LC-MS-based metabolomics reveals metabolite dynamic changes of beef after superchilling early post-mortem.

To explore the underlying mechanisms by which superchilling (SC, -3 °C within 5 h of slaughter) improves beef tenderness, an untargeted metabolomics strategy was employed. M. Longissimus lumborum (LL) muscles from twelve beef carcasses were assigned to either SC or very fast chilling (VFC, 0 °C within 5 h of slaughter) treatments, with conventional chilling (CC, 0 âˆ¼ 4 °C until 24 h post-mortem) serving as the control (6 per group). Biochemical properties and metabolites were investigated during the early post-mortem period. The results showed that the degradation of µ-calpain and caspase 3 occurred earlier in SC treated sample, which might be attributed to the accelerated accumulation of free Ca2+. The metabolomic profiles of samples from the SC and CC treatments were clearly distinguished based on partial least squares-discriminant analysis (PLS-DA) at each time point. It is noteworthy that more IMP and 4-hydroxyproline were found in the comparison between SC and CC treatments. According to the results of metabolic pathways analysis and the correlation analysis between traits related to tenderness and metabolites with significant differences (SC vs. CC), it can be suggested that the tenderization effect of the SC treatment may be related to the alteration of arginine and proline metabolism, and purine metabolism in the early post-mortem phase.

Identifying the Potential Apoptotic Metabolites in Postmortem Beef Muscle by Targeted Metabolomics.

Apoptotic cells may release specific metabolites to act as messengers during the apoptotic process. This study represents the first attempt to identify potential apoptotic metabolites in postmortem muscle. Ninety potential apoptotic metabolites in beef were selected and analyzed through targeted metabolomics, with 84 of them exhibiting significant differences over the postmortem time. Following the addition of the mitochondria-targeted antiapoptotic agent mitoquinone to postmortem muscle, metabolomic analysis revealed that 73 apoptotic metabolites still underwent significant changes, even against the backdrop of altered apoptosis. Of these 73 apoptotic metabolites, 54 exhibited similar trends at various treatment times with adding mitoquinone, including lipids (6), amino acids (27), nucleosides (11), and carbohydrate and energy metabolism (10). Mitoquinone significantly reduced the levels of most apoptotic metabolites, and inhibition of apoptosis resulted in a significant decrease in the levels of numerous apoptotic metabolites. Consequently, these apoptotic metabolites are considered complementary to apoptosis in postmortem muscle, with their increased levels potentially promoting apoptosis. Noteworthy apoptotic metabolites, such as glycerol 3-phosphate, serine, AMP, ATP, GMP, and creatine, were identified as active signaling molecules that attract and recruit phagocytes during apoptosis, assisting in recognizing apoptotic cells by phagocytes. This study provides, for the first time, insights into potential apoptotic metabolites in postmortem muscle, contributing to a better understanding of meat biochemistry.

A Solvent-Templated Porous Liquid Crystal Elastomer with Tactile Sensation beyond Reversible Actuation toward Versatile Artificial Muscles.

Liquid crystal elastomers (LCEs), as a classical two-way shape-memory material, are good candidates for developing artificial muscles that mimic the contraction, expansion, or rotational behavior of natural muscles. However, biomimicry is currently focused more on the actuation functions of natural muscles dominated by muscle fibers, whereas the tactile sensing functions that are dominated by neuronal receptors and synapses have not been well captured. Very few studies have reported the sensing concept for LCEs, but the signals were still donated by macroscopic actuation, that is, variations in angle or length. Herein, we develop a conductive porous LCE (CPLCE) using a solvent (dimethyl sulfoxide (DMSO))-templated photo-cross-linking strategy, followed by carbon nanotube (CNT) incorporation. The CPLCE has excellent reversible contraction/elongation behavior in a manner similar to the actuation functions of skeletal muscles. Moreover, the CPLCE shows excellent pressure-sensing performance by providing real-time electrical signals and is capable of microtouch sensing, which is very similar to natural tactile sensing. Furthermore, macroscopic actuation and tactile sensation can be integrated into a single system. Proof-of-concept studies reveal that the CPLCE-based artificial muscle is sensitive to external touch while maintaining its excellent actuation performance. The CPLCE with tactile sensation beyond reversible actuation is expected to benefit the development of versatile artificial muscles and intelligent robots.

