Duchenne Muscular Dystrophy (DMD) is a progressive and fatal neuromuscular disease. Cycles of myofibre degeneration and regeneration are hallmarks of the disease where immune cells infiltrate to repair damaged skeletal muscle. Benfotiamine is a lipid soluble precursor to thiamine, shown clinically to reduce inflammation in diabetic related complications. We assessed whether benfotiamine administration could reduce inflammation related dystrophic pathology. Benfotiamine (10 mg/kg/day) was fed to male mdx mice (n = 7) for 15 weeks from 4 weeks of age. Treated mice had an increased growth weight (5-7 weeks) and myofibre size at treatment completion. Markers of dystrophic pathology (area of damaged necrotic tissue, central nuclei) were reduced in benfotiamine mdx quadriceps. Grip strength was increased and improved exercise capacity was found in mdx treated with benfotiamine for 12 weeks, before being placed into individual cages and allowed access to an exercise wheel for 3 weeks. Global gene expression profiling (RNAseq) in the gastrocnemius revealed benfotiamine regulated signalling pathways relevant to dystrophic pathology (Inflammatory Response, Myogenesis) and fibrotic gene markers (Col1a1, Col1a2, Col4a5, Col5a2, Col6a2, Col6a2, Col6a3, Lum) towards wildtype levels. In addition, we observed a reduction in gene expression of inflammatory gene markers in the quadriceps (Emr1, Cd163, Cd4, Cd8, Ifng). Overall, these data suggest that benfotiamine reduces dystrophic pathology by acting on inflammatory and fibrotic gene markers and signalling pathways. Given benfotiamine's excellent safety profile and current clinical use, it could be used in combination with glucocorticoids to treat DMD patients.

Abstract Background Little is known about potential differences in the left recurrent laryngeal nerve (Lrln) and left cricoarytenoideus dorsalis (LCAD) muscle between domestic and feral horse populations. If a difference exists, feral horses may provide a useful control population for research related to recurrent laryngeal neuropathy (RLN) and increase our understanding of potential population pressures influencing the incidence RLN. Objectives The objective of this study was to compare the Lrln and LCAD of domestic and feral horses using histological and immunohistochemical techniques (IHC). Methods Sixteen horses, domestic (n = 8) and feral (n = 8), without clinical or ancillary examinations that were processed at an abattoir had the Lrln and LCAD muscle harvested immediately following death. Carcass weights were recorded. Subjective and morphometric histologic assessment were performed on Lrln sections. The LCAD was assessed for myosin heavy chain (fibre type proportion, diameter and grouping using IHC. Results Fibre‐type grouping consistent with RLN was seen in both groups. Regenerating fibre clusters were more common in domestic compared to feral horses (p = 0.04). No other histologic differences occurred between groups. Muscle fibre typing demonstrated a lower mean percentage of type IIX fibres in the feral group compared to the domestic group (p = 0.03). There was no difference in type I or IIA proportions or mean diameter of any fibre type between the groups. Conclusions The domestic population showed evidence of nerve regeneration suggesting RLN in this group, yet this was not supported by the higher proportion of type IIX muscle fibres compared to the feral population. Further evaluation to clarify the significance and wider occurrence of the differences is indicated.

Microbial communities inhabiting the gut have the ability to influence physiological processes contributing to livestock production and performance. Livestock enterprises rely on animal production traits such as growth performance for profit. Previous studies have shown that gut microbiota are correlated to growth performance and could even influence it. The aim of this study was to characterise the faecal microbial profiles of Angus steers with high and low ADG at both weaning and yearling stages by profiling 16S rRNA gene sequences from rectal faecal samples. When microbial profiles were compared in terms of relative abundances, LEfSe analysis, alpha diversity metrics, and beta diversity, at the weaning stage, few significant differences were found between the high and low ADG groups. However, at yearling stage, microbial profiles significantly differed between the high and low ADG groups. The relative abundances of eight phyla and six genera significantly differed between the two groups. Alpha diversity metrics showed a significant decrease (p = 0.001) in species richness in the high ADG group. Similarly, beta diversity analysis showed that samples clustered clearly according to high and low ADG groups at yearling stage, indicating that phylogenetic similarity between the two ADG groups was significantly reduced (p = 0.005).

