RESUMO
AIM: Striatin (Strn) is a scaffold protein expressed in cardiomyocytes (CMs) and alteration of its expression are described in various cardiac diseases. However, the alteration underlying its pathogenicity have been poorly investigated. METHODS: We studied the role(s) of cardiac Strn gene (STRN) by comparing the functional properties of CMs, generated from Strn-KO and isogenic WT mouse embryonic stem cell lines. RESULTS: The spontaneous beating rate of Strn-KO CMs was faster than WT cells, and this correlated with a larger fast INa conductance and no changes in If. Paced (2-8 Hz) Strn-KO CMs showed prolonged action potential (AP) duration in comparison with WT CMs and this was not associated with changes in ICaL and IKr. Motion video tracking analysis highlighted an altered contraction in Strn-KO CMs; this was associated with a global increase in intracellular Ca2+, caused by an enhanced late Na+ current density (INaL) and a reduced Na+/Ca2+ exchanger (NCX) activity and expression. Immunofluorescence analysis confirmed the higher Na+ channel expression and a more dynamic microtubule network in Strn-KO CMs than in WT. Indeed, incubation of Strn-KO CMs with the microtubule stabilizer taxol, induced a rescue (downregulation) of INa conductance toward WT levels. CONCLUSION: Loss of STRN alters CMs electrical and contractile profiles and affects cell functionality by a disarrangement of Strn-related multi-protein complexes. This leads to impaired microtubules dynamics and Na+ channels trafficking to the plasma membrane, causing a global Na+ and Ca2+ enhancement.
Assuntos
Cálcio , Miócitos Cardíacos , Animais , Miócitos Cardíacos/metabolismo , Camundongos , Cálcio/metabolismo , Potenciais de Ação/efeitos dos fármacos , Camundongos Knockout , Proteínas Musculares/metabolismo , Proteínas Musculares/genética , Trocador de Sódio e Cálcio/metabolismo , Trocador de Sódio e Cálcio/genética , Células-Tronco Embrionárias Murinas/metabolismo , Sódio/metabolismoRESUMO
PURPOSE: Pathological complete response (pCR) following neoadjuvant chemoradiotherapy or radiotherapy in locally advanced rectal cancer (LARC) is reached in approximately 15-30% of cases, therefore it would be useful to assess if pretreatment of 18F-FDG PET/CT and/or MRI texture features can reliably predict response to neoadjuvant therapy in LARC. METHODS: Fifty-two patients were dichotomized as responder (pR+) or non-responder (pR-) according to their pathological tumor regression grade (TRG) as follows: 22 as pR+ (nine with TRG = 1, 13 with TRG = 2) and 30 as pR- (16 with TRG = 3, 13 with TRG = 4 and 1 with TRG = 5). First-order parameters and 21 second-order texture parameters derived from the Gray-Level Co-Occurrence matrix were extracted from semi-automatically segmented tumors on T2w MRI, ADC maps, and PET/CT acquisitions. The role of each texture feature in predicting pR+ was assessed with monoparametric and multiparametric models. RESULTS: In the mono-parametric approach, PET homogeneity reached the maximum AUC (0.77; sensitivity = 72.7% and specificity = 76.7%), while PET glycolytic volume and ADC dissimilarity reached the highest sensitivity (both 90.9%). In the multiparametric analysis, a logistic regression model containing six second-order texture features (five from PET and one from T2w MRI) yields the highest predictivity in distinguish between pR+ and pR- patients (AUC = 0.86; sensitivity = 86%, and specificity = 83% at the Youden index). CONCLUSIONS: If preliminary results of this study are confirmed, pretreatment PET and MRI could be useful to personalize patient treatment, e.g., avoiding toxicity of neoadjuvant therapy in patients predicted pR-.
Assuntos
Fluordesoxiglucose F18 , Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Imagem Multimodal , Terapia Neoadjuvante , Tomografia por Emissão de Pósitrons , Neoplasias Retais/diagnóstico por imagem , Neoplasias Retais/terapia , Adulto , Feminino , Humanos , Masculino , Neoplasias Retais/patologiaRESUMO
Computer-aided diagnosis (CAD) systems are increasingly being used in clinical settings to report multi-parametric magnetic resonance imaging (mp-MRI) of the prostate. Usually, CAD systems automatically highlight cancer-suspicious regions to the radiologist, reducing reader variability and interpretation errors. Nevertheless, implementing this software requires the selection of which mp-MRI parameters can best discriminate between malignant and non-malignant regions. To exploit functional information, some parameters are derived from dynamic contrast-enhanced (DCE) acquisitions. In particular, much CAD software employs pharmacokinetic features, such as K trans and k ep, derived from the Tofts model, to estimate a likelihood map of malignancy. However, non-pharmacokinetic models can be also used to describe DCE-MRI curves, without any requirement for prior knowledge or measurement of the arterial input function, which could potentially lead to large errors in parameter estimation. In this work, we implemented an empirical function derived from the phenomenological universalities (PUN) class to fit DCE-MRI. The parameters of the PUN model are used in combination with T2-weighted and diffusion-weighted acquisitions to feed a support vector machine classifier to produce a voxel-wise malignancy likelihood map of the prostate. The results were all compared to those for a CAD system based on Tofts pharmacokinetic features to describe DCE-MRI curves, using different quality aspects of image segmentation, while also evaluating the number and size of false positive (FP) candidate regions. This study included 61 patients with 70 biopsy-proven prostate cancers (PCa). The metrics used to evaluate segmentation quality between the two CAD systems were not statistically different, although the PUN-based CAD reported a lower number of FP, with reduced size compared to the Tofts-based CAD. In conclusion, the CAD software based on PUN parameters is a feasible means with which to detect PCa, without affecting segmentation quality, and hence it could be successfully applied in clinical settings, improving the automated diagnosis process and reducing computational complexity.
