RESUMO
BACKGROUND AND AIMS: Elevated plasminogen activator inhibitor 1 (PAI-1) concentrations are a hallmark of obesity and are considered to contribute to the development of cardiovascular disease. As adipose tissue constitutes a major source for PAI-1 in obesity, we investigated the individual contribution of subcutaneous and intra-abdominal fat on PAI-1 concentrations during pronounced weight loss after bariatric surgery. METHODS AND RESULTS: Thirty-seven obese adults were examined before and 18 months after surgery. Abdominal fat distribution was determined by ultrasound, metabolic parameters and plasma PAI-1 levels by standard methods. BMI was reduced by 9.2 ± 4.9 kg/m(2), while total fat mass and visceral fat diameter (VFD) decreased by 20.7 ± 11.9 kg and 4.2 ± 2.3 cm, respectively. Concomitantly, PAI-1 levels diminished by 3.2 ± 5.6 ng/ml (all p ≤ 0.015). Change in PAI-1 levels was correlated with change in VFD (r = 0.441, p = 0.008), but not with subcutaneous fat diameter. In stepwise multiple regression analysis change in VFD was an independent predictor of change in PAI-1 concentrations. When adjusted for age and sex or total fat mass associations between PAI-1 and VFD remained significant. CONCLUSION: We demonstrate that VFD is a major determinant for PAI-1 concentrations during pronounced weight loss after bariatric surgery. Thus, significant reduction of visceral fat mass may contribute to the reduced cardiovascular morbidity and mortality after bariatric surgery by a concomitant decrease in PAI-1 concentrations.
Assuntos
Cirurgia Bariátrica , Obesidade Abdominal/sangue , Inibidor 1 de Ativador de Plasminogênio/sangue , Redução de Peso , Adulto , LDL-Colesterol/sangue , Feminino , Humanos , Modelos Lineares , Masculino , Síndrome Metabólica/diagnóstico , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
BACKGROUND: Apolipoprotein A5 (apoA5) is a recently described liver-specific protein that has been shown to influence triglyceride (TG) metabolism. ApoA5 transgenic mice display dramatically reduced TG levels, while in contrast apoA5 deficiency in humans was reported to result in marked hypertriglyceridemia. ApoA5 exerts its extracellular effects by increasing lipolysis of TG-rich lipoproteins, while in vitro data suggest additional intrahepatic effects. METHODS: In this study the authors set out to investigate a possible role of apoA5 in non-alcoholic fatty liver disease (NAFLD). We thus determined hepatic apoA5 expression in 15 obese subjects with histologically proven NAFLD undergoing bariatric surgery. In addition, the authors established a hepatic cell culture model of apoA5 knockdown by transfecting human hepatoma cells (HepG2) with apoA5 small interfering (si) RNA, and determined intracellular TG content and expression levels of key enzymes and transcription factors of intrahepatic lipid metabolism in these cells. RESULTS: Pronounced weight loss and associated histologically verified improvement of hepatic steatosis were accompanied by significant reductions of hepatic apoA5 mRNA expression levels. Significant apoA5 knockdown in HepG2 cells resulted in a marked decrease of intracellular TG content. When HepG2 cells were co-transfected with apoA5 and peroxisome proliferator-activated receptor gamma (PPARγ), reductions in hepatic TG accumulation were significantly less pronounced when compared to apoA5 siRNA transfected HepG2 cells. CONCLUSIONS: In obese subjects, hepatic apoA5 mRNA expression decreases after weight loss and improvements in hepatic steatosis. The authors' in vitro data demonstrate that apoA5 influences intrahepatic TG metabolism and that these intracellular effects of apoA5 are accompanied by changes in PPARγ mRNA expression. In summary, the data suggest that as well as several other factors, apoA5 might be involved in the pathogenesis of hepatic steatosis.