Context Heating of meat leads to structural changes reflected in the juiciness and the tenderness of the cooked meat. Aims This study aimed to characterise the effect of prolonged ageing and cooking on pork-quality traits. Methods Longissimus lumborum samples from 12 carcasses were aged 3 days (conventional ageing) or 15 days (prolonged ageing) and pork cuboids were cooked at 50–80°C for 30 min. Cooking loss, total water content (TWC), Warner–Bratzler shear force (WBSF) and shrinkage (longitudinal, transverse and estimated volume) of the pork loin cuboids were measured. Key results Prolonged ageing for 15 days reduced the WBSF of samples cooked at 50°C, and the cooking loss for samples cooked at 70°C and 80°C, relative to conventional ageing for 3 days. The WBSF of pork aged for 15 days was not different from that of pork aged for 3 days. Prolonged ageing reduced longitudinal shrinkage of cuboids, but TWC and transverse/volume shrinkage of cuboids were not affected by ageing. The diameter of cooked muscle fibre fragments was smaller in pork subjected to prolonged ageing. Conclusions Prolonged ageing was favourable for minimising cooking loss at higher cooking temperatures but was only favourable for tenderness at the lowest cooking temperature. Low pH of the samples is likely to have caused the lack of tenderisation with ageing.

Duchenne muscular dystrophy involves an absence of dystrophin, a cytoskeletal protein which supports cell structural integrity and scaffolding for signalling molecules in myocytes. Affected individuals experience progressive muscle degeneration that leads to irreversible loss of ambulation and respiratory diaphragm function. Although clinical management has greatly advanced, heart failure due to myocardial cell loss and fibrosis remains the major cause of death. We examined cardiac morphology and function in D2.B10-Dmd mdx /J (D2-mdx) mice, a relatively new mouse model of muscular dystrophy, which we compared to their wild-type background DBA/2J mice (DBA/2). We also tested whether drug treatment with a specific blocker of mitochondrial permeability transition pore opening (Debio-025), or ACE inhibition (Perindopril), had any effect on dystrophy-related cardiomyopathy. D2-mdx mice were treated for six weeks with Vehicle control, Debio-025 (20 mg/kg/day), Perindopril (2 mg/kg/day), or a combination (n = 8/group). At 18 weeks, compared to DBA/2, D2-mdx hearts displayed greater ventricular collagen, lower cell density, greater cell diameter, and greater protein expression levels of IL-6, TLR4, BAX/Bcl2, caspase-3, PGC-1α, and notably monoamine oxidases A and B. Remarkably, these adaptations in D2-mdx mice were associated with preserved resting left ventricular function similar to DBA/2 mice. Compared to vehicle, although Perindopril partly attenuated the increase in heart weight and collagen at 18 weeks, the drug treatments had no marked impact on dystrophic cardiomyopathy.

The microbial communities that inhabit the intestinal tract play an important role in modulating health and productivity. Environmental stressors can impact microbial communities, which can significantly influence host physiology. Cattle are subjected to several environmental stressors when placed on feedlots, such as transportation stress, exposure to feedlot environments and change in diet and management. Exposure to these stressors could influence host gut microbiota, which in turn, could potentially influence host health and performance. The aim of the current study was to characterise the temporal changes that occur in intestinal microbiota as a consequence of feedlot placement by profiling 16s rRNA sequences in rectal faecal samples. When faecal microbiome profiles were compared in terms of relative abundances and alpha diversity metrics, the results showed significant, observable changes in profiles 2 days post-feedlot induction. Furthermore, beta-diversity analysis indicated that the phylogenetic similarity between samples significantly decreased on day 2 (PERMANOVA, p < 0.001). These trends were suggestive of a short-term reduction in microbial diversity coupled with decreased similarity between animals. These changes warrant further investigation and could provide opportunities for improved performance, health and even welfare of feedlot cattle in future.

Duchenne muscular dystrophy (DMD) is a progressive fatal neuromuscular disorder with no cure. Therapies to restore dystrophin deficiency have been approved in some jurisdictions but long-term effectiveness is yet to be established. There is a need to develop alternative strategies to treat DMD. Resveratrol is a nutraceutical with anti-inflammatory properties. Previous studies have shown high doses (100–400 mg/kg bodyweight/day) benefit mdx mice. We treated 4-week-old mdx and wildtype mice with a lower dose of resveratrol (5 mg/kg bodyweight/day) for 15 weeks. Voluntary exercise was used to test if a lower dosage than previously tested could reduce exercise-induced damage where a greater inflammatory infiltrate is present. We found resveratrol promoted skeletal muscle hypertrophy in wildtype mice. In dystrophic muscle, resveratrol reduced exercise-induced muscle necrosis. Gene expression of immune cell markers, CD86 and CD163 were reduced; however, signalling targets associated with resveratrol’s mechanism of action including Sirt1 and NF-κB were unchanged. In conclusion, a lower dose of resveratrol compared to the dosage used by other studies reduced necrosis and gene expression of inflammatory cell markers in dystrophic muscle suggesting it as a therapeutic candidate for treating DMD.

