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This study aimed to clarify how masticatory muscle atrophy induced by botulinum toxin (BTX) injection affects cortical bone quality of the mandible using 3D modeling technology. A total of 39 young (26.9 ± 6.0 years) and 38 post-menopausal (55.3 ± 6.3 years) females were included. Computed tomography (CT) images were obtained before and after 12 months of treatment. Predictor variables were application of a stabilization splint, and/or two times of BTX injection in the bilateral temporalis and masseter muscles within a six-month interval. Outcome variables were changes in average Hounsfield units (HU) and cortical thickness of region of interest (ROI). 3D mandibular models were reconstructed using CT images, and models were used to calculate average HU and cortical thickness of ROIs, including inferior half of the lateral surface of ascending ramus, coronoid process, and temporomandibular joint condyle. Cortical bone quality at muscle insertion site was influenced by decreased muscle thickness but seemed not to be affected by decreased functional loading. Reduced functional loading seemed to influence cortical bone quality of the condyles. These effects were more remarkable in post-menopausal females. Hence, decreased masticatory muscle thickness may lead to alterations of the mandibular cortical structures, especially in post-menopausal females.

To examine the role of adipose-resident macrophages in insulin resistance, we examined the gene expression of CD68, a macrophage marker, along with macrophage chemoattractant protein-1 (MCP-1) in human subcutaneous adipose tissue using real-time RT-PCR. Both CD68 and MCP-1 mRNAs were expressed in human adipose tissue, primarily in the stromal vascular fraction. When measured in the adipose tissue from subjects with normal glucose tolerance, covering a wide range of BMI (21-51 kg/m2) and insulin sensitivity (S(I)) (0.6-8.0 x 10(-4)min(-1).microU(-1).ml(-1)), CD68 mRNA abundance, which correlated with the number of CD68-positive cells by immunohistochemistry, tended to increase with BMI but was not statistically significant. However, there was a significant inverse relation between CD68 mRNA and S(I) (r=-0.55, P=0.02). In addition, there was a strong positive relationship among adipose tissue CD68 mRNA, tumor necrosis factor-alpha (TNF-alpha) secretion in vitro (r=0.79, P<0.005), and plasma interleukin-6 (r=0.67, P < 0.005). To determine whether improving S(I) in subjects with impaired glucose tolerance (IGT) was associated with decreased CD68 expression, IGT subjects were treated for 10 weeks with pioglitazone or metformin. Pioglitazone increased S(I) by 60% and in the same subjects reduced both CD68 and MCP-1 mRNAs by >50%. Furthermore, pioglitazone resulted in a reduction in the number of CD68-positive cells in adipose tissue and reduced plasma TNF-alpha. Metformin had no effect on any of these measures. Thus, treatment with pioglitazone reduces expression of CD68 and MCP-1 in adipose tissue, apparently by reducing macrophage numbers, resulting in reduced inflammatory cytokine production and improvement in S(I).

Aims/hypothesisSubcellular localisation is an important factor in the known impact of bioactive lipids, such as diacylglycerol and sphingolipids, on insulin sensitivity in skeletal muscle; yet, the role of localised intramuscular triacylglycerol (IMTG) is yet to be described. Excess accumulation of IMTG in skeletal muscle is associated with insulin resistance, and we hypothesised that differences in subcellular localisation and composition of IMTG would relate to metabolic health status in humans.MethodsWe evaluated subcellular localisation of IMTG in lean participants, endurance-trained athletes, individuals with obesity and individuals with type 2 diabetes using LC-MS/MS of fractionated muscle biopsies and insulin clamps.ResultsInsulin sensitivity was significantly different between each group (athletes>lean>obese>type 2 diabetes; p < 0.001). Sarcolemmal IMTG was significantly greater in individuals with obesity and type 2 diabetes compared with lean control participants and athletes, but individuals with type 2 diabetes were the only group with significantly increased saturated IMTG. Sarcolemmal IMTG was inversely related to insulin sensitivity. Nuclear IMTG was significantly greater in individuals with type 2 diabetes compared with lean control participants and athletes, and total and saturated IMTG localised in the nucleus had a significant inverse relationship with insulin sensitivity. Total cytosolic IMTG was not different between groups, but saturated cytosolic IMTG species were significantly increased in individuals with type 2 diabetes compared with all other groups. There were no significant differences between groups for IMTG concentration in the mitochondria/endoplasmic reticulum.Conclusions/interpretationThese data reveal previously unknown differences in subcellular IMTG localisation based on metabolic health status and indicate the influence of sarcolemmal and nuclear IMTG on insulin sensitivity. Additionally, these studies suggest saturated IMTG may be uniquely deleterious for muscle insulin sensitivity.

