RESUMEN
OBJECTIVE: To estimate creatine concentrations in maternal plasma and urine, and establish relationships with maternal characteristics, diet and fetal growth. DESIGN: Retrospective cohort study. SETTING: Lyell McEwin Hospital, Adelaide, Australia. POPULATION: A biobank of plasma and urine samples collected at 13, 18, 30 and 36 weeks' gestation from 287 pregnant women from a prospective cohort of asthmatic and non-asthmatic women. METHODS: Creatine was measured by enzymatic analysis. Change in creatine over pregnancy was assessed using the Friedman test. Linear mixed models regression was used to determine associations between maternal factors and diet with creatine across pregnancy and between creatine with indices of fetal growth at birth. MAIN OUTCOME MEASURES: Maternal creatine concentrations, associations between maternal factors and creatine and between creatine and fetal growth parameters. RESULTS: Maternal smoking, body mass index, asthma and socio-economic status were positively and parity negatively associated with maternal plasma and/or urine creatine. Maternal urine creatine concentration was positively associated with birthweight centile and birth length. After adjustment, each µmol/l increase in maternal urinary creatine was associated with a 1.23 (95% CI 0.44-2.02) unit increase in birthweight centile and a 0.11-cm (95% CI 0.03-0.2) increase in birth length. CONCLUSIONS: Maternal factors and fetal growth measures are associated with maternal plasma and urine creatine concentrations. TWEETABLE ABSTRACT: Maternal creatine is altered by pregnancy; fetal growth measures are associated with maternal creatine concentrations.
Asunto(s)
Creatina/sangre , Creatina/orina , Desarrollo Fetal/fisiología , Trimestres del Embarazo/sangre , Trimestres del Embarazo/orina , Adulto , Asma/sangre , Asma/orina , Bancos de Muestras Biológicas , Peso al Nacer/fisiología , Femenino , Edad Gestacional , Humanos , Recién Nacido , Modelos Lineales , Paridad , Embarazo , Complicaciones del Embarazo/sangre , Complicaciones del Embarazo/orina , Estudios Prospectivos , Estudios Retrospectivos , Fumar/sangre , Fumar/orina , Clase SocialRESUMEN
The Na+ -K+ -ATPase enzyme is vital in skeletal muscle function. We investigated the effects of acute high-intensity interval exercise, before and following high-intensity training (HIT), on muscle Na+ -K+ -ATPase maximal activity, content, and isoform mRNA expression and protein abundance. Twelve endurance-trained athletes were tested at baseline, pretrain, and after 3 wk of HIT (posttrain), which comprised seven sessions of 8 x 5-min interval cycling at 80% peak power output. Vastus lateralis muscle was biopsied at rest (baseline) and both at rest and immediately postexercise during the first (pretrain) and seventh (posttrain) training sessions. Muscle was analyzed for Na+ -K+ -ATPase maximal activity (3-O-MFPase), content ([3H]ouabain binding), isoform mRNA expression (RT-PCR), and protein abundance (Western blotting). All baseline-to-pretrain measures were stable. Pretrain, acute exercise decreased 3-O-MFPase activity [12.7% (SD 5.1), P < 0.05], increased alpha1, alpha2, and alpha3 mRNA expression (1.4-, 2.8-, and 3.4-fold, respectively, P < 0.05) with unchanged beta-isoform mRNA or protein abundance of any isoform. In resting muscle, HIT increased (P < 0.05) 3-O-MFPase activity by 5.5% (SD 2.9), and alpha3 and beta3 mRNA expression by 3.0- and 0.5-fold, respectively, with unchanged Na+ -K+ -ATPase content or isoform protein abundance. Posttrain, the acute exercise induced decline in 3-O-MFPase activity and increase in alpha1 and alpha3 mRNA each persisted (P < 0.05); the postexercise 3-O-MFPase activity was also higher after HIT (P < 0.05). Thus HIT augmented Na+ -K+ -ATPase maximal activity despite unchanged total content and isoform protein abundance. Elevated Na+ -K+ -ATPase activity postexercise may contribute to reduced fatigue after training. The Na+ -K+ -ATPase mRNA response to interval exercise of increased alpha- but not beta-mRNA was largely preserved posttrain, suggesting a functional role of alpha mRNA upregulation.
