In vivo assessment of the passive stretching response of the bi-compartmental human semitendinosus muscle using shear wave elastography

The semitendinosus muscle contains distinct proximal and distal compartments arranged anatomically in-series but separated by a tendinous inscription, with each compartment innervated by separate nerve branches. Although extensively investigated in other mammals, compartment-specific mechanical properties within the human semitendinosus have scarcely been assessed in vivo. Experimental data obtained during muscle-tendon unit stretching (e.g., slack angle) can also be used to validate and/or improve musculoskeletal model estimates of semitendinosus muscle force. The purpose of this study was to investigate the passive stretching response of proximal and distal humans semitendinosus compartments to distal joint extension. Using two-dimensional shear wave elastography, we bilaterally obtained shear moduli of both semitendinosus compartments from 14 prone-positioned individuals at ten knee flexion angles (from 90° to 0° [full extension] at 10° intervals). Passive muscle mechanical characteristics (slack angle, slack shear modulus, and the slope of the increase in shear modulus) were determined for each semitendinosus compartment by fitting a piecewise exponential model to the shear modulus-joint angle curves. We found no differences between compartments or legs for slack angle, slack shear modulus, or the slope of the increase in shear modulus. We also found the experimentally determined slack angle occurred at ~15-80° higher knee flexion angles compared to estimates from two commonly used musculoskeletal models, depending on participant and model used. Overall, these findings demonstrate that passive shear modulus-joint angle curves do not differ between proximal and distal human semitendinosus compartments, and provide experimental data to improve semitendinosus force estimates derived from musculoskeletal models.

Profiling of open chromatin in developing pig (Sus scrofa) muscle to identify regulatory regions

There is very little information about how the genome is regulated in domestic pigs (Sus scrofa). This lack of knowledge hinders efforts to define and predict the effects of genetic variants in pig breeding programmes. In order to address this knowledge gap, we need to identify regulatory sequences in the pig genome starting with regions of open chromatin. We used the ‘Improved Protocol for the Assay for Transposase-Accessible Chromatin (Omni-ATAC-Seq)’ to identify putative regulatory regions in flash frozen semitendinosus muscle from 24 male piglets. We collected samples from the smallest, average, and largest sized male piglets from each litter through five developmental time points. Of the 4,661 ATAC-Seq peaks identified that represent regions of open chromatin, >50% were within 1 kb of known transcription start sites. Differential read count analysis revealed 377 ATAC-Seq defined genomic regions where chromatin accessibility differed significantly across developmental time points. We found regions of open chromatin associated with down regulation of genes involved in muscle development that were present in small sized foetal piglets but absent in large foetal piglets at day 90 of gestation. The dataset that we have generated provides: a resource for studies of genome regulation in pigs, and contributes valuable functional annotation information to filter genetic variants for use in genomic selection in pig breeding programmes.

500 Late-Breaking: Heart Rate During Exercise Is Positively Correlated with State IV Mitochondrial Respiration in the Equine Skeletal Muscle

Mitochondria are critical for oxidative phosphorylation in skeletal muscle, especially in athletic species such as the horse. Mitochondrial respiration increases with physical exercise, but the relationship between mitochondrial respiration and cardiovascular functions are not well described in the horse. The objective of this study was to determine if there is a relationship between heart rate (HR) during and after submaximal exercise tests (SETs) and skeletal muscle mitochondrial respiration in polo ponies. We hypothesized that horses with greater maximum HR and average HR during the exercise tests would have greater mitochondrial respiration in skeletal muscle. Twelve fit polo ponies (14.8 ± 1.7 years old, 10 mares and 2 geldings) were equipped with Polar equine heart rate monitors (Polar Electro Inc., Lake Success, NY) and underwent 26-minute SETs designed to mimic a polo chukker followed by a 30-minute recovery period. Muscle biopsy samples from the semitendinosus muscle were taken 2 weeks prior to the SET to determine mitochondrial oxygen consumption using the Oroboros O2k high-resolution respirometer (Oroboros Instruments, Innsbruck, Austria). Data were analyzed using the PROC CORR procedure (SAS Inst. Inc., Cary, NC). Correlations were considered strong at r > 0.6 and significant at P < 0.05. Maximum HR during SET and state IV respiration were positively correlated (P = 0.02, r = 0.68). Average HR during SET and state IV respiration were also positively correlated (P = 0.01, r = 0.72). However, correlations between maximum and average HR and state III respiration were not as strong (P ≥ 0.05, r < 0.6). These data suggest that state IV mitochondrial respiration (proton leak) in equine skeletal muscle may impact cardiac responses to high-intensity exercise. Horses with higher HR during exercise may have less efficient oxidative phosphorylation, resulting in earlier fatigue and/or greater formation of reactive oxygen species resulting from proton leak.