Assuntos
Apolipoproteínas A/fisiologia , Fígado Gorduroso/metabolismo , Adulto , Antropometria/métodos , Apolipoproteína A-V , Apolipoproteínas A/biossíntese , Apolipoproteínas A/genética , Cirurgia Bariátrica , Fígado Gorduroso/etiologia , Feminino , Regulação da Expressão Gênica/fisiologia , Técnicas de Silenciamento de Genes , Humanos , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica , Obesidade/complicações , Obesidade/metabolismo , Obesidade/cirurgia , PPAR gama/biossíntese , PPAR gama/genética , RNA Mensageiro/genética , RNA Neoplásico/genética , RNA Interferente Pequeno/genética , Transfecção , Triglicerídeos/metabolismo , Células Tumorais Cultivadas , Redução de Peso/fisiologiaRESUMO
Human immunodeficiency virus type 1 (HIV-1), in contrast to animal retroviruses such as murine leukemia virus, is not lysed by human complement. Nevertheless, HIV-1 activates complement via the classical pathway independent of antibody, and C3b deposition facilitates infection of complement receptor-bearing cells. Using gel exclusion chromatography on Sephacryl S-1000, purified virions were found to bind 125I-labeled C1q, but not 125I-labeled dimeric proenzyme C1s. Virions activated the C1 complex, reconstituted from C1q, proenzyme C1r, and 125I-labeled proenzyme C1s, to an extent comparable with that obtained with immunoglobulin G-ovalbumin immune complexes. To determine the activating viral component, recombinant viral proteins were used: in the solid phase, soluble gp41 (sgp41) (the outer membrane part of gp41, residues 539-684 of gp160) bound C1q, but not dimeric proenzyme C1s, while gp120 was ineffective. In the fluid phase, sgp41 activated the C1 complex in a dose- and time-dependent manner, more efficiently than aggregated Ig, but less efficiently than immune complexes. To localize the C1 activating site(s) in gp41, synthetic peptides (15-residue oligomers spanning amino acids 531-695 of gp160) were used. Peptides covering positions 591-605 and 601-620 and, to a lesser extent, positions 561-575, had both the ability to bind C1q and to induce C3 deposition. These data provide the first experimental evidence of a direct interaction between the C1 complex and HIV-1, and indicate that C1 binding and activation are mediated by specific sites in gp41.
Assuntos
Complemento C1/metabolismo , Via Clássica do Complemento , Proteína gp41 do Envelope de HIV/metabolismo , HIV-1/fisiologia , Sítios de Ligação , Proteína gp120 do Envelope de HIV/metabolismo , HIV-1/patogenicidade , Humanos , Polimixina B/farmacologia , Proteínas Recombinantes/metabolismoRESUMO
BACKGROUND: Subclinical inflammation in obesity is critical for development of several obesity-associated disorders. We set out to investigate the effect of pronounced weight loss on circulating chemerin levels, a chemoattractant protein that also influences adipose cell function by paracrine and autocrine mechanisms. MATERIAL AND METHODS: Thirty-two obese patients undergoing bariatric surgery were tested before and on an average of 18 months after gastric banding or gastric bypass surgery. RESULTS: Pronounced weight loss after bariatric surgery was accompanied by improvements in parameters of lipid and glucose metabolism and increased adiponectin levels. Chemoattractant chemerin significantly decreased from 175.91 +/- 24.50 to 145.53 +/- 26.44 ng mL(-1) after bariatric surgery (P < or = 0.01). Concomitantly, hs-CRP as a marker of subclinical inflammation was significantly reduced after weight reduction (P < or = 0.01). CONCLUSIONS: We hypothesize that weight-loss induced reduction in circulating chemerin might in conjunction with other factors be associated with diminished recruitment of macrophages in adipose tissue and reduction of subclinical inflammation, which again could partly explain beneficial long-term effects of weight reduction in obese subjects.
Assuntos
Cirurgia Bariátrica , Quimiocinas/sangue , Obesidade/sangue , Redução de Peso/fisiologia , Adiponectina/sangue , Adulto , Biomarcadores/sangue , Proteína C-Reativa/análise , Feminino , Humanos , Inflamação/sangue , Peptídeos e Proteínas de Sinalização Intercelular , Masculino , Obesidade/cirurgiaRESUMO
BACKGROUND AND AIMS: Several studies indicate that changes in the plasma concentrations of adipocyte-fatty acid binding protein (A-FABP), retinol binding protein-4 (RBP-4) and visfatin are associated with chronic states of insulin resistance. Recent studies have shown that postprandial lipemia induces an acute state of insulin resistance. The aim of this study was to investigate the effect of postprandial lipemia on the plasma concentrations of A-FABP, RBP-4 and visfatin. METHODS AND RESULTS: In a within-subject crossover study, we administered a standardized high-fat meal to 24 healthy subjects (12 males and 12 females). Plasma concentrations of adipocytokines were measured in the morning after an overnight fast and during postprandial lipemia, i.e. 2, 4 and 6 hours after meal ingestion (postprandial experiment). To exclude potential confounding factors affecting the adipocytokine plasma concentrations, a control experiment without meal ingestion was performed over the same time period (postabsorptive control experiment). Comparing plasma concentrations of A-FABP, RBP-4 and visfatin between the postprandial and the postabsorptive control experiments, we found no significant differences. Within either of the two experiments, a decrease of A-FABP was noted reaching, however, statistical significance only in the postprandial experiment, i.e. 2 and 4 hours after meal ingestion. CONCLUSION: Postprandial lipemia has no significant effect on the plasma concentrations of visfatin, A-FABP or RBP-4 in relation to their postabsorptive plasma profiles. We conclude that prolonged states of insulin resistance are required to affect plasma concentrations of these adipocytokines.
