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.

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.

Development of Mass Spectrometry Imaging on skeletal muscle to characterize the local pro-inflammatory and pro-resolution lipid responses in a vaccination context.

Vaccine reactogenicity is well documented at the clinical level but the mechanism involved at the local or systemic level are still poorly understood. Muscular tissue where most vaccines are administered is the first place of interaction between the vaccine formulation and the host's immune cells. So far, this site of vaccine administration is not well documented from a mechanistic standpoint. The study of early molecular events at the injection site is crucial to understand the local response to vaccines. In this paper, we report a standardized workflow, from the injection of vaccine formulations in rabbit muscle, to the analysis by desorption electrospray ionization and histology staining to understand the role of lipids involved in the inflammation and its resolution on striated muscular tissue. The analysis of lipid mediators was optimized at the site of needle insertion to allow the spatial comparison of cellular infiltrates at the injection site. We showed that lipids were distributed across the spatial tissue morphology in a time-dependent manner. The MS imaging applied to vaccinology could pave the way to a better understanding of vaccine reactogenicity and mechanism of action.

Effects of sex on meat quality traits, amino acid and fatty acid compositions, and plasma metabolome profiles in White King squabs.

The objective of this study was to investigate the effects of sex on meat quality and the composition of amino and fatty acids in the breast muscles of White King pigeon squabs. Untargeted metabolomics was also conducted to distinguish the metabolic composition of plasma in different sexes. Compared with male squabs, female squabs had greater intramuscular fat (IMF) deposition and lower myofiber diameter and hydroxyproline content, leading to a lower shear force. Female squabs also had higher monounsaturated fatty acid and lower n-6 and n-3 polyunsaturated fatty acid proportions in the breast muscle, and had greater lipogenesis capacity via upregulation of PPARγ, FAS and LPL gene expression. Moreover, female squabs had lower inosine 5'-monophosphate, essential, free and sweet-tasting amino acid contents. Furthermore, Spearman's correlations between the differential plasma metabolites and key meat parameters were assessed, and putrescine, N-acetylglutamic acid, phophatidylcholine (18:0/P-16:0) and trimethylamine N-oxide were found to contribute to meat quality. In summary, the breast meat of male squabs may have better nutritional value than that of females, but it may inferior in terms of sensory properties, which can be attributed to the lower IMF content and higher shear force value. Our findings enhance our understanding of sex variation in squab meat quality, providing a basis for future research on pigeon breeding.

Effect of housing system on carcass composition, meat quality, digestive morphometry, and leg bone dimensions of Ross 308 parent broilers.

The aim of the present study was to compare the effects of 2 Ross 308 parent broiler housing systems (SLS-slat-litter system vs. LS-litter-based system) in terms of carcass composition, meat quality traits (chemical composition, texture, physicochemical properties), as well as biometric traits of the digestive system and leg bones. The weight of the eviscerated carcass and the proportion of carcass components were determined at the end of the reproductive period (60 wk of life) following slaughter. The lengths and diameters of the individual intestinal segments, the weight of selected internal organs, the acidity (pH24) and electrical conductivity (EC24), as well as the color (L*, a*, b*) of breast and thigh muscles were assessed. The basal chemical composition of the breast and thigh muscles was also determined, texture analysis of the pectoralis major muscle and measurements of the femur and tibia of parent broilers were also carried out. The housing system differentiated the birds in terms of percentage of breast muscle (SLS-27.4% vs. LS-26.0%) and intramuscular fat content in the breast muscle (SLS-1.1% vs. LS-0.7%), spleen weight pH of the breast and thigh muscles and EC of the thigh muscles (SLS-9.3 mS/cm vs. LS-7.0 mS/cm). Differences were also found between the study groups in the color of the breast and thigh muscles. The housing system affected the results of the texture analysis of the pectoralis major muscle. The birds differed significantly (P < 0.05) in terms of gumminess (SLS-11.1 N vs. LS-16.0 N), springiness, chewiness (SLS-17.6 N × cm vs. LS-23.4 N × cm) and cohesiveness parameters. The housing system did not affect the lengths and diameters of the individual intestinal segments, except for the length of the terminal intestine. There was no significant effect of the housing system on the tibia and femur dimensions analyzed. This study provided information about differences in certain carcass characteristics, meat quality, and the digestive system of Ross 308 parent broilers in relation to the maintenance system.

Effect of packaging thickness and muscle type on ultrasound-assisted beef quality.