507 Late-Breaking: Heat Stress and Mitoq Supplementation Impact Skeletal Muscle Mitochondrial Capacities in Pigs

Heat stress can negatively impact pig health and performance but the effects of heat stress on skeletal muscle mitochondrial function are largely unknown. We hypothesized that mitochondrial function and capacity would be impaired in heat stressed (HS) compared to thermoneutral (TN) pigs but mitochondrially-targeted coenzyme Q (MitoQ) supplementation would rescue the impairment. Oxidative portions of the semitendinosus muscle were evaluated from TN and HS gilts receiving no supplementation (CON) or MitoQ for 2 d prior to and during the 24h environmental heat treatment (n = 8 per group). Mitochondrial oxidative phosphorylation (P) and electron transfer (E) capacities were determined via high resolution respirometry and mitochondrial volume density and function were quantified by citrate synthase (CS) and cytochrome c oxidase activities, respectively. Data were analyzed using linear models in SAS v9.4 with fixed effects of heat, MitoQ treatment (trt), and heat×trt interaction. There were trends for the interaction of trt and heat (P≤0.1) on integrative (per mg tissue) and intrinsic (relative to CS) P with complexes I and II (PCI+II), maximum noncoupled E (ECI+II), and E with complex II only (ECII), in which all measures were greater in HS-MitoQ than TN-MitoQ (P≤0.03), but measures did not differ due to HS in CON pigs. The contribution of leak to total E (flux control ratio, FCRLeak) was lesser in HS-MitoQ than HS-CON, TN-CON, and TN-MitoQ (P≤0.02). The FCRPCI was greater (P≤0.05) while the FCRPCI+II was lesser (P=0.01) in TN compared to HS pigs. Finally, the FCRPCI+II was greater (P=0.02) while the FCRECII tended to be lesser (P=0.09) for CON than MitoQ pigs. Neither mitochondrial volume density nor function were affected by HS or MitoQ supplementation. In total, these data indicate improved mitochondrial capacities following heat stress in pigs receiving MitoQ but no difference in mitochondrial capacities in unsupplemented, HS pigs.

Carcass Yields and Physical-Chemical Meat Quality Characteristics of Namibian Red Hartebeest (Alcelaphus buselaphus) as Influenced by Sex and Muscle

This study determined the carcass yields of red hartebeest from Namibia and compared the physical-chemical meat quality characteristics of six different muscles (biceps femoris, infraspinatus, longissimus thoracis et lumborum, semimembranosus, semitendinosus, and supraspinatus) for both males and females. Red hartebeest males were heavier (133.92 kg) than females (114.20 kg) but the average dressing percentage did not differ between the two sexes. Muscles from females had a lower mean shear force value of 3.59 kg/1.27 cm ø, compared to males (4.23 kg/1.27 cm ø). The most tender muscle was the infraspinatus of the female treatment group, while the semimembranosus of the male treatment group was the least tender muscle. Drip loss, cooking loss and L* (lightness) values were not affected by sex. The largest hue angle was observed in the semitendinosus muscle of the female treatment group (28.94°), and it was thus the lightest red muscle. The highest chroma values (17.3) were observed in the semimembranosus muscle. Muscle protein content averaged 20.5% over all treatment combinations, and the mean intra-muscular fat content for both male and female muscles was low (2.4%). The shoulder muscles, infraspinatus and supraspinatus, of the females had the highest fat content (2.7%). The results indicate that red hartebeest meat should be market according to specific muscles and that sex of the animals need not be considered during marketing.