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Adaptación Fisiológica , Ejercicio Físico/fisiología , Fatiga Muscular , Resistencia Física/fisiología , Músculo Cuádriceps/enzimología , ATPasa Intercambiadora de Sodio-Potasio/biosíntesis , Adaptación Fisiológica/genética , Inducción Enzimática , Fluoresceínas/metabolismo , Humanos , Isoenzimas/metabolismo , Masculino , Fatiga Muscular/genética , Ouabaína/metabolismo , Resistencia Física/genética , Unión Proteica , ARN Mensajero/biosíntesis , ATPasa Intercambiadora de Sodio-Potasio/genéticaRESUMEN
AIM: This study investigated the effects of endurance training status and sex differences on skeletal muscle Na+,K+-pump mRNA expression, content and activity. METHODS: Forty-five endurance-trained males (ETM), 11 recreationally active males (RAM), and nine recreationally active females (RAF) underwent a vastus lateralis muscle biopsy. Muscle was analysed for Na+,K+-pump alpha1, alpha2, alpha3, beta1, beta2 and beta3 isoform mRNA expression (real-time reverse transcription-polymerase chain reaction), content ([3H]-ouabain-binding site) and maximal activity (3-O-methylfluorescein phosphatase, 3-O-MFPase). RESULTS: ETM demonstrated lower alpha1, alpha3, beta2 and beta3 mRNA expression by 74%, 62%, 70% and 82%, respectively, than RAM (P<0.04). In contrast, [3H]-ouabain binding and 3-O-MFPase activity were each higher in ETM than in RAM, by 16% (P<0.03). RAM demonstrated a 230% and 364% higher alpha3 and beta3 mRNA expression than RAF, respectively (P<0.05), but no significant sex differences were found for alpha1, alpha2, beta1 or beta2 mRNA, [3H]-ouabain binding or 3-O-MFPase activity. No significant correlation was found between years of endurance training and either [3H]-ouabain binding or 3-O-MFPase activity. Significant but weak correlations were found between the number of training hours per week and 3-O-MFPase activity (r=0.31, P<0.02) and between incremental exercise VO2(peak)) and both [3H]-ouabain binding (r=0.33, P<0.01) and 3-O-MFPase activity (r=0.28, P<0.03). CONCLUSIONS: Isoform-specific differences in Na+,K+-pump mRNA expression were found with both training status and sex differences, but only training status influenced Na+,K+-pump content and maximal activity in human skeletal muscle.
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Regulación de la Expresión Génica , Músculo Esquelético/enzimología , Resistencia Física , Isoformas de Proteínas/genética , ARN Mensajero/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/genética , Adulto , Análisis de Varianza , Sitios de Unión , Biopsia , Estudios Transversales , Ciclofilinas/genética , Activación Enzimática , Femenino , Humanos , Masculino , Ouabaína/metabolismo , Educación y Entrenamiento Físico , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores Sexuales , ATPasa Intercambiadora de Sodio-Potasio/análisis , Factores de TiempoRESUMEN
This study investigated effects of prolonged submaximal exercise on Na+-K+-ATPase mRNA and protein expression, maximal activity, and content in human skeletal muscle. We also investigated the effects on mRNA expression of the transcription initiator gene, RNA polymerase II (RNAP II), and key genes involved in protein translation, eukaryotic initiation factor-4E (eIF-4E) and 4E-binding protein 1 (4E-BP1). Eleven subjects (6 men, 5 women) cycled at 75.5% (SD 4.8%) peak O2 uptake and continued until fatigue. A vastus lateralis muscle biopsy was taken at rest, fatigue, and 3 and 24 h postexercise. We analyzed muscle for Na+-K+-ATPase alpha1, alpha2, alpha3, beta1, beta2, and beta3, as well for RNAP II, eIF-4E, and 4E-BP1 mRNA expression by real-time RT-PCR and Na+-K+-ATPase isoform protein abundance using immunoblotting. Muscle homogenate maximal Na+-K+-ATPase activity was determined by 3-O-methylfluorescein phosphatase activity and Na+-K+-ATPase content by [3H]ouabain binding. Cycling to fatigue [54.5 (SD 20.6) min] immediately increased alpha3 (P = 0.044) and beta2 mRNA (P = 0.042) by 2.2- and 1.9-fold, respectively, whereas alpha1 mRNA was elevated by 2.0-fold at 24 h postexercise (P = 0.036). A significant time main effect was found for alpha3 protein abundance (P = 0.046). Exercise transiently depressed maximal Na+-K+-ATPase activity (P = 0.004), but Na+-K+-ATPase content was unaltered throughout recovery. Exercise immediately increased RNAP II mRNA by 2.6-fold (P = 0.011) but had no effect on eIF-4E and 4E-BP1 mRNA. Thus a single bout of prolonged submaximal exercise induced isoform-specific Na+-K+-ATPase responses, increasing alpha1, alpha3, and beta2 mRNA but only alpha3 protein expression. Exercise also increased mRNA expression of RNAP II, a gene initiating transcription, but not of eIF-4E and 4E-BP1, key genes initiating protein translation.
