Temporal analysis of PI-3kinase and MAPK signal transduction during skeletal muscle overload

Skeletal muscles can adapt to increased mechanical forces (or loading) by increasing the size and strength of the muscle. Knowledge of the molecular mechanisms by which muscle responds to increased loading may lead to the discovery of novel treatment strategies for muscle wasting and frailty. The objective of this research was to examine the temporal associations between the activation of specific signaling pathway intermediates and their potential upstream regulator(s) in response to increased muscle loading. Previous work has demonstrated that focal adhesion kinase (FAK) activity is increased in overloaded hypertrophying skeletal muscle. Thus FAK is a candidate for transducing the loading stimulus in skeletal muscle, potentially by activating phosphatidylinositol 3-kinase (PI3K) and members of the mitogen-activated protein kinase (MAPK) family. However, it was unknown if muscle overload would result in activation of PI3K or the MAPKs. Thus, this work seeks to characterized the temporal response of (1) MAPK phosphorylation (including Erk 2, p38 MAPK and JNK), (2) PI3K activity, and (3) FAK tyrosine phosphorylation in response to 24 hours of compensatory overload in the rat soleus and plantaris muscles. In both muscles, overload resulted in transient Increases in the phosphorylation state of Erk2 and JNK, which peaked within the first hour of overload and returned to baseline thereafter. In contrast, p38 MAPK phosphorylation remained elevated throughout the entire 24-hour overload period. Moreover, overload increased PI3K activity only, in the plantaris and only at 12 hours. Moreover, 24 hours of overload induced a significant increase in total protein content in the plantaris but not the soleus. Thus an increase in total muscle protein content within the 24-hour loading period was observed only in muscle exhibiting increased PI3K activity. Surprisingly, FAK tyrosine phosphorylation was not increased during the overload period in either muscle, indicating that PI3K activation and increased MAPK phosphorylation were independent of increased FAK tyrosine phosphorylation. In summary, increased PI3K activity and sustained elevation of p38 MAPK phosphorylation were associated with muscle overload, identifying these pathways as potential mediators of the early hypertrophic response to skeletal muscle overload. This suggests that stimuli or mechanisms that activate these pathways may reduce/minimize muscle wasting and frailty. ^

Respiratory rhythm generation: A modeling study of the respiratory central pattern generator and the phrenic motor neuron

The respiratory central pattern generator is a collection of medullary neurons that generates the rhythm of respiration. The respiratory central pattern generator feeds phrenic motor neurons, which, in turn, drive the main muscle of respiration, the diaphragm. The purpose of this thesis is to understand the neural control of respiration through mathematical models of the respiratory central pattern generator and phrenic motor neurons. ^ We first designed and validated a Hodgkin-Huxley type model that mimics the behavior of phrenic motor neurons under a wide range of electrical and pharmacological perturbations. This model was constrained physiological data from the literature. Next, we designed and validated a model of the respiratory central pattern generator by connecting four Hodgkin-Huxley type models of medullary respiratory neurons in a mutually inhibitory network. This network was in turn driven by a simple model of an endogenously bursting neuron, which acted as the pacemaker for the respiratory central pattern generator. Finally, the respiratory central pattern generator and phrenic motor neuron models were connected and their interactions studied. ^ Our study of the models has provided a number of insights into the behavior of the respiratory central pattern generator and phrenic motor neurons. These include the suggestion of a role for the T-type and N-type calcium channels during single spikes and repetitive firing in phrenic motor neurons, as well as a better understanding of network properties underlying respiratory rhythm generation. We also utilized an existing model of lung mechanics to study the interactions between the respiratory central pattern generator and ventilation. ^

Effect of endurance, speed and strength training on skin temperature measured by infrared thermography = Efecto del entrenamiento de resistencia, velocidad y fuerza en la temperatura de la piel a través de la termografía infrarroja