Аutosomal recessive limb-girdle muscular dystrophy type 2A: two cases in Ukraine with different age of onset

The paper reports on two cases of young women from Ukraine with autosomal recessive limb-girdle muscular dystrophy type 2A with different age of symptoms onset and an absence of any family history presented with gradual onset of proximal muscle weakness in four limbs and thinning of shoulders, arms and thighs. Both patients had elevated creatine phosphokinase level and c.550delA mutations in CAPN3 gene. Sequence analysis and deletion/duplication testing of the 159 genes from skeletal muscles disease testing panel of 5-year-old girl identified deletion of exon 8 (heterozygous) and c.550delA (p.Thr184Argfs*36) mutation (heterozygous), were in CAPN3 gene. Magnetic Resonance Imaging of soft tissue of the proximal lower extremities was performed which showed signs of symmetrical atrophic changes in the major adductor muscle, the long and short adductor muscles, the semitendinosus muscle of the thigh, as a manifestations of autosomal recessive limb-girdle muscular dystrophy type 2A. Homozygous, pathogenic variant of the defect in the CAPN3 gene c.550del (p.Thr184Argfs * 36) was identified in a 25-year-old woman. Type 2A is the most common form of limb-girdle muscular dystrophy, accounting for about 30% of cases. The autosomal recessive limb-girdle muscular dystrophy type 2A is on caused by mutations in the CAPN3 gene, and it is characterized by selective atrophy and weakness of proximal limb and girdle muscles. The age of onset of muscle weakness is extremely variable; the most common being between 8 and 15 years, although it can range between 2 and 50 years. The diagnosis can be suspected by findings on a muscle biopsy or when a doctor experienced in muscular dystrophy examines you. A serum creatine kinase blood test may also show raised levels which indicate a problem in the muscles. The diagnosis has to be confirmed by means of identifying a mutation in the CAPN3 gene which is done on a deoxyribonucleic acid sample from a blood test. To date there are no specific treatments for limb-girdle muscular dystrophy, however careful management of the symptoms of the condition can improve a person’s quality of life. Joint contractures (tightening) can occur in limb-girdle muscular dystrophy and therefore regular physiotherapy is recommended.

Comparison of Nutritional and Meat Quality Characteristics between Two Primal Cuts from Aceh Cattle in Aceh Province, Indonesia

The Aceh cattle are local Indonesian beef cattle that are farmed in Aceh Province. This type of cattle is one of the sources of meat for the Aceh people. This study aims to analyze the quality of two primal cuts (longissimus lumborum and semitendinosus muscle) from Aceh cattle based on the muscle microstructure characteristics and MSTN gene expression. This study used a sample of longissimus lumborum and semitendinosus muscles from 18 adult male Aceh cattle with the age of 2–2.5 years and a BCS of 3.24. Muscle samples were obtained shortly after the cattle were slaughtered in slaughterhouses in Banda Aceh and Aceh Besar districts. Muscle microstructure analysis was performed using the HE, Masson’s trichrome, and immunohistochemistry staining methods, while the MSTN gene expression analysis was performed using the qPCR method. The analysis of the physical quality of meat includes pH, meat color, fat color, cooking loss, water holding capacity, and WBSF value. The results showed that the area of LL muscle fibers was smaller than that of ST with relatively the same diameter. Both muscles were dominated by fast fibers with a percentage of 82.37% (LL muscle) and 91.80% (ST muscle). The area and composition of the type of muscle fibers are the main factors that influence the tenderness of Aceh beef. A higher distribution of collagen was found in ST muscles than in LL muscles. MSTN gene expression in both muscle types was relatively the same. Aceh cattle have large muscle fibers and are dominated by fast fibers with a high percentage, resulting in a low level of the tenderness of Aceh beef. However, the level of tenderness of Aceh beef is still in accordance with the cooking preparation of original and favorite cuisine of Aceh people.