Assuntos
Proteínas de Ligação a Ácido Graxo/sangue , Alimentos , Hiperlipidemias/sangue , Nicotinamida Fosforribosiltransferase/sangue , Proteínas Plasmáticas de Ligação ao Retinol/análise , Adipocinas/sangue , Adulto , Glicemia/análise , HDL-Colesterol/sangue , Estudos Cross-Over , Gorduras na Dieta/administração & dosagem , Jejum , Ácidos Graxos não Esterificados/sangue , Feminino , Humanos , Insulina/sangue , Masculino , Triglicerídeos/sangueRESUMO
Non-alcoholic fatty liver disease (NAFLD) comprises a disease spectrum ranging from simple steatosis and steatohepatitis to cirrhosis. Based on its strongest risk factors namely visceral obesity and insulin resistance, NAFLD is thought to be the hepatic manifestation of the metabolic syndrome and is considered to be the most common liver disorder in Western countries. Pathophysiological mechanisms include an enlarged pool of fatty acids, subclinical inflammation, oxidative stress and imbalances of various adipocytokines such as adiponectin. Accordingly, targets for therapeutic interventions are miscellaneous: amelioration of obesity by pharmacological, surgical or lifestyle intervention has been evaluated with success in numerous, but not all studies. Some efficacy was reported for metformin and short-term glitazone treatment. In a large recently reported trial, vitamin E supplementation improved biochemical and histological markers in subjects with non-alcoholic steatohepatitis. Blockade of the endocannabinoid system has been proposed to be a promising target in NAFLD; however, very recently the cannabinoid receptor blocker rimonabant has been withdrawn because of central nervous system toxicity. Cytoprotective therapies and statins have been mainly ineffective in NAFLD. New but so far insufficiently studied therapeutic approaches include inhibitors of the renin-angiotensin system as well as incretin mimetics respectively.
Assuntos
Fármacos Gastrointestinais/uso terapêutico , Fármacos Antiobesidade/uso terapêutico , Cirurgia Bariátrica/métodos , Dieta , Terapia por Exercício/métodos , Fígado Gorduroso/diagnóstico , Fígado Gorduroso/etiologia , Fígado Gorduroso/terapia , Humanos , Estilo de Vida , Hepatopatia Gordurosa não AlcoólicaRESUMO
BACKGROUND: Weight loss induced by bariatric surgery is an effective method to reverse obesity and comorbidities. The aim of this prospective weight loss study was to investigate changes of body fat distribution in relation to adiponectin and its isoforms and further to investigate the influence of both body fat distribution and adiponectin on the degree of liver steatosis. DESIGN: Fifteen severely obese female patients (body mass index 43.1 +/- 4.1, mean age 34.5 +/- 8.6 years) were examined before and after surgical treatment. Grading of fatty liver disease and the subcutaneous and visceral fat diameters were determined by abdominal ultrasonography. Metabolic parameters were determined using standard methods; serum total adiponectin and its isoforms were detected by enzyme immuno assay (EIA). RESULTS: Mean weight loss was 28.3 kg, which was mostly due to a loss in fat mass, accompanied by an increase in total adiponectin and the high molecular weight (HMW) adiponectin isoform. Visceral adipose tissue (VAT) diameter was highly correlated with liver steatosis, even more strongly than the parameters of liver function. In addition, liver steatosis correlated negatively with HMW adiponectin and binary logistic regression revealed that changes in fat mass, HMW adiponectin and alanine aminotransferase (ALT) were the best predictors for changes in the degree of hepatic steatosis. CONCLUSIONS: Our results suggest that circulating HMW adiponectin is associated with both VAT and liver steatosis. In summary, the major findings were that the VAT diameter is highly correlated with liver steatosis, even stronger than the parameters of liver function and the association of HMW adiponectin with liver steatosis was better than with total adiponectin.