Isolation of Nuclei from Human Intermuscular Adipose Tissue and Downstream Single-Nuclei RNA Sequencing.

Intermuscular adipose tissue (IMAT) is a relatively understudied adipose depot located between muscle fibers. IMAT content increases with age and BMI and is associated with metabolic and muscle degenerative diseases; however, an understanding of the biological properties of IMAT and its interplay with the surrounding muscle fibers is severely lacking. In recent years, single-cell and nuclei RNA sequencing have provided us with cell type-specific atlases of several human tissues. However, the cellular composition of human IMAT remains largely unexplored due to the inherent challenges of its accessibility from biopsy collection in humans. In addition to the limited amount of tissue collected, the processing of human IMAT is complicated due to its proximity to skeletal muscle tissue and fascia. The lipid-laden nature of the adipocytes makes it incompatible with single-cell isolation. Hence, single nuclei RNA sequencing is optimal for obtaining high-dimensional transcriptomics at single-cell resolution and provides the potential to uncover the biology of this depot, including the exact cellular composition of IMAT. Here, we present a detailed protocol for nuclei isolation and library preparation of frozen human IMAT for single nuclei RNA sequencing. This protocol allows for the profiling of thousands of nuclei using a droplet-based approach, thus providing the capacity to detect rare and low-abundant cell types.

Changes in the amount of nucleotide sugars in aged mouse tissues.

Aging affects tissue glycan profiles, which may alter cellular functions and increase the risk of age-related diseases. Glycans are biosynthesized by glycosyltransferases using the corresponding nucleotide sugar, and the availability of nucleotide sugars affects glycosylation efficiency. However, the effects of aging on nucleotide sugar profiles and contents are yet to be elucidated. Therefore, this study aimed to investigate the effects of aging on nucleotide sugars using a new LC-MS/MS method. Specifically, the new method was used to determine the nucleotide sugar contents of various tissues (brain, liver, heart, skeletal muscle, kidney, lung, and colon) of male C57BL/6NCr mice (7- or 26-month-old). Characteristic age-associated nucleotide sugar changes were observed in each tissue sample. Particularly, there was a significant decrease in UDP-glucuronic acid content in the kidney of aged mice and a decrease in the contents of several nucleotide sugars, including UDP-N-acetylgalactosamine, in the brain of aged mice. Additionally, there were variations in nucleotide sugar profiles among the tissues examined regardless of the age. The kidneys had the highest concentration of UDP-glucuronic acid among the seven tissues. In contrast, the skeletal muscle had the lowest concentration of total nucleotide sugars among the tissues; however, CMP-N-acetylneuraminic acid and CDP-ribitol were relatively enriched. Conclusively, these findings may contribute to the understanding of the roles of glycans in tissue aging.

The Meat Standards Australia carcass grading site affects assessment of marbling and prediction of meat-eating quality in growing European beef cattle.

The lack of consumer feedback on beef eating quality contributes to reduced beef consumption in Europe. The Meat Standards Australia (MSA) grading scheme can assess the palatability of beef carcasses usually graded at the 10th thoracic vertebrae. However, the European beef industry relies on late-maturing breeds usually cut at the 5th vertebrae due to commercial reasons. Data from 55 young bulls and heifers of late-maturing breeds were collected in an Italian slaughterhouse following the MSA guidelines at both carcass grading sites and sides. Intramuscular fat levels were assessed through two scores and used with other variables to feed the MSA model, which predicts the MSA index, the meat-eating quality scores (MQ4) for 5 muscles and for each carcass grading site × side combination. The scores were analyzed using a mixed linear model. A correlation analysis was conducted to predict the variables measured at the 10th site using their correspondent at the 5th carcass grading site. A stepwise regression was conducted to understand the weight of each measured variable on marbling and MQ4 scores measured both at 5th and 10th carcass grading sites. Results showed significantly higher value for the studied traits at the 5th carcass grading site, while carcass side had no significant impact. The equations had high predictive capability and MSA marbling score played a key role in explaining the variability across carcass grading sites. The differences in marbling and MQ4 scores between the carcass grading sites suggest considering this factor if the MSA grading system will be applied to Europe.