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Ejercicio Físico/fisiología , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/genética , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Adulto , Femenino , Regulación Enzimológica de la Expresión Génica , Humanos , Masculino , Músculo Esquelético/enzimología , Ouabaína/metabolismo , Unión Proteica , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Subunidades de Proteína/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
We investigated whether depressed muscle Na(+)-K(+)-ATPase activity with exercise reflected a loss of Na(+)-K(+)-ATPase units, the time course of its recovery postexercise, and whether this depressed activity was related to increased Na(+)-K(+)-ATPase isoform gene expression. Fifteen subjects performed fatiguing, knee extensor exercise at approximately 40% maximal work output per contraction. A vastus lateralis muscle biopsy was taken at rest, fatigue, 3 h, and 24 h postexercise and analyzed for maximal Na(+)-K(+)-ATPase activity via 3-O-methylfluorescein phosphatase (3-O-MFPase) activity, Na(+)-K(+)-ATPase content via [(3)H]ouabain binding sites, and Na(+)-K(+)-ATPase alpha(1)-, alpha(2)-, alpha(3)-, beta(1)-, beta(2)- and beta(3)-isoform mRNA expression by real-time RT-PCR. Exercise [352 (SD 267) s] did not affect [(3)H]ouabain binding sites but decreased 3-O-MFPase activity by 10.7 (SD 8)% (P < 0.05), which had recovered by 3 h postexercise, without further change at 24 h. Exercise elevated alpha(1)-isoform mRNA by 1.5-fold at fatigue (P < 0.05). This increase was inversely correlated with the percent change in 3-O-MFPase activity from rest to fatigue (%Delta3-O-MFPase(rest-fatigue)) (r = -0.60, P < 0.05). The average postexercise (fatigue, 3 h, 24 h) alpha(1)-isoform mRNA was increased 1.4-fold (P < 0.05) and approached a significant inverse correlation with %Delta3-O-MFPase(rest-fatigue) (r = -0.56, P = 0.08). Exercise elevated alpha(2)-isoform mRNA at fatigue 2.5-fold (P < 0.05), which was inversely correlated with %Delta3-O-MFPase(rest-fatigue) (r = -0.60, P = 0.05). The average postexercise alpha(2)-isoform mRNA was increased 2.2-fold (P < 0.05) and was inversely correlated with the %Delta3-O-MFPase(rest-fatigue) (r = -0.68, P < 0.05). Nonsignificant correlations were found between %Delta3-O-MFPase(rest-fatigue) and other isoforms. Thus acute exercise transiently decreased Na(+)-K(+)-ATPase activity, which was correlated with increased Na(+)-K(+)-ATPase gene expression. This suggests a possible signal-transduction role for depressed muscle Na(+)-K(+)-ATPase activity with exercise.
Asunto(s)
Ejercicio Físico , Fatiga/enzimología , Músculo Esquelético/enzimología , ARN Mensajero/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/genética , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Adulto , Volumen Sanguíneo , Femenino , Humanos , Masculino , Concentración Osmolar , Potasio/sangre , Factores de TiempoRESUMEN
1. The nucleoside intermediate 5'-aminoimidazole-4-carboxyamide-ribonucleoside (AICAR) activates skeletal muscle AMP-activated protein kinase (AMPK) and increases glucose uptake. The AMPK phosphorylates neuronal nitric oxide synthase (nNOS)mu in skeletal muscle fibres. There is evidence that both AMPK and nNOSmu may be involved in the regulation of contraction-stimulated glucose uptake. 2. We examined whether both AICAR- and contraction-stimulated glucose uptake were mediated by NOS in rat skeletal muscle. 3. Rat isolated epitrochlearis muscles were subjected in vitro to electrically stimulated contractions for 10 min and/or incubated in the presence or absence of AICAR (2 mmol/L) or the NOS inhibitor NG-monomethyl-L-arginine (L-NMMA; 100 micromol/L). 4. Muscle contraction significantly (P < 0.05) altered the metabolic profile of the muscle. In contrast, AICAR and L-NMMA had no effect on the metabolic profile of the muscle, except that AICAR increased muscle 5'-aminoimidazole-4-carboxyamide-ribonucleotide (ZMP) and AICAR content. Nitric oxide synthase inhibition caused a small but significant (P < 0.05) reduction in basal 3-O-methylglucose transport, which was observed in all treatments. 5'-Aminoimidazole-4-carboxyamide-ribonucleoside significantly increased (P < 0.05) glucose transport above basal, with NOS inhibition decreasing this slightly (increased by 209% above basal compared with 184% above basal with NOS inhibition). Contraction significantly increased glucose transport above basal, with NOS inhibition substantially reducing this (107% increase vs 31% increase). 5'-Aminoimidazole-4-carboxyamide-ribonucleoside plus contraction in combination were not additive on glucose transport. 5. These results suggest that NO plays a role in basal glucose uptake and may regulate contraction-stimulated glucose uptake. However, NOS/nitric oxide do not appear to be signalling intermediates in AICAR-stimulated skeletal muscle glucose uptake.
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Aminoimidazol Carboxamida/análogos & derivados , Aminoimidazol Carboxamida/farmacología , Inhibidores Enzimáticos/farmacología , Glucosa/metabolismo , Músculo Esquelético/metabolismo , Óxido Nítrico Sintasa/antagonistas & inhibidores , Ribonucleótidos/farmacología , 3-O-Metilglucosa/metabolismo , Animales , Transporte Biológico Activo/efectos de los fármacos , Técnicas In Vitro , Masculino , Contracción Muscular/fisiología , Músculo Esquelético/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , omega-N-Metilarginina/farmacologíaRESUMEN
Characterization of expression of, and consequently also the acute exercise effects on, Na(+),K(+)-ATPase isoforms in human skeletal muscle remains incomplete and was therefore investigated. Fifteen healthy subjects (eight males, seven females) performed fatiguing, knee extensor exercise at approximately 40% of their maximal work output per contraction. A vastus lateralis muscle biopsy was taken at rest, fatigue and 3 and 24 h postexercise, and analysed for Na(+),K(+)-ATPase alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) mRNA and crude homogenate protein expression, using Real-Time RT-PCR and immunoblotting, respectively. Each individual expressed gene transcripts and protein bands for each Na(+),K(+)-ATPase isoform. Each isoform was also expressed in a primary human skeletal muscle cell culture. Intense exercise (352 +/- 69 s; mean +/-s.e.m.) immediately increased alpha(3) and beta(2) mRNA by 2.4- and 1.7-fold, respectively (P < 0.05), whilst alpha(1) and alpha(2) mRNA were increased by 2.5- and 3.5-fold at 24 h and 3 h postexercise, respectively (P < 0.05). No significant change occurred for beta(1) and beta(3) mRNA, reflecting variable time-dependent responses. When the average postexercise value was contrasted to rest, mRNA increased for alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) isoforms, by 1.4-, 2.2-, 1.4-, 1.1-, 1.0- and 1.0-fold, respectively (P < 0.05). However, exercise did not alter the protein abundance of the alpha(1)-alpha(3) and beta(1)-beta(3) isoforms. Thus, human skeletal muscle expresses each of the Na(+),K(+)-ATPase alpha(1), alpha(2), alpha(3), beta(1), beta(2) and beta(3) isoforms, evidenced at both transcription and protein levels. Whilst brief exercise increased Na(+),K(+)-ATPase isoform mRNA expression, there was no effect on isoform protein expression, suggesting that the exercise challenge was insufficient for muscle Na(+),K(+)-ATPase up-regulation.