Assuntos
Adiponectina/sangue , Distribuição da Gordura Corporal , Fígado Gorduroso/metabolismo , Gordura Intra-Abdominal/metabolismo , Obesidade/patologia , Redução de Peso , Adiponectina/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Obesidade/metabolismo , Obesidade/terapia , Estudos Prospectivos , Isoformas de Proteínas/sangue , Resultado do TratamentoRESUMO
BACKGROUND: Adiponectin is an insulin-sensitizing, antiatherogenic and anti-inflammatory adipocytokine that circulates in three isoforms: a trimer [low-molecular weight (LMW)], a hexamer (trimer-dimer) of medium molecular weight (MMW) and a multimeric high molecular weight (HMW) isoform. Evidence is accumulating that HMW adiponectin is the active isoform of the adipocytokine. We investigated the impact of adipose tissue and insulin sensitivity on adiponectin isoform distribution. MATERIALS AND METHODS: One hundred and eighty-seven normolipidaemic, non-diabetic lean or obese subjects with or without insulin resistance participating in the Salzburg Atherosclerosis Prevention program in subjects at High Individual Risk (SAPHIR) were included in the study. Insulin sensitivity was determined by the short insulin tolerance test and the homeostasis model assessment (HOMA) index. Serum adiponectin isoform distribution was determined by an enzyme immunoassay. RESULTS: Total adiponectin as well as HMW/total adiponectin ratio was significantly increased in female subjects. Circulating total adiponectin levels were lowest in obese patients due to reduced concentrations of HMW adiponectin. As determined by stepwise regression analysis, besides age and high density lipoprotein (HDL) cholesterol, visceral fat area and waist-to-hip ratio predicted concentrations of HMW adiponectin, while insulin sensitivity had no influence on either total adiponectin or its isoforms. CONCLUSIONS: Our results underline that determination of adiponectin isoforms are more useful than measurement of total adiponectin in clinical settings. Our data suggest that adiponectin concentrations are strongly associated with visceral fat area but not with insulin sensitivity. Thus, we hypothesize that insulin resistance is a consequence rather than the cause of hypoadiponectinaemia in obese subjects.
Assuntos
Adiponectina/sangue , Tecido Adiposo/patologia , Resistência à Insulina , Obesidade/sangue , Adiponectina/química , Adulto , Idoso , Índice de Massa Corporal , Colesterol/sangue , Feminino , Galactose/análogos & derivados , Humanos , Imunoensaio , Masculino , Pessoa de Meia-IdadeRESUMO
PURPOSE: The aim of the study was to determine the influence of valproic acid (VPA) treatment on leptin, the soluble leptin receptor (sOB-R), the sOB-R/leptin ratio, body composition and insulin resistance in epileptic children. METHODS: A cross-sectional cohort study was conducted at the Medical University Innsbruck, Austria. Children >6 years with idiopathic epilepsy and antiepileptic drug therapy since at least six months were eligible. Leptin concentration, the sOB-R, the sOB-R/leptin ratio, body composition and glucose homeostasis were determined. RESULTS: 87 children (median [range] age 12.8 years [6.0-18.6]) were on treatment with VPA, 55 (12.3 years [6.4-18.3]) on other AEDs, comprising the non-VPA group. VPA-treated children had higher leptin concentrations, body-mass-index standard-deviation score (SDS), body fat (each p<0.001), serum insulin concentrations (p=0.014) and homeostasis model assessment (HOMA) index (p=0.009), as well as a lower sOB-R/leptin ratio (p<0.001) when compared to the non-VPA group. Overweight VPA-treated children showed lower sOB-R concentrations and a lower sOB-R/leptin ratio (each p<0.001) as well as higher body fat and leptin levels (each p<0.001) compared to lean VPA-treated children. CONCLUSION: VPA monotherapy was associated with higher body weight, body fat and serum leptin concentrations as well as impaired glucose homeostasis. Low sOB-R concentrations and a low sOB-R/leptin ratio in overweight VPA-treated patients might contribute to disturbances in glucose homeostasis and to the development of the metabolic syndrome in these children later in life.
Assuntos
Anticonvulsivantes/farmacologia , Composição Corporal/efeitos dos fármacos , Epilepsia/metabolismo , Leptina/sangue , Receptores para Leptina/sangue , Ácido Valproico/farmacologia , Adolescente , Antropologia Física/métodos , Anticonvulsivantes/uso terapêutico , Índice de Massa Corporal , Criança , Estudos Transversais , Relação Dose-Resposta a Droga , Epilepsia/tratamento farmacológico , Feminino , Humanos , Masculino , Fatores Sexuais , Ácido Valproico/uso terapêuticoRESUMO
OBJECTIVE: Evaluation of the influence of intradialytic parenteral nutrition (IDPN) in patients suffering from Malnutrition-Inflammation Complex Syndrome (MICS) on nutritional status, inflammation, adipocytokines and serum lipids. SUBJECTS: Six patients with MICS were assigned to IDPN, whereas six patients matched for age, sex, body mass index (BMI) and co-morbidity without malnutrition served as controls. Patients were recruited from Outpatient Dialysis Unit, Medical University Innsbruck and from Dialysis Unit, Hospital Feldkirch. RESULTS: In all patients with IDPN, dry body weight increased during the interventional period whereas body weight remained stable in patients without IDPN. Tumor necrosis factor (TNF)-alpha levels were higher in patients with MICS compared with controls at all time points. Total cholesterol, LDL- and HDL-levels significantly increased during dialysis at all time points in controls but not in patients with MICS. Albumin, C-reactive protein, interleukin-6 (IL-6), soluble interleukin-2 receptor (sIL-2R) and adipocytokines did not differ between patients and controls during the study period. CONCLUSIONS: IDPN in patients with MICS increases body weight despite not influencing inflammatory status. Furthermore, IDPN does not induce a pro-atherogenic lipid composition enhancing the risk for atherosclerosis. Thus, IDPN is a safe and effective treatment of malnutrition in patients with MICS.