Assuntos
Algoritmos , Diagnóstico por Computador/métodos , Interpretação de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Neoplasias da Próstata/diagnóstico , Idoso , Meios de Contraste , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , SoftwareRESUMO
To explore contrast (C) and homogeneity (H) gray-level co-occurrence matrix texture features on T2-weighted (T2w) Magnetic Resonance (MR) images and apparent diffusion coefficient (ADC) maps for predicting prostate cancer (PCa) aggressiveness, and to compare them with traditional ADC metrics for differentiating low- from intermediate/high-grade PCas. The local Ethics Committee approved this prospective study of 93 patients (median age, 65 years), who underwent 1.5 T multiparametric endorectal MR imaging before prostatectomy. Clinically significant (volume ≥0.5 ml) peripheral tumours were outlined on histological sections, contoured on T2w and ADC images, and their pathological Gleason Score (pGS) was recorded. C, H, and traditional ADC metrics (mean, median, 10th and 25th percentile) were calculated on the largest lesion slice, and correlated with the pGS through the Spearman correlation coefficient. The area under the receiver operating characteristic curve (AUC) assessed how parameters differentiate pGS = 6 from pGS ≥ 7. The dataset included 49 clinically significant PCas with a balanced distribution of pGS. The Spearman ρ and AUC values on ADC were: -0.489, 0.823 (mean); -0.522, 0.821 (median); -0.569, 0.854 (10th percentile); -0.556, 0.854 (25th percentile); -0.386, 0.871 (C); 0.533, 0.923 (H); while on T2w they were: -0.654, 0.945 (C); 0.645, 0.962 (H). AUC of H on ADC and T2w, and C on T2w were significantly higher than that of the mean ADC (p = 0.05). H and C calculated on T2w images outperform ADC parameters in correlating with pGS and differentiating low- from intermediate/high-risk PCas, supporting the role of T2w MR imaging in assessing PCa biological aggressiveness.
Assuntos
Imagem de Difusão por Ressonância Magnética/métodos , Interpretação de Imagem Assistida por Computador/métodos , Neoplasias da Próstata/diagnóstico , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de TumoresRESUMO
Proton magnetic resonance spectroscopy ((1)H-MRS) is largely exploited in clinical settings to non-invasively investigate chemical compounds in human tissues. Applications of (1)H-MRS in oncology field are connected to the detection of abnormal levels of choline compounds in more active tumours, providing useful information for cancer diagnosis and treatment monitoring. Since benign lesions may also show presence of a choline peak, implementing absolute evaluation will help differentiating benign from malignant tumours. An external reference procedure was described to provide choline quantification in standard unit of measurements. Spectra were acquired on a 1.5 T scanner using both phantoms and healthy volunteers with a PRESS sequence. The implemented quantification procedure used metabolite and noise measurements on the spectrum to remove large part of scanner settings contributing to metabolites of interest. A standard quantification was also used to compare performances of the noise-based method. In vitro quantification had accuracy and precision in the range (95-99)% and (5-13)%, respectively. When applied to in vivo studies on healthy volunteers, the method provided very close values of choline concentration, more exactly (1.73 ± 0.24) mmol/l. The method proposed can quantify the proper choline content in phantoms as well as in human structures, as brain. The method is ease of use, computational costless and it can be rapidly calibrated and implemented in any centre.
