin treatment. Young and old rats (3 and 8 months old) were treated with or without 5 micrograms/d leptin. Leptin treatment increased PPAR gamma expression by 70-80% (P < 0.01) in both age groups. Leptin treatment decreased the expression of UCP2 (P < 0.01) in young rats, whereas it increased UCP2 expression (P < 0.01) in old rats. Leptin treatment also decreased TNF alpha expression by 40% (P < 0.01) in young rats but did not influence its expression in old rats. The basal level of expression of PPAR gamma was greater in 3-month-old rats than in 8-month-old rats. The basal level of UCP2 and TNF alpha expression was not different between the two age groups. These immunoblotting data were further confirmed by in situ immunocytochemical analysis. The present study suggests that expression of PPAR gamma may be directly involved in the leptin-induced adipocyte apoptosis signal pathway, whereas UCP2 and TNF alpha may play roles in the leptin-induced lipolysis process.

Adipose Tissue↗
PubMed1998

Endogenous activation of apoptosis in bursal lymphocytes: inhibition by phorbol esters and protein synthesis inhibitors.

The bursa of Fabricius represents the primary immune organ where immature B cells undergo maturational changes in avian species. Isolation of bursal lymphocytes for analysis in cell culture results in the rapid endogenous activation of apoptosis. After 2 h of incubation, over 45% of the lymphocytes were shown to be undergoing apoptosis and by 6 h 80% were undergoing apoptosis as demonstrated by a terminal deoxynucleotidyltransferase-mediated dUTP-fluorescein isothiocynate nick end-labeling flow-cytometric analysis. These results were corroborated by a propidium iodide-staining flow-cytometric assay and by an agarose gel electrophoresis DNA fragmentation assay that demonstrated internucleosomal DNA cleavage of genomic DNA in apoptotic bursal lymphocytes. Endogenous activation of apoptosis in bursal lymphocytes could be inhibited in a dose-dependent fashion with the phorbol ester, phorbol 12-myristate 13-acetate, but not the phorbol ester antagonist 4 alpha-phorbol 12,13-didecanoate. In addition, apoptosis could be inhibited in a dose-dependent fashion with inhibitors of protein translation, cycloheximide, and puromycin, as well as the transcriptional inhibitor actinomycin D. These results suggest that endogenous activation of bursal lymphocyte apoptosis may be mediated by the protein kinase C signal transduction pathway and activation of this process appears to be dependent upon de novo protein biosynthesis.

Animals↗
PubMed1998

Brain administration of leptin causes deletion of adipocytes by apoptosis.

Leptin, produced in adipocytes, works through the central nervous system (CNS) to modulate food intake and energy expenditure, resulting in rapid loss of body fat depots. It is now shown that this process includes adipocyte apoptosis. Adipocyte deletion by apoptosis occurred after intracerebroventricular (i.c.v.) administration of leptin in rats. Adipose tissue of leptin-treated rats demonstrated characteristic features of apoptosis, including internucleosomal fragmentation of genomic DNA, elevated levels of DNA strand breaks and a reduction in total DNA content and cellular volume. These apoptotic features were absent in control and pair-fed rats and in other tissues of leptin-treated rats.

Adipocytes↗
PubMed1993

Absence of endonuclease activation during acute cell death in renal proximal tubules.

The role of endonuclease and poly(ADP-ribose) polymerase activation in various types of cell injuries and death to rabbit renal proximal tubule suspensions was examined. Proximal tubules were exposed to the mitochondrial inhibitor antimycin A (0.1 microM), the protonophore carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone (FCCP, 1 microM), the calcium ionophore ionomycin (5 microM), or the oxidant t-butyl hydroperoxide (TBHP, 0.5 mM) in the absence or presence of the endonuclease inhibitor aurintricarboxylic acid or the poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide. Lactate dehydrogenase (LDH) release was used as a marker of cell death and analysis of genomic DNA for internucleosomal cleavage was used as a marker of endonuclease activation. Aurintricarboxylic acid and 3-aminobenzamide had no effect on the proximal tubule LDH release produced by 1 h exposure to antimycin A, FCCP, or ionomycin, or 2 h exposure to TBHP. Furthermore, there was no evidence of DNA fragmentation with any compound prior to or after cell death began. As a positive control, proximal tubules exposed to digitonin in the absence of metabolic substrates resulted in the chelator-inhibitable fragmentation of DNA, indicating that the endonuclease is present in proximal tubules. These results show that endonuclease activation did not occur prior to or after cell death began. Furthermore, these results suggest that endonuclease and poly(ADP-ribose) polymerase activation do not play a role in this model of acute renal proximal tubule cell injury and death induced by agents that cause oxidative stress, mitochondrial dysfunction, or increases in cytosolic free calcium.

Animals↗
PubMed1993

Glucocorticoid-prolactin interactions in Nb2 lymphoma cells: antiproliferative versus anticytolytic effects.