High-intensity ultrasound (HIU) can modify muscle structure, leading to improvements in tenderness. However, factors such as packing type and muscle complexity may attenuate the acoustic cavitation. In this research, the effect of packing thickness (40.6-70 µm) on the quality of bovine Gluteus medius and Biceps femoris treated with HIU (37 kHz, 90 W/cm2, 40 min) was evaluated. The hardness of G. medius decreased significantly as the thickness of the packing bag decreased. The wide interfibrillar and intermyofibrillar spaces corroborated the tenderizing effect. These effects are related to damage of cell structure and changes in the collagen content (3.37 ± 0.1 µg/mL). In addition, the HIU decrease the variability in the water holding capacity of the muscle produced by the use of low thickness bags during storage. The trained sensory panel described the sonicated samples in 50.8 µm bags as less hard and juicier. Contrarily, in B. femoris no significant effects were reported in the variables evaluated. B. femoris is a white muscle, with a high amount of collagen (3.59 ± 0.1 µg/mL) and little intramuscular fat. Consequently, the effect of the HIU on muscle quality is associated with the composition of the muscle fibers and the thickness of the packing bag. HIU application is recommended to improve the quality of leg muscles whenever low-thickness bags (50.8 µm or less) are used.

Novel needle-probe single-fiber reflectance spectroscopy to quantify sub-surface myoglobin forms in beef psoas major steaks during retail display.

Meat discoloration starts at the interface between the bright red oxymyoglobin layer and the interior deoxymyoglobin layer. Currently, limited tools are available to characterize myoglobin forms formed within the sub-surface of meat. The objective was to demonstrate a needle-probe based single-fiber reflectance (SfR) spectroscopy approach for characterizing sub-surface myoglobin forms of beef psoas major muscles during retail storage. A 400-µm fiber was placed in a 17-gauge needle, and the assembly was inserted into the muscle at five depths of 1 mm increment and 1 cm lateral shift. Metmyoglobin content increased at all depths during display and content at 1 mm was greater compared to that of 2 to 5 mm depth. The a* values decreased (P < 0.05) during retail display aligning with the sub-surface formation of metmyoglobin. In summary, the results suggest that needle-probe SfR spectroscopy can determine interior myoglobin forms and characterize meat discoloration.

Assessment of mitochondrial DNA copy number variation relative to nuclear DNA quantity between different tissues.

Mitochondrial DNA is a widely tested genetic marker in various fields of research and diagnostics. Nonetheless, there is still little understanding on its abundance and quality within different tissues. Aiming to obtain deeper knowledge about the content and quality of mtDNA, we investigated nine tissues including blood, bone, brain, hair (root and shaft), cardiac muscle, liver, lung, skeletal muscle, and buccal mucosa of 32 deceased individuals using two real-time quantitative PCR-based assays with differently sized mtDNA and nDNA targets. The results revealed that the quantity of nDNA is a weak surrogate to estimate mtDNA quantities among tissues of an individual, as well as tissues across individuals. Especially hair showed extreme variation, depicting a range of multiple magnitudes of mtDNA molecules per hair fragment. Furthermore, degradation can lead to fewer fragments being available for PCR. The results call for parallel determination of the quantity and quality of mtDNA prior to downstream genotyping assays.

The effect of modified atmosphere packaging at an early postmortem stage on lamb meat quality during subsequent aging.

This study explores the influence of modified atmosphere packaging (MAP) on fresh lamb meat quality with respect to gas concentration, rigor state, and post-mortem aging time. A comparison was done for the quality characteristics of lamb Longissimus thoracis lumborum chops that had been packaged separately in air, 75%O2  + 25%CO2 MAP or 50%O2  + 50%CO2 MAP at 1, 6, and 24 h post-mortem and then stored for 6, 12, 24, 72, and 144 h post-mortem, and the quality of lamb chops had been evaluated at each post-mortem period separately. Chops packaged at 1 and 6 h post-mortem in MAP had reduced pH decline, less purge loss, and enhanced redness at early post-mortem storage times. Lamb color stability was evidently greater in 75%O2  + 25%CO2 MAP than in 50%O2  + 50%CO2 MAP during the early storage period when a* and R630/R580 were taken into account. Shear force values were lowest in lambs packaged at 1 h post-mortem with 75%O2  + 25%CO2 MAP at 12 h post-mortem and then increased until 72 h post-mortem, suggesting that rigor has been delayed by such a high O2 MAP. Thus, fresh lamb quality was maintained most effectively when packaged at 1 h post-mortem in 75%O2  + 25%CO2 MAP for consumption at 12 h post-mortem. The exact mechanisms and optimization of MAP based on Chinese retail conditions should be considered in future studies. PRACTICAL APPLICATION: In this study, three slaughter patterns in the meat industry involving boning immediately after dressing (hot-boning) and chilling for a short period (warm-boning) or overnight (cold-boning) are considered, as well as the behavior of non-immediate consumption after purchase. Modified atmosphere packaging provides an effective preservation of early post-mortem muscles with enhanced color stability, water holding capacity, and texture during refrigerated storage. This could provide new insights into how to process lamb muscles in the early post-mortem period to improve and stabilize lamb quality.

