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Objective: Sarcoplasmic proteins include proteins that play critical roles in biological processes of living organisms. How seasons influence biological processes and meat quality of postmortem muscles through the regulation of protein phosphorylation remain to be investigated. In this study, the phosphorylation of sarcoplasmic proteins in pork longissimus muscle was investigated in four seasons.Methods: Sarcoplasmic proteins were extracted from 40 pork carcasses (10 for each season) and analyzed through ProQ Diamond staining for phosphorylation labeling and Sypro Ruby staining for total protein labeling. The pH of muscle, contents of glycogen and ATP were measured at 45 min, 3 h, and 9 h postmortem and the water (P2b, P21, and P22) was measured at 3 h and 9 h.Results: A total of 21 bands were detected. Band 8 (heat shock cognate 71 kDa protein; heat shock 70 kDa protein 1B) had higher phosphorylation level in summer than that in other seasons at 45 min postmortem. The phosphorylation levels of 3 Bands were significantly different between fast and normal pH decline groups (p21) and positive association with free water (P22).Conclusion: The phosphorylation levels of sarcoplasmic proteins involved in energy metabolism and heat stress response at early postmortem time differed depending on the seasons. These proteins include heat shock protein 70, pyruvate kinase, phosphoglucomutase-1, glucose-6-phosphate isomerase, and carbonic anhydrase 3. High temperatures in summer might result in the phosphorylation of those proteins, leading to pH decline and low water holding capacity.

BACKGROUND The abnormal ultimate pH (pHu ) in postmortem muscles affect the meat quality and results in substantial economic losses. Dark, firm, and dry (DFD) meat linked with the higher postmortem pHu values and exhibited many quality issues such as dark color, tough texture and shorter shelf life. This research aimed to investigate the effect of protein phosphorylation on variations in beef pHu in order to explore the possible mechanisms underlying DFD meat formation. RESULTS Glycogen and lactate contents were higher, while L* and a* were lower in high pHu beef. Shear force was higher in intermediate pHu group. Global phosphorylation of sarcoplasmic proteins was higher in low pHu samples on day 1 and of myofibrillar proteins was higher in intermediate pHu meat on days 1 and 2 postmortem. Sarcoplasmic protein bands with different phosphorylation levels were identified as containing some glycometabolism and stress response proteins and phosphorylated myofibrillar protein bands were identified sarcomeric and metabolic proteins. CONCLUSIONS Phosphorylation of multiple proteins of glycolytic pathway and contractile machinery may play critical roles in development of DFD beef. © 2021 Society of Chemical Industry.

NEW FINDINGS What is the main observation in this case? The main observation of this case report is that blood flow-restricted exercise can cause myofibrils to have an aberrant wave-like appearance that is accompanied by irregular pockets of sarcoplasm in the intermyofibrillar space, while traditional forms of damage to the Z-discs and contractile elements are not as apparent. What insights does it reveal? Our findings indicate that blood flow restriction-mediated fluid pooling might cause alterations in skeletal muscle ultrastructure after exercise that might be directly related to myofibre swelling. ABSTRACT The acute effects of blood flow-restricted (BFR) exercise training on skeletal muscle ultrastructure are poorly understood owing to inconsistent findings and the use of largely imprecise systemic markers for indications of muscle damage. The purpose of this study was to compare myofibrillar ultrastructure before and 30 min after normal and BFR resistance exercise using transmission electron microscopy in a single individual to evaluate the feasibility of this more nuanced approach. One apparently healthy male with 13 years of resistance exercise completed six sets of both BFR [30% of one-repetition maximum (1-RM)] and normal non-occluded (70% of 1-RM) unilateral angled leg press on the contralateral leg, as a control, after assessment of 1-RM 72 h before. Vastus lateralis muscle biopsies were collected before and 30 min after each exercise session. The lengths and widths of 250 sarcomeres and the sarcoplasmic area were assessed via 20 individual transmission electron photomicrographs. Analysis revealed that BFR training (1.769 ± 0.12 μm) increased sarcomere length when compared with normal exercise (1.64 ± 0.17 μm; P