Assuntos
Colina/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Espectroscopia de Ressonância Magnética/normas , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Imagens de Fantasmas , Padrões de Referência , Adulto JovemRESUMO
Dynamic contrast-enhanced study in magnetic resonance imaging (DCE-MRI) is an important tool in oncology to visualize tissues vascularization and to define tumour aggressiveness on the basis of an altered perfusion and permeability. Pharmacokinetic models are generally used to extract hemodynamic parameters, providing a quantitative description of the contrast uptake and wash-out. Empirical functions can also be used to fit experimental data without the need of any assumption about tumour physiology, as in pharmacokinetic models, increasing their diagnostic utility, in particular when automatic diagnosis systems are implemented on the basis of an MRI multi-parametric approach. Phenomenological universalities (PUN) represent a novel tool for experimental research and offer a simple and systematic method to represent a set of data independent of the application field. DCE-MRI acquisitions can thus be advantageously evaluated by the extended PUN class, providing a convenient diagnostic tool to analyse functional studies, adding a new set of features for the classification of malignant and benign lesions in computer aided detection systems. In this work the Tofts pharmacokinetic model and the class EU1 generated by the PUN description were compared in the study of DCE-MRI of the prostate, evaluating complexity of model implementation, goodness of fitting results, classification performances and computational cost. The mean R² obtained with the EU1 and Tofts model were equal to 0.96 and 0.90, respectively, and the classification performances achieved by the EU1 model and the Tofts implementation discriminated malignant from benign tissues with an area under the receiver operating characteristic curve equal to 0.92 and 0.91, respectively. Furthermore, the EU1 model has a simpler functional form which reduces implementation complexity and computational time, requiring 6 min to complete a patient elaboration process, instead of 8 min needed for the Tofts model analysis.
Assuntos
Meios de Contraste/farmacocinética , Imageamento por Ressonância Magnética , Modelos Biológicos , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/metabolismoRESUMO
OBJECTIVES: To determine whether proton magnetic resonance spectroscopy (1H-MRS) can help differentiate between benign and malignant soft tissue lesions, and to assess if there is a correlation between 1H-MRS data and the mitotic index. METHODS: MR measurements were performed in 43 patients with soft tissue tumours >15 mm in diameter. Six cases were excluded for technical failure. Examinations were performed at 1.5 T using a single-voxel point resolved spectroscopy sequence (PRESS) with TR/TE = 2000/150 ms. The volume of interest was positioned within the lesion avoiding inclusion of necrotic regions. In all patients, a histological diagnosis was obtained and the corresponding mitotic index was also computed. 1H-MRS results and histopathological findings were compared using the chi-squared test and correlation coefficient. RESULTS: Choline was detected in 18/19 patients with malignant tumours and in 3/18 patients with benign lesions. The three benign lesions included one desmoid tumour, one ossificans myositis and one eccrine spiradenoma. Choline was not detected in 15 patients with benign lesions or in one patient with dermatofibrosarcoma protuberans. Resulting 1H-MRS sensitivity and specificity were 95% and 83% respectively. CONCLUSIONS: Absence of choline peak is highly predictive of benign tumours suggesting that 1H-MRS can help to differentiate malignant from benign tumours. KEY POINTS: ⢠1H-MRS may allow differentiation between benign and malignant soft tissue lesions ⢠Absence of choline peak is highly predictive of benign soft tissue lesions ⢠Malignant tumours with a mitotic index >2/10 HPF had a positive choline peak ⢠A choline peak may still be identified in some benign tumours.
Assuntos
Biomarcadores Tumorais/análise , Colina/análise , Diagnóstico por Computador/métodos , Espectroscopia de Ressonância Magnética/métodos , Neoplasias de Tecidos Moles/diagnóstico , Neoplasias de Tecidos Moles/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prótons , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
AIM: The primary purpose of this study was to compare energy expenditure among resistance exercise protocols using maximally explosive or slow contractions versus recreational in trained and untrained men. METHODS: Seven trained (21.9±2.1 yrs) and seven untrained men (20.1±2.2 yrs) performed three nearly identical exercise protocols, and a no-exercise (CONTROL) session in a randomly assigned, counterbalanced order. Subjects performed three sets of squats, dumbbell-row, deadlift, bench press, lat-pulldown, shoulder press, arm curls and dips using either recreational (REC), 2s (SLOW) or maximally explosive contractions (MAX). Expired air was collected continuously for 15 min before, ~37-43 min during, and 2 hr postexercise. Finger prick samples (25 µL) were collected and analyzed for blood lactate (BL) (mmol.L-1) before, immediately after, and during 120 min of recovery. RESULTS: Rates of energy expenditure were significantly (P≤0.05) greater for MAX than SLOW and REC during all exercises and +5 min after exercise in trained men, and MAX was greater than REC during all exercises except deadlift in untrained men. In trained men, total kcal were significantly greater (P≤0.05) with MAX (507±48) compared to REC (431±47), but not in untrained. Conversely, BL was significantly greater (P≤0.05) after SLOW compared to REC in trained and untrained men, while BL was only greater after MAX versus REC in trained men. CONCLUSION: For whole-body resistance exercise programs, maximally explosive contractions optimize energy expenditure in trained men, but slow contractions are recommended for untrained exercisers. Therefore, contraction intensity should be considered a program design variable for exercise prescriptions aimed to improve general health and fitness.