Beef-on-dairy: Meat quality of veal and prediction of intramuscular fat using the Q-FOM™ Beef camera at the 5th-6th thoracic vertebra.

This study aims to describe the meat quality of young Holstein (HOL) beef-on-dairy heifers and bulls sired by Angus (ANG, n = 109), Charolais (CHA, n = 101) and Danish Blue (DBL, n = 127), and to investigate the performance of the handheld vision-based Q-FOM™ Beef camera in predicting the intramuscular fat concentration (IMF%) in M. longissimus thoracis from carcasses quartered at the 5th-6th thoracic vertebra. The results showed significant differences between crossbreeds and sexes on carcass characteristics and meat quality. DBL × HOL had the highest EUROP conformation scores, whereas ANG × HOL had darker meat with higher IMF% (3.52%) compared to CHA × HOL (2.99%) and DBL × HOL (2.51%). Bulls had higher EUROP conformation scores than heifers, and heifers had higher IMF% (3.70%) than bulls (2.31%). These findings indicate the potential for producing high-quality meat from beef-on-dairy heifers and ANG bulls. The IMF% prediction model for Q-FOM performed well with R2 = 0.91 and root mean squared error of cross validation, RMSECV = 1.33%. The performance of the prediction model on the beef-on-dairy veal subsample ranging from 0.9 to 7.4% IMF had lower accuracy (R2 = 0.48) and the prediction error (RMSEveal) was 1.00%. When grouping beef-on-dairy veal carcasses into three IMF% classes (2.5% IMF bins), 62.6% of the carcasses were accurately predicted. Furthermore, Q-FOM IMF% predictions and chemically determined IMF% were similar for each combination of sex and crossbreed, revealing a potential of Q-FOM IMF% predictions to be used in breeding, when aiming for higher meat quality.

Effect of pulsed electric field (PEF) on NaCl diffusion in beef and consequence on meat quality.

The impact of various field strength (2, 3, 4 kV/cm) and treatment time (60s and 90s) combinations on NaCl content and diffusion coefficient of beef were evaluated in the current study. Weight change, water content, water holding capacity, and texture of beef after brining were also explored. The results demonstrated pulsed electric field (PEF) pre-treatment significantly increased NaCl uptake when the brining time was 150 min (P < 0.05). The maximum NaCl content increased by 19.50% and the diffusion coefficient increased by 58.50%. Relatively mild PEF (60s) could improve beef qualities, but longer treatment time (90s) was detrimental to these qualities. Meanwhile, more complete myofibrillar structure and lower lipid oxidation extent were observed in the samples treated by PEF, contributing to the higher a* values. In conclusion, short processing time (60s) and high field strength (4 kV/cm) treatment is a potential strategy for meat brining acceleration and quality improvement in practical industrial production.

Effect of a novel steak fabrication method by trimming subcutaneous and intermuscular fats on palatability and calorie content of beef ribeye steaks.

This study investigated the effects of a novel steak fabrication method, involving removal of both subcutaneous and intermuscular fats between the longissimus thoracis (LT) and spinalis dorsi (SD) muscles before cooking, on beef ribeye steak palatability and calorie content. Canada AA (n = 10) and AAA (n = 10) ribeyes were fabricated into steaks either with (fat-on) or without (fat-off) subcutaneous and intermuscular fats. Fat-on steaks had shorter cooking times and lower cooking losses than fat-off steaks (P < 0.001), regardless of quality grade. There were treatment × quality grade interactions for initial (P < 0.01) and sustainable juiciness (P < 0.05) of the LT samples and initial juiciness (P < 0.05) of the SD samples, with the fat-on AA samples being more juicy than the fat-off AA samples, but the AAA treatments were not different from each other. Regardless of quality grade, fewer panelist responses indicated livery flavour (P < 0.05) for fat-on compared to fat-off LT samples. Regardless of fat-on/fat-off treatment, AA compared to AAA samples had more responses for bloody/serumy (P < 0.01) and unidentified off-flavours (P < 0.05) in LT samples. However, AAA samples had more responses for metallic off-flavour in LT (P = 0.059) and SD (P < 0.05). There were no differences in calorie content between fat-on compared to fat-off steaks (P > 0.1) regardless of quality grade and muscle type, or between AA and AAA steaks regardless of cooking with fat-on or off (P > 0.1). Maintaining the subcutaneous and intermuscular fats while cooking will improve ribeye steak palatability without increasing calorie content, especially for leaner steaks.

