Prospective study of HDR brachytherapy (BT), external beam radiotherapy (EBRT) and androgen deprivation therapy (ADT): 10-years experience of an MRI-guided approach.

OBJECTIVES: To evaluate the predictive factors for biochemical failure and distant metastases in a prospective cohort of patients with localized prostate cancer treated with the combination of HDR BT and EBRT. METHODS AND MATERIALS: Patients with intermediate (IR) or high-risk (HR) prostate adenocarcinoma received a single fraction of HDR of 15 Gy combined with RT of 37.5 Gy in 15 fractions. ADT duration was used depending on risk-group. Descriptive analyses were performed. Univariate and multivariate Hazard Ratios were obtained. Finally, the Kaplan-Meier model was used to describe the survival of the events of interest. RESULTS: 309 patients were treated prospectively (199 were IR and 110 HR). Median age was 72 years; 58.3 % were MRI stage ≤ T2c, 34.1 % T3a and 7.6 % T3b; ISUP-grade 1-3 in 78.9 % and ISUP 4-5 in 21.1 %. 71.8 % of patients had ≤ 50 % positive-cores in biopsy and 28.2 % had > 50 %. Median pre-treatment PSA was 9.9 ng/mL. After a median follow-up of 88 months, 41 patients presented biochemical failure and 18 developed distant metastases. Multivariate cox-regression analyses found that MR-T3b Stage (HR 3.88, p = 0.001) and ADT use (HR 3.99, p = 0.03) were the only predictive factors for biochemical failure and the number of positive cores (>50 %) the only independent predictive factor of distant metastases (HR 4.36, p = 0.002). CONCLUSIONS: Patients with mpMRI evidence of invasion of the SV and involvement of more than 50% of the cores in the prostate biopsy are patients with a higher risk of presenting a biochemical recurrence or developing metastasis due to their prostate cancer, respectively.

Radiological Structured Report Integrated with Quantitative Imaging Biomarkers and Qualitative Scoring Systems.

The benefits of structured reporting (SR) in radiology are well-known and have been widely described. However, there are limitations that must be overcome. Radiologists may be reluctant to change the conventional way of reporting. Error rates could potentially increase if SR is used improperly. Interruption of the visual search pattern by keeping the eyes focused on the report rather than the images may increase reporting time. Templates that include unnecessary or irrelevant information may undermine the consistency of the report. Last, the lack of support for multiple languages may hamper the adaptation of the report to the target audience. This work aims to mitigate these limitations with a web-based structured reporting system based on templates. By including field validators and logical rules, the system avoids reporting mistakes and allows to automatically calculate values and radiological qualitative scores. The system can manage quantitative information from imaging biomarkers, combining this with qualitative radiological information usually present in the structured report. It manages SR templates as plugins (IHE MRRT compliant and compatible with RSNA's Radreport templates), ensures a seamless integration with PACS/RIS systems, and adapts the report to the target audience by means of natural language extracts generated in multiple languages. We describe a use case of SR template for prostate cancer including PI-RADS 2.1 scoring system and imaging biomarkers. For the time being, the system comprises 24 SR templates and provides service in 37 hospitals and healthcare institutions, endorsing the success of this contribution to mitigate some of the limitations of the SR.

Phenomenological modelling and simulation of cell clusters in 3D cultures.

Cell clustering and aggregation are fundamental processes in the development of several tissues and the progression of many diseases. The formation of these aggregates also has a direct impact on the oxygen concentration in their surroundings due to cellular respiration and poor oxygen diffusion through clusters. In this work, we propose a mathematical model that is capable of simulating cell cluster formation in 3D cultures through combining a particle-based and a finite element approach to recreate complex experimental conditions. Cells are modelled considering cell proliferation, cell death and cell-cell mechanical interactions. Additionally, the oxygen concentration profile is calculated through finite element analysis using a reaction-diffusion model that considers cell oxygen consumption and diffusion through the extracellular matrix and the cell clusters. In our model, the local oxygen concentration in the medium determines both cell proliferation and cell death. Numerical predictions are also compared with experimental data from the literature. The simulation results indicate that our model can predict cell clustering, cluster growth and oxygen distribution in 3D cultures. We conclude that the initial cell distribution, cell death and cell proliferation dynamics determine the size and density of clusters. Moreover, these phenomena are directly affected by the oxygen transport in the 3D culture.