Assuntos
Metabolismo Energético/fisiologia , Contração Muscular/fisiologia , Músculo Esquelético/fisiologia , Esforço Físico/fisiologia , Aptidão Física/fisiologia , Análise de Variância , Área Sob a Curva , Composição Corporal , Dieta , Humanos , Masculino , Força Muscular/fisiologia , Consumo de Oxigênio/fisiologia , Adulto JovemRESUMO
Dynamic contrast enhancement in magnetic resonance imaging (DCE-MRI) is a promising tool for the clinical diagnosis of tumors, whose implementation may be improved through the use of suitable hemodynamic models. If one prefers to avoid assumptions about the tumor physiology, empirical fitting functions may be adopted. For this purpose, in this paper we discuss the exploitation of a recently proposed phenomenological universalities (PUN) formalism. In fact, we show that a novel PUN class may be used to describe the time-signal intensity curves in both healthy and tumoral tissues, discriminating between the two cases and thus potentially providing a convenient diagnostic tool. The proposed approach is applied to analysis of the DCE-MRI data relative to a study group composed of ten patients with spine tumors.
Assuntos
Meios de Contraste , Aumento da Imagem/métodos , Imageamento por Ressonância Magnética/métodos , Humanos , Processamento de Imagem Assistida por Computador , Neoplasias da Medula Espinal/diagnóstico , Fatores de TempoRESUMO
Using a standardized rat model of contusive spinal cord injury (SCI; [Gorio A, Gokmen N, Erbayraktar S, Yilmaz O, Madaschi L, Cichetti C, Di Giulio AM, Vardar E, Cerami A, Brines M (2002) Recombinant human erythropoietin counteracts secondary injury and markedly enhances neurological recovery from experimental spinal cord trauma. Proc Natl Acad Sci U S A 99:9450-9455]), we previously showed that the administration of recombinant human erythropoietin (rhEPO) improves both tissue sparing and locomotory outcome. In the present study, to better understand rhEPO-mediated effects on chronic astrocyte response to SCI in rat, we have used immunocytochemical methods combined with confocal and electron microscopy to investigate, 1 month after injury, the effects of a single rhEPO administration on the expression of a) aquaporin 4 (AQP4), the main astrocytic water channel implicated in edema development and resolution, and two molecules (dystrophin and syntrophin) involved in its membrane anchoring; b) glial fibrillary acidic protein (GFAP) and vimentin as markers of astrogliosis; c) chondroitin sulfate proteoglycans of the extracellular matrix which are upregulated after SCI and can inhibit axonal regeneration and influence neuronal and glial properties. Our results show that rhEPO administration after SCI modifies astrocytic response to injury by increasing AQP4 immunoreactivity in the spinal cord, but not in the brain, without apparent modifications of dystrophin and syntrophin distribution. Attenuation of astrogliosis, demonstrated by the semiquantitative analysis of GFAP labeling, was associated with a reduction of phosphacan/RPTP zeta/beta, whereas the levels of lecticans remained unchanged. Finally, the relative volume of a microvessel fraction was significantly increased, indicating a pro-angiogenetic or a vasodilatory effect of rhEPO. These changes were consistently associated with remarkable reduction of lesion size and with improvement in tissue preservation and locomotor recovery, confirming previous observations and underscoring the potentiality of rhEPO for the therapeutic management of SCI.
Assuntos
Astrócitos/metabolismo , Contusões/metabolismo , Eritropoetina/farmacologia , Traumatismos da Medula Espinal/metabolismo , Animais , Aquaporina 4/metabolismo , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Contusões/patologia , Distrofina/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Gliose/tratamento farmacológico , Gliose/patologia , Imuno-Histoquímica , Técnicas In Vitro , Indicadores e Reagentes , Masculino , Microscopia Confocal , Microscopia Imunoeletrônica , Atividade Motora/fisiologia , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes , Traumatismos da Medula Espinal/patologia , Vimentina/metabolismoRESUMO