Assuntos
Adipocinas/sangue , Inflamação/terapia , Lipídeos/sangue , Desnutrição/terapia , Estado Nutricional , Nutrição Parenteral/métodos , Idoso , Idoso de 80 Anos ou mais , Proteína C-Reativa/metabolismo , Colesterol/sangue , Estudos de Coortes , Feminino , Humanos , Inflamação/sangue , Inflamação/imunologia , Falência Renal Crônica/sangue , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Masculino , Desnutrição/sangue , Necessidades Nutricionais , Projetos Piloto , Estudos Prospectivos , Diálise Renal/métodos , Albumina Sérica/metabolismo , Resultado do Tratamento , Aumento de PesoRESUMO
OBJECTIVE: Obesity is associated with increased morbidity and mortality from atherosclerotic disease. Lipid abnormalities contribute to the increased relative risk in obese subjects. Cholesteryl ester transfer protein (CETP) mass is increased in these patients and might mediate the atherogenic lipoprotein pattern observed in obesity. METHODS AND RESULTS: Twenty-one morbidly obese, middle-aged, female subjects participated in this prospective study. Subjects were examined before and 1 year after surgical treatment. Fat mass was determined by body impedance analysis; CETP mass, by ELISA; CETP activity, by exogenous substrate assay; and LDL particle diameter, by gradient gel electrophoresis. Mean weight loss after 1 year was 28.7 kg; mean fat mass loss was 22.6 kg. Mean CETP mass decreased from 1.81 to 1.32 microg/mL (P=0.008); mean CETP activity decreased from 244 to 184 nmol x mL(-1) x h(-1) (P=0.004); and in parallel, the mean diameter of LDL particles increased (256.8 to 258.4 A, P=0.04). CONCLUSIONS: We conclude that weight loss is associated with a pronounced decrease in CETP mass and activity and a consistent increase in LDL particle diameter. After 1 year of this prospective study in morbidly obese subjects undergoing weight loss by surgical treatment, it has been determined that some features of the atherogenic lipoprotein profile can be reversed.
Assuntos
Arteriosclerose/sangue , Arteriosclerose/etiologia , Proteínas de Transporte/sangue , Glicoproteínas , Lipoproteínas/sangue , Obesidade Mórbida/sangue , Obesidade Mórbida/complicações , Adulto , Apolipoproteínas/sangue , Arteriosclerose/dietoterapia , Índice de Massa Corporal , Proteínas de Transporte/metabolismo , Proteínas de Transferência de Ésteres de Colesterol , Feminino , Humanos , Lipoproteínas LDL/sangue , Pessoa de Meia-Idade , Obesidade Mórbida/dietoterapia , Estudos Prospectivos , Redução de PesoRESUMO
Human immunodeficiency virus type 1 activates the complement cascade via the classical pathway by direct binding of C1q through specific sites in the TM surface protein, gp41. In this paper we investigated the divalent cation dependence of the interaction between HIV-1 gp41 and C1q or gp120. A solid phase radioimmunoassay was used to investigate the interaction between a recombinant soluble form of HIV-1 gp41 (rsgp41) and C1q and an enzyme linked immunoassay was used to investigate the interaction between rsgp41 and gp120. The interaction between C1q and rsgp41, but not between C1q and immune complexes, was dependent upon the presence of calcium. Calcium could not be replaced by larger cations such as strontium, barium, lead or smaller ions such as magnesium and manganese. Zinc increased binding to 22% of binding achieved with calcium. The interaction between rsgp41 and gp120 was not dependent upon the presence of divalent ions. Thus, calcium is required for the interaction between rsgp41 and C1q, whereas the interaction between rsgp41 and gp120 is independent of divalent cations.