Muscle of dark and normal beef differs metabolically.

Reduction in muscle glycogen triggered by adverse antemortem handling events alters postmortem energy metabolism and results in a high ultimate pH and dark, firm and dry beef, often referred to as 'dark-cutting'. However, the relationship between atypical dark (AT) beef, postmortem energy metabolism and underlying tissue characteristics remains somewhat unclear. Cattle harvested in the US and Canada representing normal (pH < 5.6), AT dark (pH 5.6-5.8) and dark cutting (DC; pH > 5.8) beef were analyzed for tissue characteristics related to energy metabolism. Results show AT dark beef is more oxidative but similar to normal beef in glycolytic potential and nucleotide abundance. Mitochondria DNA content (P < 0.05, Canada; P < 0.005, US) and oxidative enzymes for DC and AT dark beef were greater (P < 0.01; Canada and US) compared to normal beef. Myoglobin tracked (P < 0.01) with color classification. These findings show both DC and AT beef are inherently more oxidative and raise the possibility that more oxidative muscle may be more prone to develop dark beef.

Effect of carnosine synthesis precursors in the diet on jejunal metabolomic profiling and biochemical compounds in slow-growing Korat chicken.

The slow-growing Korat chicken (KR) has been developed to provide an alternative breed for smallholder farmers in Thailand. Carnosine enrichment in the meat can distinguish KR from other chicken breeds. Therefore, our aim was to investigate the effect of enriched carnosine synthesis, obtained by the ß-alanine and L-histidine precursor supplementation in the diet, on changes to metabolomic profiles and biochemical compounds in slow-growing KR jejunum tissue. Four hundred 21-day-old female KR chickens were divided into 4 experimental groups: a group with a basal diet, a group with a basal diet supplemented with 1.0% ß-alanine, 0.5% L-histidine, and a mix of 1.0% ß-alanine and 0.5% L-histidine. The feeding trial lasted 70 d. Ten randomly selected chickens from each group were slaughtered. Metabolic profiles were analyzed using proton nuclear magnetic resonance spectroscopy. In total, 28 metabolites were identified. Significant changes in the concentrations of these metabolites were detected between the groups. Partial least squares discriminant analysis was used to distinguish the metabolites between the experimental groups. Based on the discovered metabolites, 34 potential metabolic pathways showed differentiation between groups, and 8 pathways (with impact values higher than 0.05, P < 0.05, and FDR < 0.05) were affected by metabolite content. In addition, biochemical changes were monitored using synchrotron radiation-based Fourier transform infrared microspectroscopy. Supplementation of ß-alanine alone in the diet increased the ß-sheets and decreased the α-helix content in the amide I region, and supplementation of L-histidine alone in the diet also increased the ß-sheets. Furthermore, the relationship between metabolite contents and biochemical compounds were confirmed using principal component analysis (PCA). Results from the PCA indicated that ß-alanine and L-histidine precursor group was highly positively correlated with amide I, amide II, creatine, tyrosine, valine, isoleucine, and aspartate. These findings can help to understand the relationships and patterns between the spectral and metabolic processes related to carnosine synthesis.

Different effects of dietary ß-hydroxy-ß-methylbutyrate on composition of fatty acid and free amino acid, and fatty metabolism in the different muscles of broilers.