La termografía infrarroja (TI) es una técnica no invasiva y de bajo coste que permite, con el simple acto de tomar una fotografía, el registro sin contacto de la energía que irradia el cuerpo humano (Akimov & Son’kin, 2011, Merla et al., 2005, Ng et al., 2009, Costello et al., 2012, Hildebrandt et al., 2010). Esta técnica comenzó a utilizarse en el ámbito médico en los años 60, pero debido a los malos resultados como herramienta diagnóstica y la falta de protocolos estandarizados (Head & Elliot, 2002), ésta se dejó de utilizar en detrimento de otras técnicas más precisas a nivel diagnóstico. No obstante, las mejoras tecnológicas de la TI en los últimos años han hecho posible un resurgimiento de la misma (Jiang et al., 2005, Vainer et al., 2005, Cheng et al., 2009, Spalding et al., 2011, Skala et al., 2012), abriendo el camino a nuevas aplicaciones no sólo centradas en el uso diagnóstico. Entre las nuevas aplicaciones, destacamos las que se desarrollan en el ámbito de la actividad física y el deporte, donde recientemente se ha demostrado que los nuevos avances con imágenes de alta resolución pueden proporcionar información muy interesante sobre el complejo sistema de termorregulación humana (Hildebrandt et al., 2010). Entre las nuevas aplicaciones destacan: la cuantificación de la asimilación de la carga de trabajo físico (Čoh & Širok, 2007), la valoración de la condición física (Chudecka et al., 2010, 2012, Akimov et al., 2009, 2011, Merla et al., 2010), la prevención y seguimiento de lesiones (Hildebrandt et al., 2010, 2012, Badža et al., 2012, Gómez Carmona, 2012) e incluso la detección de agujetas (Al-Nakhli et al., 2012). Bajo estas circunstancias, se acusa cada vez más la necesidad de ampliar el conocimiento sobre los factores que influyen en la aplicación de la TI en los seres humanos, así como la descripción de la respuesta de la temperatura de la piel (TP) en condiciones normales, y bajo la influencia de los diferentes tipos de ejercicio. Por consiguiente, este estudio presenta en una primera parte una revisión bibliográfica sobre los factores que afectan al uso de la TI en los seres humanos y una propuesta de clasificación de los mismos. Hemos analizado la fiabilidad del software Termotracker, así como su reproducibilidad de la temperatura de la piel en sujetos jóvenes, sanos y con normopeso. Finalmente, se analizó la respuesta térmica de la piel antes de un entrenamiento de resistencia, velocidad y fuerza, inmediatamente después y durante un período de recuperación de 8 horas. En cuanto a la revisión bibliográfica, hemos propuesto una clasificación para organizar los factores en tres grupos principales: los factores ambientales, individuales y técnicos. El análisis y descripción de estas influencias deben representar la base de nuevas investigaciones con el fin de utilizar la TI en las mejores condiciones. En cuanto a la reproducibilidad, los resultados mostraron valores excelentes para imágenes consecutivas, aunque la reproducibilidad de la TP disminuyó ligeramente con imágenes separadas por 24 horas, sobre todo en las zonas con valores más fríos (es decir, zonas distales y articulaciones). Las asimetrías térmicas (que normalmente se utilizan para seguir la evolución de zonas sobrecargadas o lesionadas) también mostraron excelentes resultados pero, en este caso, con mejores valores para las articulaciones y el zonas centrales (es decir, rodillas, tobillos, dorsales y pectorales) que las Zonas de Interés (ZDI) con valores medios más calientes (como los muslos e isquiotibiales). Los resultados de fiabilidad del software Termotracker fueron excelentes en todas las condiciones y parámetros. En el caso del estudio sobre los efectos de los entrenamientos de la velocidad resistencia y fuerza en la TP, los resultados muestran respuestas específicas según el tipo de entrenamiento, zona de interés, el momento de la evaluación y la función de las zonas analizadas. Los resultados mostraron que la mayoría de las ZDI musculares se mantuvieron significativamente más calientes 8 horas después del entrenamiento, lo que indica que el efecto del ejercicio sobre la TP perdura por lo menos 8 horas en la mayoría de zonas analizadas. La TI podría ser útil para cuantificar la asimilación y recuperación física después de una carga física de trabajo. Estos resultados podrían