Individual Muscle use in Hamstring Exercises by Soccer Players Assessed using Functional MRI.

This study used functional magnetic resonance imaging (fMRI) to compare individual muscle use in exercises aimed at preventing hamstring injuries. Thirty-six professional soccer players were randomized into 4 groups, each performing either Nordic hamstring, flywheel leg curl, Russian belt or conic-pulley exercise. MRIs were performed before and immediately after a bout of 4 sets of 8 repetitions. Pre-post exercise differences in contrast shift (T2) were analyzed for the long (BFLh) and short head (BFSh) of biceps femoris, semitendinosus (ST), semimembranosus (SM) and gracilis (GR) muscles. Flywheel leg curl increased (P<0.001) T2 of GR (95%), ST (65%), BFSh (51%) and BFLh (14%). After the Nordic hamstring, GR (39%), ST (16%) and BFSh (14%) showed increased T2 (P<0.001). Russian belt and conic-pulley exercise produced subtle (P<0.02) T2 increases of ST (9 and 6%, respectively) and BFLh (7 and 6%, respectively). Russian belt increased T2 of SM (7%). Among exercises examined, flywheel leg curl showed the most substantial hamstring and GR muscle use. However, no single exercise executed was able to increase T2 of all hamstring and synergist muscles analyzed. It is therefore suggested that multiple exercises must be carried out to bring in, and fully activate all knee flexors and hip extensors.

A New Surgical Model of Skeletal Muscle Injuries in Rats Reproduces Human Sports Lesions.

Skeletal muscle injuries are the most common sports-related injuries in sports medicine. In this work, we have generated a new surgically-induced skeletal muscle injury in rats, by using a biopsy needle, which could be easily reproduced and highly mimics skeletal muscle lesions detected in human athletes. By means of histology, immunofluorescence and MRI imaging, we corroborated that our model reproduced the necrosis, inflammation and regeneration processes observed in dystrophic mdx-mice, a model of spontaneous muscle injury, and realistically mimicked the muscle lesions observed in professional athletes. Surgically-injured rat skeletal muscles demonstrated the longitudinal process of muscle regeneration and fibrogenesis as stated by Myosin Heavy Chain developmental (MHCd) and collagen-I protein expression. MRI imaging analysis demonstrated that our muscle injury model reproduces the grade I-II type lesions detected in professional soccer players, including edema around the central tendon and the typically high signal feather shape along muscle fibers. A significant reduction of 30% in maximum tetanus force was also registered after 2 weeks of muscle injury. This new model represents an excellent approach to the study of the mechanisms of muscle injury and repair, and could open new avenues for developing innovative therapeutic approaches to skeletal muscle regeneration in sports medicine.

Facial Video-Based Photoplethysmography to Detect HRV at Rest.

Our aim is to demonstrate the usefulness of photoplethysmography (PPG) for analyzing heart rate variability (HRV) using a standard 5-min test at rest with paced breathing, comparing the results with real RR intervals and testing supine and sitting positions. Simultaneous recordings of R-R intervals were conducted with a Polar system and a non-contact PPG, based on facial video recording on 20 individuals. Data analysis and editing were performed with individually designated software for each instrument. Agreement on HRV parameters was assessed with concordance correlations, effect size from ANOVA and Bland and Altman plots. For supine position, differences between video and Polar systems showed a small effect size in most HRV parameters. For sitting position, these differences showed a moderate effect size in most HRV parameters. A new procedure, based on the pixels that contained more heart beat information, is proposed for improving the signal-to-noise ratio in the PPG video signal. Results were acceptable in both positions but better in the supine position. Our approach could be relevant for applications that require monitoring of stress or cardio-respiratory health, such as effort/recuperation states in sports.

Ordered transcriptional factor recruitment and epigenetic regulation of tnf-alpha in necrotizing acute pancreatitis.