Assuntos
Cálcio/fisiologia , Complemento C1q/metabolismo , Proteína gp120 do Envelope de HIV/metabolismo , Proteína gp41 do Envelope de HIV/metabolismo , HIV-1/metabolismo , Cátions Bivalentes , Ensaio de Imunoadsorção Enzimática , Humanos , Ligação Proteica/fisiologia , Radioimunoensaio , Proteínas Recombinantes/metabolismoRESUMO
OBJECTIVE: To further characterize cell surface proteins binding to recombinant soluble (rs) forms of the transmembrane glycoproteins gp41 of HIV-1 (rsgp41) and gp36 of HIV-2 (rsgp36). METHODS: Various human and murine cell lines of different lineages were surface-labelled with 125I. rsgp41 and rsgp36 were bound to CnBr-Sepharose and used as an affinity matrix for the surface-labelled cell lysates. The bound cell surface proteins were separated on sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing conditions. A rabbit serum was produced against one of the cell surface proteins and flow cytometry used to compare the results with those obtained from affinity chromatography. RESULTS: We have confirmed and extended the results obtained by Qureshi et al. [1]. In addition to the 44kD protein, we identified cell surface proteins with molecular weights of 98 and 106 kD binding with high affinity to both rsgp41 and rsgp36. We have demonstrated differences between human and murine cell lines in the expression of the cell surface proteins that interact with rsgp41 and rsgp36. Furthermore, a correlation between the level of rsgp41 and rsgp36 binding proteins, detected either by affinity chromatography or by reactivity with an antiserum directed against one of the cell surface binding components was shown. CONCLUSIONS: Three cell surface proteins, with molecular weights of 44, 98 and 106 kD, bind with high affinity to rs forms of gp41 and gp36. Their expression decreases from a T-lymphoid cell line, to a monoblastoid cell line, to a cell line representing mature monocytes. Human T-cell lymphotropic virus-infected cell lines show a predominance of the 44 kD protein. There are species-specific differences, in that murine cell lines lack the 44 kD protein.
Assuntos
Produtos do Gene env/metabolismo , Antígenos HIV/metabolismo , Proteína gp41 do Envelope de HIV/metabolismo , HIV-1/metabolismo , HIV-2/metabolismo , Animais , Linhagem Celular , Humanos , Cinética , Proteínas de Membrana/química , Proteínas de Membrana/metabolismo , Camundongos , Peso Molecular , Ligação Proteica , Proteínas Recombinantes/metabolismo , Solubilidade , Especificidade da Espécie , Produtos do Gene env do Vírus da Imunodeficiência HumanaRESUMO
Self-administration of anabolic-androgenic steroids to increase muscular strength and lean body mass has been used widely among athletes. Flow mediated dilatation (FMD) determined by ultrasound of the brachial artery is accepted as both an in vivo index of endothelial function and an indicator for future atherosclerosis. FMD was calculated in 20 male non-smoking body builders in different phases of their training cycle and in six male non-smoking control athletes. Ultrasound studies of the brachial artery were performed according to the protocol of Celermajer et al. Of the entire training cycle, work-out phase was training phase without actual intake of anabolic-androgenic steroids over 8 weeks; build-up phase included actual intake of anabolic-androgenic steroids; and competition phase consisted of 8 weeks post intake of anabolic-androgenic steroids. Baseline characteristics did not differ between body builder groups except for a higher weight in competition phase body builders. Hormonal analysis revealed suppressed luteinizing hormone and follicle stimulating hormone levels in build-up phase body builders. The lipid profiles showed a marked reduction of HDL-C in build-up phase body builders. FMD was reduced in body builders of all phases when compared to control athletes (work-out phase: 2.5+/-2.7%; build-up phase: 2.1+/-3.0%; competition phase: 0.4+/-2.9% vs. 10.9+/-4.4%, P<0.05 by pairwise comparison using Scheffe's test for work-out phase, build-up phase and competition phase vs. control athletes). The glyceryl trinitrate-induced vasodilatation was diminished, though not statistically significantly, in body builders when compared with control athletes. The differences in FMD persisted after adjustment for vessel size. Our data indicate that intake of anabolic-androgenic steroids is associated with both an atherogenic blood lipid profile and endothelial dysfunction and thus may pose an increased risk of atherosclerosis.