The objective of this study was to examine the relationship between meat quality attributes and the changes of sarcoplasmic protein acetylation and myofibrillar protein acetylation in lamb longissimus thoracis et lumborum muscles at different postmortem phases. Protein acetylation, color, pH, shear force, myofibril fragmentation index and cooking loss were measured. The total level of acetylated sarcoplasmic proteins showed a negative relation with pH, a positive relation with a*, b* and cooking loss at the pre-rigor phase. Sarcoplasmic proteins acetylation affected postmortem pH by regulating glycolysis, which in turn affects color and cooking loss. The total level of acetylated myofibrillar proteins showed a positive relation with shear force at the pre-rigor phase. Myofibrillar proteins acetylation affected meat tenderness by regulating muscle contraction. This study indicated that acetylation played a regulatory role of meat color, water-holding capacity, and tenderization process at early postmortem.

The protein-bound heterocyclic amines (HAs) and their generation pattern are still unclear. Generation of sarcoplasmic (SP)- and myofibrillar protein (MP)-bound HAs under different heating conditions was investigated in chemical model systems using ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The results showed that eight types (387.01 ± 37.50 ng/100 mg MP) and six types (452.06 ± 33.32 ng/100 mg SP) of protein-bound HAs were generated at 180 °C/40 min, respectively. MP system can generate either more types or amounts of bound HAs at 90-150 °C. The main categories of protein-bound HAs in two systems were almost the same: β-carbolines, α-carbolines, pyridines, and quinolines, among which nonpolar HAs dominated by β-carbolines (at least 69.2%) are most easily formed. Principal component analysis revealed no significant difference in bound HAs at 90-120 °C but showed a significant increase at 150-180 °C. The higher the temperature, the more significant the differences between samples with different durations at the same temperature.

A 63-year old male presented with a 20-year history of facial weakness and several years of nasal regurgitation and dysphonia. Examination revealed bilateral facial weakness with nasal speech. Serum creatine kinase was 918U/L. Neurophysiological studies suggested a myopathy and biopsy of the left vastus lateralis showed serpentine basophilic inclusions in the sarcoplasm and strong oxidative enzyme activity suggesting mitochondria accumulation. The muscle MRI showed selective fatty replacement within semitendinosus, gastrocnemius and soleus indicative of a desminopathy. A heterozygous missense variant c.17C>G (p.Ser6Trp) was identified within DES, predicted to be pathogenic in silico and previously described in a family with distal limb weakness. There are no previous case reports of desminopathy presenting with facial weakness, to our knowledge. Diagnosis was suggested following myoimaging of clinically unaffected muscles. Our study highlights the importance of muscle MRI in the diagnostic evaluation of muscle disease and further expands the known phenotypic heterogeneity of desminopathies.

Rationale: We recently discovered pivotal contributions of stress kinase JNK2 (c-Jun N-terminal kinase isoform 2) in increased risk of atrial fibrillation through enhanced diastolic sarcoplasmic reticulum (SR) calcium (Ca 2+ ) leak via RyR2 (ryanodine receptor isoform 2). However, the role of JNK2 in the function of the SERCA2 (SR Ca 2+ -ATPase), essential in maintaining SR Ca 2+ content cycling during each heartbeat, is completely unknown. Objective: To test the hypothesis that JNK2 increases SERCA2 activity SR Ca 2+ content and exacerbates an arrhythmic SR Ca 2+ content leak-load relationship. Methods and Results: We used confocal Ca 2+ imaging in myocytes and HEK-RyR2 (ryanodine receptor isoform 2-expressing human embryonic kidney 293 cells) cells, biochemistry, dual Ca 2+ /voltage optical mapping in intact hearts from alcohol-exposed or aged mice (where JNK2 is activated). We found that JNK2, but not JNK1 (c-Jun N-terminal kinase isoform 1), increased SERCA2 uptake and consequently elevated SR Ca 2+ content load. JNK2 also associates with and phosphorylates SERCA2 proteins. JNK2 causally enhances SERCA2-ATPase activity via increased maximal rate, without altering Ca 2+ affinity. Unlike the CaMKII (Ca 2+ /calmodulin-dependent kinase II)-dependent JNK2 action in SR Ca 2+ leak, JNK2-driven SERCA2 function was CaMKII independent (not prevented by CaMKII inhibition). With CaMKII blocked, the JNK2-driven SR Ca 2+ loading alone did not significantly raise leak. However, with JNK2-CaMKII–driven SR Ca 2+ leak present, the JNK2-enhanced SR Ca 2+ uptake limited leak-induced reduction in SR Ca 2+ , normalizing Ca 2+ transient amplitude, but at a higher arrhythmogenic SR Ca 2+ leak. JNK2-specific inhibition completely normalized SR Ca 2+ handling, attenuated arrhythmic Ca 2+ activities, and alleviated atrial fibrillation susceptibility in aged and alcohol-exposed myocytes and intact hearts. Conclusions: We have identified a novel JNK2-induced activation of SERCA2. The dual action of JNK2 in CaMKII-dependent arrhythmic SR Ca 2+ leak and a CaMKII-independent uptake exacerbates atrial arrhythmogenicity, while helping to maintain normal levels of Ca 2+ transients and heart function. JNK2 modulation may be a novel therapeutic target for atrial fibrillation prevention and treatment.