Tauhe expression of the critical initiator cytokine TNF-alpha was strongly upregulated in vivo in acute necrotic pancreatitis (AP) in rodents and in vitro in TNF-alpha activated acinar AR42J cells. Upregulation of tnf-alpha, inos, icam-1 and il-6 occurred both in TNF-alpha receptor 1 and 2 knock-out mice, but not in TNF-alpha knock-out mice, in cerulein-induced acute pancreatitis. Chromatin immunoprecipitation analysis showed that transcriptional factors (ELK-1, SP1, NF-kappaB and EGR-1) and chromatin modification complexes (HDAC1, HDAC2, GCN5, PCAF and CBP) were recruited and/or released from the promoter in a strictly ordered mechanism. Activation of tnf-alpha gene was also accompanied by an ordered increased level of histone H3K9, H3K14 and H3K18-acetylation and H3K4 methylation, as well as H4K5 acetylation. A better knowledge of the molecular mechanisms that control tnf-alpha gene regulation will provide deeper understanding of the initiation and development of the inflammatory processes occurring in acute pancreatitis triggered by TNF-alpha cytokine.

Strength and endurance in elite football players.

We aimed to improve the physical capacity of a top-level elite football team during its pre-season by implementing a maximal strength and high-intensity endurance training program. 21 first league elite football players (20-31 yrs, height 171-194 cm, mass 58.8-88.1 kg) having recently participated in the UEFA Champions' League, took part in the study. Aerobic interval-training at 90-95% of maximal heart rate and half-squats strength training with maximum loads in 4 repetitions × 4 sets were performed concurrently twice a week for 8 weeks. The players were not familiar with maximal strength training as part of their regular program. Maximal oxygen uptake (VO(2max)) increased 8.6% (1.7-16.6) (p<0.001), from 60.5 (51.7-67.1) to 65.7 (58.0-74.5) mL · kg (-1) · min (-1) whereas half-squat one repetition maximum increased 51.7% (13.3-135.3) (p<0.001), from 116 (85-150) to 176 (160-210) kg. The 10-m sprint time also improved by 0.06 s (0.02-0.16) (p<0.001); while counter movement jump improved 3.0 cm (0.1-6.2) (p<0.001), following the training program. The concurrent strength and endurance training program together with regular football training resulted in considerable improvement of the players' physical capacity and so may be successfully introduced to elite football players.

PAP1 signaling involves MAPK signal transduction.

Pancreatitis-associated protein 1 (PAP1) belongs to the Reg family of secretory proteins. Several important biological roles have been attributed to PAP1 but the signaling pathways activated by this protein remain only partially understood. Here, we describe the intracellular pathways triggered by PAP1 in a pancreatic acinar cell line. Taking advantage of the fact that PAP1 induces its own transcription, we performed ChIP assays to analyze the recruitment of transcriptional factors on its promoter. Our results show that PAP1 increased the transactivation activity of pap1 and the binding on its promoter of the nuclear factors C/EBPbeta, P-CREB, P-ELK1, EGR1, STAT3, and ETS2, which are downstream targets of MAPK signaling. p44/42, p38, and JNK MAPKs activity increased after PAP1 treatment. In addition, pharmacological inhibition of these kinases markedly inhibited the induction of pap1 mRNA. Taken together, these results indicated that the mechanism of PAP1 action involves the activation of the MAPK superfamily.

Comparison of Omega Wave System and Polar S810i to detect R-R intervals at rest.

The present study was performed to compare R-R interval data and heart rate variability indices obtained from the Polar S810i and the Omega Wave Sport System for a total of 96 adults in a supine position. Data were simultaneously recorded with the Polar S810i and the Omega Wave Sport System and processed by unique software. Bland-Altman analysis for the R-R intervals shows minimal bias for free and paced breathing. No significant differences were observed for heart rate variability indices derived from the signal from both devices, except for the power of the high frequency band and the acceleration changes index during either free or paced breathing. Coefficients of correlations were all above 0.96. These data suggest that both systems are valid to record R-R interval signals and to obtain a valid analysis of heart rate variability. However, Omega Wave Sport System enables data to be collected without any artifacts making the analysis of heart rate variability easier than the analysis of Polar S810i. Nevertheless, Polar S810i continues to be more practical in clinical and applied situations due to the affordability of the device.

EPDR1 up-regulation in human colorectal cancer is related to staging and favours cell proliferation and invasiveness.

