RESUMO
Cell death often occurs after hypoxic/ischemic injury to the central nervous system. Changes in levels of the anti-apoptotic Bcl-X(L) protein may be a determining factor in hypoxia-induced neuronal apoptosis. The transcription factor NF-kappa B regulates bcl-x gene expression. In this study, we examined the role of NF-kappa B in the regulation of bcl-x in hypoxia-induced cell death. Rat hippocampus and basal forebrain tissues were collected at different time points after hypoxia (7%O(2), 93% N(2) for 10 or 20 min). We found that 1) hypoxia induced apoptosis in the hippocampus and basal forebrain; 2) the NF-kappa B dimers c-Rel/p50 and p50/p50 bound to the bcl-x promoter NF-kappa B sequence (CS4) in the hippocampus, but only p50/p50 bound to the CS4 sequence in the basal forebrain and hypoxia-induced differential binding patterns of c-Rel/p50 and p50/p50 correlated with the bcl-x expression pattern in the hippocampus; 3) the hypoxia-induced patterns of binding of c-Rel/p50 to the bcl-x promoter CS4 sequence were different from those to the IgG-kappa B enhancer sequence, whereas those of p50/p50 were similar to both sequences; 4) nuclear protein levels of c-Rel, but not p50, correlated with the c-Rel/p50 DNA binding patterns to the bcl-x CS4 site; and 5) there were differential responses to hypoxia among the different NF-kappa B protein subunits. These results suggest that there is a tissue-specific regulation of bcl-x gene expression by NF-kappa B in hypoxia-induced cell death in the hippocampus. The absence of these regulating features in the basal forebrain may account for the early appearance of apoptosis in response to hypoxia as compared with that in hippocampus.
Assuntos
Hipocampo/metabolismo , Hipóxia Encefálica/metabolismo , NF-kappa B/metabolismo , Prosencéfalo/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Apoptose/fisiologia , Masculino , Regiões Promotoras Genéticas/fisiologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Transcrição Gênica/fisiologia , Proteína bcl-XRESUMO
Chronic oxidative stress has been hypothesized to be a major contributor to the aging process. The continued exposure to reactive oxygen species (ROS) generated by oxidative metabolism or environmental sources can damage critical cellular structures and be responsible for some age-related pathology. The exposure of rodents to 100% oxygen, isobaric hyperoxia, increases ambient ROS levels and significantly increases apoptosis in brain. The deleterious effects of ROS also include increased lipid peroxidation, protein oxidation, and DNA damage. Although differences in the relative amounts of oxidative stress in young and old brains have been observed, the mechanisms responsible for impaired aging-associated DNA repair processes have not been characterized. We measured DNA levels of the DNA repair enzyme apurinic/apyrimidinic endonuclease (APE/Ref-1) protein by Western blot analysis in the brains of young (3-month) and old (30-month) male rats exposed to isobaric hyperoxia. Given that APE/Ref-1 is the rate-limiting enzyme in the repair pathway of apurinic/apyrimidinic sites generated in DNA by oxidative damage, we assumed that APE/Ref-1 protein levels were a good reflection of ongoing DNA base excision repair. Isobaric hyperoxia stimulated APE/Ref-1 expression in the hippocampus and basal forebrain of young rats experiencing 100% oxygen for 6 hr, while aged rats showed no significant changes in APE/Ref-1 protein levels in all brain areas at any time tested (0-48 hr) after hyperoxia. Differences in the stress-induced levels of expression of DNA repair enzymes may contribute to apoptotic increases and pathology associated with the aging process.