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Ejercicio Físico/fisiología , Isoenzimas/genética , Músculo Esquelético/fisiología , ATPasa Intercambiadora de Sodio-Potasio/genética , Adulto , Células Cultivadas , Femenino , Humanos , Masculino , Fibras Musculares Esqueléticas/citología , Fibras Musculares Esqueléticas/enzimología , Músculo Esquelético/citología , ARN Mensajero/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Regulación hacia Arriba/fisiologíaRESUMEN
This study investigated the effects of riluzole (Ril), creatine (Cr) and a combination of these treatments on the onset and progression of clinical signs and neuropathology in an animal model of familial amyotrophic lateral sclerosis, the G93A transgenic mouse (n=13-17 per group). The onset of clinical signs was delayed (P<0.05) by about 12 days in all treatment groups compared with control; however, no differences occurred between treatments. All animals were killed at 199 days of age. At the end of the experimental period the severity of clinical signs was less (P<0.05) with all treatments compared with control. Again no differences between treatments were observed. The treatments had no effect on the number of neurons in ventral horns of the lumbar region of the spinal cord. Transgenic mice ingesting Cr displayed elevated (P<0.05) total Cr levels in cerebral hemispheres (5%) and spinal cord (8%), but not skeletal muscles. These data demonstrate that treatment with Ril and Cr were both effective in delaying disease onset and clinical disability. To the age of killing, no additional benefit was conferred by co-administration of Ril and Cr.
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Esclerosis Amiotrófica Lateral/tratamiento farmacológico , Células del Asta Anterior/efectos de los fármacos , Creatina/farmacología , Fármacos Neuroprotectores/farmacología , Riluzol/farmacología , Superóxido Dismutasa/deficiencia , Edad de Inicio , Esclerosis Amiotrófica Lateral/enzimología , Esclerosis Amiotrófica Lateral/genética , Animales , Células del Asta Anterior/enzimología , Células del Asta Anterior/patología , Muerte Celular/efectos de los fármacos , Muerte Celular/genética , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/enzimología , Corteza Cerebral/fisiopatología , Creatina/uso terapéutico , Modelos Animales de Enfermedad , Sinergismo Farmacológico , Quimioterapia Combinada , Femenino , Masculino , Ratones , Ratones Transgénicos , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/enzimología , Fármacos Neuroprotectores/uso terapéutico , Riluzol/uso terapéutico , Superóxido Dismutasa/genética , Resultado del TratamientoRESUMEN
The present study examined the validity and reliability of measuring the expression of various genes in human skeletal muscle using quantitative real-time RT-PCR on a GeneAmp 5700 sequence detection system with SYBR Green 1 chemistry. In addition, the validity of using some of these genes as endogenous controls (i.e., housekeeping genes) when human skeletal muscle was exposed to elevated total creatine levels and exercise was also examined. For all except 28S, linear relationships between the logarithm of the starting RNA concentrations and the cycle threshold (C(T)) values were established for beta-actin, beta2-microglobulin (beta2M), cyclophilin (CYC), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a linear response between C(T) values and the logarithm of a given amount of starting cDNA for all the genes tested. The overall intra-assay coefficient of variance for these genes was 1.3% and 21% for raw C(T) values and the linear value of 2(-C(T)), respectively. Interassay variability was 2.3% for raw C(T) values and 34% for the linear value of 2(-C(T)). We also examined the expression of various housekeeping genes in human skeletal muscle at days 0, 1, and 5 following oral supplementation with either creatine or a placebo employing a double-blind crossover study design. Treatments were separated by a 5-wk washout period. Immediately following each muscle sampling, subjects performed two 30-s all-out bouts on a cycle ergometer. Creatine supplementation increased (P < 0.05) muscle total creatine content above placebo levels; however, there were no changes (P > 0.05) in C(T) values across the supplementation periods for any of the genes. Nevertheless, 95% confidence intervals showed that GAPDH was variable, whereas beta-actin, beta2M, and CYC were the least varying genes. Normalization of the data to these housekeeping genes revealed variable behavior for beta2M with more stable expressions for both beta-actin and CYC. We conclude that, using real-time RT-PCR, beta-actin or CYC may be used as housekeeping genes to study gene expression in human muscle in experiments employing short-term creatine supplementation combined with high-intensity exercise.