A systematic review of image segmentation methodology, used in the additive manufacture of patient-specific 3D printed models of the cardiovascular system.

BACKGROUND: Shortcomings in existing methods of image segmentation preclude the widespread adoption of patient-specific 3D printing as a routine decision-making tool in the care of those with congenital heart disease. We sought to determine the range of cardiovascular segmentation methods and how long each of these methods takes. METHODS: A systematic review of literature was undertaken. Medical imaging modality, segmentation methods, segmentation time, segmentation descriptive quality (SDQ) and segmentation software were recorded. RESULTS: Totally 136 studies met the inclusion criteria (1 clinical trial; 80 journal articles; 55 conference, technical and case reports). The most frequently used image segmentation methods were brightness thresholding, region growing and manual editing, as supported by the most popular piece of proprietary software: Mimics (Materialise NV, Leuven, Belgium, 1992-2015). The use of bespoke software developed by individual authors was not uncommon. SDQ indicated that reporting of image segmentation methods was generally poor with only one in three accounts providing sufficient detail for their procedure to be reproduced. CONCLUSIONS AND IMPLICATION OF KEY FINDINGS: Predominantly anecdotal and case reporting precluded rigorous assessment of risk of bias and strength of evidence. This review finds a reliance on manual and semi-automated segmentation methods which demand a high level of expertise and a significant time commitment on the part of the operator. In light of the findings, we have made recommendations regarding reporting of 3D printing studies. We anticipate that these findings will encourage the development of advanced image segmentation methods.

Speech rate in Parkinson's disease: A controlled study. / Estudio controlado del ritmo del habla en la enfermedad de Parkinson.

INTRODUCTION: Speech disturbances will affect most patients with Parkinson's disease (PD) over the course of the disease. The origin and severity of these symptoms are of clinical and diagnostic interest. PURPOSE: To evaluate the clinical pattern of speech impairment in PD patients and identify significant differences in speech rate and articulation compared to control subjects. Speech rate and articulation in a reading task were measured using an automatic analytical method. PATIENTS: A total of 39 PD patients in the 'on' state and 45 age-and sex-matched asymptomatic controls participated in the study. None of the patients experienced dyskinesias or motor fluctuations during the test. RESULTS: The patients with PD displayed a significant reduction in speech and articulation rates; there were no significant correlations between the studied speech parameters and patient characteristics such as L-dopa dose, duration of the disorder, age, and UPDRS III scores and Hoehn & Yahr scales. CONCLUSION: Patients with PD show a characteristic pattern of declining speech rate. These results suggest that in PD, disfluencies are the result of the movement disorder affecting the physiology of speech production systems.

Resonancia magnética en cardiopatías congénitas. Aplicaciones clínicas y nuevos avances / Magnetic resonance imaging in congenital heart diseases. Clinincal utility and recent developments

La resonancia magnética cardíaca es una técnica diagnóstica emergente que ha irrumpido en el campo de las cardiopatías congénitas, permitiendo un salto cualitativo en la evaluación de la antomía y función cardiovascular. Es, además, una tecnica dinámica en constante desarrollo. Cada vez son más las nuevas secuencias y parámetros hemodinámicos que puede evaluar y que abren nuevas puertas a la investigación y, por tanto, al cnocimiento de la fisiopatología cardiovascular en el campo de las cardiopatias congénitas. En este artículo de revisión se evaluarán las aplicaciones clínicas más habituales y las nuevas líneas de investigación que se están desarrollando (AU)

Manetic Resonance Imaging (MRI) is an emerging tool in the field of congenital heart diseases. It has allowed a qualitative step forward in the evaluation of cardiovascular anatomy and physiology. MRI sequences and novel parameters to evaluate cardiovascular haemodynamics is increasing. It opens the door to new research and knowledge of the pathophysiological events in the field of congenital heart diseases. In this review, we summarize state-of-the-art in MRI and the most recent research areas (AU)

Generating anatomical models of the heart and the aorta from medical images for personalized physiological simulations.