Assuntos
Envelhecimento/metabolismo , Apoptose , Carbono-Oxigênio Liases/biossíntese , Reparo do DNA , DNA Liase (Sítios Apurínicos ou Apirimidínicos) , Hipocampo/enzimologia , Oxigênio/toxicidade , Prosencéfalo/enzimologia , Animais , Carbono-Oxigênio Liases/genética , DNA/análise , Indução Enzimática , Masculino , Estresse Oxidativo , Ratos , Ratos Endogâmicos BN , Ratos Endogâmicos F344 , Espécies Reativas de OxigênioRESUMO
Apoptosis in the central nervous system (in contrast to necrosis) is an endogenous cell suicide mechanism triggered in response to biological factors and genotoxic stimuli often resulting from oxidative stress. Excessive neural apoptosis may result in longterm brain dysfunction. A significant proportion of prematurely born infants are exposed to high oxygen and nutritional regimens deficient in antioxidant precursors. Such infants frequently display cognitive deficits when studied in later childhood. Studies in cell culture have characterized a close relationship between oxidative stress, glutathione availability and cell death. Here, we assessed this relationship in rat brain, as a model approximation of the situation that occurs in human infants. Two day old rats were exposed to an atmosphere of 95% oxygen and treated with buthionine sulfoximine (BSO), a glutathione synthesis inhibitor. Control groups consisted of rat-pups kept in air, air plus BSO, or oxygen alone. At the end of 5 days of treatment, brains were harvested, dissected and nerve growth factor protein (NGF), glutathione, and extent of apoptosis were measured. Hyperoxia induced a decrease in NGF protein while BSO induced a decrease in glutathione concentrations. Animals treated with both hyperoxia and BSO had a dramatic increase in the extent of brain apoptosis detected. We conclude from these studies that the brains of animals exposed to both oxidative stress and limited antioxidant protection are liable to pro-apoptotic changes. Increased cell death via apoptosis reflecting changes in neurotrophin and glutathione homeostasis may represent the mechanism responsible for the induction of the longterm cognitive deficits observed in some preterm infants.
Assuntos
Animais Recém-Nascidos/crescimento & desenvolvimento , Apoptose , Encéfalo/crescimento & desenvolvimento , Glutationa/antagonistas & inibidores , Oxigênio/administração & dosagem , Animais , Peso Corporal , Encéfalo/citologia , Encéfalo/metabolismo , Butionina Sulfoximina/farmacologia , Catarata/induzido quimicamente , Cerebelo/citologia , Cerebelo/metabolismo , Corpo Estriado/citologia , Corpo Estriado/metabolismo , Inibidores Enzimáticos/farmacologia , Feminino , Glutationa/metabolismo , Hipocampo/citologia , Hipocampo/metabolismo , Fatores de Crescimento Neural/metabolismo , Gravidez , Prosencéfalo/citologia , Prosencéfalo/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
BACKGROUND: N-acetylcysteine (NAC) may protect against oxidative injury by providing cysteine for glutathione (GSH) biosynthesis or by direct reactions with electrophiles. We have recently shown that hyperoxic exposure of rats prior to liver perfusion is associated with significant decreases in hepatic GSH and significant changes in biliary amino acid concentrations. We hypothesized that NAC administration during hyperoxic exposure would prevent depletion of hepatic GSH by providing cysteine for GSH biosynthesis. METHODS: NAC was administered during two conditions known to induce GSH depletion: hyperoxic exposure and biochemical inhibition of GSH synthesis using buthionine sulfoximine (BSO). After 48 hours, GSH concentrations in bile, liver and perfusate and biliary amino acid concentrations were determined using isolated perfused liver preparations. RESULTS: Administration of NAC to rats maintained in normoxic or hyperoxic conditions, prior to liver perfusion, resulted in dose-dependent increases in GSH concentrations in bile, liver and perfusate, increases in bile flow rates and changes in biliary amino acid concentrations. When BSO was given concurrently with NAC in normal or hyperoxic conditions, these effects were not observed, and oxidant stress was evident. CONCLUSIONS: NAC prevents oxidant stress during hyperoxic exposure, most likely by supplying cysteine as a precursor for GSH synthesis.