The results of the study of the histological structure of the locomotor axial (skeletal) muscle tissue of the beaked redfish Sebastes mentella are presented and its ontogenetic changes are considered. Its skeletal muscles compose of glycolytic (white) and oxidative (red) fibers. Data show the quantitative distribution of these fibers along the body of sexually mature fish in the cranio-caudal direction, their ratio in the trunk and tail parts. The diameter of these two types of fibers reaches significant sizes, as in a number of deep-sea slow-growing species (> 300 μm - white fibers, > 150 μm - red fibers). The density of glycolytic fibers is higher than that of red ones (80,7±2,0 versus 54,7±1,5 %), which is due to the different content of adipose and connective tissue (endomysium). The variability of white fibers in most fish size groups is significantly lower than that of red ones. A feature that is not noted in other deep-sea fish species with a low growth rate is the relatively frequent occurrence of red fibers with stratification of sarcoplasm into two zones (layers) - external and internal ("double" structures), the nature of which is not yet clear.

In order to reduce the immunoreactivity of sarcoplasmic calcium-binding protein (SCP), site-directed mutations were used to replace key amino acids in the conformational epitopes and calcium-binding sites. The mutant SCPs (mSCPs) were expressed in Escherichia coli, and their immunoreactivities were analyzed using iELISA and basophil activation assays. Furthermore, the structural changes of mSCPs were determined from the circular dichroism spectra. The iELISA results showed that mSCPs could effectively inhibit the binding of wild-type SCP (wtSCP) to sensitive serum, with inhibition rates that reached 90%. Moreover, mSCPs could downregulate the expression levels of CD63 and CD203c on the basophil surface. Compared with wtSCP, the peak values were significantly changed, and the calcium binding ability was impaired, which explained the decline in immunoreactivities of the mSCPs. All of the data confirmed that this approach was effective in reducing the immunoreactivity of SCP and could be applied to other shellfish allergens.

ObjectiveTo identify the characteristic pathologic features of dermatomyositis (DM) associated with anti-Mi-2 autoantibodies (anti-Mi-2 DM).MethodsWe reviewed 188 muscle biopsies from patients (1) pathologically diagnosed with DM through the sarcoplasmic expression for the myxovirus-resistant protein A and (2) serologically positive for 1 of 5 DM-specific autoantibodies (DMSAs) (anti-Mi-2, n = 30; other DMSAs, n = 152) or negative for all 5 DMSAs (n = 6). We then compared the histopathologic and immunohistochemical features of patients with anti-Mi-2 DM to those with non-Mi-2 DM and patients with anti-synthetase syndrome (ASS) (n = 212) using thettest, Fisher exact test, and a logistic regression model.ResultsPatients with anti-Mi-2 DM showed significantly higher severity scores in muscle fiber and inflammatory domains than non-Mi-2 DM patients. The presence of perifascicular necrosis, increased perimysial alkaline phosphatase activity, and sarcolemmal membrane attack complex deposition was more frequent in patients with anti-Mi-2 DM (p< 0.01). After Bonferroni correction, there were no significant differences in the percentages of the features mentioned above between the patients with anti-Mi-2 DM and those with ASS (p> 0.01).ConclusionPerifascicular necrosis and perimysial pathology, features previously reported in ASS, are common in patients with anti-Mi-2 DM. Our findings not only assist in differentiating anti-Mi-2 DM from other DM subtypes but also suggest the possibility of an overlapping mechanism between anti-Mi-2 DM and ASS.Classification of EvidenceThis study provides Class II evidence that the muscle biopsies of DM patients with anti-Mi-2 autoantibodies are more likely to demonstrate higher severity scores in muscle fiber and inflammatory domains.