ser muy útiles para entender mejor el complejo sistema de termorregulación humano, y por lo tanto, para utilizar la TI de una manera más objetiva, precisa y profesional con visos a mejorar las nuevas aplicaciones termográficas en el sector de la actividad física y el deporte Infrared Thermography (IRT) is a safe, non-invasive and low-cost technique that allows the rapid and non-contact recording of the irradiated energy released from the body (Akimov & Son’kin, 2011; Merla et al., 2005; Ng et al., 2009; Costello et al., 2012; Hildebrandt et al., 2010). It has been used since the early 1960’s, but due to poor results as diagnostic tool and a lack of methodological standards and quality assurance (Head et al., 2002), it was rejected from the medical field. Nevertheless, the technological improvements of IRT in the last years have made possible a resurgence of this technique (Jiang et al., 2005; Vainer et al., 2005; Cheng et al., 2009; Spalding et al., 2011; Skala et al., 2012), paving the way to new applications not only focused on the diagnose usages. Among the new applications, we highlighted those in physical activity and sport fields, where it has been recently proven that a high resolution thermal images can provide us with interesting information about the complex thermoregulation system of the body (Hildebrandt et al., 2010), information than can be used as: training workload quantification (Čoh & Širok, 2007), fitness and performance conditions (Chudecka et al., 2010, 2012; Akimov et al., 2009, 2011; Merla et al., 2010; Arfaoui et al., 2012), prevention and monitoring of injuries (Hildebrandt et al., 2010, 2012; Badža et al., 2012, Gómez Carmona, 2012) and even detection of Delayed Onset Muscle Soreness – DOMS- (Al-Nakhli et al., 2012). Under this context, there is a relevant necessity to broaden the knowledge about factors influencing the application of IRT on humans, and to better explore and describe the thermal response of Skin Temperature (Tsk) in normal conditions, and under the influence of different types of exercise. Consequently, this study presents a literature review about factors affecting the application of IRT on human beings and a classification proposal about them. We analysed the reliability of the software Termotracker®, and also its reproducibility of Tsk on young, healthy and normal weight subjects. Finally, we examined the Tsk thermal response before an endurance, speed and strength training, immediately after and during an 8-hour recovery period. Concerning the literature review, we proposed a classification to organise the factors into three main groups: environmental, individual and technical factors. Thus, better exploring and describing these influence factors should represent the basis of further investigations in order to use IRT in the best and optimal conditions to improve its accuracy and results. Regarding the reproducibility results, the outcomes showed excellent values for consecutive images, but the reproducibility of Tsk slightly decreased with time, above all in the colder Regions of Interest (ROI) (i.e. distal and joint areas). The side-to-side differences (ΔT) (normally used to follow the evolution of some injured or overloaded ROI) also showed highly accurate results, but in this case with better values for joints and central ROI (i.e. Knee, Ankles, Dorsal and Pectoral) than the hottest muscle ROI (as Thigh or Hamstrings). The reliability results of the IRT software Termotracker® were excellent in all conditions and parameters. In the part of the study about the effects on Tsk of aerobic, speed and strength training, the results of Tsk demonstrated specific responses depending on the type of training, ROI, moment of the assessment and the function of the considered ROI. The results showed that most of muscular ROI maintained warmer significant Tsk 8 hours after the training, indicating that the effect of exercise on Tsk last at least 8 hours in most of ROI, as well as IRT could help to quantify the recovery status of the athlete as workload assimilation indicator. Those results could be very useful to better understand the complex skin thermoregulation behaviour, and therefore, to use IRT in a more objective, accurate and professional way to improve the new IRT applications for the physical activity and sport sector.