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Creatina/farmacología , Suplementos Dietéticos , Genes/efectos de los fármacos , Músculo Esquelético/química , Músculo Esquelético/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Actinas/genética , Actinas/metabolismo , Adulto , Análisis de Varianza , Sistemas de Computación/estadística & datos numéricos , Creatina/metabolismo , Estudios Cruzados , Ciclofilinas/biosíntesis , Ciclofilinas/genética , Ciclofilinas/metabolismo , ADN Complementario/biosíntesis , ADN Complementario/genética , ADN Complementario/metabolismo , Método Doble Ciego , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Variación Genética/genética , Gliceraldehído-3-Fosfato Deshidrogenasas/biosíntesis , Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Humanos , Masculino , Músculo Esquelético/metabolismo , Proyectos Piloto , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/estadística & datos numéricos , Microglobulina beta-2/biosíntesis , Microglobulina beta-2/genética , Microglobulina beta-2/metabolismoRESUMEN
We have estimated the reliability of performance in a commonly employed exercise test consisting of repeated sprints on a cycle ergometer. Eight recreationally active young men completed a practice trial and three more trials at 3- to 6-day intervals. Each trial consisted of two bouts of 30-s maximal-effort cycling on an electromagnetically braked cycle ergometer; the bouts were separated by 4 min of rest. The typical (standard) errors of measurement for peak and mean power between trials 2 to 4 were 2.5 and 1.7% respectively for the first bout and 1.9 and 1.8% for the second bout. These errors are substantially less than those in previous reliability studies of single 30-s sprint tests, probably because of differences in quality of ergometer. The typical errors for the difference between bouts (i.e., fatigue) for peak power and mean power were 3.0 and 2.5%, respectively. Typical errors for the average of the two bouts were 1.6 and 1.2% for peak and mean power respectively, which are small enough to give adequate precision for moderate treatment effects in studies with modest sample sizes.
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Ciclismo/fisiología , Prueba de Esfuerzo/normas , Adulto , Ergometría , Prueba de Esfuerzo/instrumentación , Humanos , Masculino , Reproducibilidad de los Resultados , Programas InformáticosRESUMEN
The cellular role of creatine (Cr) and Cr phosphate (CrP) has been studied extensively in neural, cardiac and skeletal muscle. Several studies have demonstrated that alterations in the cellular total Cr (Cr + CrP) concentration in these tissues can produce marked functional and/or structural change. The primary aim of this review was to critically evaluate the literature that has examined the regulation of cellular total Cr content. In particular, the review focuses on the regulation of the activity and gene expression of the Cr transporter (CreaT), which is primarily responsible for cellular Cr uptake. Two CreaT genes (CreaT1 and CreaT2) have been identified and their chromosomal location and DNA sequencing have been completed. From these data, putative structures of the CreaT proteins have been formulated. Transcription products of the CreaT2 gene are expressed exclusively in the testes, whereas CreaT1 transcripts are found in a variety of tissues. Recent research has measured the expression of the CreaT1 protein in several tissues including neural, cardiac and skeletal muscle. There is very little information available about the factors regulating CreaT gene expression. There is some evidence that suggests the intracellular Cr concentration may be involved in the regulatory process but there is much more to learn before this process is understood. The activity of the CreaT protein is controlled by many factors. These include substrate concentration, transmembrane Na+ gradients, cellular location, and various hormones. It is also likely that transporter activity is influenced by its phosphorylation state and by its interaction with other plasma membrane proteins. The extent of CreaT protein glycosylation may vary within cells, the functional significance of which remains unclear.
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Creatina/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Empalme Alternativo , Animales , Transporte Biológico , Regulación de la Expresión Génica , Humanos , Cinética , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/genética , Especificidad de Órganos , Fosfocreatina/metabolismo , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismoRESUMEN
OBJECTIVES: Compare the discrimination of risk-adjustment models for primary cesarean delivery derived from medical record data and birth certificate data and determine if the two types of models yield similar hospital profiles of risk-adjusted cesarean delivery rates. DATA SOURCES/STUDY SETTING: The study involved 29,234 women without prior cesarean delivery admitted for labor and delivery in 1993-95 to 20 hospitals in northeast Ohio for whom data abstracted from patient medical records and data from birth certificates could be linked. STUDY DESIGN: Three pairs of multivariate models of the risk of cesarean delivery were developed using (1) the full complement of variables in medical records or birth certificates; (2) variables that were common to the two sources; and (3) variables for which agreement between the two data sources was high. Using each of the six models, predicted rates of cesarean delivery were determined for each hospital. Hospitals were classified as outliers if observed and predicted rates of cesarean delivery differed (p < .05). PRINCIPAL FINDINGS: Discrimination of the full medical record and birth certificate models was higher (p < .001) than the discrimination of the more limited common and reliable variable models. Based on the full medical record model, six hospitals were classified as statistical (p < .01) outliers (three high and three low). In contrast, the full birth certificate model identified five low and four high outliers, and classifications differed for seven of the 20 hospitals. Even so, the correlation between adjusted hospital rates was substantial (r = .71). Interestingly, correlations between the full medical record model and the more limited common (r = .84) and reliable (r = .88) variable birth certificate models were higher, and differences in classification of hospital outlier status were fewer. CONCLUSION: Birth certificates can be used to develop cesarean delivery risk-adjustment models that have excellent discrimination. However, using the full complement of birth certificate variables may lead to biased hospital comparisons. In contrast, limiting models to data elements with known reliability may yield rankings that are more similar to rankings based on medical record data.