This work represents the first ultrastructural study of the tegument of a freshwater teleost-infecting aporocotylid blood fluke and the first detailed observations of the surface sensory receptors of any aporocotylid. The work was carried out on Sanguinicola inermis from the branchial arteries of the pike, Esox lucius. Light and scanning electron microscopy observations failed to show the presence of protruding surface spines in S. inermis. Transmission electron microscopy studies revealed, deep beneath the distal cytoplasm of the tegument, sporadic, membrane-bound sarcoplasmic processes, each containing a single spine with the typical crystalline structure of digenean spines. On rare occasions, the spine may protrude above the body surface, but in such cases the distal tegumental cytoplasm is extremely flattened. This finding supports a unique, deep origin of aporocotylid spines when compared to those of other digenean families. This character may well prove to be an autapomorphy of the Aporocotylidae, and the position of the base of aporocotylid spines within sarcoplasmic processes supports their origin as a muscle derivative. Unlike other digenean groups, the distal region of the cytoplasmic processes, connecting sunken tegumental perikarya with the syncytial tegumental cytoplasm, is strengthened by peripheral microtubules. Three types of ciliated and three types of non-ciliated sensory receptors were distinguished in the tegument of S. inermis. One of the distinctive features of the ciliated receptors is the unusual position of the basal region of the cilium within a tegumental outgrowth. Such a position has previously been described for certain receptors types in larval and adult schistosomatids and larval diplostomids, suggesting that this feature, in adult worms, might prove to be an apomorphic trait for some or all groups of blood flukes.

Significance Selenoprotein N (SEPN1) is a type II transmembrane protein of the endoplasmic reticulum (ER) that senses luminal calcium through an EF-hand domain. On calcium depletion, a SEPN1 oligomer, prevalent under basal calcium concentrations, dissociates to generate a monomeric polypeptide that has enhanced redox trapping potential for its target the calcium pump, SERCA2, as well as for many additional interactors, indicating enhanced reductase activity. Our studies not only support that SEPN1 is one of the long-sought reductases of the ER, but also identify a feedback mechanism through which SEPN1 senses the luminal calcium level to modulate downstream signal transduction. Our results suggest that SEPN1 regulates the SERCA-mediated replenishment of ER calcium stores, a crucial mechanism for excitation-contraction coupling in skeletal muscle., The endoplasmic reticulum (ER) is the reservoir for calcium in cells. Luminal calcium levels are determined by calcium-sensing proteins that trigger calcium dynamics in response to calcium fluctuations. Here we report that Selenoprotein N (SEPN1) is a type II transmembrane protein that senses ER calcium fluctuations by binding this ion through a luminal EF-hand domain. In vitro and in vivo experiments show that via this domain, SEPN1 responds to diminished luminal calcium levels, dynamically changing its oligomeric state and enhancing its redox-dependent interaction with cellular partners, including the ER calcium pump sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA). Importantly, single amino acid substitutions in the EF-hand domain of SEPN1 identified as clinical variations are shown to impair its calcium-binding and calcium-dependent structural changes, suggesting a key role of the EF-hand domain in SEPN1 function. In conclusion, SEPN1 is a ER calcium sensor that responds to luminal calcium depletion, changing its oligomeric state and acting as a reductase to refill ER calcium stores.

Consisting of 25 to 30% of protein in carp, water-soluble sarcoplasmic proteins lost in wash water, have been recovered and freeze-dried into a protein-rich powder. Study objectives were to evaluate protein quality and safety of a silver carp sarcoplasm derived protein powder (CSP) compared to commercial protein supplements, casein, and whey. In vivo protein quality assessment of CSP showed a lower (P < 0.05) protein digestibility corrected amino acid score compared to the commercial protein sources. Despite greater (P < 0.05) fecal amino acid excretion in casein-fed rats, there were no significant differences in liver and muscle amino acid profiles. All low (10% kcal) protein diets supported growth with the normal range. However, whey protein supplementation resulted in greater (P < 0.05) adiposity. CSP, casein, or whey-fed rats showed no differences in major organ weights, renal damage biomarkers, or bone indices. Collectively, results indicated CSP was safe with protein quality comparable to casein. PRACTICAL APPLICATION: As much as 40 percent of protein in fish can be lost due to sarcoplasmic protein solubilization in processing wash water. Silver carp sarcoplasm protein powder may have similar commercial potential as a sustainable and nutritious alternative to whey and casein proteins. This project aimed to verify the protein quality and safety of this economical protein source.