We investigated the effect of a single administration of recombinant human erythropoietin (rhEPO) on the preservation of the ventral white matter of rats at 4 weeks after contusive spinal cord injury (SCI), a time at which functional recovery is significantly improved in comparison to the controls [Gorio A, Necati Gokmen N, Erbayraktar S, Yilmaz O, Madaschi L, Cichetti C, Di Giulio AM, Enver Vardar E, Cerami A, Brines M (2002) Recombinant human erythropoietin counteracts secondary injury and markedly enhances neurological recovery from experimental spinal cord trauma. Proc Natl Acad Sci U S A 99:9450-9455; Gorio A, Madaschi L, Di Stefano B, Carelli S, Di Giulio AM, De Biasi S, Coleman T, Cerami A, Brines M (2005) Methylprednisolone neutralizes the beneficial effects of erythropoietin in experimental spinal cord injury. Proc Natl Acad Sci U S A 102:16379-16384]. Specifically, we examined, by morphological and cytochemical methods combined with light, confocal and electron microscopy, i) myelin preservation, ii) activation of adult oligodendrocyte progenitors (OPCs) identified for the expression of NG2 transmembrane proteoglycan, iii) changes in the amount of the chondroitin sulfate proteoglycans neurocan, versican and phosphacan and of their glycosaminoglycan component labeled with Wisteria floribunda lectin, and iv) ventral horn density of the serotonergic plexus as a marker of descending motor control axons. Injured rats received either saline or a single dose of rhEPO within 30 min after SCI. The results showed that the significant improvement of functional outcome observed in rhEPO-treated rats was associated with a better preservation of myelin in the ventral white matter. Moreover, the significant increase of both the number of NG2-positive OPCs and the labeling for Nogo-A, a marker of differentiated oligodendrocytes, suggested that rhEPO treatment could result in the generation of new myelinating oligodendrocytes. Sparing of fiber tracts in the ventral white matter was confirmed by the increased density of the serotonergic plexus around motor neurons. As for chondroitin sulfate proteoglycans, only phosphacan, increased in saline-treated rats, returned to normal levels in rhEPO group, probably reflecting a better maintenance of glial-axolemmal relationships along nerve fibers. In conclusion, this investigation expands previous studies supporting the pleiotropic neuroprotective effect of rhEPO on secondary degenerative response and its therapeutic potential for the treatment of SCI and confirms that the preservation of the ventral white matter, which contains descending motor pathways, may be critical for limiting functional deficit.
Assuntos
Eritropoetina/farmacologia , Fibras Nervosas Mielinizadas/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Traumatismos da Medula Espinal/tratamento farmacológico , Medula Espinal/efeitos dos fármacos , Degeneração Walleriana/tratamento farmacológico , Animais , Antígenos/efeitos dos fármacos , Antígenos/metabolismo , Axônios/metabolismo , Axônios/ultraestrutura , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Proteoglicanas de Sulfatos de Condroitina/efeitos dos fármacos , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Eritropoetina/uso terapêutico , Masculino , Microscopia Eletrônica de Transmissão , Proteínas da Mielina/efeitos dos fármacos , Proteínas da Mielina/metabolismo , Bainha de Mielina/efeitos dos fármacos , Bainha de Mielina/metabolismo , Bainha de Mielina/ultraestrutura , Fibras Nervosas Mielinizadas/metabolismo , Fibras Nervosas Mielinizadas/patologia , Regeneração Nervosa/efeitos dos fármacos , Regeneração Nervosa/fisiologia , Vias Neurais/efeitos dos fármacos , Vias Neurais/metabolismo , Fármacos Neuroprotetores/uso terapêutico , Proteínas Nogo , Oligodendroglia/efeitos dos fármacos , Oligodendroglia/metabolismo , Oligodendroglia/ultraestrutura , Proteoglicanas/efeitos dos fármacos , Proteoglicanas/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes de Fusão/farmacologia , Proteínas Recombinantes de Fusão/uso terapêutico , Serotonina/metabolismo , Medula Espinal/metabolismo , Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/metabolismo , Traumatismos da Medula Espinal/fisiopatologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Células-Tronco/ultraestrutura , Resultado do Tratamento , Degeneração Walleriana/fisiopatologia , Degeneração Walleriana/prevenção & controleRESUMO
Brain inflammation is a common event in the pathogenesis of several neurological diseases. It is unknown whether leukocyte/endothelium interactions are sufficient to promote homing of blood-borne cells into the brain compartment. The role of mononuclear cells and endothelium was analyzed in a new experimental model, the isolated guinea-pig brain maintained in vitro by arterial perfusion. This preparation allows one to investigate early steps of brain inflammation that are impracticable in vivo. We demonstrate by confocal microscopy analysis that in vitro co-perfusion of pro-inflammatory agents and pre-activated fluorescent mononuclear cells induced endothelial expression of selectins and intracellular adhesion molecule-1 in correspondence of arrested mononuclear cells, and correlates with a moderate increase in blood-brain barrier permeability. Separate perfusion of pro-inflammatory agents and mononuclear cells induced neither mononuclear cell adhesion nor adhesion molecule expression. We demonstrate that co-activation of mononuclear cells and cerebral endothelium is an essential requirement for cell arrest and adhesion in the early stages of experimental cerebral inflammation.