Selenium (Se) is an essential trace element for humans and animals. Appropriate amount of Se in the body can prevent a variety of diseases. However, Se deficiency leads to pathological changes such as skeletal muscle necrosis and pancreatic atrophy in livestock and poultry. Se preparations are widely used in the prevention and treatment of Se-deficient disease, but there is no unified standard of medication, and the safe dose range of Se is narrow. Therefore, it is of great significance to study the pharmacokinetics of low-Se ducklings and to formulate drug administration schemes. In the present study, eighty 1-day-old healthy ducklings were randomly selected, and fed with low-Se diet to 30 days of age (blood Se content ≦ 0.03 μg/mL). After the low Se duckling models were duplicated, blood samples and tissues of livers, pancreases, and thigh muscles were collected at different time points to detect Se content following oral administration of 0.1% sodium selenite (Na 2 SeO 3 ) at 0.8 mg/kg BW, and the pharmacokinetics parameters were automatically calculated by MCPKP program. The results showed that pharmacokinetics characteristics of Na 2 SeO 3 in blood, livers, and pancreases of ducklings were consistent with the first-order absorption and two-compartment open models; in thigh muscles was consistent with the first-order absorption and one compartment with a lag time open model. The primary kinetic parameters of Na 2 SeO 3 in blood: the half-life of absorption was 5.9026 h, the time of reaching maximum concentration was 23.03 h, and the half-life of elimination was 131.13 h. The absorption of Na 2 SeO 3 in livers was the quickest, pancreases and thigh muscles were in order of becoming slower, and the elimination of Na 2 SeO 3 in thigh muscles was the quickest, livers and pancreases were in order of becoming slower. The administration parameters of multi-dose were calculated according to the kinetic of single-dose: loading dose (D*) was 1.7046 mg/kg BW, maintenance dose (D 0 ) was 0.8 mg/kg BW, and dosing interval (τ) was 120 h. The results of this study can supplement and improve the theoretical system of Se metabolic kinetics, and provide experimental basis for the prevention and treatment of Se deficiency disease by rational drug use.

Background: This study evaluates whether the initial amount of dietary protein intake could influence the combined effect of high-intensity interval training (HIIT) and citrulline (CIT), or HIIT alone, on body composition, muscle strength, and functional capacities in obese older adults. Methods: Seventy-three sedentary obese older men and women who completed a 12-week elliptical HIIT program with double-blinded randomized supplementation of CIT or placebo (PLA) were divided into four groups according to their initial protein intake (CIT–PROT+: n = 21; CIT–PROT−: n = 19; PLA–PROT+: n = 19; PLA–PROT−: n = 14). Body composition (fat and fat-free masses), handgrip (HSr) strength, knee extensor (KESr) strength, muscle power, and functional capacities were measured pre-intervention and post-intervention. Results: Following the intervention, the four groups improved significantly regarding all the parameters measured. For the same initial amount of protein intake, the CIT–PROT− group decreased more gynoid fat mass (p = 0.04) than the PLA–PROT− group. The CIT–PROT+ group increased more KESr (p = 0.04) than the PLA–PROT+ group. In addition, the CIT–PROT− group decreased more gynoid FM (p = 0.02) and improved more leg FFM (p = 0.02) and HSr (p = 0.02) than the CIT–PROT+ group. Conclusion: HIIT combined with CIT induced greater positive changes than in the PLA groups. The combination seems more beneficial in participants consuming less than 1 g/kg/d of protein, since greater improvements on body composition and muscle strength were observed.