Background Fortetropin is a proteo-lipid complex made from fertilized egg yolk and, in young men, has been shown to increase lean body mass. Methods The purpose of this study was to examine the effects of 21 days of Fortetropin supplementation on the fractional synthetic rate (FSR) of muscle protein in 10 healthy, older men and 10 women (66.4 ± 4.5 y). We used 2H2O labeling to measure FSR of multiple muscle protein ontologies. D3-creatine dilution was used to determine muscle mass at baseline. Subjects ingested 70% 2H2O for 21 day and saliva samples were collected to determine body 2H2O enrichment. A microbiopsy was obtained from the m. vastus lateralis on Day 21. Subjects were randomly assigned to Fortetropin (19.8 g/d) or placebo (cheese powder, 19.8 g/d). Results Restricting kinetic data to proteins with ≥2 peptides measured in at least 4 subjects per group resulted in 117 proteins meeting these criteria. The mean FSR for a majority of proteins in several muscle gene ontologies was higher in the Fortetropin group compared to placebo (32/38 myofibril proteins, 33/44 sarcoplasmic proteins, and 12/17 mitochondrial proteins) and this proportion was significantly different between groups using a binomial test and were independent of sex or baseline muscle mass. Conclusions The overall magnitude of the difference in muscle protein FSR of Fortetropin from placebo was 18%, with multiple gene ontologies affected. While these results should be confirmed in larger cohorts, they suggest that Fortetropin supplementation is effective for promoting muscle protein synthesis in older people.

The article presents the research of the protein composition of meat of wild animals (elk and wild boar) and domestic animals (beef), by their isolation and separation methods due to their different solubility in different solutions, conducted in the laboratory of Biological Safety of Food Systems of the Department of Meat and Dairy Products Technology of Mari State University. The total proteins or protein fractions were extracted from a homogenized chilled meat sample. Then, the colorless proteins in the extract were transferred to the colored product, adding a biuret reagent, and studied the D optical density of separate protein fractions on the photoelectric concentration colorimeter PCC-2, determining the absorption of emitting at a wavelength of 540 nm. The protein content of the samples was expressed in the mass fraction of the protein fraction in the sample in g/ 100g of meat and in percentages. The calculation was based on the previously found linear calibration dependence of the optical density on the concentration of the standard protein. Using sequential extraction of muscle proteins with water, saline and alkaline it is possible to obtain different protein fractions. Differentiation of proteins by solubility (water-soluble, soluble in salts, alkaline-soluble, albumins, globulins), fractional composition (sarcoplasmic, myo????ibrillar and connective tissue), peculiarities of the biochemical structure (high-grade and inferior). Protein extracts of muscle tissue of wild and domestic animals obtained during sequential extraction are used to quantify the protein content of a certain fraction, as well as to research the properties of proteins.

This study addressed the hypothesis that cardiac Sirtuin 1 (Sirt1) deficiency alters cardiomyocyte Ca2+ and Na+ regulation, leading to cardiac dysfunction and arrhythmogenesis. We used mice with cardiac‐specific Sirt1 knockout (Sirt1−/−). Sirt1flox/flox mice were served as control. Sirt1−/− mice showed impaired cardiac ejection fraction with increased ventricular spontaneous activity and burst firing compared with those in control mice. The arrhythmic events were suppressed by KN93 and ranolazine. Reduction in Ca2+ transient amplitudes and sarcoplasmic reticulum (SR) Ca2+ stores, and increased SR Ca2+ leak were shown in the Sirt1−/− mice. Electrophysiological measurements were performed using patch‐clamp method. While L‐type Ca2+ current (I Ca, L) was smaller in Sirt1−/− myocytes, reverse‐mode Na+/Ca2+ exchanger (NCX) current was larger compared with those in control myocytes. Late Na+ current (I Na, L) was enhanced in the Sirt1−/− mice, alongside with elevated cytosolic Na+ level. Increased cytosolic and mitochondrial reactive oxygen species (ROS) were shown in Sirt1−/− mice. Sirt1−/− cardiomyocytes showed down‐regulation of L‐type Ca2+ channel α1c subunit (Cav1.2) and sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2a (SERCA2a), but up‐regulation of Ca2+/calmodulin‐dependent protein kinase II and NCX. In conclusions, these findings suggest that deficiency of Sirt1 impairs the regulation of intracellular Ca2+ and Na+ in cardiomyocytes, thereby provoking cardiac dysfunction and arrhythmogenesis.