Assuntos
Acetilcisteína/uso terapêutico , Glutationa/deficiência , Hiperóxia/complicações , Acetilcisteína/administração & dosagem , Aminoácidos/metabolismo , Animais , Bile/metabolismo , Glutationa/metabolismo , Fígado/metabolismo , Masculino , Proteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
BACKGROUND: Hepatic stores of glutathione may be depleted by hyperoxic exposure or poor nutritional status. We studied the effects of hyperoxia or hepatic glutathione depletion on bile flow rates, and on biliary concentrations of glutathione and amino acids. METHODS: Glutathione depletion was induced in vivo by 1) hyperoxic exposure (O2) for 48 hours, 2) inhibition of glutathione synthesis by treatment with buthionine sulfoximine (BSO), 3) a combination of BSO + O2, or 4) inhibition of cysteine synthesis by propargyglycine (PPG). Livers were then isolated and perfused. RESULTS: Glutathione concentrations in bile, liver, and perfusate were significantly decreased by all treatments. Bile flow was significantly decreased in groups treated with BSO or O2 + BSO, and perfusate LDH was increased by O2 + BSO or PPG. Significant changes in biliary amino acid concentrations included decreased sulfur-containing amino acids and increased branched-chain amino acids in groups treated with BSO, PPG, or O2; and increased essential amino acids in groups treated with O2 or PPG. CONCLUSION: Oxygen exposure or inhibition of glutathione synthesis results in significant decreases in hepatic, perfusate and biliary glutathione concentrations, and increases in biliary amino acids. A decrease in bile flow rate was associated only with the most severe glutathione depletion.
Assuntos
Glutationa/metabolismo , Hiperóxia/metabolismo , Fígado/metabolismo , Alcinos/farmacologia , Aminoácidos Sulfúricos/análise , Animais , Bile/efeitos dos fármacos , Bile/metabolismo , Butionina Sulfoximina/farmacologia , Inibidores Enzimáticos/farmacologia , Glicina/análogos & derivados , Glicina/farmacologia , L-Lactato Desidrogenase/metabolismo , Fígado/efeitos dos fármacos , Masculino , Técnicas de Cultura de Órgãos , Oxigênio/administração & dosagem , Perfusão , Ratos , Ratos Sprague-DawleyRESUMO
The production of IL-10 by human neonatal blood mononuclear leukocytes (BML) stimulated with lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-alpha), antibodies to CD3, or phorbol 12-myristate 13-acetate (PMA) was measured. The production of IL-10 by neonatal BML cultured with LPS or TNF-alpha was approximately 20 and approximately 15%, respectively, of adult BML. The combination of human recombinant TNF-alpha and LPS failed to augment IL-10 production in neonatal BML. The decreased production of IL-10 by neonatal leukocytes was not due to an autocrine feedback mechanism because only low concentrations of IL-10 were found in newborn sera. A connection with TNF-alpha could not be ruled out, because TNF-alpha production by LPS-stimulated newborn BML and the expression of TNF-alpha receptors on newborn monocytes were reduced. Mean +/- SD of concentrations of IL-10 in supernatants from adult and neonatal BML after stimulation with antibodies to human CD3 for 48 or 72 h were 914 +/- 386 and 178 +/- 176 pg/mL, respectively (p < 0.0001). In experiments with enriched populations of neonatal T cells, the addition of PMA failed to augment IL-10 production. This suggested that newborn T cells may be in a different state of activation than adult T cells Thus, IL-10 production in neonatal monocytes and T cells is reduced and this study suggests that the reduction may be secondary in part to regulatory processes involving TNF-alpha and its receptors.
Assuntos
Recém-Nascido/sangue , Interleucina-10/biossíntese , Leucócitos Mononucleares/metabolismo , Receptores do Fator de Necrose Tumoral/biossíntese , Linfócitos T/metabolismo , Fator de Necrose Tumoral alfa/biossíntese , Adolescente , Adulto , Envelhecimento/sangue , Anticorpos Monoclonais , Complexo CD3/imunologia , Sangue Fetal/citologia , Sangue Fetal/metabolismo , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Modelos Lineares , Lipopolissacarídeos/farmacologia , Pessoa de Meia-Idade , Valores de Referência , Estimulação QuímicaRESUMO
The glutathione precursor cysteine is not contained in most total parenteral nutrition (TPN) formulations, and premature infants may not be capable of synthesizing cysteine because of a deficiency of cystathionase. Glutathione depletion may have negative effects on host defense against oxidative damage. Several studies have suggested that glutathione depletion induces ornithine decarboxylase activity and increases in polyamine concentrations. Since an inverse relationship between polyamine and glutathione concentrations has been suggested, the concentrations of both of these compounds may be altered in premature infants receiving TPN. We measured glutathione and polyamine concentrations of the small intestine and colon of prematurely delivered newborn rabbits administered TPN for 7 days after birth with or without added cysteine (75 or 150 mg kg-1 day-1). Maternally reared kits were also studied. Total glutathione concentrations in the gastrointestinal tract were significantly lower in kits administered cysteine-free TPN than in kits receiving cysteine or who were maternally reared. Polyamine concentrations did not differ among groups. Glutathione depletion of the small intestine and colon does occur during cysteine-free parenteral nutrition and may compromise intestinal defense against oxidant damage.