The anatomy and motion of the heart and the aorta are essential for patient-specific simulations of cardiac electrophysiology, wall mechanics and hemodynamics. Within the European integrated project euHeart, algorithms have been developed that allow to efficiently generate patient-specific anatomical models from medical images from multiple imaging modalities. These models, for instance, account for myocardial deformation, cardiac wall motion, and patient-specific tissue information like myocardial scar location. Furthermore, integration of algorithms for anatomy extraction and physiological simulations has been brought forward. Physiological simulations are linked closer to anatomical models by encoding tissue properties, like the muscle fibers, into segmentation meshes. Biophysical constraints are also utilized in combination with image analysis to assess tissue properties. Both examples show directions of how physiological simulations could provide new challenges and stimuli for image analysis research in the future.

Amylin effect in extrapancreatic tissues participating in glucose homeostasis, in normal, insulin-resistant and type 2 diabetic state.

Amylin is co-secreted with insulin, responds to the same stimuli, is anorectic, lowers body weight by reducing fat mass, and is proposed for diabetes treatment. We examined the effect of a 3-day constant infusion of close to physiological doses of amylin in Wistar rats, on glucotransporter expression, glycogen content (G), glycogen synthase a activity (GSa) and glucose transport (GT), in liver, muscle and fat from insulin resistant (IR) and type 2 diabetic (T2D) models, compared to normal (N) animals; plasma glucose and insulin were measured. Plasma insulin in IR was higher than in N or T2D, and amylin normalized the value. In both, IR and T2D, liver G was lower than normal, accompanied by GLUT-2, mRNA and protein, higher and lower, respectively, than in N; amylin normalized G in both groups, without changes in GLUT-2, except for an mRNA increase in T2D. In IR and T2D, muscle GSa was reduced, together with respective over- and under-GLUT-4 expression; amylin induced only a trend toward GSa normalization in both groups. In isolated adipocytes, GT and GLUT-4 in IR and T2D were lower and higher, respectively, than in N; after amylin, not only GT was normalized in both groups but also the response to insulin was much more pronounced, including that in N, without major changes in GLUT-4. This suggests that the beneficial effect of amylin in states running with altered glucose homeostasis could occur by partially acting on the hexose metabolism of the liver and mainly on that of the adipose tissue.

Characteristics of palytoxin-induced cytotoxicity in neuroblastoma cells.

Cation fluxes appear to play a key role in palytoxin-induced signal. There are other cellular targets that have not been described as well as the biochemical signaling cascades that transmit palytoxin-stimulated signals remain to be clarified. Since modifications of cations, mainly calcium, are generally associated to cell death or apoptosis, we wanted to further evaluate the effect of palytoxin on cell death. Then, in vitro cytotoxic effects of palytoxin were characterized on human neuroblastoma cells. By using several techniques, we studied markers of cell death and apoptosis, such as cell detachment, mitochondrial membrane potential, caspases, DNA damage, LDH leakage, propidium iodide uptake, F-actin depolymerization and inhibition of cellular proliferation. Results show that palytoxin triggers a series of toxic responses; it inhibits cell proliferation, induces cell rounding, detachment from the substratum and F-actin disruption. Among the apoptotic markers studied we only detected fall in mitochondrial membrane potential. Neither caspases activation nor chromatin condensation or DNA fragmentation were observed in palytoxin-treated cells.

Induction and reversibility of insulin resistance in rats exposed to exogenous D-fructose.