The finding of novel molecular markers for prediction or prognosis of invasiveness in colorectal cancer (CRC) constitutes an appealing challenge. Here we show the up-regulation of EPDR1 in a prospective cohort of 101 CRC patients, in a cDNA array of 43 patients and in in silico analyses. EPDR1 encodes a protein related to ependymins, a family of glycoproteins involved in intercellular contacts. A thorough statistical model allowed us to conclude that the gene is significantly up-regulated in tumour tissues when compared with normal mucosa. These results agree with those obtained by the analysis of three publicly available databases. EPDR1 up-regulation correlates with the TNM staging parameters, especially T and M. Studies with CRC cell lines revealed that the methylation of a CpG island controls EPDR1 expression. siRNA knocking-down and overexpression of the gene following transient plasmid transfection, showed that EPDR1 favours cell proliferation, migration, invasiveness and adhesion to type I collagen fibres, suggesting a role in epithelial to mesenchymal transition. Both statistical and functional analysis correlated EPDR1 overexpression with invasiveness and dissemination of tumour cells, supporting the inclusion of EPDR1 in panels of genes used to improve molecular subtyping of CRC. Eventually, EPDR1 may be an actionable target.

Fast and slow myosins as markers of muscle injury.

OBJECTIVE: The diagnosis of muscular lesions suffered by athletes is usually made by clinical criteria combined with imaging of the lesion (ultrasonography and/or magnetic resonance) and blood tests to detect the presence of non-specific muscle markers. This study was undertaken to evaluate injury to fast and slow-twitch fibres using specific muscle markers for these fibres. METHODS: Blood samples were obtained from 51 non-sports people and 38 sportsmen with skeletal muscle injury. Western blood analysis was performed to determine fast and slow myosin and creatine kinase (CK) levels. Skeletal muscle damage was diagnosed by physical examination, ultrasonography and magnetic resonance and biochemical markers. RESULTS: The imaging tests were found to be excellent for detecting and confirming grade II and III lesions. However, grade I lesions were often unconfirmed by these techniques. Grade I lesions have higher levels of fast myosin than slow myosin with a very small increase in CK levels. Grade II and III lesions have high values of both fast and slow myosin. CONCLUSIONS: The evaluation of fast and slow myosin in the blood 48 h after the lesion occurs is a useful aid for the detection of type I lesions in particular, since fast myosin is an exclusive skeletal muscle marker. The correct diagnosis of grade I lesions can prevent progression of the injury in athletes undergoing continual training sessions and competitions, thus aiding sports physicians in their decision making.

Detection of muscle gap by L-BIA in muscle injuries: clinical prognosis.

Sport-related muscle injury classifications are based basically on imaging criteria such as ultrasound (US) and magnetic resonance imaging (MRI) without consensus because of a lack of clinical prognostics for return-to-play (RTP), which is conditioned upon the severity of the injury, and this in turn with the muscle gap (muscular fibers retraction). Recently, Futbol Club Barcelona's medical department proposed a new muscle injury classification in which muscle gap plays an important role, with the drawback that it is not always possible to identify by MRI. Localized bioimpedance measurement (L-BIA) has emerged as a non-invasive technique for supporting US and MRI to quantify the disrupted soft tissue structure in injured muscles. OBJECTIVE: To correlate the severity of the injury according to the gap with the RTP, through the percent of change in resistance (R), reactance (Xc) and phase-angle (PA) by L-BIA measurements in 22 muscle injuries. MAIN RESULTS: After grouping the data according to the muscle gap (by MRI exam), there were significant differences in R between grade 1 and grade 2f (myotendinous or myofascial muscle injury with feather-like appearance), as well as between grade 2f and grade 2g (myotendinous or myofascial muscle injury with feather and gap). The Xc and PA values decrease significantly between each grade (i.e. 1 versus 2f, 1 versus 2g and 2f versus 2g). In addition, the severity of the muscle gap adversely affected the RTP with significant differences observed between 1 and 2g as well as between 2f and 2g. SIGNIFICANCE: These results show that L-BIA could aid MRI and US in identifying the severity of an injured muscle according to muscle gap and therefore to accurately predict the RTP.