Assuntos
Animais Recém-Nascidos/metabolismo , Cisteína/administração & dosagem , Glutationa/metabolismo , Mucosa Intestinal/metabolismo , Nutrição Parenteral Total , Poliaminas/metabolismo , Animais , Peso Corporal , Colo/metabolismo , Cisteína/farmacologia , Idade Gestacional , Intestino Delgado/metabolismo , CoelhosRESUMO
BACKGROUND: TNF-alpha mediates the hepatic response to sepsis by mechanisms which are not well understood. TNF-alpha is known to stimulate the hepatocellular uptake of specific amino acids in vivo; however, little is known about the direct effects of TNF-alpha on hepatic amino acid or glutathione homeostasis, which is a potential factor in the acute hepatic response to sepsis. METHODS: Using the isolated perfused rat liver, we characterized the effects of TNF-alpha on the secretion of amino acids and glutathione into bile and perfusate. Livers taken from adult male Sprague-Dawley rats were perfused with TNF-alpha at a dose of 1 or 2 micrograms. Bile and perfusate were collected for the quantitation of amino acid and glutathione concentrations. RESULTS: Administration of 2 micrograms TNF-alpha resulted in significant increases in biliary and perfusate concentrations of branched chain, gluconeogenic, and total amino acids. TNF-alpha was also associated with dose-related increases in oxygen uptake, and greater biliary concentrations of glutathione. CONCLUSION: TNF-alpha has direct effects upon hepatic amino acid metabolism, which represent some of the early events involved in the mechanism of response to sepsis.
Assuntos
Aminoácidos/metabolismo , Fígado/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologia , Animais , Relação Dose-Resposta a Droga , Masculino , Perfusão , Ratos , Ratos Sprague-DawleyRESUMO
Deficiencies in the quantitative expression of CD45RA and CD45RO on CD4+ and CD8+ T cells and in a population of CD45RA(low)CD45RO(low) T cells in blood from term newborn infants were found by flow cytometry. The relative frequencies of CD45RO on CD4+ T cells from adults and newborn infants were 72 and 58%, respectively. However, in newborn infants greater than 70% of T cells expressing CD45RO also expressed CD45RA. In addition, the quantitative expression of CD45RA and CD45RO on newborn T cells was significantly less than that found on adult blood T cells.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Antígenos Comuns de Leucócito/biossíntese , Subpopulações de Linfócitos T/imunologia , Adolescente , Adulto , Sangue Fetal/citologia , Citometria de Fluxo , Humanos , Recém-Nascido , Antígenos Comuns de Leucócito/sangue , Antígenos Comuns de Leucócito/químicaRESUMO
Total parenteral nutrition (TPN) has been adapted as a standard for providing nutrition to ill term and preterm infants. The availability of tyrosine in amino acid preparations utilized for TPN is limited and may potentiate a tyrosine-deficient state. Phenlyalanine hydroxylase activity, responsible for catalyzing tyrosine synthesis, has been suggested to be decreased in fetal and neonatal animals. Parenterally nourished premature rabbits (n = 16) and suckled rabbits (n = 19) were studied in order to compare growth parameters and amino acids in the plasma and brain, as well as whole brain catecholamine concentrations. Influx velocities into the brain of amino acids were also determined in these two groups. The preterm rabbit's average birth weight (42.6 +/- 6.0) was less than that of term rabbits (56.7 +/- 8.7, P < 0.005). Significantly lower concentrations of the catecholamine precursor tyrosine were found in both the plasma and brain of the parenterally nourished animals compared to the suckled animals. Tyrosine is reduced in the brain in TPN-supported animals reflecting both low tyrosine intake and increased plasma concentrations of large neutral amino acids that compete for uptake at the blood-brain barrier. However, no difference was observed between the two groups in their brain catecholamine concentrations. The seven-day parenterally nourished rabbit appears to be tyrosine-deficient but no evident effects on brain catecholamine concentrations were seen. The effects and impact of a tyrosine-deficient state might better be evaluated by regional evaluation of catecholaminergic areas of the brain or over a longer period of parenteral nutrition.