Long-term exposure of normal rats to a fructose-enriched diet or drinking water is currently used as an animal model for experimental insulin resistance. The present study deals with a comparison between rats given access to either a fructose-enriched diet or fructose-enriched drinking water. In both situations, a decrease in food intake and body weight gain, and the induction of insulin resistance with intolerance to D-glucose despite increased secretory response to the aldohexose of insulin-producing cells were documented. Moreover, the rats exposed to exogenous D-fructose displayed a lesser sensitivity to overnight fasting than control animals, in terms of the alteration of glucose homeostasis and reduction of the ratio between plasma insulin and D-glucose concentration. It is also shown that the fructose-induced insulin resistance, as assessed in a hyperinsulinemic-euglycemic clamp, represents a phenomenon reversed within 15-30 days after removal of the keto-hexose from the drinking water.

In vitro approaches to evaluate palytoxin-induced toxicity and cell death in intestinal cells.

Palytoxin isolated from the genus Palythoa is the most potent marine toxin known. The aim of the present study was to quantify palytoxin-induced cellular injury in the human intestinal cell line Caco-2. Cellular damage was measured by evaluating cell proliferation, cell membrane permeability, cell morphology and apoptotic markers. Furthermore, changes in F-actin were studied after exposure of cells to increasing amounts of palytoxin. The results show that cell proliferation decreased in a concentration-dependent manner with a mean IC(50) value of about 0.1 nM. A noticeable increase of cell detachment correlated with cell rounding and F-actin depolymerization was observed in palytoxin-treated cells. Moreover LDH was released from the cells in a dose and time dependent manner, although under these conditions there was no propidium iodide uptake. On the other hand, palytoxin impaired mitochondrial activity but other apoptotic markers, such as DNA fragmentation or caspases activation, were not observed. The results obtained in this paper suggest that the effects of palytoxin in Caco-2 cells were very potent and unspecific, since a primary necrosis and a secondary apoptosis seem to occur under these conditions.

Glucose intolerance associated to insulin resistance and increased insulin secretion in rats depleted in long-chain omega3 fatty acids.

An intragastric D-glucose tolerance test was performed, after overnight starvation, in female rats depleted in long-chain polyunsaturated omega3 fatty acids (omega3D rats) and control rats of same age and gender. The plasma D-glucose and insulin concentrations, insulinogenic index, and HOMA for insulin resistance were all higher, after overnight starvation, in omega3D rats than in control animals. Over the 120-minute period following the intragastric administration of D-glucose, the area under the curve for the same four variables was also higher in omega3D rats than in control animals. In addition to visceral obesity, liver steatosis, hypertension, and cardiac hypertrophy, the omega3D rats thus display further features of the metabolic syndrome, namely glucose intolerance and insulin resistance, despite hyperinsulinemia.

Effects of diet supplementation with olive oil and guar upon fructose-induced insulin resistance in normal rats.

Exposure of normal rats to fructose-containing drinking water represents a current model of insulin resistance. The major aim of the present study was to assess the possible effect of diet supplementation with either olive oil or guar upon the metabolic consequences of exposure to exogenous fructose. For this purpose, the changes in body weight, plasma D-glucose and insulin concentrations, and D-glucose infusion rate during a hyperinsulinemic-euglycemic clamp were measured after 65 days exposure to exogenous fructose and either olive oil- or guar-enriched diet. The results were compared to those previously collected in control animals exposed for the same period to either tap water or the fructose-containing drinking water and a standard diet. Diet supplementation with olive oil or guar failed to affect the increase in the insulinogenic index and the decrease in insulin sensitivity and fasted/fed ratio for plasma insulin concentration caused by exogenous fructose. In the rats exposed to exogenous fructose, the olive oil-fed rats differed from other animals by the absence of a decrease in food intake and body weight gain, whilst the guar-fed rats differed from other animals in a lower plasma D-glucose concentration in fed state and an absence, at day 65, of a higher plasma D-glucose concentration than that at day 0 measured in after overnight fasting state. These findings argue in favour of guar, rather than olive oil, to oppose the effect of exogenous fructose on glucose homeostasis.

High-speed gel microelectrophoresis, a new and easy approach for detection of PCR-amplified microbial DNA from environmental and clinical samples in microgels using conventional equipment.