The functional state of RyR depends on the intracellular calcium concentration and on the oxidation state of its protein components in some particular sites and of some sentinel amino acids. In addition to the regulation of the RyR channel by exogenous substances (caffeine, ryanodine), ions environmental situations (oxidative state), other components, such as some endogenous proteins present in the sarcoplasm and/or in muscle membranes that are able to determine changes in Ca2+ channel activity. Among these, calmodulin and S-100A could determine modifications in the status of RyR channel in the skeletal muscle. The currently available data can be justified the use of a simplified S-100/CaM and RyR interaction model for the regulation of Ca2+ release in skeletal muscle. Under resting conditions, the CaM/S100A1 binding domain on RyR1 is predominantly dependent on S100A1. Vice versa when the intracellular Ca2+ concentration becomes high as well as during repetitive (tetanus) stimulation, the Ca-CaM bond becomes dominant, shifting S100A1 from RyR1 and promoting channel inactivation. This may be one of the mechanism of muscle fatigue.

Aims Nakajo-Nishimura syndrome (NNS) is an autosomal recessive disease caused by biallelic mutations in the PSMB8 gene that encodes the immunoproteasome subunit β5i. There have been only a limited number of reports on the clinicopathological features of the disease in genetically confirmed cases. Methods We studied clinical and pathological features of three NNS patients who all carry the homozygous p.G201V mutations in PSMB8. Patients' muscle specimens were analysed with histology and immunohistochemistry. Results All patients had episodes of typical periodic fever and skin rash, and later developed progressive muscle weakness and atrophy, similar to previous reports. Oral corticosteroid was used for treatment but showed no obvious efficacy. On muscle pathology, lymphocytes were present in the endomysium surrounding non-necrotic fibres, as well as in the perimysium perivascular area. Nearly all fibres strongly expressed MHC-I in the sarcolemma. In the eldest patient, there were abnormal protein aggregates in the sarcoplasm, immunoreactive to p62, TDP-43 and ubiquitin antibodies. Conclusions These results suggest that inflammation, inclusion pathology and aggregation of abnormal proteins underlie the progressive clinical course of the NNS pathomechanism.

Hypertension is an important risk factor for cardiovascular diseases. Besides cardiovascular system, it could cause damage to liver. It has been shown that endoplasmic reticulum stress (ERS) plays a crucial role in the pathogenesis of hypertension. ERS inhibitor tauroursodeoxycholic-acid (TUDCA) has favorable effects on various pathologies including cardiovascular, metabolic and hepatic diseases. In this study, the hepatoprotective effect and mechanism of TUDCA were investigated in the deoxycorticosterone acetate (DOCA)-salt-induced hypertension. Male Wistar rats were used and divided into four groups: Control, DOCA, TUDCA and DOCA + TUDCA. Hypertension was induced by DOCA-salt administration for twelve weeks after the unilateral nephrectomy. TUDCA was given for the last 4 weeks. Systolic blood pressure was measured by using tail-cuff method. At the end of the treatment, liver was isolated and weighed. The expressions of various proteins and histopathological evaluation were examined in the liver. TUDCA markedly decreased systolic blood pressure in the hypertensive animals. Hypertension caused increase in the expressions of glucose-regulated protein-78 (GRP78), matrix metalloproteinase-2 (MMP-2) and phospho-inhibitor kappa B-alpha (p-I kappa B-alpha) and the decrease in the expression of sarcoplasmic/endoplasmic reticulum Ca2+-ATPase2 (SERCA2) and phospho-extracellular signal-regulated kinase (p-ERK) in the liver. Alterations in these protein expressions were not detected in the TUDCA-treated hypertensive group. Also, hepatic balloon degeneration, inflammation and fibrosis were observed in the hypertensive group. TUDCA improved inflammation and fibrosis in the hypertensive liver. Our findings indicate that the detrimental effect of DOCA-salt-induced hypertension on the liver was defended by the inhibition of ERS. Hepatic ERS and its treatment should be taken into consideration for therapeutic approaches to hypertension.