Viral mediated expression of insulin-like growth factor I blocks the aging-related loss of skeletal muscle function

During the aging process, mammals lose up to a third of their skeletal muscle mass and strength. Although the mechanisms underlying this loss are not entirely understood, we attempted to moderate the loss by increasing the regenerative capacity of muscle. This involved the injection of a recombinant adeno-associated virus directing overexpression of insulin-like growth factor I (IGF-I) in differentiated muscle fibers. We demonstrate that the IGF-I expression promotes an average increase of 15% in muscle mass and a 14% increase in strength in young adult mice, and remarkably, prevents aging-related muscle changes in old adult mice, resulting in a 27% increase in strength as compared with uninjected old muscles. Muscle mass and fiber type distributions were maintained at levels similar to those in young adults. We propose that these effects are primarily due to stimulation of muscle regeneration via the activation of satellite cells by IGF-I. This supports the hypothesis that the primary cause of aging-related impairment of muscle function is a cumulative failure to repair damage sustained during muscle utilization. Our results suggest that gene transfer of IGF-I into muscle could form the basis of a human gene therapy for preventing the loss of muscle function associated with aging and may be of benefit in diseases where the rate of damage to skeletal muscle is accelerated.

Early alterations in serum creatine kinase and total cholesterol following high intensity eccentric muscle actions

Aim. The purpose of this experiment was to assess the levels of muscle soreness, serum total cholesterol (TC) and creatine kinase (CK) in the first 48 hours following fatiguing eccentric exercise performed with the triceps brachii. Methods. Eleven untrained male college students performed a total of 50 eccentric elbow extensions in 8 sets (6x7 and 2x4) with a load equal to 85% of their maximal concentric elbow extension strength. Isometric elbow extension strength, muscle soreness and circumference, and serum CK and TC concentrations were measured before, immediately after, and 2, 24 and 48 hours after the exercise. Results. Statistically reliable changes in isometric strength, serum CK and TC, muscle soreness and upper arm circumference occurred within the first 48 hours following eccentric exercise. Serum TC concentrations exhibited a very rapid (within 2 hours) reduction from pre-exercise values after eccentric exercise to a relatively stable concentration of approximately 85% of baseline. Conclusion. These results suggest that serum TC concentration may follow the time-course of reductions in force generating capacity more closely than other biochemical markers of muscle damage.

Differential expression of muscle damage in humans following acute fast and slow velocity eccentric exercise

We sought to determine if the velocity of an acute bout of eccentric contractions influenced the duration and severity of several common indirect markers of muscle damage. Subjects performed 36 maximal fast (FST, n=8: 3.14 rad center dot s(-1)) or slow (SLW, n=7: 0.52 rad center dot s(-1)) velocity isokinetic eccentric contractions with the elbow flexors of the non-dominant arm. Muscle soreness, limb girth, plasma creatine kinase (CK) activity, isometric torque and concentric and eccentric torque at 0.52 and 3.14 rad center dot s(-1) were assessed prior to and for several days following the eccentric bout. Peak plasma CK activity was similar in SLW (4030 +/- 1029 U center dot l(-1)) and FST (5864 +/- 2664 U center dot l(-1)) groups, (p > 0.05). Both groups experienced similar decrement in all strength variables during the 48 hr following the eccentric bout. However, recovery occurred more rapidly in the FST group during eccentric (0.52 and 3.14 rad center dot s(-1)) and concentric (3.14 rad center dot s(-1)) post-testing. The severity of muscle soreness was similar in both groups. However, the FST group experienced peak muscle soreness 48 hr later than the SLW group (24 hr vs. 72 hr). The SLW group experienced a greater increase in upper arm girth than the FST group 20 min, 24 hr and 96 hr following the eccentric exercise bout. The contraction velocity of an acute bout of eccentric exercise differentially influences the magnitude and time course of several indirect markers of muscle damage.

Age-related differences in rapid muscle activation after rate of force development training of the elbow flexors

In young adults, improvements in the rate of force development as a result of resistance training are accompanied by increases in neural drive in the very initial phase of muscle activation. The purpose of this experiment was to determine if older adults also exhibit similar adaptations in response to rate of force development (RFD) training. Eight young (21-35 years) and eight older (60-79 years) adults were assessed during the production of maximum rapid contractions, before and after four weeks of progressive resistance training for the elbow flexors. Young and older adults exhibited significant increases (P< 0.01) in peak RFD, of 25.6% and 28.6% respectively. For both groups the increase in RFD was accompanied by an increase in the root mean square (RMS) amplitude and in the rate of rise (RER) in the electromyogram (EMG) throughout the initial 100 ms of activation. For older adults, however, this training response was only apparent in the brachialis and brachioradialis muscles. This response was not observed in surface EMG recorded from the biceps brachii muscle during either RFD testing or throughout training, nor was it observed in the pronator teres muscle. The minimal adaptations observed for older adults in the bifunctional muscles biceps brachii and pronator teres are considered to indicate a compromise of the neural adaptations older adults might experience in response to resistance training.