Interaction between N-terminal domain of H4 and DNA is regulated by the acetylation degree.

To study whether the acetylation of one or more of the four acetylatable lysines of histone H4 affects its binding to DNA, we have designed a protection experiment with a model system consisting in phage lambda DNA as substrate, StuI as restriction endonuclease and histone H4 with different degrees of acetylation as the protective agent. It can be deduced from the experimental data that the protection afforded by the histone is not dependent on the number of positive charges lost by acetylation. Thus, non-acetylated H4 and mono-acetylated H4 cause similar protection, while di-acetylation of the histone seems to be the crucial step in significantly weakening the interaction between H4 and DNA. This is confirmed by the results obtained in protection experiments carried out using H4 peptide (1-24) with different degrees of acetylation as the protecting agent. As restriction enzyme can imitate any trans-acting factor with sequence recognition, the di-acetylated isoform of histone H4 can be the starting point, through acetylation, to unmask DNA sequences, allowing the accessibility of regulatory factors to DNA in the chromatin.

Effects of muscle injury severity on localized bioimpedance measurements.

Muscle injuries in the lower limb are common among professional football players. Classification is made according to severity and is diagnosed with radiological assessment as: grade I (minor strain or minor injury), grade II (partial rupture, moderate injury) and grade III (complete rupture, severe injury). Tetrapolar localized bioimpedance analysis (BIA) at 50 kHz made with a phase-sensitive analyzer was used to assess damage to the integrity of muscle structures and the fluid accumulation 24 h after injury in 21 injuries in the quadriceps, hamstring and calf, and was diagnosed with magnetic resonance imaging (MRI). The aim of this study was to identify the pattern of change in BIA variables as indicators of fluid [resistance (R)] and cell structure integrity [reactance (Xc) and phase angle (PA)] according to the severity of the MRI-defined injury. The % difference compared to the non-injured contralateral muscle also measured 24-h after injury of R, Xc and PA were respectively: grade I (n = 11; -10.4, -17.5 and -9.0%), grade II (n = 8; -18.4, -32.9 and -16.6%) and grade III (n = 2; -14.1, -52.9 and -43.1%), showing a greater significant decrease in Xc (p < 0.001). The greatest relative changes were in grade III injuries. However, decreases in R, that indicate fluid distribution, were not proportional to the severity of the injury. Disruption of the muscle structure, demonstrated by the localized determination of Xc, increased with the severity of muscle injury. The most significant changes 24 h after injury was the sizeable decrease in Xc that indicates a pattern of disrupted soft tissue structure, proportional to the severity of the injury.

Histone acetyltransferases during the cell cycle and differentiation of Physarum polycephalum.

The dynamic state of histone acetylation is maintained by histone acetyltransferases (HATs) and deacetylases. Cellular fractionation of plasmodia of Physarum polycephalum and partial purification of subcellular fractions by chromatography revealed the existence of a cytoplasmic B-type and four nuclear A-type HATs. The cytoplasmic B-enzyme was highly specific for histone H4, causing di-acetylation of H4 in vitro. The nuclear enzymes (HAT-A1 to HAT-A4) accepted all core histones as substrates, but differed by the preference for certain histone species. Enzymes were analyzed during the naturally synchronous cell cycle of macroplasmodia. Each of the enzymes had its individual cell cycle activity pattern, indicating diverse functions in nuclear metabolism. When growing plasmodia were induced to undergo differentiation into dormant sclerotia, an additional enzyme (HAT-AS) appeared at a late stage of sclerotization which correlated with differentiation-specific histone synthesis and acetylation in the absence of DNA replication. When dormant sclerotia were induced to reenter the cell cycle, a further enzyme form (HAT-AG) appeared during a short time period prior to the first post-germination mitosis. This enzyme had a strong preference for H2B, correlating with the overproportional in vivo acetate incorporation in H2B. Both differentiation-associated HATs were undetectable in growing plasmodia. The results demonstrate that different functions of core histone acetylation are based on multiple enzyme forms that are independently regulated during the cell cycle. Transitions from one developmental stage into another are accompanied by specific enzyme forms. With respect to recent data in the literature it may be assumed that these HAT-forms are subunits of a HAT-complex whose composition changes during the cell cycle and differentiation.