AIMS: Microelectrophoresis allows the detection of DNA bands using minimal amounts of sample in a short time, but commonly requires the use of special equipment which is not available in all laboratories. This fact has limited the application of this technique in microbiology despite its advantages. In this work, we describe a new approach to perform gel microelectrophoresis, named high-speed gel microelectrophoresis (HSGME), and its application for rapid detection of bacteria, protozoa and viruses in clinical, vegetal and environmental samples. METHODS AND RESULTS: Aliquots of 0.4-1 microl of PCR product were loaded in 2 cm 1% agarose microgels and electrophoresed at high voltage (125 V cm(-1)) in conventional submarine horizontal mini-slabs. By using HSGME, single-DNA bands obtained after specific-PCR useful in diagnosis of different diseases caused by micro-organisms were detected in 5 min. CONCLUSIONS: HSGME is a rapid and easy procedure applicable to detection of microbial genes, which is carried out using conventional equipment and thus can be performed in any research and diagnostic laboratory. SIGNIFICANCE AND IMPACT OF THE STUDY: The performance of HSGME saves up to 90% time, material and energy costs, as well as laboratory hazardous wastes including carcinogenic agents used for visualizing DNA bands.

Short-term and long-term effects of guar on postprandial plasma glucose, insulin and glucagon-like peptide 1 concentration in healthy rats.

Ingestion of guar gum decreases postprandial glycemia and insulinemia and improves sensitivity to insulin in diabetic patients and several animal models of diabetes. The aim of the present study was to compare the short-term and long-term effects of guar on plasma insulin and glucagon-like peptide 1 concentration in healthy rats. In the short-term experiments, the concomitant intragastric administration of glucose and guar reduced the early increment in plasma glucose, insulin and glucagon-like peptide 1 concentration otherwise induced by glucose alone. Comparable findings were made after twelve days of meal training exposing the rats to either a control or guar-enriched diet for fifteen minutes. Mean plasma glucose concentrations were lower while mean insulin concentrations were higher in the guar group than in the controls according to intragastric glucose tolerance tests conducted in overnight fasted rats maintained for 19 to 36 days on either the control or guar-enriched diet. The intestinal content of glucagon-like peptide 1 at the end of the experiments was also lower in the guar group. Changes in body weight over 62 days of observation were comparable in the control and guar rats. Thus, long-term intake of guar improves glucose tolerance and insulin response to glucose absorption, without improving insulin sensitivity, in healthy rats.

El GLP-1: acción biológica y posibles efectos terapéuticos / GLP-1: biological action and possible therapeutic effects

El GLP-1 (glucagon-like peptide-1) es una hormona con carácter de incretina que contribuye al control de la homeostasis de la glucosa, y que por su acción insulinotrópica, insulinotrófica y también insulinomimética, se está considerando para el tratamiento de la diabetes mellitus tipo 2. In vitro, el GLP-1 tiene efectos anabólicos sobre el metabolismo hepático de la glucosa en la rata normal y diabética, y sobre el del músculo y la grasa de la rata y del hombre; en el tejido adiposo, el GLP-1 es, además, lipolítico y lipogénico. En estos 3 tejidos extrapancreáticos, el GLP-1 parece actuar a través de receptores específicos, distintos en estructura y/o vía de señalización del pancreático, sobre los que se ha propuesto un inositolfosfoglicano como posible segundo mensajero. Por otro lado, la respuesta secretora de la célula ß al GLP-1, modulada en condiciones normales por la concentración extracelular de glucosa, está alterada en la diabetes tipo 2, debido, posiblemente, a la imposibilidad de la célula para reconocer a la hexosa; sin embargo, nutrientes no glucídicos, capaces de sortear los defectos específicos de reconocimiento de la célula ß diabética hacia la glucosa, como son los ésteres de ácidos dicarboxílicos, potencian y/o prolongan su acción insulinotrópica. Además, el GLP-1 no sólo ejerce un papel regulador de la ingestión de alimentos, induciendo sensación de saciedad, sino que también parece tener acciones beneficiosas en síndromes de carácter neurodegenerativo central y periférico. En cualquier caso, el valor del papel potencial terapéutico del GLP-1 no sólo en la diabetes sino también en la obesidad y en las enfermedades cardíacas y nerviosas, es un hecho que no se debe ignorar, cuyo mecanismo de acción concreto, no del todo conocido, requiere aclaración