Statins are the most commonly used drugs worldwide. Besides a significant decrease in cardiovascular diseases (CVDs) risk, the use of statins is also connected with a broad beneficial pleiotropic effect. At the same time, it is burdened with different side effects. The most common ones are muscle issues (from mild myalgia to rhabdomyolysis). The mechanisms of many of them are still unclear. Therefore, an analytical method for the determination of simvastatin (SIM) and its main metabolite (the hydroxy acid form - SIMA) in muscle tissue was developed. Muscle samples were homogenized with ammonium acetate buffer using the bead mill homogenizer, and then statins were extracted with a mixture of methanol and ethanol. Prepared samples were analyzed with liquid chromatography (using a reverse-phase column with a gradient elution) combined with the mass spectrometer which was operated in a multiple reaction monitoring mode. The assay was linear over a 0.1-5 ng/mL range for both statin forms. Inter- and intra-day precision and accuracy were characterized. The method was considered precise (with the following relative standard deviation values: 6.0-6.9% for SIM, and 8.1-12.9% for SIMA) and accurate (with the following mean accuracies: 91.4-100.1% for SIM, and 102.2-115.4% for SIMA). The extraction efficiency was evaluated by recovery determination (76% for SIM, and 99% for SIMA). Moreover, the matrix effect was calculated with the following results: 87% for SIM, and 139% for SIMA. The proposed method was applied for SIM and SIMA determination in skeletal muscle tissues obtained from statin-treated patients. The obtained results proved that the method may be a useful tool for explaining muscle effects related to statin therapy.

The objective of this study was to determine the palatability of various beef cuts from 3 USDA quality grades. Five different beef subprimals from USDA Prime, Choice, and Select (n = 10/quality grade) carcasses were utilized for the study, including: strip loins, inside rounds, bottom rounds, shoulder clods, and chuck rolls. Subprimals were fabricated into 9 retail cuts, which contained the following beef muscles: longissimus lumborum (LL); longissimus thoracis, complexus, and spinalis dorsi (LCS); infraspinatus (IF); serratus ventralis (SV); triceps brachii (TB); teres major (TM); adductor (AD); semimembranosus (SM); and biceps femoris (BF). The pH and percentage of fat, moisture, protein, and collagen was determined for each muscle on a raw basis. Additionally, cooked steak measurements included Warner-Bratzler shear force (WBSF) and slice shear force (SSF). Consumer and trained sensory panelists evaluated palatability traits of each cut and quality grade combination. A quality grade × muscle interaction was determined for trained panelists assessment of overall tenderness (P = 0.03), SSF (P = 0.02), proximate composition (P < 0.01), and pH (P < 0.01). In all objective and subjective measurements of tenderness, the LCS was the most tender (P < 0.05), while cuts from the round (BF, AD, and SM) were among the toughest and least juicy (P < 0.05). Conversely, consumers and trained sensory panelists identified the LCS, IF, and SV to be juicier (P < 0.05) than all others. The TB, TM, and LL were perceived by consumers most often as being everyday quality. The LCS was found by consumers to be the most acceptable (P < 0.05) across all attributes, with the SM being the least (P < 0.05) acceptable muscle. For each muscle, fat percentage was the greatest (P < 0.05) in Prime cuts. Slice shear force determined Prime IF, LL, and SV to be more tender (P < 0.05) than Choice and Select. No SSF differences (P > 0.05) were found among quality grades for the AD, BF, and SM. The WBSF values decreased (P < 0.05) across all muscles, as quality grade increased (Prime < Choice < Select). The results of this study indicate that muscles from the chuck may be utilized to provide consumers with a positive eating experience. Meanwhile, muscles from the round are likely to provide consumers with a lower quality eating experience.