Ultraviolet-visible (UV-Vis) absorption spectra are routinely collected as part of high-performance liquid chromatography (HPLC) analysis systems and can be used to identify chemical reaction products by comparison to reference spectra. Here, we present UV-adVISor as a new computational tool for predicting UV-Vis spectra from a molecule’s structure alone. UV-Vis prediction was approached as a sequence-to-sequence problem. We utilized Long-Short Term Memory and attention-based neural networks with Extended Connectivity Fingerprint diameter 6 or molecule SMILES to generate predictive models for UV-spectra. We have produced two spectrum datasets (Dataset I, N = 949 and Dataset II, N = 2222) using different compound collections and spectrum acquisition methods to train, validate, and test our models. We evaluated the prediction accuracy of the complete spectra by the correspondence of wavelengths of absorbance maxima and with a series of statistical measures (the best test set median model parameters are in parentheses for Model II), including RMSE (0.064), R2 (0.71), and dynamic time warping (DTW, 0.194) of the entire spectrum curve. Scrambling molecule structures with experimental spectra during training resulted in a degraded R2, confirming the utility of the approaches for prediction. UV-adVISor is able to provide fast and accurate predictions for libraries of compounds.

The effect of thermal protein denaturation on the structure and quality of muscles of different fibre types is not well understood. Unaged masseter (100% type I fibres) and cutaneous trunci (93% type II fibres) muscles (N = 10) were assessed for their characteristics, protein denaturation, cooking loss, Warner- Bratzler shear force (WBSF) and shrinkage after heating at 50 °C - 85 °C with a rate of 5 °C/ min. Raw masseter had a higher pH, collagen and water content, shorter sarcomere, comparable fibre diameter, and shorter and wider fragments upon homogenization, than cutaneous trunci. In cutaneous trunci, at 55 °C - 60 °C, the lower transition temperature of myosin and the greater cumulative enthalpy resulted in greater cooking loss in muscle cuboids, and greater transverse, longitudinal and volume shrinkage in fibres and fibre fragments, than in masseter. Protein denaturation explained 71% variability in fibre fragment volume and 58% in cooking loss of both muscles, as well as 47% variability in WBSF of masseter.

Duchenne muscular dystrophy (DMD) is a lethal muscle wasting disorder caused by mutations in the DMD gene that leads to the absence or severe reduction of dystrophin protein in muscle. The mdx mouse, also dystrophin deficient, is the model most widely used to study the pathology and test potential therapies, but the phenotype is milder than human DMD. This limits the magnitude and range of histological damage parameters and molecular changes that can be measured in pre-clinical drug testing. We used 3 weeks of voluntary wheel running to exacerbate the mdx phenotype. In mdx mice, voluntary exercise increased the amount of damaged necrotic tissue and macrophage infiltration. Global gene expression profiling revealed that exercise induced additional and larger gene expression changes in mdx mice and the pathways most impacted by exercise were all related to immune function or cell-extracellular matrix (ECM) interactions. When we compared the matrisome and inflammation genes that were dysregulated in mdx with those commonly differentially expressed in DMD, we found the exercised mdx molecular signature more closely resembled that of DMD. These gene expression changes in the exercised mdx model thus provide more scope to assess the effects of pre-clinical treatments. Our gene profiling comparisons also highlighted upregulation of ECM proteins involved in innate immunity pathways, proteases that can release them, downstream receptors and signaling molecules in exercised mdx and DMD, suggesting that the ECM could be a major source of pro-inflammatory molecules that trigger and maintain the immune response in dystrophic muscle.

The changes in secondary structure of proteins with heating were characterised and compared for bovine masseter (fibre type I) and cutaneous trunci (fibre type II) muscles by Differential Scanning Calorimetry (DSC) and Fourier Transform InfraRed (FTIR) microspectroscopy. Heating led to a decrease in alpha helices, and an increase in aggregated strands, random coils and aromatic side chains in the muscle fibres of both muscles. In the intramuscular connective tissue (IMCT) of both muscles, a decrease in alpha helix, turn and unordered structures was complemented with an increase in aggregated strands. At temperatures < 60 degrees C, the greater thermal denaturation of proteins in cutaneous trunci than in masseter (FTIR), supported by a myosin associated peak at 55.8 degrees C for cutaneous trunci and no peak for masseter (DSC), indicates that myosin in type II fibres is more sensitive to thermal denaturation than myosin in type I fibres and this should be considered in thermal meat processing.