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Cesárea/estadística & datos numéricos , Auditoría Médica/métodos , Servicio de Ginecología y Obstetricia en Hospital/clasificación , Pautas de la Práctica en Medicina/estadística & datos numéricos , Ajuste de Riesgo , Adulto , Certificado de Nacimiento , Estudios de Cohortes , Recolección de Datos , Femenino , Investigación sobre Servicios de Salud , Humanos , Registros Médicos , Modelos Estadísticos , Servicio de Ginecología y Obstetricia en Hospital/estadística & datos numéricos , Ohio/epidemiología , Embarazo , Revisión de Utilización de RecursosRESUMEN
AMP-activated protein kinase (AMPK) is a metabolic stress-sensing protein kinase responsible for coordinating metabolism and energy demand. In rodents, exercise accelerates fatty acid metabolism, enhances glucose uptake, and stimulates nitric oxide (NO) production in skeletal muscle. AMPK phosphorylates and inhibits acetyl-coenzyme A (CoA) carboxylase (ACC) and enhances GLUT-4 translocation. It has been reported that human skeletal muscle malonyl-CoA levels do not change in response to exercise, suggesting that other mechanisms besides inhibition of ACC may be operating to accelerate fatty acid oxidation. Here, we show that a 30-s bicycle sprint exercise increases the activity of the human skeletal muscle AMPK-alpha1 and -alpha2 isoforms approximately two- to threefold and the phosphorylation of ACC at Ser(79) (AMPK phosphorylation site) approximately 8.5-fold. Under these conditions, there is also an approximately 5.5-fold increase in phosphorylation of neuronal NO synthase-mu (nNOSmu;) at Ser(1451). These observations support the concept that inhibition of ACC is an important component in stimulating fatty acid oxidation in response to exercise and that there is coordinated regulation of nNOSmu to protect the muscle from ischemia/metabolic stress.
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Acetil-CoA Carboxilasa/metabolismo , Complejos Multienzimáticos/metabolismo , Contracción Muscular , Músculo Esquelético/enzimología , Óxido Nítrico Sintasa/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Transducción de Señal , Proteínas Quinasas Activadas por AMP , Adulto , Biopsia , Activación Enzimática , Ejercicio Físico/fisiología , Ácidos Grasos/metabolismo , Femenino , Glucosa/metabolismo , Humanos , Isoenzimas/metabolismo , Masculino , Óxido Nítrico Sintasa de Tipo I , Oxidación-Reducción , Consumo de Oxígeno , FosforilaciónRESUMEN
There has been recent interest in the potential performance and metabolic effects of carbohydrate ingestion during exercise lasting approximately 1 h. In this study, 13 well-trained men ingested in randomized order either a 6% glucose solution (CHO trial) or a placebo (Con trial) during exercise to exhaustion at 83+/-1% peak oxygen uptake. In six subjects, vastus lateralis muscle was sampled at rest, at 32 min, and at exhaustion, and in six subjects, glucose kinetics was determined by infusion of [6,6-(2)H]glucose in both trials and ingestion of [6-(3)H]glucose in the CHO trial. Of the 84 g of glucose ingested during exercise in the CHO trial, only 22 g appeared in the peripheral circulation. This resulted in a small (12 g) but significant (P<0.05) increase in glucose uptake without influencing carbohydrate oxidation, muscle glycogen use, or time to exhaustion (CHO: 68.1+/-4.1 min; Con: 69.6+/-5.5 min). Decreases in muscle phosphocreatine content and increases in muscle inosine monophosphate and lactate content during exercise were similar in the two trials. Although endogenous glucose production during exercise was partially suppressed in the CHO trial, it remained significantly above preexercise levels throughout exercise. In conclusion, only 26% of the ingested glucose appeared in the peripheral circulation. Glucose ingestion increased glucose uptake and partially reduced endogenous glucose production but had no effect on carbohydrate oxidation, muscle metabolism, or time to exhaustion during exercise at 83% peak oxygen uptake.
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Glucosa/biosíntesis , Glucosa/farmacocinética , Músculo Esquelético/metabolismo , Resistencia Física/fisiología , Adenosina Difosfato/metabolismo , Adenosina Monofosfato/metabolismo , Adenosina Trifosfato/metabolismo , Adulto , Glucosa/metabolismo , Frecuencia Cardíaca/fisiología , Hemoglobinas , Humanos , Inosina Monofosfato/metabolismo , Insulina/metabolismo , Ácido Láctico/metabolismo , Oxidación-Reducción , Consumo de Oxígeno/fisiología , Fosfocreatina/metabolismo , Resistencia Física/efectos de los fármacos , Volumen Plasmático/fisiología , TritioRESUMEN
The effects of sprint training on muscle metabolism and ion regulation during intense exercise remain controversial. We employed a rigorous methodological approach, contrasting these responses during exercise to exhaustion and during identical work before and after training. Seven untrained men undertook 7 wk of sprint training. Subjects cycled to exhaustion at 130% pretraining peak oxygen uptake before (PreExh) and after training (PostExh), as well as performing another posttraining test identical to PreExh (PostMatch). Biopsies were taken at rest and immediately postexercise. After training in PostMatch, muscle and plasma lactate (Lac(-)) and H(+) concentrations, anaerobic ATP production rate, glycogen and ATP degradation, IMP accumulation, and peak plasma K(+) and norepinephrine concentrations were reduced (P<0.05). In PostExh, time to exhaustion was 21% greater than PreExh (P<0.001); however, muscle Lac(-) accumulation was unchanged; muscle H(+) concentration, ATP degradation, IMP accumulation, and anaerobic ATP production rate were reduced; and plasma Lac(-), norepinephrine, and H(+) concentrations were higher (P<0.05). Sprint training resulted in reduced anaerobic ATP generation during intense exercise, suggesting that aerobic metabolism was enhanced, which may allow increased time to fatigue.