Helical structures are ubiquitous in nature and impart unique mechanical properties and multifunctionality 1 . So far, synthetic architectures that mimic these natural systems have been fabricated by winding, twisting and braiding of individual filaments 1 – 7 , microfluidics 8 , 9 , self-shaping 1 , 10 – 13 and printing methods 14 – 17 . However, those fabrication methods are unable to simultaneously create and pattern multimaterial, helically architected filaments with subvoxel control in arbitrary two-dimensional (2D) and three-dimensional (3D) motifs from a broad range of materials. Towards this goal, both multimaterial 18 – 23 and rotational 24 3D printing of architected filaments have recently been reported; however, the integration of these two capabilities has yet to be realized. Here we report a rotational multimaterial 3D printing (RM-3DP) platform that enables subvoxel control over the local orientation of azimuthally heterogeneous architected filaments. By continuously rotating a multimaterial nozzle with a controlled ratio of angular-to-translational velocity, we have created helical filaments with programmable helix angle, layer thickness and interfacial area between several materials within a given cylindrical voxel. Using this integrated method, we have fabricated functional artificial muscles composed of helical dielectric elastomer actuators with high fidelity and individually addressable conductive helical channels embedded within a dielectric elastomer matrix. We have also fabricated hierarchical lattices comprising architected helical struts containing stiff springs within a compliant matrix. Our additive-manufacturing platform opens new avenues to generating multifunctional architected matter in bioinspired motifs. A 3D printing platform comprising a rotational multimaterial printhead is demonstrated, enabling the fabrication of helically architected filaments and lattices with programmable subvoxel control.

Abstract Context Impaired lipid metabolism is linked with obesity-associated insulin resistance, which may be reversed by caloric restriction (CR). Objective In a secondary analysis of a randomized controlled trial, we compared the effects of intermittent fasting (IF) and CR on markers of lipid metabolism in muscle. Design Seventy-six women (body mass index, 25-40 kg/m2) were randomly assigned to 1 of 3 diets for 8 weeks and provided foods at 70% (CR70 and IF70) or 100% (IF100) of energy requirements. IF groups ate breakfast prior to a 24-hour fast on 3 nonconsecutive days per week. On nonfasting days, IF70 ate at 100% and IF100 ate at 145% of energy requirements to achieve the prescribed target. Weight, body composition, insulin sensitivity by clamp, nonesterified fatty acids (NEFAs), β-hydroxybutyrate (BHB), and markers of lipid metabolism and oxidative stress in muscle by quantitative polymerase chain reaction were measured at baseline and week 8 following a 12-hour overnight fast (all groups) and 24-hour fast (IF groups). Results IF70 resulted in greater weight and fat loss and reduced NEFAs vs CR70 and IF100 after an overnight fast. IF70 and IF100 induced a greater reduction only in mRNA levels of antioxidant enzymes glutathione peroxidase 1 (GPX1), superoxide dismutase 1, soluble (SOD1), and SOD2 vs CR70. Fasting for 24 hours increased NEFAs and BHB in IF groups, but impaired insulin sensitivity and increased PLIN5 mRNA levels. Conclusions In comparison to CR, IF did not increase markers of lipid metabolism in muscle, but reduced expression of antioxidant enzymes. However, fasting-induced insulin resistance was detected, alongside increased PLIN5 expression, potentially reflecting transient lipid storage.

DNA methylation is an important epigenetic mechanism involved in expression of genes in many biological processes including muscle growth and development. Its effects on economically important traits are evinced from reported significant differences in meat quality traits between Japanese black (Wagyu) and Chinese Red Steppes cattle, thus presenting a unique model for analyzing the effects of DNA methylation on these traits. In the present study, we performed whole genome DNA methylation analysis in the two breeds by whole genome bisulfite sequencing (WGBS). Overall, 23150 differentially methylated regions (DMRs) were identified which were located in 8596 genes enriched in 9922 GO terms, of which 1046 GO terms were significantly enriched (p<0.05) including lipid translocation (GO: 0034204) and lipid transport (GO: 0015914). KEGG analysis showed that the DMR related genes were distributed among 276 pathways. Correlation analysis found that 331 DMRs were negatively correlated with the expression levels of differentially expressed genes (DEGs) with 21 DMRs located in promoter regions. Our results identified novel candidate DMRs and DEGs correlated with meat quality traits, which will be valuable for future genomic and epigenomic studies of muscle development and for marker assisted selection of meat quality traits.