The interaction between the immunosuppressive effects of glucocorticoids and the mitogenic effects of prolactin (PRL) were examined in Nb2 lymphoma cells, a pre-T cell line. The synthetic glucocorticoid, dexamethasone (Dex), caused a concentration-dependent (6.25-200 nM) inhibition of basal and ovine PRL (oPRL)-stimulated Nb2 cell proliferation. Although Dex was antiproliferative, the steroid had no effect on cell viability in the presence of PRL. However, when PRL was omitted from the medium, Dex increased the proportion of dead Nb2 cells by 24 hr in a concentration (25-200 nM)-dependent fashion without affecting total cell number. The antiproliferative and cytolytic effects of Dex were mimicked by other corticosteroids (cortisol, corticosterone, aldosterone, and deoxycorticosterone) in the expected order of glucocorticoid potency, but not by other steroids (17-beta-estradiol, progesterone, testosterone, 5-alpha-dihydrotestosterone, and dehydroepiandrosterone) or triiodothyronine. In addition, the antiproliferative and cytolytic effects of glucocorticoids were antagonized by the glucocorticoid receptor antagonist RU 486. Since corticosteroid-induced cytolysis was apparent only in the absence of mitogen, the anticytolytic effects of oPRL were tested. In the presence of Dex (100 nM), oPRL (25-1600 pg/ml) caused a concentration-dependent inhibition of cytolysis without changing cell number. Other lactogenic hormones (human growth hormone, human placental lactogen, rat PRL), but not trophic nonlactogenic hormones (rat growth hormone, human chorionic gonadotropin, ACTH), also inhibited Dex (100 nM)-induced cytolysis. Agarose gel electrophoresis of DNA extracted from Nb2 cells revealed that within 12 hr, 100 nM Dex induced DNA fragmentation, indicative of programmed cell death or apoptosis. Coincubation of cells with Dex and oPRL (1 ng/ml) inhibited Dex-induced fragmentation of Nb2 cell genomic DNA. These studies reveal a complex interaction between glucocorticoids and PRL in Nb2 cells. Although a glucocorticoid receptor-mediated antiproliferative effect is evident, PRL (at concentrations that usually stimulate cell proliferation) has the capacity to protect the cell against glucocorticoid-receptor-mediated induction of apoptosis.

Analysis of Variance↗
PubMed1993

Programmed cell death in avian thymocytes: role of the apoptotic endonuclease.

Apoptosis, or programmed cell death, is a physiological process whereby a target cell dies in response to a specific signal. A prominent model system used to study this process is the glucocorticoid-mediated killing of immature thymocytes. Following glucocorticoid treatment, apoptotic thymocytes undergo a series of distinct morphological alteration including cellular shrinkage, blebbing of the cytoplasmic membrane, and chromatin condensation. The chromatin condensation that occurs during apoptosis is associated with a characteristic endonuclease activity that degrades the genome at internucleosomal sites. To study this characteristic endonuclease activity further, nuclear extracts were prepared from thymocytes of glucocorticoid-treated chicks and nuclease activity present in the protein extracts was analyzed using chicken red blood cell nuclei as a substrate. Using this in vitro assay system, it was demonstrated that the avian endonuclease activity degrades chromatin at internucleosomal sites and can be inhibited by EDTA and zinc ions. Current efforts are focused on purifying the avian apoptotic endonuclease and further characterizing this nuclease activity.

Animals↗
PubMed1992

A biochemical hallmark of apoptosis: internucleosomal degradation of the genome.

Apoptosis, or programmed cell death, is an endogenous cellular process whereby an external signal activates a metabolic pathway that results in cell death. This form of cell death appears to be a common feature in many biological processes where cell deletion is a mechanism for altering tissue structure and function. Historically, apoptosis has been studied using histological techniques; however, more recent interest has focused on analyzing this process at the biochemical level. A biochemical hallmark of apoptosis is a characteristic form of DNA degradation in which the genome is cleaved at internucleosomal sites, generating a 'ladder' of DNA fragments when analyzed by agarose gel electrophoresis. A number of assay systems have been developed to study this nuclease activity. For example, nuclease activity has been analyzed by measuring the release of endogenous DNA from apoptotic cells, by flow cytometric analysis of apoptotic cells and by analyzing in situ apoptotic nuclease activity in polyacrylamide gels containing DNA. Use of these assay systems has enabled investigators to study the signal transduction pathways that mediate apoptosis and to characterize the endonuclease itself. Future biochemical studies in this field will focus on isolating the genes and gene products that mediate apoptosis.

Animals↗
PubMed1992

Characterization of apoptosis-like endonuclease activity in avian thymocytes.

In the current study the internucleosomal DNA cleavage activity associated with apoptosis was investigated in avian thymocytes. Thymocyte nuclear proteins from glucocorticoid-treated chickens were incubated with chicken red blood cell (cRBC) nuclei, and DNA degradation was analyzed by agarose gel electrophoresis and fluorescence-activated flow cytometry. The thymocyte nuclear extract contained an endonuclease activity that degraded cRBC chromatin at internucleosomal sites as detected by agarose gel electrophoresis. Flow cytometry analysis of cRBC nuclei that were treated with thymocyte nuclear proteins demonstrated a loss of cellular DNA as a function of the amount of added nuclease activity. Furthermore, it was demonstrated that the thymocyte nuclear extract contained a nuclease activity that was capable of degrading radiolabelled naked 32P-DNA into acid soluble DNA fragments. All three assay methods demonstrate that the thymocyte nuclease activity can be inhibited by EDTA, zinc ions and the nuclease inhibitor aurintricarboxylic acid. Based on the analysis of cofactor requirement of this nuclease activity and its susceptibility to inhibitors, the endonuclease activity present in avian apoptotic thymocytes appears to be identical to the mammalian counterpart.

Animals↗