Glycinergic and GABAergic synaptic activity differentially regulate motoneuron survival and skeletal muscle innervation

GABAergic and glycinergic synaptic transmission is proposed to promote the maturation and refinement of the developing CNS. Here we provide morphological and functional evidence that glycinergic and GABAergic synapses control motoneuron development in a region-specific manner during programmed cell death. In gephyrin-deficient mice that lack all postsynaptic glycine receptor and some GABA(A) receptor clusters, there was increased spontaneous respiratory motor activity, reduced respiratory motoneuron survival, and decreased innervation of the diaphragm. In contrast, limb-innervating motoneurons showed decreased spontaneous activity, increased survival, and increased innervation of their target muscles. Both GABA and glycine increased limb-innervating motoneuron activity and decreased respiratory motoneuron activity in wild-type mice, but only glycine responses were abolished in gephyrin-deficient mice. Our results provide genetic evidence that the development of glycinergic and GABAergic synaptic inputs onto motoneurons plays an important role in the survival, axonal branching, and spontaneous activity of motoneurons in developing mammalian embryos.

Strength and endurance training lead to different post exercise glucose profiles in diabetic participants using a continuous subcutaneous glucose monitoring system

Background Although both strength training (ST) and endurance training (ET) seem to be beneficial in type 2 diabetes mellitus (T2D), little is known about post-exercise glucose profiles. The objective of the study was to report changes in blood glucose (BG) values after a 4-month ET and ST programme now that a device for continuous glucose monitoring has become available. Materials and methods Fifteen participants, comprising four men age 56.5 +/- 0.9 years and 11 women age 57.4 +/- 0.9 years with T2D, were monitored with the MiniMed (Northridge, CA, USA) continuous glucose monitoring system (CGMS) for 48 h before and after 4 months of ET or ST. The ST consisted of three sets at the beginning, increasing to six sets per week at the end of the training period, including all major muscle groups and ET performed with an intensity of maximal oxygen uptake of 60% and a volume beginning at 15 min and advancing to a maximum of 30 min three times a week. Results A total of 17 549 single BG measurements pretraining (619.7 +/- 39.8) and post-training (550.3 +/- 30.1) were recorded, correlating to an average of 585 +/- 25.3 potential measurements per participant at the beginning and at the end of the study. The change in BG-value between the beginning (132 mg dL(-1)) and the end (118 mg dL(-1)) for all participants was significant (P = 0.028). The improvement in BG-value for the ST programme was significant (P = 0.02) but for the ET no significant change was measured (P = 0.48). Glycaemic control improved in the ST group and the mean BG was reduced by 15.6% (Cl 3-25%). Conclusion In conclusion, the CGMS may be a useful tool in monitoring improvements in glycaemic control after different exercise programmes. Additionally, the CGMS may help to identify asymptomatic hypoglycaemia or hyperglycaemia after training programmes.

Sitting posture affects pelvic floor muscle activity in parous women: An observational study

Question Do different sitting postures require different levels of pelvic floor and abdominal muscle activity in healthy women? Design Observational study. Participants Eight parous women with no pelvic floor dysfunction. Outcome measures Bilateral activity of pelvic floor muscles (assessed vaginally) and two abdominal muscles, obliquus internus abdominis and obliquus externus abdominis, during three sitting postures. Results There was a significant increase in pelvic floor muscle activity from slump supported sitting (mean 7.2% maximal voluntary contraction, SD 4.8) to both upright unsupported sifting (mean 12.6% maximal voluntary contraction, SD 7.8) (p = 0.01) and very tall unsupported sitting (mean 24.3% maximal voluntary contraction, SD 14.2) (p = 0.004). Activity in both abdominal muscles also increased but did not reach statistical significance. Conclusion Both unsupported sitting postures require greater pelvic floor muscle activity than the supported sitting posture.