In the study, 336 broiler chickens were selected to explore dietary effects of different ß-hydroxy-ß-methylbutyrate (HMB) levels (0 (control), 0.05, 0.10, and 0.15%) on the compositions of fatty acids and free amino acids, and lipid metabolism in the different muscles of broilers. In the breast muscle, dietary HMB supplementation hardly affected the free amino acid composition (P > 0.05). Compared to the control group, dietary 0.10 and 0.15% HMB supplementation decreased the content of C18:1n9c and thus the monounsaturated fatty acid (MUFA), and dietary 0.15% HMB supplementation increased the sum of saturated fatty acids (SFA) (P < 0.05). Moreover, compared to the control group, dietary 0.05 and 0.10% HMB increased the mRNA expression of proliferator activated receptor-γ and the activity of fatty acid synthase (FAS), and dietary 0.10% HMB increased the acetyl-CoA carboxylase activity (P < 0.05). In the leg muscle, dietary 0.10 and 0.15% HMB increased the MUFA content and decreased the polyunsaturated fatty acid (PUFA) content, the PUFA to SFA ratio, the mRNA expression of sterol regulatory element binding proteins-1c, and the activities of acyl-CoA oxidase 1 and acetyl-CoA synthetase (P < 0.05). Moreover, dietary 0.10% HMB decreased the activities of hydroxy-3-methylglutaryl-CoA synthase 1 and FAS in comparison to the control group (P < 0.05). Dietary 0.05% HMB decreased the contents of essential amino acids and nonessential amino acids (NEAA), and dietary 0.15% HMB decreased the NEAA content (P < 0.05). In summary, dietary 0.10% HMB supplementation had superior efficiency on lipogenesis in the breast muscle of broilers. However, dietary HMB supplementation, especially at the level of 0.05 and 0.15%, decreased meat nutritional values and the lipogenesis in leg muscles.

Validation of protein biological markers of lamb meat quality characteristics based on the different muscle types.

This work investigated the ability of 8 potential biomarkers (phosphoglycerate kinase-1 (PGK1), pyruvate kinase-M2 (PKM2), phosphoglucomutase-1 (PGM1), ß-enolase (ENO3, myosin-binding protein-C (MYBPC1), myosin regulatory light chain-2 (MYLPF), troponin C-1 (TNNC1) and troponin I-1 (TNNI1)) to characterize meat quality by analyzing their relative abundance and enzymatic activity. Two different meat quality groups (Quadriceps femoris (QF) and Longissimus thoracis (LT) muscles) were selected at 24 h postmortem from 100 lamb carcasses. The relative abundance of PKM2, PGK1, PGM1, ENO3, MYBPC1, MYLPF, and TNNI1 was significantly different between LT and QF muscle groups (P < 0.01). Moreover, PKM, PGK, PGM, and ENO activity in LT muscle group was significantly lower than that in QF muscle (P < 0.05). Suggesting that PKM2, PGK1, PGM1, ENO3, MYBPC1, MYLPF, and TNNI1 can be used as robust biomarkers of lamb meat quality, providing the reference for understanding the molecular mechanism of postmortem meat quality formation in future.

Robustness of tissue oxygenation estimates by continuous wave space-resolved near infrared spectroscopy.

Significance: Continuous wave near infrared spectroscopy (CW-NIRS) is widely exploited in clinics to estimate skeletal muscles and brain cortex oxygenation. Spatially resolved spectroscopy (SRS) is generally implemented in commercial devices. However, SRS suffers from two main limitations: the a priori assumption on the spectral dependence of the reduced scattering coefficient [µs'(λ)] and the modeling of tissue as homogeneous. Aim: We studied the accuracy and robustness of SRS NIRS. We investigated the errors in retrieving hemodynamic parameters, in particular tissue oxygen saturation (StO2), when µs'(λ) was varied from expected values, and when layered tissue was considered. Approach: We simulated hemodynamic variations mimicking real-life scenarios for skeletal muscles. Simulations were performed by exploiting the analytical solutions of the photon diffusion equation in different geometries: (1) semi-infinite homogeneous medium and constant µs'(λ); (2) semi-infinite homogeneous medium and linear changes in µs'(λ); (3) two-layered media with a superficial thickness s1=5, 7.5, 10 mm and constant µs'(λ). All simulated data were obtained at source-detector distances ρ=35, 40, 45 mm, and analyzed with the SRS approach to derive hemodynamic parameters (concentration of oxygenated and deoxygenated hemoglobin, total hemoglobin concentration, and tissue oxygen saturation, StO2) and their relative error. Results: Variations in µs'(λ) affect the estimated StO2 (up to ±10%), especially if changes are different at the two wavelengths. However, the main limitation of the SRS method is the presence of a superficial layer: errors strongly larger than 20% were retrieved for the estimated StO2 when the superficial thickness exceeds 5 mm. Conclusions: These results highlight the need for more sophisticated strategies (e.g., the use of multiple short and long distances) to reduce the influence of superficial tissues in retrieving hemodynamic parameters and warn the SRS users to be aware of the intrinsic limitation of this approach, particularly when exploited in the clinical environment.