Assuntos
Aminoácidos/metabolismo , Catecolaminas/metabolismo , Nutrição Parenteral Total , Tirosina/metabolismo , Aminoácidos/sangue , Animais , Animais Recém-Nascidos , Barreira Hematoencefálica , Idade Gestacional , Peso Molecular , Coelhos , Tirosina/sangueRESUMO
BACKGROUND: Administration of parenteral nutrition (PN) that has been irradiated with light is associated with hepatic dysfunction in rats in vivo. Using the isolated perfused rat liver, we report the in vitro hepatic response to a light-exposed amino acid-vitamin (AAV) solution, compared with a light-protected solution. METHODS: The amino acid-vitamin solution (3 g Aminosyn and 2.5 mL MVI-12 added to buffer) was placed under a lamp in a beaker that was covered completely with foil (light-protected) or with a transparent wrap (light-exposed) for 24 hours before liver perfusion. Livers from adult male rats were isolated and perfused with buffer for 30 minutes, with the AAV solution for 60 minutes, and again with buffer for 30 minutes. RESULTS: Infusion with the AAV solution resulted in decreases in bile flow rates. Compared with light-protected groups, light-exposure was associated with significantly lower bile flow rates, significant increases in biliary concentrations of oxidized glutathione (GSSG), and significantly decreased biliary concentrations of free amino acids, including the glutathione precursors glutamate and glycine. CONCLUSIONS: Perfusion of the isolated rat liver with an AAV solution that has been irradiated with light for 24 hours results in a decrease in bile flow rates and an increase in biliary GSSG concentrations, suggesting oxidant stress. Consideration should be given to protecting solutions from light in the clinical setting.
Assuntos
Aminoácidos/farmacologia , Luz , Fígado/efeitos dos fármacos , Vitaminas/farmacologia , Aminoácidos/administração & dosagem , Aminoácidos/efeitos da radiação , Animais , Bile/química , Bile/metabolismo , Glutationa/metabolismo , Técnicas In Vitro , Infusões Intravenosas , L-Lactato Desidrogenase/metabolismo , Fígado/metabolismo , Masculino , Nutrição Parenteral , Perfusão , Ratos , Ratos Sprague-Dawley , Fatores de Tempo , Vitaminas/administração & dosagem , Vitaminas/efeitos da radiação , gama-Glutamiltransferase/metabolismoRESUMO
OBJECTIVES: The objectives of this investigation were: (1) to determine degree of elevation of serum C-reactive protein (CRP) in uncomplicated acute otitis media (AOM); (2) to compare serum CRP levels in bacterial and viral otitis media; and (3) to determine whether a single serum CRP level, obtained early in the course of AOM, could be used to differentiate between viral and bacterial otitis media. DESIGN AND METHODS: Sera were obtained from otherwise healthy infants and children with AOM who were 3 months to 7 years of age between 1989 and 1991. Tympanocentesis, bacterial and viral studies of the middle ear fluids, virologic studies of nasal wash specimens, measurements of serum antibody titers to respiratory viruses, blood counts, and quantitation of serum CRP concentrations were performed. After the initial tympanocentesis, an oral antibiotic was given for the next 10 days. The patients were clinically reevaluated over next 4 weeks. OUTCOME MEASURES: Serum CRP concentrations were compared among subjects with AOM who were divided into four groups based on the results of bacteriologic and virologic studies: group I, Bacterial infection (n = 82); group II, bacterial and viral infections (n = 69); group III, viral infection (n = 12); and group IV, no identifiable pathogen (n = 22). RESULTS: There was no statistical difference in serum CRP values among the four groups. The ranges of CRP were less than 0.6 to 22.8, less than 0.6 to 17.8, less than 0.6 to 2.0, and less than 0.6 to 6.8 mg/dL in groups I through IV, respectively. However, when CRP values in bacteria-positive cases were compared with CRP concentrations in bacteria-negative cases (1.58 +/- 3.16 vs 0.64 +/- 1.24 mg/dL), the difference was statistically significant. Furthermore, a significantly higher proportion of bacteria-positive cases had serum CRP concentrations greater than 2 mg/dL, compared with those in bacteria-negative cases. There was no correlation between initial CRP values and clinical findings and/or the clearance of bacteria from the middle ear. After 10 days of antibiotic treatment, CRP values returned to normal (< 0.6 mg/dL) in all cases. CONCLUSION: In AOM, the range of serum CRP varied from less than 0.6 to 22.8 mg/dL. High CRP values (> 2.0 mg/dL) were associated with 22% of cases of bacterial AOM but only with 6% of nonbacterial AOM. High levels of serum CRP were found to be very specific in detecting bacterial AOM, and no cases of viral AOM without a concurrent bacterial infection were found to exhibit high serum levels of CRP.