Assuntos
Circulação Cerebrovascular/fisiologia , Endotélio Vascular/fisiopatologia , Inflamação/fisiopatologia , Animais , Sobrevivência Celular , Modelos Animais de Doenças , Endotélio Vascular/patologia , Cobaias , Técnicas In Vitro , Inflamação/patologia , Microscopia ConfocalRESUMO
AIM: This investigation determined the effects of 84 days of bedrest on the composition of myosin heavy chain (MHC) in single skeletal muscle fibres with and without a resistance-training countermeasure programme. METHODS: Muscle biopsies were obtained from the m. vastus lateralis (VL) and m. soleus (SOL) before and after 84 days of bedrest. While control (BR) subjects (VL n = 9; SOL n = 3) refrained from exercise, BRE subjects (VL n = 8; SOL n = 3) performed knee extensor and plantar flexor resistance exercise every third day. Approximately 110 fibres per sample were analysed for MHC composition using SDS-PAGE. RESULTS: BR-VL had 16 and 14% decreases (P < 0.05) in MHC I and IIa fibres, respectively. There were 10% increases (P < 0.05) in MHC I/IIa, IIa/IIx, I/IIa/IIx, and a approximately 30% increase (P < 0.05) in total hybrid fibres. BRE-VL showed a 15% reduction (P < 0.05) in MHC I fibres, no change in MHC IIa fibres, and a 13% increase (P < 0.05) in total hybrids. BR-SOL had a 19% decrease (P < 0.05) in MHC I fibres with a 22% increase in total hybrids. BRE-SOL showed no change in MHC composition across all fibre types. CONCLUSION: These data suggest that the exercise countermeasures programme prevented MHC shifts in the SOL and mitigated MHC shifts in the VL. Furthermore, in the VL it appears that the resistance training programme employed in this investigation during bedrest, emphasized the use of MHC IIa phenotype muscle fibres.
Assuntos
Repouso em Cama , Exercício Físico/fisiologia , Músculo Esquelético/metabolismo , Cadeias Pesadas de Miosina/metabolismo , Adulto , Eletroforese em Gel de Poliacrilamida , Decúbito Inclinado com Rebaixamento da Cabeça/fisiologia , Humanos , Articulação do Joelho/fisiologia , Masculino , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/fisiologia , Simulação de Ausência de PesoRESUMO
The binding of Lycopersicon esculentum lectin (LEA) to the vascular endothelium was studied in the central nervous system of rat, mouse and guinea pig at different developmental ages, and in a gliosarcoma model. Our observations showed that LEA consistently stained the entire vascular tree in the spinal cord and in the brain of all animal species at all developmental ages investigated. In the tumor model, the staining of the vascular network was very reproducible, enabled an easy identification of vascular profiles and displayed a higher efficiency when compared to two other commonly used vascular marker (EHS laminin and PECAM-1). Moreover, our results showed that LEA staining was comparable in both vibratome and paraffin sections and could be easily combined with other markers in double labeling experiments. These observations indicate that LEA staining may represent an effective and versatile endothelial marker for the study of the vasculature of the central nervous system in different animal species and experimental conditions.
Assuntos
Biomarcadores Tumorais/análise , Neoplasias do Sistema Nervoso Central/irrigação sanguínea , Sistema Nervoso Central/irrigação sanguínea , Endotélio Vascular/química , Gliossarcoma/irrigação sanguínea , Lectinas de Plantas/metabolismo , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Biomarcadores Tumorais/metabolismo , Endotélio Vascular/metabolismo , Feminino , Cobaias , Camundongos , Gravidez , RatosRESUMO
The effects of exercise on the molecular nature of secreted human growth hormone (GH) or its biological activity are not well understood. Plasma from women (average age 23.6 yr, n = 35), drawn before and after an acute heavy resistance exercise test, was fractionated by size exclusion chromatography into three size classes, namely, > 60 kDa (fraction A), 30-60 kDa (fraction B), and < 30 kDa (fraction C), before GH assay. Concentrations of GH in these fractions, as well as in unfractioned plasma, were measured by the Nichols immunoradiometric assay, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) polyclonal competitive RIA, Diagnostic Systems Laboratory's immunofunctional assay (measures dimerization-capable species), and the rat tibial bioassay. Significantly increased circulating GH concentrations of two- to fourfold were observed when immunoassays in unfractionated plasma samples were used, but they showed no significant change with use of the rat tibial bioassay. Significant exercise-induced increases in GH were found in fractions B and C but not in fraction A. Because chemical reduction of the samples before GH immunoassay significantly increased GH concentrations in fractions B and C (Nichols and NIDDK kits) after exercise, it is concluded that exercise may specifically increase release of disulfide-linked hormone molecules and/or fragments. Finally, because most of the GH released after exercise was able to dimerize the GH receptor in vitro, it is also concluded that these forms have the two intact binding sites required to initiate signal transduction in target cells.