Background As myosin heavy chain (MyHC) profile of muscle fibres is heavily influenced by neural input, changes in MyHC expression are expected in horses clinically affected with recurrent laryngeal neuropathy (RLN) yet, this has not been thoroughly investigated. Objectives To describe the changes in MyHC and fibre diameter in left cricoarytenoideus dorsalis (L-CAD) muscle of horses with clinical signs of RLN. Study design Observational cohort study. Methods Immunohistochemistry was used to assess the MyHC-based fibre-type proportion, size and grouping in the L-CAD of 10 Thoroughbred horses, five clinically affected with RLN and five unaffected controls based on resting endoscopic examination. The Mann-Whitney U test was used to compare the two groups. Results Compared to controls (of mean age 3.0 +/- 1.7 years) which only expressed type I, IIA and IIX MyHC, the L-CAD of affected horses (of mean age 2.8 +/- 0.8 years) had obvious fibre-type grouping, and despite apparent compensatory hypertrophy of a small number of fibres, a decrease in overall fibre diameter (median difference -35.2 mu m, 95% CI -47.4 to -7.9, P = .02) and diameter of type IIA fibres (median difference -46.8 mu m, 95% CI -52.1 to -5.0, P = .03) was observed. Anti-fast MyHC (MY32) cross-immunoreacted with embryonic-MyHC. Whereas MY32-positive fibres were identified as type IIX in controls, in affected horses these fibres were less than 50 mu m diameter with internal nuclei and were MYH3-positive for embryonic myosin indicating depletion of type IIX fibres, yet active regeneration and fibre renewal. Main limitations Small sample size that did not include subclinical cases. Fibre size and appearance rather than staining colour were relied upon to differentiate embryonic from type IIX MyHC. Conclusions Horses clinically affected with RLN have overall atrophy of fibres, loss of IIX fibres and expression of embryonic myosin indicating regenerative capacity. Despite hypertrophy of some remaining fibres, the overall decline in the bulk of fibres, including those most fatigue-resistant, may be the critical change that results in failure to maintain arytenoid abduction during exercise although direct comparison to subclinical cases is needed to confirm this.

Arising from the ablation of the cytoskeletal protein dystrophin, Duchenne Muscular Dystrophy (DMD) is a debilitating and fatal skeletal muscle wasting disease underpinned by metabolic insufficiency. The inability to facilitate adequate energy production may impede calcium (Ca 2+ ) buffering within, and the regenerative capacity of, dystrophic muscle. Therefore, increasing the metabogenic potential could represent an effective treatment avenue. The aim of our study was to determine the efficacy of adenylosuccinic acid (ASA), a purine nucleotide cycle metabolite, to stimulate metabolism and buffer skeletal muscle damage in the mdx mouse model of DMD. Dystrophin-positive control (C57BL/10) and dystrophin-deficient mdx mice were treated with ASA (3000 µg.mL −1 ) in drinking water. Following the 8-week treatment period, metabolism, mitochondrial density, viability and superoxide (O 2 − ) production, as well as skeletal muscle histopathology, were assessed. ASA treatment significantly improved the histopathological features of murine DMD by reducing damage area, the number of centronucleated fibres, lipid accumulation, connective tissue infiltration and Ca 2+ content of mdx tibialis anterior. These effects were independent of upregulated utrophin expression in the tibialis anterior. ASA treatment also increased mitochondrial viability in mdx flexor digitorum brevis fibres and concomitantly reduced O 2 − production, an effect that was also observed in cultured immortalised human DMD myoblasts. Our data indicates that ASA has a protective effect on mdx skeletal muscles.

Automated peptide and oligonucleotide synthesizers enabled a revolution in molecular biology and helped pave the way to modern synthetic biology. Similarly, fully automated synthetic chemistry could herald a new wave of innovation in biology and materials sciences by greatly facilitating access to known and novel molecules. Here, we report on an automated multistep chemical synthesizer, AutoSyn, that makes milligram-to-gram-scale amounts of virtually any drug-like small molecule in a matter of hours and demonstrate its versatility with the synthesis of ten known drugs. Of the FDA-approved small-molecule drugs for which we were able to compute a synthetic route, 87% are predicted to be synthesizable on AutoSyn. Moreover, AutoSyn enables digital synthesis protocols that ensure the reproducibility and transferability of synthesis protocols from one lab to another.