DNA methylation and histone acetylation of rat methionine adenosyltransferase 1A and 2A genes is tissue-specific.

Methionine adenosyltransferase (MAT) catalyzes the biosynthesis of S-adenosylmethionine (AdoMet). In mammals MAT activity derives from two separate genes which display a tissue-specific pattern of expression. While MAT1A is expressed only in the adult liver, MAT2A is expressed in non-hepatic tissues. The mechanisms behind the selective expression of these two genes are not fully understood. In the present report we have evaluated MAT1A and MAT2A methylation in liver and in other tissues, such as kidney, by methylation-sensitive restriction enzyme digestion of genomic DNA. Our data indicate that MAT1A is hypomethylated in liver and hypermethylated in non-expressing tissues. The opposite situation is found for MAT2A. Additionally, histones associated to MAT1A and MAT2A genes showed enhanced levels of acetylation in expressing tissues (two-fold for MAT1A and 3.5-fold for MAT2A liver and kidney respectively). These observations support a role for chromatin structure and its modification in the tissue-specific expression of both MAT genes.

Enzymes involved in the dynamic equilibrium of core histone acetylation of Physarum polycephalum.

DEAE-Sepharose chromatography of extracts from plasmodia of the myxomycete Physarum polycephalum revealed the presence of multiple histone acetyltransferases and histone deacetylases. A cytoplasmic histone acetyltransferase B, specific for histone H4, and two nuclear acetyltransferases A1 and A2 were identified; A1 acetylates all core histones with a preference for H3 and H2A, whereas A2 is specific for H3 and also slightly for H2B. Two histone deacetylases, HD1 and HD2, could be discriminated. They differ with respect to substrate specificity and pH dependence. For the first time the substrate specificity of histone deacetylases was determined using HPLC-purified individual core histone species. The order of acetylated substrate preference is H2A much greater than H3 greater than or equal to H4 greater than H2B for HD1 and H3 greater than H2A greater than H4 for HD2, respectively; HD2 is inactive with H2B as substrate. Moreover histone deacetylases are very sensitive to butyrate, since 2 mM butyrate leads to more than 50% inhibition of enzyme activity.

Histone deacetylase. A key enzyme for the binding of regulatory proteins to chromatin.

Core histones can be modified by reversible, posttranslational acetylation of specific lysine residues within the N-terminal protein domains. The dynamic equilibrium of acetylation is maintained by two enzyme activities, histone acetyltransferase and histone deacetylase. Recent data on histone deacetylases and on anionic motifs in chromatin- or DNA-binding regulatory proteins (e.g. transcription factors, nuclear proto-oncogenes) are summarized and united into a hypothesis which attributes a key function to histone deacetylation for the binding of regulatory proteins to chromatin by a transient, specific local increase of the positive charge in the N-terminal domains of nucleosomal core histones. According to our model, the rapid deacetylation of distinct lysines in especially H2A and H2B would facilitate the association of anionic protein domains of regulatory proteins to specific nucleosomes. Therefore histone deacetylation (histone deacetylases) may represent a unique regulatory mechanism in the early steps of gene activation, in contrast to the more structural role of histone acetylation (histone acetyltransferases) for nucleosomal transitions during the actual transcription process.

Cardiorespiratory response to exercise in elite Sherpa climbers transferred to sea level.

Himalayan Sherpas are well known for their extraordinary adaptation to high altitude and some of them for their outstanding physical performance during ascents to the highest summits. To cast light on this subject, we evaluated the cardiorespiratory response during exercise at sea level of six of the most acknowledged Sherpa climbers, mean age (+/- SD) 37 (+/- 7) yr old. Continuous electrocardiogram and breath-by-breath pulmonary gas exchange until exhaustion were obtained by following the Bruce protocol. We detected a maximal oxygen uptake (VO2max) of 66.7 (+/- 3.7) mL-min-1.kg-1, maximal cardiac frequency of 199 (+/- 7) beats.min-1, and ventilatory anaerobic threshold at 62 (+/- 4) % of VO2max. These factors could help to explain the greater performance level shown by several elite climbers of this ethnic group. The high functional reserve demonstrated by this very select group of highlanders could be associated with natural selection and with special physiological adaptations probably induced by long-training in a hostile environment.