Attachment of Shiga toxigenic Escherichia coli to beef muscle and adipose tissue

Shiga toxigenic Escherichia coli (STEC) serotypes are important foodborne pathogens that cause gastrointestinal disease worldwide. An understanding of how STEC strains attach to surfaces may provide insight into the potential persistence of and contamination with STEC in food environments. The initial attachment of a selection of STEC serotypes to beef muscle and adipose tissue was evaluated for isolates grown in planktonic and sessile culture. Initial experiments were performed to determine whether attachment differed among STEC strains and between the two modes of growth. Viable counts were obtained for loosely and strongly attached cells, and the strength of attachment (S-r) was calculated. All bacterial isolates grown in sessile culture attached in higher numbers to muscle and adipose tissue than did bacteria in planktonic cultures. For all attachment assays performed, mean concentrations for loosely attached cells were consistently higher than concentrations for strongly attached cells. The mean concentrations for strongly attached bacteria for planktonic and sessile cultures were significantly higher (P < 0.05) on adipose than on muscle tissue. However, some strains of STEC, particularly those from sessile culture, did not differ in their attachment to muscle or adipose tissue. S-r values were not significantly different (P > 0.05) among STEC isolates for all assays. No correlation was found between bacterial hydrophobicity and surface charge values (previously determined) and production of surface structures, viable counts, and S-r values. STEC grown in planktonic and sessile culture seems to behave differently with respect to attachment to muscle and adipose tissue. Cells in sessile culture may have a greater potential to strongly attach to meat surfaces.

Involuntary glottal closure during inspiration in muscle tension dysphonia

Objective/Hypothesis: The purpose of this study was to examine respiratory function in a group of patients with muscle tension dysphonia (MTD) Design: Cross-sectional analytical study. Methods: Participants included 15 people with a diagnosis of MTD referred to speech pathology for management of their voice disorder, fiberoptic evidence of glottal or supraglottic constriction during phonation with or without posterior chink, or bowing combined and deviation in perceptual voice quality. A second group of 15 participants with no history of voice disorder served as healthy controls,. Baseline pulmonary function test measures included forced expiratory volume in the first second (FEV1), FVC, FEF25 to 75, FIF50, FEV1/FVC, ratio and FEF50/FIF50 ratio. Hypertonic saline challenge test measures included FEV1 and FIF50 after provocation, close response slope, and provocation dose. Results: Compared with healthy controls, participants with MTD demonstrated a higher prevalence of glottal constriction during inspiration after provocation with nebulized hypertonic saline as demonstrated by a reduction in FIF50 after the hypertonic saline challenge. There was no significant difference between the MTD and healthy control groups in baseline pulmonary function testing. Participants with MTD demonstrated a higher prevalence than healthy controls of abnormal glottic closure during inspiration similar to paradoxical vocal fold movement (PVFM). This suggests that they either had previously undiagnosed coexisting PVFM or that the condition of MTD could be expanded to include descriptions of aberrant glottic function during respiration. This study enhances the understanding of PVFM and MTD by combining research advances made in the fields of otolaryngology and respiratory medicine.

Loss of skeletal muscle in cancer: biochemical mechanisms

Patients with cancer often undergo a specific loss of skeletal muscle mass, while the visceral protein reserves are preserved. This condition known as cachexia reduces the quality of life and eventually results in death through erosion of the respiratory muscles. Nutritional supplementation or appetite stimulants are unable to restore the loss of lean body mass, since protein catabolism is increased mainly as a result of the activation of the ATP-ubiquitin-dependent proteolytic pathway. Several mediators have been proposed. An enhanced protein degradation is seen in skeletal muscle of mice administered tumour necrosis factor (TNF), which appears to be mediated by oxidative stress. There is some evidence that this may be a direct effect and is associated with an increase in total cellular-ubiquitin-conjugated muscle proteins. Another cytokine, interleukin-6 (IL-6), may play a role in muscle wasting in certain animal tumours, possibly through both lysosomal (cathepsin) and non-lysosomal (proteasome) pathways. A tumour product, proteolysis-inducing factor (PIF) is produced by cachexia-inducing murine and human tumours and initiates muscle protein degradation directly through activation of the proteasome pathway. The action of PIF is blocked by eicosapentaenoic acid (EPA), which has been shown to attenuate the development of cachexia in pancreatic cancer patients. When combined with nutritional supplementation EPA leads to accumulation of lean body mass and prolongs survival. Further knowledge on the biochemical mechanisms of muscle protein catabolism will aid the development of effective therapy for cachexia.