Assuntos
Infecções Bacterianas/diagnóstico , Proteína C-Reativa/análise , Otite Média/microbiologia , Viroses/diagnóstico , Doença Aguda , Infecções Bacterianas/sangue , Criança , Pré-Escolar , Diagnóstico Diferencial , Feminino , Humanos , Lactente , Masculino , Otite Média/virologia , Sensibilidade e Especificidade , Viroses/sangueRESUMO
The concentrations of immunoreactive IL-10 in the aqueous fraction of 20 specimens of human milk obtained during the first 80 h of lactation and stored at -60 degrees C ranged from 66 to 9301 pg/mL (mean +/- SD, 3304 +/- 3127 pg/mL). IL-10 was present also in the lipid layer of milk. Gel filtration revealed that IL-10 was located in a high molecular weight fraction, where certain other cytokines in human milk have been found. In addition, immunoreactive IL-10 in milk increased after treatment with sodium taurocholate. Bioactive IL-10 was demonstrated by the finding that human milk inhibited [3H]thymidine uptake by human blood lymphocytes and that inhibition was partly overcome by concomitant incubation with antibodies to human IL-10. IL-10 mRNA but no protein product was found in cultured human mammary epithelial cells. Some IL-10 was associated with preparations of human milk leukocytes, but the data did not suggest that the cells were producing the cytokine. Bioactive IL-10 in a possible protected compartment suggests that IL-10 in human milk may have immunomodulating, antiinflammatory effects on the alimentary tract of the recipient infant.
Assuntos
Citocinas/biossíntese , Interleucina-10/análise , Leite Humano/química , Adulto , Sequência de Bases , Feminino , Humanos , Dados de Sequência Molecular , TemperaturaRESUMO
We studied the effects of a transient elevation in biliary pressure on biliary glutathione and amino acids in rats. Other biliary solutes monitored were total bile salt, Pi, which is a putative marker of paracellular leakage, and glucose, which is reabsorbed from the biliary tract. Experiments were carried out on anesthetized rats intraduodenally infused with taurocholate to maintain bile flow during a 2-hr basal period, a 4-hr pressure period during which the bile duct cannula was elevated until bile flow decreased to 1/3 the basal rate, and a 2-hr period after release of hydrostatic biliary pressure. We found that pressure treatment caused biliary concentrations of glutathione to progressively decrease by 80%, while biliary Pi rapidly rose approximately 3- to 4-fold, bile salt gradually increased approximately 3-fold, and biliary glucose concentration progressively rose 15-fold. HPLC analysis of monobromobimane-derivatized biliary thiols indicated that the decline in biliary glutathione was not accompanied by an increase in its breakdown products, cysteine and cysteinylglycine. Pressure treatment led to four patterns of change in biliary amino acid concentrations: (1) increases of 29 to 76% for the basic amino acids lysine and arginine, which have very low bile/plasma ratios of about 0.1; (2) no change for the more water soluble amino acids with bile/plasma ratios close to 1.0, e.g., histidine and urea; (3) modest decreases of 16 to 48% for a variety of amino acids including serine, glutamate, and glycine; and (4) marked, progressive decreases of > 50% for aromatic and branched chain amino acids. By 2 hr after release of pressure, only the alterations in biliary glucose and some amino acids, particularly the branched chains, persisted. This is the first report of cholestasis-induced alterations in biliary amino acids.