Glucagon-like peptide-1 (GLP-1) is an incretin hormone that contributes to the control of glucose homeostasis. Because of its insulinotropic, insulinotrophic and insulinomimetic actions, it is being considered for the treatment of type 2 diabetes mellitus. In vitro, GLP-1 has anabolic effects on glucose metabolism in the liver of normal and diabetic rats and in rat and human skeletal muscle and fat; moreover, in fat tissue, GLP-1 is also lipolytic and lipogenic. In these three extrapancreatic tissues, GLP-1 seems to act through specific receptors, distinct in structure and/or signaling pathway from pancreatic one, for which an inositolphosphoglycan has been proposed as a possible second messenger. The secretory response of ß cells to GLP-1, modulated in normal conditions by the extracellular glucose concentration, is altered in type 2 diabetes mellitus, possibly due to the inability of the cell to recognize the hexose; however, non-glucose nutrients able to bypass the specific hexose recognition defects of the diabetic ß cell, such as dicarboxylic acid esters, potentiate and/or prolong the insulinotropic action of GLP-1. Moreover, GLP-1 not only plays a regulatory role in food intake, by inducing satiety, but also seems to have beneficial effects on central and peripheral neurodegenerative syndromes. The value of the potential therapeutic role of GLP-1, not only in diabetes but also in obesity and cardiac and central nervous system diseases, should not be ignored, and its mechanism of action, which is not clearly understood, requires elucidation

Effects of an olive oil-enriched diet on glucagon-like peptide 1 release and intestinal content, plasma insulin concentration, glucose tolerance and pancreatic insulin content in an animal model of type 2 diabetes.

In the light of a recent study conducted in normal rats, the present investigations were aimed at exploring the immediate and long-term effects of an olive oil-enriched diet (OO diet) on GLP-1 release and intestinal content, plasma insulin concentration, glucose tolerance and pancreatic insulin content in adult rats that had been injected with streptozotocin during the neonatal period (STZ rats). The OO diet, when compared to a standard diet, increased the immediate GLP-1 response in meal-trained rats, but decreased GLP-1 content in the intestinal tract after 50 days. Over 50 days, the body weight gain was lower in the rats fed the OO diet compared to standard diet. In the former, however, no improvement of glucose tolerance or insulin response during an oral glucose tolerance test was observed. Thus, a paradoxical lowering of the insulinogenic index, i. e. the paired ratio between plasma insulin and glucose concentration, was recorded during the oral glucose tolerance test in rats fed either standard or OO diet. Moreover, the insulin content of the pancreas was equally low in the STZ rats fed either standard or OO diet. These findings will be discussed in the framework of possible differences in the pathophysiology of B-cell dysfunction in most patients with type-2 diabetes and the present animal model of non-insulin-dependent diabetes.

Plasma malonaldehyde (MDA) and total antioxidant status (TAS) during lactation in dairy cows.

Metabolic profiles are widely used to monitor health, reproductive status and nutritional status. In the last few years, the evaluation of oxidative stress has contributed increasingly to our knowledge of the fundamental mechanisms involved in metabolic disorders, especially important in dairy cows, in which lactation imposes great physiological demands on the body's homeostatic mechanisms. The aim of the present study was to evaluate oxidative status in healthy cows during lactation (from lactation onset to peak lactation) using two parameters: (i) plasma levels of malondialdehyde (MDA) and (ii) total antioxidant status (TAS). Our results confirm that nutrition can influence the characteristic metabolic changes occurring between lactation onset and peak lactation. In addition, the combination of MDA and TAS can provide complementary information about the metabolic status of the cow. Thus, the proper metabolic adaptation to the onset of lactation, showed by metabolic profiles, contrasts with the high levels of free radicals which cause lipid peroxidation and high MDA values (68.99+/-33.64 microm/L) which is maintained only for a short period of time. In this moment, the antioxidant system can cope efficiently with lipoperoxide production. The most remarkable fact was the great inter-individual variations observed in MDA that might be studied in further investigations. When the animal reaches peak lactation, metabolic status is stabilized, and this is reflected by antioxidant status with mean values of 28.87+/-5.33 microm/L for MDA and 0.154+/-0.002 mmol/L for TAS values.