A deep learning-based approach for fully automated segmentation and quantitative analysis of muscle fibers in pig skeletal muscle.

Muscle fiber properties exert a significant influence on pork quality, with cross-sectional area (CSA) being a crucial parameter closely associated with various meat quality indicators, such as shear force. Effectively identifying and segmenting muscle fibers in a robust manner constitutes a vital initial step in determining CSA. This step is highly intricate and time-consuming, necessitating an accurate and automated analytical approach. One limitation of existing methods is their tendency to perform well on high signal-to-noise ratio images of intact, healthy muscle fibers but their lack of validation on more complex image datasets featuring significant morphological changes, such as the presence of ice crystals. In this study, we undertake the fully automatic segmentation of muscle fiber microscopic images stained with myosin adenosine triphosphate (mATPase) activity using a deep learning architecture known as SOLOv2. Our objective is to efficiently derive accurate measurements of muscle fiber size and distribution. Tests conducted on actual images demonstrate that our method adeptly handles the intricate task of muscle fiber segmentation, yielding quantitative results amenable to statistical analysis and displaying reliability comparable to manual analysis.

Effect of accelerated aging on shelf-stability, product loss, sensory and biochemical characteristics in 2 lower quality beef cuts.

The aim of this study was to determine the impact of accelerated aging (AA) on shelf stability, product loss, sensory and biochemical characteristics of 2 lower quality beef cuts. Triceps brachii (TB) and semimembranosus (SM) were collected and fabricated from 10 USDA Choice beef carcasses and assigned to 1 of 6 treatments: 3 d cooler aged (control), 21 d cooler aged, AA 49 °C for 2 h, AA 49 °C for 3 h, AA 54 °C for 2 h, and AA 54 °C for 3 h. The results showed that AA can decrease APC counts on steak surface and in purge and redness, but increase lightness and product loss of the steaks (P < 0.01). Lower shear force was also found for AA steaks compared to those from the control (P < 0.01), with the AA 54 °C treatments being comparable to 21 d cooler aging. However, the trained sensory panel determined AA steaks were less juicy and flavorful than those from the control and 21 d cooler aged samples (P < 0.05). There was no off-flavor detected in AA steaks though lipid oxidation was higher in AA samples than those in the control steaks (P < 0.01). The AA treatments stimulated cathepsin activity (P < 0.05), which may have enhanced the solubilization of stromal proteins and led to a different troponin-T degradation pattern compared to those from the 21 d aged samples (P < 0.01). Although AA is an economical and time-efficient method to increase tenderness of lower-quality beef cuts, further research is needed to determine strategies to mitigate the decrease in juiciness from AA treatments.

Comparative evaluation of different modalities for measuring in vivo carnosine levels.

Carnosine is an endogenous di-peptide (ß-alanine -L- histidine) involved in maintaining tissue homeostasis. It is most abundant in skeletal muscle where its concentration has been determined in biopsy samples using tandem mass spectrometry (MS-MS). Carnosine levels can also be assessed in intact leg muscles by proton magnetic resonance spectroscopy (1H-MRS) or in blood and urine samples using mass spectrometry. Nevertheless, it remains uncertain how carnosine levels from these distinct compartments are correlated with each other when measured in the same individual. Furthermore, it is unclear which measurement modality might be most suitable for large-scale clinical studies. Hence, in 31 healthy volunteers, we assessed carnosine levels in skeletal muscle, via 1H-MRS, and in erythrocytes and urine by MS-MS. While muscle carnosine levels were higher in males (C2 peak, p = 0.010; C4 peak, p = 0.018), there was no sex-associated difference in urinary (p = 0.433) or erythrocyte (p = 0.858) levels. In a linear regression model adjusted for age, sex, race, and diet, there was a positive association between erythrocyte and urinary carnosine. However, no association was observed between 1H-MRS and erythrocytes or urinary measures. In the relationship between muscle versus urinary and erythrocyte measures, females had a positive association, while males did not show any association. We also found that 1H-MRS measures were highly sensitive to location of measurement. Thus, it is uncertain whether 1H-MRS can accurately and reliably predict endogenous carnosine levels. In contrast, urinary and erythrocyte carnosine measures may be stable and in greater synchrony, and given financial and logistical concerns, may be a feasible alternative for large-scale clinical studies.