Assuntos
Aminoácidos/metabolismo , Bile/metabolismo , Sistema Biliar/fisiopatologia , Glutationa/metabolismo , Animais , Ácidos e Sais Biliares/metabolismo , Cromatografia Líquida de Alta Pressão , Glucose/metabolismo , Masculino , Fosfatos/metabolismo , Pressão , Ratos , Ratos Sprague-Dawley , Compostos de Sulfidrila/metabolismo , Ureia/metabolismoRESUMO
Despite the fact that breastfeeding is the most appropriate form of nutrition for the healthy term infant, the rate of initiation in the U.S. is declining. One demographic factor associated with this low rate is ethnicity and so in this study we measured acculturation (one aspect of ethnicity) into the U.S. and its relationship to the successful initiation of breastfeeding in a sample of women recruited approximately 2 months prenatally in a U.S.-Mexico border city. Interviews were administered in English or Spanish by bilingual interviewers prenatally (n = 906), natally (n = 788), and postnatally (n = 715). Acculturation (measured with a 20 item instrument) was strongly related to the intent to (p < 0.001) or the successful initiation of breastfeeding (p < 0.001). Marital status (p = 0.014) and education (p = 0.002) were related to breastfeeding prenatally and natally. Initiation of breastfeeding was highest among those women least acculturated (52.9%) and lowest in those most acculturated (36.1%) indicating an inhibiting effect of acculturation. To improve the rate of initiation of breastfeeding in the U.S. (a national health goal) intervention programs must consider cultural factors.
Assuntos
Aculturação , Aleitamento Materno/etnologia , Americanos Mexicanos , Adolescente , Adulto , Aleitamento Materno/psicologia , Aleitamento Materno/estatística & dados numéricos , Feminino , Educação em Saúde/métodos , Humanos , Americanos Mexicanos/psicologia , Fatores Socioeconômicos , Estados Unidos/epidemiologiaRESUMO
P2-fractions were isolated from rat brain, and used to study net taurine transport. The fractions were incubated in increasing concentrations of [3H]taurine and the intraterminal concentration measured by liquid scintillation and amino acid analysis. The membrane potential of the isolated fractions was estimated using 86Rb+ as a marker for intracellular K+. Taurine was synthesized in the P2-fraction when incubated in taurine free medium. At external taurine concentrations below 370 microM a significant amount of the endogenous taurine was released to the incubation medium. Net taurine uptake into the P2-fraction was achieved at external taurine concentrations exceeding 370 microM. The taurine antagonist 6-aminomethyl-3-methyl-4H, 1, 2, 4-benzothiadiazine-1, 1-dioxide (TAG) competitively inhibited taurine and [3H]taurine transport into the P2-fraction. As the external concentration of taurine was increased, the accumulation of 86Rb+ into the P2-fraction was facilitated. This indicated an increasing hyperpolarization of the neuronal membrane as taurine transport shifted from release towards uptake. TAG reduced the hyperpolarization that paralleled taurine accumulation, in a dose dependent manner. Our results indicate that relatively low transmembranal gradients of taurine may be maintained by an electrogenic taurine transporter having a large transport capacity. Such a transporter may well serve the needs of osmotic regulation, i.e. to transport large amounts of taurine in any direction across the neuronal membrane.