Assuntos
Exercício Físico/fisiologia , Hormônio do Crescimento Humano/sangue , Adulto , Animais , Bioensaio/métodos , Cromatografia em Gel/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Teste de Esforço , Feminino , Lâmina de Crescimento/efeitos dos fármacos , Lâmina de Crescimento/fisiologia , Hormônio do Crescimento Humano/farmacologia , Humanos , Hipofisectomia , Ensaio Imunorradiométrico/métodos , Ratos , Ratos Sprague-Dawley , Análise de Regressão , TíbiaRESUMO
PURPOSE: This investigation was designed to examine the influence of creatine (Cr) supplementation on acute cardiovascular, renal, temperature, and fluid-regulatory hormonal responses to exercise for 35 min in the heat. METHODS: Twenty healthy men were matched and then randomly assigned to consume 0.3 g.kg(-1) Cr monohydrate (N = 10) or placebo (N = 10) for 7 d in a double-blind fashion. Before and after supplementation, both groups cycled for 30 min at 60-70% VO2(peak) immediately followed by three 10-s sprints in an environmental chamber at 37 degrees C and 80% relative humidity. RESULTS: Body mass was significantly increased (0.75 kg) in Cr subjects. Heart rate, blood pressure, and sweat rate responses to exercise were not significantly different between groups. There were no differences in rectal temperature responses in either group. Sodium, potassium, and creatinine excretion rates obtained from 24-h and exercise urine collection periods were not significantly altered in either group. Serum creatinine was elevated in the Cr group but within normal ranges. There were significant exercise-induced increases in cortisol, aldosterone, renin, angiotensin I and II, atrial peptide, and arginine vasopressin. The aldosterone response was slightly greater in the Cr (263%) compared with placebo (224%) group. Peak power was greater in the Cr group during all three 10-s sprints after supplementation and unchanged in the placebo group. There were no reports of adverse symptoms, including muscle cramping during supplementation or exercise. CONCLUSION: Cr supplementation augments repeated sprint cycle performance in the heat without altering thermoregulatory responses.
Assuntos
Regulação da Temperatura Corporal/efeitos dos fármacos , Creatina/farmacologia , Exercício Físico/fisiologia , Adulto , Aldosterona , Pressão Sanguínea/fisiologia , Índice de Massa Corporal , Regulação da Temperatura Corporal/fisiologia , Água Corporal/fisiologia , Creatina/administração & dosagem , Creatinina/sangue , Creatinina/urina , Frequência Cardíaca/fisiologia , Hormônios/metabolismo , Temperatura Alta , Humanos , Masculino , Natriurese/fisiologia , Esforço Físico/fisiologia , Volume Plasmático/fisiologia , Potássio/urina , Sudorese/fisiologia , Urina/fisiologiaRESUMO
PURPOSE: The effects of resistance training programs on strength, power, and military occupational task performances in women were examined. METHODS: Untrained women aged (mean +/- SD) 23 +/- 4 yr were matched and randomly placed in total- (TP, N = 17 and TH, N = 18) or upper-body resistance training (UP, N = 18 and UH, N = 15), field (FLD, N = 14), or aerobic training groups (AER, N = 11). Two periodized resistance training programs (with supplemental aerobic training) emphasized explosive exercise movements using 3- to 8-RM training loads (TP, UP), whereas the other two emphasized slower exercise movements using 8- to 12-RM loads (TH, UH). The FLD group performed plyometric and partner exercises. Subjects were tested for body composition, strength, power, endurance, maximal and repetitive box lift, 2-mile loaded run, and U.S. Army Physical Fitness Tests before (T0) and after 3 (T3) and 6 months of training (T6). For comparison, untrained men (N = 100) (MEN) were tested once. RESULTS: Specific training programs resulted in significant increases in body mass (TP), 1-RM squat (TP, TH, FLD), bench press (all except AER), high pull (TP), squat jump (TP, TH, FLD), bench throw (all except AER), squat endurance (all except AER), 1-RM box lift (all except aerobic), repetitive box lift (all), push-ups (all except AER), sit-ups (all except AER), and 2-mile run (all). CONCLUSIONS: Strength training improved physical performances of women over 6 months and adaptations in strength, power, and endurance were specific to the subtle differences (e.g., exercise choice and speeds of exercise movement) in the resistance training programs (strength/power vs strength/hypertrophy). Upper- and total-body resistance training resulted in similar improvements in occupational task performances, especially in tasks that involved upper-body musculature. Finally, gender differences in physical performance measures were reduced after resistance training in women, which underscores the importance of such training for physically demanding occupations.