Participation of protein kinases in the stimulant action of GLP-1 on 2-deoxy-D-glucose uptake by normal rat skeletal muscle.

Several protein kinases were recently proposed for involvement in GLP-1-stimulated D-glucose transport in skeletal muscle from both normal subjects and type 2 diabetic patients. This study was mainly aimed at investigating the effect of potential inhibitors of distinct protein kinases and protein phosphatase-1 upon insulin- and GLP-1-stimulated 2-deoxy-D-glucose net uptake by normal rat skeletal muscle. The basal uptake of the D-glucose analog was decreased by wortmannin--a phosphatidylinositol-3-kinase inhibitor--, PD98059--a mitogen-activated protein kinases inhibitor--, and TNFalpha--a protein phosphatase-1 inhibitor--, but not by either rapamycin--a p70s6 kinase inhibitor--, or H-7--, a protein kinase C inhibitor--. The enhancing action of both insulin and GLP-1 upon 2-deoxy-D-glucose transport was abolished by PD98059 and H-7, but largely unaffected by TNFalpha. Wortmannin and rapamycin preferentially affected the response to GLP-1 and insulin, respectively. These findings thus document both analogies and dissimilarities in the participation of the concerned enzymes in the stimulant action of insulin versus GLP-1 upon D-glucose transport in normal rat skeletal muscle.

Cell signalling of glucagon-like peptide-1 action in rat skeletal muscle.

Glucagon-like peptide-1 (GLP-1), an incretin with glucose-dependent insulinotropic and insulin-independent antidiabetic properties, has insulin-like effects on glucose metabolism in extrapancreatic tissues participating in overall glucose homeostasis. These effects are exerted through specific receptors not associated with cAMP, an inositol phosphoglycan being a possible second messenger. In rat hepatocytes, activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB), protein kinase C (PKC) and protein phosphatase 1 (PP-1) has been shown to be involved in the GLP-1-induced stimulation of glycogen synthase. We have investigated the role of enzymes known or suggested to mediate the actions of insulin in the GLP-1-induced increase in glycogen synthase a activity in rat skeletal muscle strips. We first explored the effect of GLP-1, compared with that of insulin, on the activation of PI3K, PKB, p70s6 kinase (p70s6k) and p44/42 mitogen-activated protein kinases (MAPKs) and the action of specific inhibitors of these kinases on the insulin- and GLP-1-induced increment in glycogen synthase a activity. The study showed that GLP-1, like insulin, activated PI3K/PKB, p70s6k and p44/42. Wortmannin (a PI3K inhibitor) reduced the stimulatory action of insulin on glycogen synthase a activity and blocked that of GLP-1, rapamycin (a 70s6k inhibitor) did not affect the action of GLP-1 but abolished that of insulin, PD98059 (MAPK inhibitor) was ineffective on insulin but blocked the action of GLP-1, okadaic acid (a PP-2A inhibitor) and tumour necrosis factor-alpha (a PP-1 inhibitor) were both ineffective on GLP-1 but abolished the action of insulin, and Ro 31-8220 (an inhibitor of some PKC isoforms) reduced the effect of GLP-1 while completely preventing that of insulin. It was concluded that activation of PI3K/PKB and MAPKs is required for the GLP-1-induced increment in glycogen synthase a activity, while PKC, although apparently participating, does not seem to play an essential role; unlike in insulin signaling, p70s6k, PP-1 and PP-2A do not seem to be needed in the action of GLP-1 upon glycogen synthase a activity in rat muscle.