Assuntos
Anabolizantes/efeitos adversos , Velocidade do Fluxo Sanguíneo , Endotélio Vascular/fisiologia , Vasodilatação , Levantamento de Peso/fisiologia , Adulto , Anabolizantes/administração & dosagem , Artéria Braquial/diagnóstico por imagem , Artéria Braquial/fisiologia , Colesterol/sangue , Hormônio Foliculoestimulante/sangue , Humanos , Hormônio Luteinizante/sangue , Masculino , Nitroglicerina/farmacologia , Radioimunoensaio , Testosterona/sangue , Ultrassonografia , Vasodilatadores/farmacologiaRESUMO
The hormone leptin (OB) and its receptor (OB-R) are key homeostatic regulators of mammalian body weight. Two predominant isoforms of OB-R are expressed by alternative splicing: the long form, OB-RL, with full signalling capacity is highly expressed in the hypothalamus and the short, signalling-defective form, OB-Rs, is ubiquitously expressed. In a previous study we detected expression of OB-RL and OB-Rs in human syncytiotrophoblast cells using in situ hybridization and immunohistochemistry (Bodner et al., 1999). The aim of this study was to investigate leptin receptor isoform expression and phosphorylation in paired, syncytial, microvillous and basal membranes from human term placenta by Western blot analysis. Both the OB-RL and the OB-Rs isoforms were detected in the syncytial membrane preparations. The OB-RL isoform was observed exclusively in microvillous membranes, whereas the OB-Rs isoform was found in both microvillous and basal membrane preparations. No significant differences were observed between syncytial membranes from normal and type 1 diabetic pregnancies. To test the phosphorylation capacity of the OB-R isoforms, microvillous and basal membrane vesicles loaded with ATP were stimulated with leptin and the phosphorylation status of the OB-R at the tyrosine 985 (Y985) was determined. A single band at the molecular weight corresponding to the molecular weight of the OB-RL isoform was detected exclusively in the ATP-loaded microvillous vesicles. We conclude that the long form OB-RL is expressed exclusively in the microvillous membrane of the syncytiotrophoblast and is capable of being phosphorylated, suggesting that it has signal transduction capacity.
Assuntos
Membranas Intracelulares/metabolismo , Placenta/metabolismo , Receptores de Superfície Celular/metabolismo , Adolescente , Adulto , Western Blotting , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Fosforilação , Gravidez , Gravidez em Diabéticas/metabolismo , Isoformas de Proteínas/metabolismo , Receptores para LeptinaRESUMO
In the present study, we investigated the expression and localization of leptin receptors in human term placentae. On human term placenta tissue slices, digoxigenin-UTP labelled RNA-probe detected the long form of the leptin receptor ObR(L)mRNA in syncytiotrophoblasts of the villi, whereas the haematological subtype of the leptin receptor ObR/B219.1 was detected in blood cells of the intervillous space and fetal vessels. Immunohistochemistry, with two polyclonal antibodies to the N-terminus recognizing ObR(L)and ObR(S)of the leptin receptors and one to the C-terminus recognizing the long form of the leptin receptor ObR(L), localized leptin receptor protein at the apical membrane of the syncytiotrophoblasts. Our results show that the long form of the leptin receptor ObR(L)is expressed in human term placentae. We localized the long form of leptin receptor mRNA to the cytoplasm of syncytiotrophoblasts and leptin receptor proteins in human term placentae to the apical membrane of syncytiotrophoblasts. We conclude that in term placentae, leptin could mediate a growth promoting effect in the fetoplacental unit through the long form of the leptin receptor localized in the syncytiotrophoblasts. In contrast, the haematological subtype of the leptin receptor is not expressed in placental cells, but solely by blood cells in the intervillous space and fetal vessels.
Assuntos
Proteínas de Transporte/análise , Placenta/química , Receptores de Superfície Celular , Receptores de Citocinas/análise , Cesárea , Feminino , Humanos , Imuno-Histoquímica , Hibridização In Situ , Gravidez , Receptores para Leptina , Transdução de Sinais/fisiologiaRESUMO
Paroxysmal nocturnal haemoglobinuria (PNH) is an acquired clonal stem cell disorder resulting in insufficient and defective haematopoesis associated frequently with aplastic anaemia (AA). A deficiency of the glycosyl phosphatidylinositol (GPI)-anchored complement activation regulatory proteins CD55 and CD59 is responsible for an increased sensitivity of erythrocytes to complement attack leading to chronic intravascular haemolysis with haemoglobinuria. In this study we investigated the effects of complement activation caused by anti-thymocyte globulin (ATG) treatment on the PNH clone in a patient affected with the PNH/AA-syndrome. Fluid phase complement components C3, C4, C6 and terminal complement complex (TCC) were assayed by ELISA. CD55, CD59 and cell-associated TCC were monitored by flow cytometry. ATG treatment resulted in profound systemic complement activation which led to a decrease in the levels of native C3 and C4 to 65% and 40%, respectively, of the original levels on day 5 and of C6 and TCC to 61% and 23%, respectively, on day 10. A return to pre-treatment levels was observed for C3 by day 15, for C6 by day 30 and for C4 by day 90. Flow cytometry revealed that the deficiency in the GPI-anchored protein was restricted to granulocytes, while lymphocytes remained unaffected. Cell-bound TCC increased by 1.67-fold and 2.37-fold on day 5 and day 10, respectively, decreasing to 1.40-fold and 1.30-fold on day 15 and day 30, respectively. The percentage of PNH granulocytes as identified by the absence of the CD55- and CD59-antigens exhibited a temporary decrease from 72% on day 0 to 65% on day 5 and 59% on day 10 and returned thereafter to the original percentage of 70% by day 15 and exceeding this level to 76% on day 30 and 79% on day 90. We report profound activation of the classical pathway of the complement cascade and the terminal complement complex by the globulin leading to a transient decrease of the PNH clone, presumably due to subsequent lysis of the PNH cells devoid of complement regulatory proteins.