This study examines the effects of ageing (1, 14 days), cathepsin inhibition (No or Yes) and temperature (25-90 degrees C) on the shrinkage of fibre fragments from three bovine muscles (semitendinosus, biceps femoris and psoas major) during heating. Shrinkage was quantified using light microscopy images. Muscle fibres (except in psoas major) had greater transverse shrinkage, and less longitudinal shrinkage in aged than in unaged muscles at temperatures >= 60-75 degrees C. In addition, cathepsin inhibition during heating at >= 65-90 degrees C caused greater transverse shrinkage in semitendinosus fibres, and reduced longitudinal shrinkage for all muscles. At temperatures >= 75 degrees C, the longitudinal and transverse shrinkage of the fibres was correlated for all muscles. Ageing of biceps femoris increases volume shrinkage on a fibre level, and hence potentially cooking loss, while cathepsin activity in the semitendinosus reduces volume shrinkage. In conclusion, cathepsin activity and ageing influence the shrinkage that occurs during heating and these factors should be explored further to enable optimisation of thermal meat processing.

Meat shrinks and assumes an irregular shape during heating due to the varying distribution of connective tissue and extracellular spaces. The terrestrial 3D laser scanning technology is proposed as an alternative method to manual measurements to estimate the volume of irregularly shaped meat cuboids and to predict the cooking loss based on the heating induced volume shrinkage. Cuboids from aged pork loins (longissimus lumborum, n = 12) were heated at 50, 60, 70 or 80 °C for 30 min. Two methods of 3D reconstruction and volume estimation of the pork cuboids by laser scanning were used; without a base scan (laser-B) and with inclusion of a base scan (laser+B), as well as two methods based on manual caliper measurements of all twelve edges (caliper-12) or of three edges in each direction (caliper-3). Both laser scanning methods (Laser+B and Laser-B) resulted in greater volume estimates for the raw samples than the caliper-12 and caliper-3 measurements (38.3, 39.4 compared to 33.9, 34.8 cm3, respectively). Cooking loss across the different temperatures could be best predicted by the caliper-based perimeter shrinkage (r = 0.94, P < 0.001), followed by the caliper-based volume shrinkage estimates (r = 0.67 and 0.64 for caliper-12 and caliper-3, respectively, p < 0.001), while volume shrinkage measured by laser scanning had low (laser-B, r = 0.35, P < 0.05) or no correlation (laser+B, r = 0.17, P > 0.05) with the cooking loss. 3D laser scanning technologies can be considered by the food industry for 3D reconstruction and volume estimation, however improvements are needed in the data processing to allow for better predictability of meat quality attributes.

This study aimed to quantify the effect of muscle, ageing and cooking temperature on the texture, cooking loss and shrinkage of cooked beef. Cuboids from unaged (1 day post mortem) and aged (14 days post mortem) semitendinosus, biceps femoris and psoas major muscles, from both sides of five beef carcasses, were cooked at four different cooking temperatures (50, 60, 70 and 80 °C) for 30 min. and their Warner–Bratzler shear force (WBSF), cooking loss and shrinkage (longitudinal and transverse) were quantified. The WBSF was reduced by ageing in the muscles at the specific cooking temperatures: psoas major (cooked at 50, 60 and 80 °C), semitendinosus (70 and 80 °C) and biceps femoris (80 °C). The cooking loss was 3% greater in aged compared to unaged muscles. The longitudinal shrinkage was greatest in psoas major at 80 °C amongst the muscle types and it was reduced by ageing in psoas major (70 and 80 °C) and biceps femoris (80 °C). The transverse shrinkage was reduced by ageing only in biceps femoris, across all temperatures; and the diameter of homogenized fibre fragments from semitendinosus and biceps femoris was reduced more by cooking at 50 °C in unaged compared to aged condition. WBSF was related to transverse shrinkage, and cooking loss was related to longitudinal shrinkage. The effect of muscle type on the physical changes occurring during cooking of beef is dependent on ageing and cooking temperature.

We carried out laboratory experiments to study the decomposition of carbon fibers at high temperature and studied the material used in charring carbon phenolic ablators for planetary probe heatshields. Porous plug samples were exposed to controlled rates of molecular oxygen, carbon dioxide, and carbon monoxide at furnace-heated flow-tube reactor temperatures between 700 and 1500 K and pressures between 2000 and 6200 Pa. We used calibrated mass spectroscopy to make time-resolved quantitative measurements of decomposition products of the gases downstream of the test sample. Absolute and differential pressure measurements were used to determine the high-temperature permeability of the material and to monitor changes during material decomposition, and we characterized the microstructure of the porous sample using scanning electron microscopy. We found substantial carbon fiber oxidation for O-2 flows, resulting in increasing CO/CO2 production ratios with increasing temperature. The CO2 was shown to react with carbon fibers following a Boudouard reaction at temperatures above 1200 K. (C) 2019 Elsevier Ltd. All rights reserved.