Assuntos
Benzotiadiazinas/farmacologia , Encéfalo/metabolismo , Taurina/metabolismo , Animais , Transporte Biológico/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Feminino , Cinética , Ratos , Ratos Sprague-Dawley , Rubídio/metabolismo , Radioisótopos de Rubídio , Contagem de Cintilação , Taurina/antagonistas & inibidores , TrítioRESUMO
The intravenous infusion of amino acid solutions has been associated with cholestatic liver injury in hospitalized patients and in laboratory animals. In the isolated rat liver, we recently showed that the acute decrease in bile flow, previously reported by other investigators, is dose related, reversible, and associated with dose-related increases in total biliary amino acid concentrations. In the present study, we characterized the effects of graded infusions of amino acid solutions, with and without taurocholate, on biliary secretion of individual amino acids and glutathione, an important regulator of bile flow. Livers from young adult male rats were perfused with an amino acid solution for 1 hour and allowed to recover for 30 minutes. Infusion of the amino acid solution was associated with dose-related increases in biliary concentrations of most amino acids included in the amino acid solution. Infusion of amino acid solutions resulted in a decreased bile/perfusate ratio of most amino acids, which were secreted into bile in amounts approximating their calculated uptake from the infusate. The inclusion of taurocholate in the infusate was associated with lower biliary concentrations of each individual amino acid and significant decreases in biliary total, reduced, and oxidized glutathione. Further investigation of the relationship between these changes in biliary amino acids and glutathione concentrations and the cholestasis associated with infusion of amino acid solutions may provide insights into the mechanism by which amino acids induce such cholestasis.
Assuntos
Aminoácidos/administração & dosagem , Aminoácidos/metabolismo , Ácidos e Sais Biliares/metabolismo , Glutationa/metabolismo , Fígado/efeitos dos fármacos , Aminoácidos/farmacologia , Animais , Bile/química , Ductos Biliares/metabolismo , Relação Dose-Resposta a Droga , Infusões Intravenosas , Fígado/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Ácido Taurocólico/metabolismoRESUMO
BACKGROUND: In 1983 the Department of Pediatrics at the University of Texas Medical Branch at Galveston established a faculty development program to address faculty needs for continuing education and improved resources for research. At first a part-time coordinator was hired; then, in 1985, a full-time, faculty-level science communicator provided help with strategic planning of projects and intensive review of grant proposals and journal articles. Faculty participation in the program was voluntary. METHOD: Pre- and post-intervention data for 1983-1992 included numbers of faculty using the program, faculty evaluations of the program, grant dollars awarded, counts of grant submissions and awards, and numbers of published articles. RESULTS: The review services were used heavily for grant proposals (75% of the department's proposals), but were used lightly for research articles (18% of publications). Grant funding quadrupled from 1983 to 1988; although funding peaked in 1988, it thereafter remained at three to four times the 1983 level. In contrast, the mean number of publications per faculty per year dropped between 1983 and 1990. CONCLUSION: The program provided valuable assistance to the faculty in writing grant proposals, and it helped to generate critically needed resources. However, the program's failure to increase the publication productivity of the faculty suggests that despite financial pressures, similar programs should use their influence and resources to promote a balance between scholarly publication and grant acquisition.
Assuntos
Docentes de Medicina , Apoio à Pesquisa como Assunto , Desenvolvimento de Pessoal , Apoio ao Desenvolvimento de Recursos HumanosRESUMO
Previous studies have demonstrated a strong association between breastfeeding incidence and ethnic background, indicating the presence of strong cultural influences on infant-feeding patterns within ethnic groups. This study analyzed the relationship of acculturation into the United States, one aspect of ethnicity, to the initiation of breastfeeding among a sample of 213 women recruited approximately 2 months prenatally in the United States-Mexico border city of Brownsville, Texas. An in-depth, structured interview was conducted at that time (n = 213), at the time of birth (n = 207), and 2 to 3 weeks postnatally (n = 206). In addition, male partners of the mothers were interviewed prenatally when available (n = 119). Acculturation scores and demographic characteristics were tested relative to breastfeeding initiation. The degree of acculturation, age, and marital status were associated significantly with breastfeeding initiation (with breastfeeding continued at 2 to 3 weeks postnatally), but occupational status, education, and income were not. Breastfeeding incidence was highest among older, married, or low-acculturated, Anglo mothers. These data suggest that women being assimilated into the United States are inhibited in the initiation of breastfeeding. These findings suggest that to be successful, intervention programs to increase breastfeeding incidence must consider the cultural adaptation of different groups as they meld with the larger population.