Assuntos
Anemia Aplástica/sangue , Anemia Aplástica/terapia , Soro Antilinfocitário/uso terapêutico , Ativação do Complemento/imunologia , Hemoglobinúria Paroxística/sangue , Hemoglobinúria Paroxística/terapia , Síndromes Mielodisplásicas/sangue , Síndromes Mielodisplásicas/terapia , Anemia Aplástica/imunologia , Citometria de Fluxo , Granulócitos/patologia , Hemoglobinúria Paroxística/imunologia , Humanos , Contagem de Leucócitos , Síndromes Mielodisplásicas/imunologiaRESUMO
OBJECTIVE: To determine whether fetal leptin levels correlate with fetal weight and whether such correlation is direct or indirect via insulin or human placental lactogen (hPL), respectively. DESIGN: Cross-sectional study of offspring at term (n=175) with over-representation of large-for-gestational age (LGA; n=70) and small-for-gestational age (SGA; n=23) cases in a population of Caucasian women with no pregnancy pathology. METHODS: Fetal cord blood was collected after delivery. In several cases (n=62) paired mother-fetus blood samples were obtained. Leptin, insulin and hPL levels were measured by RIA. Anthropometric data (birth weight, body mass index, placental weight) were recorded. RESULTS AND CONCLUSIONS: Maternal insulin, hPL and leptin levels were higher than fetal concentrations. Cord blood leptin levels positively correlated with the anthropometric data with stronger correlations in female (0.54Assuntos
Peso ao Nascer
, Sangue Fetal
, Idade Gestacional
, Insulina/sangue
, Leptina/sangue
, Feminino
, Humanos
, Recém-Nascido
, Masculino
, Lactogênio Placentário/sangue
, Período Pós-Parto/sangue
, Gravidez
RESUMO
Undernourishment in utero appears to be associated with persisting changes in the metabolic, endocrine, and immune functions. In this study, we determined the influence of birth weight on the lipoprotein profile and cholesteryl ester transfer protein (CETP), which promotes a proatherogenic lipoprotein profile in plasma by determining the chemical, physical, and biologic properties of the respective lipoprotein particles. Triglyceride (TG) concentrations were highest and high-density lipoprotein (HDL)(2)-cholesterol levels were lowest in small for gestational age (SGA) neonates. CETP-mass was determined by enzyme-linked immunosorbent assay (ELISA) and CETP-activity by using exogenous lipoproteins. Cholesteryl ester transfer was determined as transfer of radiolabeled cholesteryl esters (CE) from HDL to apolipoprotein B-containing lipoproteins. CETP mass was lowest and cholesteryl ester transfer was highest in SGA neonates. CETP-activity did not differ among the neonates. Our results suggest that increased and decreased nourishment in utero affects the lipoprotein profile and CETP in neonates. High TG and low HDL(2) levels in SGA neonates might result from increased cholesteryl ester transfer and, may in part, explain the increased risk of coronary heart disease (CHD) of small for gestational age neonates in later life.
Assuntos
Proteínas de Transporte/sangue , Glicoproteínas , Recém-Nascido/sangue , Lipoproteínas/sangue , Adulto , Peso ao Nascer , Proteínas de Transporte/análise , Colesterol/sangue , Proteínas de Transferência de Ésteres de Colesterol , Ésteres do Colesterol/sangue , Ésteres do Colesterol/metabolismo , Feminino , Sangue Fetal , Idade Gestacional , Humanos , Triglicerídeos/sangueRESUMO
The discovery of leptin, the product of the obese (ob)-gene, has broadened the horizons of research on energy balance. This hormone, produced and secreted by adipose tissue and some placental cells, finds its way to the hypothalamus, where it binds to the leptin receptors and signals satiety through the neuroendocrine axis. The fact that adipose tissue is not merely a storage depot, but also an important endocrine tissue, has revived the interest in the "lipostatic" theory of body fat regulation and has initiated many research efforts in the field of obesity, anorexia nervosa, bulimia, reproduction and haematology.