Diagnostics of inflammaging in relation to sarcopenia.

One of the theories about aging focuses on the immune response and relates to the activation of subclinical and chronic inflammation. This study was designed to investigate the relationship between inflammation and sarcopenia and to evaluate the influence of lifestyle on the inflammatory profile. Finally, therapeutic strategies to counteract the pathophysiological effect of skeletal muscle aging were also indicated. One hundred seventy-three individuals aged 71.5 ± 6.8 years were divided into two groups: sarcopenia and probable sarcopenia (n = 39) and no sarcopenia (n = 134). Sarcopenia was assessed according to the algorithm of the European Working Group on Sarcopenia in the older adults 2. C-reactive protein (CRP) (p = 0.011) and CRP/albumin ratio (p = 0.030) as well as IL-1ß (p = 0.002), cfDNA (p < 0.001) and bilirubin levels (p = 0.002) were significantly higher in the sarcopenia group as opposed to the no sarcopenia group. No significant differences were observed between groups in the concentration of TNFα (p = 0.429) and IL-6 (p = 0.300). An inverse correlation was found between gait speed and cfDNA (rs = -0.234, p < 0.01) and IL-1ß (rs = -0.263, p < 0.01). The ROC analysis of cfDNA, CRP, IL-1ß and bilirubin ranged from 0.6 to 0.7, which confirms the association between sarcopenia and inflammatory mediators and indicates high clinical usefulness of cfDNA and bilirubin in sarcopenia prediction. We also indicated a link between inflammation and fitness level in the older adult thereby providing evidence that lifestyle exercise should be a key therapeutic strategy in sarcopenia prevention.

Injection of l-arginine or l-lysine before freezing delays the emulsifying and gelling properties deterioration of myofibrillar proteins of frozen porcine Longissimus lumborum muscle.

This study aimed to investigate the effects of injecting l-arginine and l-lysine solution before freezing and after thawing on the emulsifying and gelling properties of myofibrillar proteins (MPs) of frozen porcine longissimus dorsi. The results showed that the pre-freezing injections were more effective in alleviating the decrease in emulsifying properties of MPs compared with the post-thawing injections, as evidenced by higher emulsion creaming index, oil droplet size, interfacial absorptive protein amount, and viscoelasticity. Additionally, the pre-freezing injections could effectively mitigate the damage to the gelling properties of MPs, as evidenced by the formation of a homogeneous and compact gel network with stronger water retention, strength and chemical forces, as well as a higher proportion of non-flowing water, whereas the post-thawing injections could not. These results demonstrated that the injection of l-arginine and l-lysine solution before freezing could delay freezing-induced damage to the emulsifying and gelling properties of MPs, keeping the processing characteristics of frozen porcine.

Effects of linseed oil supplementation duration on fatty acid profile and fatty acid metabolism-related genes in the muscles of Chinese crested white ducks.

Meat rich in polyunsaturated fatty acids is considered beneficial to health. Supplementing the diet with linseed oil promotes the deposition of polyunsaturated fatty acids (PUFAs) in poultry, a conclusion that has been confirmed multiple times in chicken meat. However, fewer studies have focused on the effects of dietary fatty acids on duck meat. Therefore, this study aims to evaluate the effects of the feeding time of a linseed oil diet on duck meat performance and gene expression, including meat quality performance, plasma biochemical indicators, fatty acid profile, and gene expression. For this study, we selected 168 Chinese crested ducks at 28 days old and divided them into three groups, with 56 birds in each group. The linseed oil content in the different treatment groups was as follows: the control group (0% flaxseed oil), the 14d group (2% linseed oil), and the 28d group (2% linseed oil). Ducks in the two experimental groups were fed a linseed oil diet for 28 and 14 days at 28 and 42 days of age, respectively. The results showed that linseed oil had no negative effect on duck performance (slaughter rate, breast muscle weight, and leg muscle weight) or meat quality performance (pH, meat color, drip loss, and shear force) (P > 0.05). The addition of linseed oil in the diet increased plasma total cholesterol and high-density lipoprotein cholesterol levels (P < 0.05), while decreasing triglyceride content (P < 0.05). Furthermore, the supplementation of linseed oil for four weeks affected the composition of muscle fatty acids. Specifically, levels of α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid were increased (P < 0.05), while eicosatetraenoic acid content was negatively correlated with flaxseed oil intake (P < 0.05). qRT-PCR analysis further revealed that the expression of FATP1, FABP5, and ELOVL5 genes in the breast muscle, as well as FABP3 and FADS2 genes in the thigh muscle, increased after four weeks of linseed oil supplementation (P < 0.05). However, after two weeks of feeding, CPT1A gene expression inhibited fatty acid deposition, suggesting an increase in fatty acid oxidation (P < 0.05). Overall, the four-week feeding time may be a key factor in promoting the deposition of n-3 PUFAs in duck meat. However, the limitation of this study is that it remains unknown whether longer supplementation time will continue to affect the deposition of n-3 PUFAs. Further experiments are needed to explain how prolonged feeding of linseed oil will affect the meat quality traits and fatty acid profile of duck meat.