Assuntos
Militares , Ocupações , Resistência Física , Levantamento de Peso , Adolescente , Adulto , Feminino , Humanos , Fatores Sexuais , Análise e Desempenho de TarefasRESUMO
Circadian rhythms of serum testosterone concentrations in men have been shown, in general, to be highest in the morning and lowest in the evening. Thus, the purpose of this investigation was to determine the effects of acute resistance exercise upon the waking circadian rhythm of salivary testosterone over 2 days (with or without resistance exercise). The subjects included ten resistance-trained men (with at least 1 year of lifting experience) with the following characteristics [mean (SD)]: age 21.6 (1.1) years; height 177.8 (9.5) cm; body mass 80.5 (11.5) kg; percent body fat 7.9 (1.7)%. A matched, randomized, crossover study design was used such that each subject was tested under both the resistance exercise and control (no exercise) conditions. The resistance exercise protocol consisted of ten exercises performed for three sets of ten repetitions maximum with 2 min of rest between sets. Saliva sample 1 was collected at 0615 hours and resistance exercise began immediately afterwards at approximately 0620 hours, and sample 2 was collected at 0700 hours, which corresponded approximately to a mid-exercise (or control) time point. Saliva samples were then obtained every hour on the hour from 0800 hours until 2200 hours. No significant differences were observed between the exercise and resting conditions for salivary testosterone, with the exception of a significant decrease at 0700 hours during the resistance exercise protocol. The results of this investigation indicate that resistance exercise does not affect the circadian pattern of salivary testosterone secretion over a 16-h waking period in resistance-trained men.
Assuntos
Ritmo Circadiano/fisiologia , Exercício Físico/fisiologia , Testosterona/metabolismo , Levantamento de Peso/fisiologia , Adulto , Estudos Cross-Over , Humanos , Masculino , Saliva/química , Testosterona/análiseRESUMO
PURPOSE: The purpose of this investigation was to determine the long-term training adaptations associated with low-volume circuit-type versus periodized high-volume resistance training programs in women. METHODS: 34 healthy, untrained women were randomly placed into one of the following groups: low-volume, single-set circuit (SSC; N = 12); periodized high-volume multiple-set (MS; N = 12); or nonexercising control (CON) group (N = 10). The SSC group performed one set of 8-12 repetitions to muscular failure 3 d x wk(-1). The MS group performed two to four sets of 3-15 repetitions with periodized volume and intensity 4 d x wk(-1). Muscular strength, power, speed, endurance, anthropometry, and resting hormonal concentrations were determined pretraining (T1), after 12 wk (T2), and after 24 wk of training (T3). RESULTS: 1-RM bench press and leg press, and upper and lower body local muscular endurance increased significantly (P < or = 0.05) at T2 for both groups, but only MS showed a significant increase at T3. Muscular power and speed increased significantly at T2 and T3 only for MS. Increases in testosterone were observed for both groups at T2 but only MS showed a significant increase at T3. Cortisol decreased from T1 to T2 and from T2 to T3 in MS. Insulin-like growth factor-1 increased significantly at T3 for SSC and at T2 and T3 for MS. No changes were observed for growth hormone in any of the training groups. CONCLUSION: Significant improvements in muscular performance may be attained with either a low-volume single-set program or a high-volume, periodized multiple-set program during the first 12 wk of training in untrained women. However, dramatically different training adaptations are associated with specific domains of training program design which contrast in speed of movement, exercise choices and use of variation (periodization) in the intensity and volume of exercise.
Assuntos
Resistência Física/fisiologia , Levantamento de Peso/fisiologia , Adaptação Fisiológica , Adulto , Análise de Variância , Composição Corporal , Feminino , Hormônios/sangue , Humanos , Hidrocortisona/sangue , RadioimunoensaioRESUMO
PURPOSE: The purpose of this study was to investigate the comprehensive physiological alterations that take place during the combination of bench-step aerobics (BSA) and resistance exercise training. METHODS: Thirty-five healthy, active women were randomly assigned to one of four groups that either a) performed 25 min of BSA only (SA25); b) performed a combination of 25 min of BSA and a multiple-set upper and lower body resistance exercise program (SAR); c) performed 40 min of BSA only (SA40); or d) served as a control group (C), only performing activities of daily living. Direct assessments for body composition, aerobic fitness, muscular strength, endurance, power, and cross-sectional area were performed 1 wk before and after 12 wk of training. RESULTS: All training groups significantly improved peak VO(2) (3.7 to 5.3 mL O(2).kg(-1).min(-1)), with the greatest improvement observed in the SAR group (P = 0.05). Significant reductions in preexercise heart rates (8-9 bpm) and body fat percent (5--6%) were observed in all training groups after training. Significant reductions in resting diastolic blood pressure were observed for the SAR and SA40 groups (6.7 and 5.8 mm Hg, respectively). Muscular strength and endurance only improved significantly in the SAR group (21 and 11% respectively). All groups demonstrated increased lower body power (11--14%), but only the SAR group significantly improved upper body power (32%). Thigh muscle cross-sectional areas measured via magnetic resonance imaging (MRI) increased primarily for the SAR group. CONCLUSION: BSA is an exercise modality effective for improving physical fitness and body composition in healthy women. The addition of resistance exercise appears to enhance the total fitness profile by improving muscular performances, muscle morphology, and cardiovascular fitness greater than from performing BSA alone. Therefore, the inclusion of both modalities to an exercise program is most effective for improving total body fitness and a woman's health profile.