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Adaptación Fisiológica/fisiología , Ejercicio Físico/fisiología , Músculo Esquelético/metabolismo , Potasio/sangre , Carrera/fisiología , Equilibrio Ácido-Base/fisiología , Adenosina Trifosfato/biosíntesis , Adulto , Umbral Anaerobio/fisiología , Dióxido de Carbono/sangre , Epinefrina/sangre , Glucógeno/metabolismo , Glucólisis/fisiología , Frecuencia Cardíaca/fisiología , Humanos , Concentración de Iones de Hidrógeno , Ácido Láctico/metabolismo , Masculino , Norepinefrina/sangre , Oxígeno/sangre , Consumo de Oxígeno/fisiología , Resistencia Física/fisiología , Protones , Intercambio Gaseoso Pulmonar/fisiologíaRESUMEN
PURPOSE: This study examined the effect of training status and relative exercise intensity on physiological responses to endurance exercise in humans. METHODS: Seven endurance trained (TR: peak oxygen uptake [VO2peak] = 65.8 +/- 2.4 mL x kg(-1) min(-1)) and six untrained (UT: VO2peak = 46.2 +/- 1.9 mL x kg(-1) x min(-1)) men cycled for 60 min, either at a work rate corresponding to approximately 70% VO2peak or approximately 95% lactate threshold (LT). RESULTS: The work rate and relative exercise intensity (i.e., % VO2peak) for UT 95% LT were lower (P < 0.01) than for all of the other trials. Although the work rate for UT 70% VO2peak was lower (P < 0.001) than for TR 70% VO2peak and TR 95% LT, average heart rate (HR) for the trial was higher (P < 0.01) throughout exercise in UT 70% VO2peak compared with all of the other trials. Plasma lactate and ammonia concentrations were greater (P < 0.01) during exercise in UT 70% VO2peak compared with all of the other trials. There was a tendency (P = 0.077) for plasma hypoxanthine to be greater at 60 min in UT 70% VO2peak compared with the other trials. At no time were any of the plasma metabolite concentrations different between the UT 95% LT, TR 95% LT and TR 70% VO2peak trials. CONCLUSIONS: These data demonstrate that HR and plasma markers of metabolic stress were greater in UT compared with TR when exercise was performed at 70% VO2peak but were similar during exercise at 95% LT.
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Ejercicio Físico/fisiología , Resistencia Física/fisiología , Adulto , Amoníaco/sangre , Biomarcadores/sangre , Frecuencia Cardíaca , Humanos , Ácido Láctico/sangre , Masculino , Consumo de OxígenoRESUMEN
The relationship between changes in the muscle total adenine nucleotide pool (TAN = ATP + ADP + AMP) and IMP during and after 30 s of sprint cycling was examined. Skeletal muscle samples were obtained from the vastus lateralis muscle of seven untrained men (23. 9 +/- 2.3 yr, 74.4 +/- 3.6 kg, and 55.0 +/- 2.9 ml. kg(-1). min(-1) peak oxygen consumption) before and immediately after exercise and after 5 and 10 min of passive recovery. The exercise-induced increase in muscle IMP was linearly related to the decrease in muscle TAN (r = -0.97, P < 0.01), and the slope of this relationship (-0.83) was not different from 1.0 (P > 0.05), indicating a 1:1 stoichiometric relationship. This interpretation must be treated cautiously, because all subjects displayed a greater decrease in TAN compared with the increase in IMP content, and the TAN + IMP + inosine + hypoxanthine content was lower (P < 0.05) immediately after exercise compared with during rest. During the first 5 min of recovery, the increase in TAN was not correlated with the decrease in IMP (r = -0.18, P > 0.05). In all subjects, the magnitude of TAN increase was higher than the magnitude of IMP decrease over this recovery period. In contrast, the increase in TAN was correlated with the decrease in IMP throughout the second 5 min of recovery (r = -0.80, P < 0.05), and it was a 1:1 stoichiometric relationship (slope = -1.12). These data indicate that a small proportion of the TAN pool was temporarily lost from the muscle purine stores during sprinting but was rapidly recovered after exercise.