Double-headed binding of myosin II to F-actin shows the effect of strain on head structure.

Force production in muscle is achieved through the interaction of myosin and actin. Strong binding states in active muscle are associated with Mg·ADP bound to the active site; release of Mg·ADP allows rebinding of ATP and dissociation from actin. Thus, Mg·ADP binding is positioned for adaptation as a force sensor. Mechanical loads on the lever arm can affect the ability of myosin to release Mg·ADP but exactly how this is done is poorly defined. Here we use F-actin decorated with double-headed smooth muscle myosin fragments in the presence of Mg·ADP to visualize the effect of internally supplied tension on the paired lever arms using cryoEM. The interaction of the paired heads with two adjacent actin subunits is predicted to place one lever arm under positive and the other under negative strain. The converter domain is believed to be the most flexible domain within myosin head. Our results, instead, point to the segment of heavy chain between the essential and regulatory light chains as the location of the largest structural change. Moreover, our results suggest no large changes in the myosin coiled coil tail as the locus of strain relief when both heads bind F-actin. The method would be adaptable to double-headed members of the myosin family. We anticipate that the study of actin-myosin interaction using double-headed fragments enables visualization of domains that are typically noisy in decoration with single-headed fragments.

Molecular mechanisms contributing to the development of beef sensory texture and flavour traits and related biomarkers: Insights from early post-mortem muscle using label-free proteomics.

Beef sensory quality comprises a suite of traits, each of which manifests its ultimate phenotype through interaction of muscle physiology with environment, both in vivo and post-mortem. Understanding variability in meat quality remains a persistent challenge, but omics studies to uncover biological connections between natural variability in proteome and phenotype could provide validation for exploratory studies and offer new insights. Multivariate analysis of proteome and meat quality data from Longissimus thoracis et lumborum muscle samples taken early post-mortem from 34 Limousin-sired bulls was conducted. Using for the first-time label-free shotgun proteomics combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), 85 proteins were found to be related with tenderness, chewiness, stringiness and flavour sensory traits. The putative biomarkers were classified in five interconnected biological pathways; i) muscle contraction, ii) energy metabolism, iii) heat shock proteins, iv) oxidative stress, v) regulation of cellular processes and binding. Among the proteins, PHKA1 and STBD1 correlated with all four traits, as did the GO biological process 'generation of precursor metabolites and energy'. Optimal regression models explained a high level (58-71%) of phenotypic variability with proteomic data for each quality trait. The results of this study propose several regression equations and biomarkers to explain the variability of multiple beef eating quality traits. Thanks to annotation and network analyses, they further suggest protein interactions and mechanisms underpinning the physiological processes regulating these key quality traits. SIGNIFICANCE: The proteomic profiles of animals with divergent quality profiles have been compared in numerous studies; however, a wide range of phenotypic variation is required to better understand the mechanisms underpinning the complex biological pathways correlated with beef quality and protein interactions. We used multivariate regression analyses and bioinformatics to analyse shotgun proteomics data to decipher the molecular signatures involved in beef texture and flavour variations with a focus on multiple quality traits. We developed multiple regression equations to explain beef texture and flavour. Additionally, potential candidate biomarkers correlated with multiple beef quality traits are suggested, which could have utility as indicators of beef overall sensory quality. This study explained the biological process responsible for determining key quality traits such as tenderness, chewiness, stringiness, and flavour in beef, which will provide support for future beef proteomics studies.