The impact of ultrasound treatment on glycolytic enzymes when applied to crude extracts from early post-mortem bovine muscle.

The rate of pH decline post - mortem and its interaction with temperature influences the final tenderness of meat, and therefore, the manipulation of the rate of pH decline is a strategy of interest in order to obtain consistent high quality meat. Ultrasound is a potential early post - mortem carcass intervention, which may alter the rate of glycolysis based on its ability to alter enzyme activity. In this study, homogenates (prepared from early post-mortem Longissimus thoracis et lumborum muscle) were subjected to different ultrasound intensities (0 %/60 %/100 % amp) and treatment durations (15/ 30 min). The effect of these treatments on the inherent activity of the glycolytic enzymes was investigated using an in vitro glycolytic buffer model system. It was found that ultrasound treatment intensity and duration had a significant interactive effect on the rate of pH decline, and on reducing sugars and lactic acid concentrations, specifically following the 100 % amp ultrasound for 30 min treatment and between 30 and 240 min incubation. No significant differences in pH or metabolites content were observed between treatments after 1440 min of incubation. No effect of ultrasound intensity or treatment duration was observed on the degradation of glycogen. Under the reported conditions of this trial, it can be concluded that the application of ultrasound has limited potential to have an impact on the glycolytic pathways in bovine muscle.

Exploration of the shelf-life difference between chilled beef and pork with similar initial levels of bacterial contamination.

This study compared the shelf-life of beef and pork longissimus lumborum muscles (loins) that had the same initial bacterial loads and were held under the same chilled storage conditions. To identify the underlying pathways, comparisons were conducted from the perspective of the spoilage indicators; protease/lipase activity, and the volatile organic compounds (VOC) generated over 28 d of chilled storage. The initial total viable microbial count (TVC) on Day 0 for both type of meat was 4.3 log10 CFU/g. It was found that the TVC of beef and pork did not differ throughout the total chilled storage period and both ultimately exceeded 7 log10 CFU/g after 28 d. Based on total volatile basic nitrogen (TVB-N) guidelines, pork was spoilt after 21 d of chilled storage and therefore 7 d earlier than beef. Changes in the concentration of VOC spoilage biomarkers, including 1-octen-3-ol, 1-octanol, nonanal, and others, confirmed that pork had a shorter shelf-life than beef. An important reason for the difference in shelf-life between the two types of meat was that pork had a higher protease activity, although the beef had higher levels of total lipase activity. These findings help us understand the differences in the spoilage process of raw meat from different species and explore specific measures to control the spoilage of beef or pork.

Pork belly quality variation and its association with fatness level.

Belly is a very popular pork cut composed of different layers of fat and muscle tissue. This work aims to investigate the effect of belly fatness on the morphological, mechanical (firmness) and compositional characteristics of fresh pork bellies and the distribution of the fat within the belly slice. A total of 182 bellies, selected to ensure variability of fatness, sexes and genotypes, were scanned by computed tomography (CT) to determine the fat content which, together with the genotype, led to the formation of 5 classes: F1 class below 26%, F2 class from 26% to 33.9%, and F3 class above 33.9% of fat content from common commercial crossbred pigs; F4 class with an average fatness of 47.3% from pure Duroc pigs; and last, F5 class with 62.6% average fat content from Iberian×Duroc pigs. The distribution of the fat in the central belly slice obtained by CT revealed important differences by region although the fat content was proportional to the overall fatness of the belly. Both belly weight and belly firmness increased with higher fatness. In bellies from common commercial pigs, an increase of SFA and MUFA and a decrease of PUFA as fatness increased was observed. This study highlights variations in belly characteristics among different fat classes, indicating considerable differences in the quality of bellies currently available in the market. This may influence producers and consumers acceptability such that fat content could be considered as a quality criterion to pre-classify bellies and better match the raw product with its final destination.