Differences between lactate concentration of samples from ear lobe and the finger tip.

Blood lactate concentrations in capillary samples obtained from the ear lobe or from the finger tip are used indistinctly, since they are considered equivalents. The aim of the study reported in this paper was to verify whether that assumption is valid due to the practical implications which any possible differences between these two sampling sites would have in the planning and assessing of an athletic training program. Twenty six healthy male athletes competing in different sports at the national level (9 rowers, 7 cyclists and 10 runners) were studied during the performance of a graded exercise test up to the point of exhaustion, on specific ergometers. In each group, capillary blood samples were obtained simultaneously from both the ear lobe and the finger tip at three different times during the test: 1) in resting conditions; 2) when exercising at a submaximal work load and 3) seven minutes after the point of exhaustion. Significant differences were found between the blood lactate concentrations of samples obtained from the ear lobe and from the finger tip (p < 0.001). The method error of repeated measurements for lactate concentrations from paired samples obtained in resting conditions was 27%, when exercising at a submaximal work load, 16% and at maximal work load, 3%. Capillary blood samples collected from the finger tip consistently showed higher values in lactate concentration than those obtained, at the same time, from the ear lobe.

Localized BIA identifies structural and pathophysiological changes in soft tissue after post-traumatic injuries in soccer players.

Localized bioimpedance (BIA) was measured with a single frequency phase-sensitive analyzer at 50 kHz in three post-traumatic types of injuries on four professional soccer players: (1) myositis ossificans, (2) intramuscular seroma and (3) trochanteric (hip) bursitis. Normal reference value (no injury) was obtained from the contra lateral not injured limb at a mirror-like location of the injury. The relative variations resistance (R) and reactance (Xc) at the time of injury was confronted with the not injured values. Relative variations between acute measurements and post medication ones on intramuscular seroma and bursitis have been computed. In intramuscular seroma and trochanteric bursitis we have obtained a percent of change between injury data and after medical intervention. On myositis ossificans, localized BIA showed a 7-8 % decrease in Xc whereas the percent of change of R was negligible (1 %). These percent of changes are in concordance with histological evidence. In the case of a presence of seroma or the lower thigh and trochanteric bursitis, the soft tissue cavity accumulates fluid. Post-injury localized BIA, relative with respect to non-injured side, confirmed sizeable soft tissue destruction evidenced by 50 % decrease of Xc and 24-31 % decrease of R due to interstitial fluid accumulation. Once the seroma and the blood in the bursitis was removed the localized the immediate post-injury BIA parameters increased as follows: a) intramuscular seroma + 10 % on R and + 74 % of Xc; b) trochanteric bursitis + 20 % of R and +24 % of Xc. Localized BIA other than classifying soft tissue injuries, can be useful to understand the pathophysiology and structural impairments of other kind of injuries and to understand their behavior.

Can local corticosteroid injection in the retrocalcaneal bursa lead to rupture of the Achilles tendon and the medial head of the gastrocnemius muscle?

PURPOSE: The purpose of the study is to explain the cause-effect relationship in three patients who reported combined ruptures of the Achilles tendon and the gastrosoleus complex 6 months after they had received corticosteroids injections for the management of retrocalcaneal bursitis. METHODS: Three cryopreserved cadavers (three men, three left legs) were examined to assess the anatomic connection between the retrocalcaneal bursa and the Achilles tendon (distal and anterior fibers). Blue triptan medium contrast was injected. RESULTS: An unexpected connection between the retrocalcaneal bursa and the anterior fibers of the Achilles tendon was found in all instances. CONCLUSIONS: Local corticosteroid injection of the retrocalcaneal bursa may help the symptoms of retrocalcanear bursitis, but pose a risk of Achilles tendon rupture. This risk-benefit has to be taken into account when corticosteroid injections are prescribed to professional and high-level athletes.