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Nucleótidos de Adenina/metabolismo , Ejercicio Físico/fisiología , Músculo Esquelético/metabolismo , Adenosina Difosfato/metabolismo , Adenosina Monofosfato/metabolismo , Adenosina Trifosfato/metabolismo , Adulto , Amoníaco/metabolismo , Humanos , Hipoxantina/metabolismo , Inosina/metabolismo , Inosina Monofosfato/metabolismo , Pierna , MasculinoRESUMEN
The present study was undertaken to examine the effect of carbohydrate ingestion on plasma and muscle ammonia (NH(3) denotes ammonia and ammonium) accumulation during prolonged exercise. Eleven trained men exercised for 2 h at 65% peak pulmonary oxygen consumption while ingesting either 250 ml of an 8% carbohydrate-electrolyte solution every 15 min (CHO) or an equal volume of a sweet placebo. Blood glucose and plasma insulin levels during exercise were higher in CHO, but plasma hypoxanthine was lower after 120 min (1.7 +/- 0.3 vs. 2.6 +/- 0.1 micromol/l; P < 0. 05). Plasma NH(3) levels were similar at rest and after 30 min of exercise in both trials but were lower after 60, 90, and 120 min of exercise in CHO (62 +/- 9 vs. 76 +/- 9 micromol/l; P < 0.05). Muscle NH(3) levels were similar at rest and after 30 min of exercise but were lower after 120 min of exercise in CHO (1.51 +/- 0.21 vs. 2.07 +/- 0.23 mmol/kg dry muscle; P < 0.05; n = 5). These data are best explained by carbohydrate ingestion reducing muscle NH(3) production from amino acid degradation, although a small reduction in net AMP catabolism within the contracting muscle may also make a minor contribution to the lower tissue NH(3) levels.
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Amoníaco/metabolismo , Carbohidratos de la Dieta/farmacología , Ejercicio Físico/fisiología , Nucleótidos de Adenina/metabolismo , Adulto , Sangre/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Músculo Esquelético/metabolismo , Consumo de Oxígeno/efectos de los fármacos , Intercambio Gaseoso Pulmonar/efectos de los fármacosRESUMEN
OBJECTIVE: To examine the relationship of in-hospital and 30-day mortality rates and the association between in-hospital mortality and hospital discharge practices. DATA SOURCES/STUDY SETTING: A secondary analysis of data for 13,834 patients with congestive heart failure who were admitted to 30 hospitals in northeast Ohio in 1992-1994. DESIGN: A retrospective cohort study was conducted. DATA COLLECTION: Demographic and clinical data were collected from patients' medical records and were used to develop multivariable models that estimated the risk of in-hospital and 30-day (post-admission) mortality. Standardized mortality ratios (SMRs) for in-hospital and 30-day mortality were determined by dividing observed death rates by predicted death rates. PRINCIPAL FINDINGS: In-hospital SMRs ranged from 0.54 to 1.42, and six hospitals were classified as statistical outliers (p <.05); 30-day SMRs ranged from 0.63 to 1.73, and seven hospitals were outliers. Although the correlation between in-hospital SMRs and 30-day SMRs was substantial (R = 0.78, p < .001), outlier status changed for seven of the 30 hospitals. Nonetheless, changes in outlier status reflected relatively small differences between in-hospital and 30-day SMRs. Rates of discharge to nursing homes or other inpatient facilities varied from 5.4 percent to 34.2 percent across hospitals. However, relationships between discharge rates to such facilities and in-hospital SMRs (R = 0.08; p = .65) and early post-discharge mortality rates (R = 0.23; p = .21) were not significant. CONCLUSIONS: SMRs based on in-hospital and 30-day mortality were relatively similar, although classification of hospitals as statistical outliers often differed. However, there was no evidence that in-hospital SMRs were biased by differences in post-discharge mortality or discharge practices.
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Insuficiencia Cardíaca/mortalidad , Mortalidad Hospitalaria , Hospitales/normas , Alta del Paciente/estadística & datos numéricos , Indicadores de Calidad de la Atención de Salud/normas , Anciano , Anciano de 80 o más Años , Femenino , Investigación sobre Servicios de Salud , Mortalidad Hospitalaria/tendencias , Hospitales/clasificación , Humanos , Masculino , Análisis Multivariante , Ohio/epidemiología , Acampadores DRG , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Factores de Riesgo , Factores de TiempoRESUMEN
The influence of the number of sprint bouts on purine loss was examined in nine men (age 24.8 +/- 1.6 yr, weight 76 +/- 3.9 kg, peak O(2) consumption 3.87 +/- 0.16 l/min) who performed either one (B1), four (B4), or eight (B8) 10-s sprints on a cycle ergometer, 1 wk apart, in a randomized order. Forearm venous plasma inosine, hypoxanthine (Hx), and uric acid concentrations were measured at rest and during 120 min of recovery. Urinary inosine, Hx, and uric acid excretion were also measured before and 24 h after exercise. During the first 120 min of recovery, plasma inosine and Hx concentrations, and urinary Hx excretion rate, were progressively higher (P < 0.05) with an increasing number of sprint bouts. Plasma uric acid concentration was higher (P < 0.05) in B8 compared with B1 and B4 after 45, 60, and 120 min of recovery. Total urinary excretion of purines (inosine + Hx + uric acid) was higher (P < 0. 05) at 2 h of recovery after B8 (537 +/- 59 micromol) compared with the other trials (B1: 270 +/- 76; B4: 327 +/- 59 micromol). These results indicate that the loss of purine from the body was enhanced by increasing the number of intermittent 10-s sprint bouts.