Effect of different levels of Lactobacillus added to diets on fat deposition and meat quality of Sunit lambs.

In this study, we examined the impact of adding Lactobacillus to the diet on fat distribution and meat quality of Sunit lambs. For 90 days, twenty-four lambs (19.31 ± 0.47 kg) were fed diets that contained 0 (NP), 6 (P1), 12 (P2), or 24 (P3) g of Lactobacillus casei/d. The results suggested that dietary supplementation with Lactobacillus decreased serum triglyceride in Sunit lambs (P < 0.001). The loin muscle area displayed notable increases in the P1 group (P < 0.05). Meanwhile, tail and visceral fat deposition of lambs were reduced when Lactobacillus was added to the diet (P < 0.05). Compared with the NP group, the values of shear force and cooking loss of in the P1 group exhibited a significant reduction, and intramuscular fat content increased significantly (P < 0.05). Additionally, the P1 group showed an increase in polyunsaturated fatty acids and a decrease in saturated fatty acids in the longissimus thoracis and biceps femoris muscles (P < 0.05). The P1 group showed downregulation of protein kinase AMP-activated catalytic subunit alpha 2 (AMPKα2) and carnitine palmitoyltransferase 1B expression in the longissimus thoracis muscle (P < 0.05). However, there was an upregulation of acetyl-CoA carboxylase (ACC), sterol regulatory element binding transcription factor 1 (SREBF1), and fatty acid synthase (FASN) expression (P < 0.05). In conclusion, feeding Sunit lambs 6 g/d of Lactobacillus as a dietary supplement may be a valuable way to improve fat distribution and meat quality. The AMPK/ACC and AMPK/SREBF1/FASN signaling pathways may be involved in this outcome.

Comparing the effects of packaging normal-pH and atypical dark-cutting beef in modified atmosphere conditions on surface color.

Limited studies have determined the effects of modified atmospheric packaging (MAP) on atypical dark-cutting beef surface color. The objective was to compare the impacts of using vacuum packaging, carbon monoxide (CO-MAP), and HiOx-MAP (high­oxygen) on the retail color of normal-pH and atypical dark-cutting beef aged 14 d. Atypical dark-cutting beef (pH 5.63) had numerically greater (P > 0.05) pH than normal-pH beef (pH 5.56). Atypical dark-cutting steaks were darker in color (lower L* values; P < 0.05) than normal-pH steaks. Atypical dark-cutting steaks had greater (P < 0.05) oxygen consumption, lower (P < 0.05) relative oxygenation, and less inter-muscle bundle space (P < 0.05) than normal-pH steaks. There were no differences (P > 0.05) in redness between normal-pH and atypical dark-cutting steaks when packaged in HiOx-MAP. Although a minimal increase in pH was observed in atypical dark-cutting beef, steaks in CO-MAP had lower redness than normal-pH steaks.

Dietary iron or inulin supplementation alters iron status, growth performance, intramuscular fat and meat quality in finisher pigs.

Forty LW × L pigs (20 boars and 20 gilts) (51.1 ± 0.41 kg) were allocated to a 2 × 2 × 2 factorial design with the respective factors being supplemental organic iron (Fe, 0 and 500 mg/kg), inulin (In, 0 and 50 g/kg) and sex (boars and gilts). After 5 weeks the animals were transported to an abattoir before slaughter and collection of samples. Serum iron was increased by supplemental Fe (28.4 v. 30.9 µmol/L, P = 0.05), although there was an interaction (P = 0.03) such that pigs fed diets with In had lower serum Fe concentrations than those without In (26.8 v. 32.3 µmol/L). Boars had lower (P < 0.01) haemoglobin (116 vs 125), haematocrit (36.7 v. 39.7%) and erythrocyte (6.6 v. 7.1 × 106/mL) concentrations than gilts. Dietary In increased liveweight gain (795 v. 869 g/d, P < 0.02) and carcass weight (62.9 v. 65.2 kg, P < 0.02). Dietary Fe or In supplementation did not improve muscle Longissimus thoracis et lumborum (LTL) total Fe concentration (P > 0.05). Muscle non-heme Fe concentration was higher in Fe-supplemented pigs (P < 0.04) and gilts (P < 0.05) than their counterparts. Muscle heme Fe concentration was greater (3.04 vs 2.51, P < 0.05) in boars than in gilts. The LTL marbling score was greater (P < 0.01) for In-supplemented pigs, and the response was more notable when Fe and In were fed together. These data show that dietary supplementation of Fe increased serum Fe and muscle non-heme Fe concentrations. Supplementation of In at 5% in the diet of finisher pigs improved liveweight gain and the marbling score of pork.

Conformational fingerprinting with Raman spectroscopy reveals protein structure as a translational biomarker of muscle pathology.

Neuromuscular disorders are a group of conditions that can result in weakness of skeletal muscles. Examples include fatal diseases such as amyotrophic lateral sclerosis and conditions associated with high morbidity such as myopathies (muscle diseases). Many of these disorders are known to have abnormal protein folding and protein aggregates. Thus, easy to apply methods for the detection of such changes may prove useful diagnostic biomarkers. Raman spectroscopy has shown early promise in the detection of muscle pathology in neuromuscular disorders and is well suited to characterising the conformational profiles relating to protein secondary structure. In this work, we assess if Raman spectroscopy can detect differences in protein structure in muscle in the setting of neuromuscular disease. We utilise in vivo Raman spectroscopy measurements from preclinical models of amyotrophic lateral sclerosis and the myopathy Duchenne muscular dystrophy, together with ex vivo measurements of human muscle samples from individuals with and without myopathy. Using quantitative conformation profiling and matrix factorisation we demonstrate that quantitative 'conformational fingerprinting' can be used to identify changes in protein folding in muscle. Notably, myopathic conditions in both preclinical models and human samples manifested a significant reduction in α-helix structures, with concomitant increases in ß-sheet and, to a lesser extent, nonregular configurations. Spectral patterns derived through non-negative matrix factorisation were able to identify myopathy with a high accuracy (79% in mouse, 78% in human tissue). This work demonstrates the potential of conformational fingerprinting as an interpretable biomarker for neuromuscular disorders.

Comprehensive lipidomic analysis revealed the effects of fermented Morus alba L. intake on lipid profile in backfat and muscle tissue of Yuxi black pigs.

Mulberry leaf is a widely used protein feed and is often used as a strategy to reduce feed costs and improve meat quality in the livestock industry. However, to date, there is a lack of research on the improvement of meat quality using mulberry leaves, and the exact mechanisms are not yet known. The results showed that fermented mulberry leaves significantly reduced backfat content but had no significant effect on intramuscular fat (IMF). Lipidomic analysis showed that 98 and 303 differential lipid molecules (p < 0.05) were identified in adipose and muscle tissues, respectively, including triglycerides (TG), phosphatidylcholine, phosphatidylethanolamine, sphingolipids, and especially TG; therefore, we analysed the acyl carbon atom number of TG. The statistical results of acyl with different carbon atom numbers of TG in adipose tissue showed that the acyl group containing 13 carbon atoms (C13) in TG was significantly upregulated, whereas C15, C16, C17, and C23 were significantly downregulated, whereas in muscle tissue, the C12, C19, C23, C25, and C26 in TG were significantly downregulated. Acyl changes in TG were different for different numbers of carbon atoms in different tissues. We found that the correlations of C (14-18) in adipose tissue were higher, but in muscle tissue, the correlations of C (18-26) were higher. Through pathway enrichment analysis, we identified six and four metabolic pathways with the highest contributions of differential lipid metabolites in adipose and muscle tissues respectively. These findings suggest that fermented mulberry leaves improve meat quality mainly by inhibiting TG deposition by downregulating medium- and short-chain fatty acids in backfat tissue and long-chain fatty acids in muscle tissue.