The present work aims to clarify the mechanism of gel weakening (modori phenomenon) on surimi-based products and provide information on how to prevent it from happening in threadfin bream. We investigated the distribution of endogenous proteinases in the normal and belly muscles and their effects on the quality of the surimi gel. There was severe degradation of myofibrillar proteins in the belly muscle during heating at 50 °C, which was not found in the normal muscle. The surimi gel prepared from the belly muscle also showed a low gel strength as compared to that of the normal muscle. The myofibrillar proteins in the belly muscle were found to be degraded by a sarcoplasmic serine proteinase (SSP). The SSP activity was detected in the belly muscle and hepatopancreas but not in the normal muscle, while the SSP mRNA was detected only in the hepatopancreas. It is suggested that SSP in the hepatopancreas leaks to the belly muscle during post-harvest storage and is responsible for the modori phenomenon. These results imply that high-quality surimi gel of the threadfin bream can be produced by preventing protease leakage to the muscle during post-harvest storage.

INTRODUCTION Protein ingestion and the ensuing hyperaminoacidemia stimulates skeletal muscle protein synthesis in the postexercise period. This response facilitates muscle remodeling, which is important during intensified training. The aim of this study was to determine whether supplementation with α-lactalbumin (LA), with high leucine and tryptophan contents, would improve responses to short periods of intensified aerobic training compared with supplementation with an isonitrogenous quantity of collagen peptides (CP). METHODS Endurance-trained participants (5 male, 6 female, 24 ± 4 yr, V˙O2 = 53.2 ± 9.1 mL·kg·min, peak power output = 320 ± 48 W; means ± SD) consumed a controlled diet (1.0 g·kg·d protein) and refrained from habitual training for 11 d while taking part in this double-blind randomized, crossover trial. The two intervention phases, which consisted of brief intensified training (4 × 4-min cycling intervals at 70% of peak power output on 3 consecutive days) combined with the ingestion of LA or CP supplements after exercise (20 g) and before sleep (40 g), were separated by 4 d of washout without protein supplementation (i.e., the control phase). In response to each phase, myofibrillar (MyoPS), sarcoplasmic protein synthesis (SarcPS) rates (via H2O ingestion) and parameters of sleep quality were measured. RESULTS LA ingestion increased plasma leucine (P < 0.001) and tryptophan concentrations (P < 0.001) relative to CP. Intensified training increased MyoPS and SarcPS above the washout phase in LA- and CP-supplemented phases (P < 0.01), with increases being 13% ± 5% and 5% ± 7% greater with LA than CP for MyoPS (P < 0.01) and SarcPS, respectively (P < 0.01). CONCLUSIONS Despite an isonitrogenous diet, protein synthesis was enhanced to a greater extent when trained participants consumed LA compared with CP during intensified aerobic training, suggesting that protein quality is an important consideration for endurance-trained athletes aiming to augment adaption to exercise training.

Meat color is important for consumer acceptability, with excessively dark meat often associated with consumer rejection. It is determined chromatically by pigment content (measured by hue and chroma) and achromatically by scattering of light by the microstructure (measured by lightness), the latter of which has received minimal research focus. This review discusses the individual components of the meat microstructure that cause differences in achromatic contributions to color. Differences in achromatic light scattering between light and dark extremes of meat color are most likely explained by structural attributes within the muscle cell. These differences are proposed to arise from variations in (a) transverse shrinkage of the structural lattice of the myofilaments, myofibrils, and muscles fibers, (b) longitudinal shrinkage of the sarcomere, and (c) different protein composition of the surrounding medium (sarcoplasm and extracellular space). These are discussed at a mechanistic level, in relation to six parameters of the muscle cell: (a) protein surface charge altering the myofilament spacing, (b) protein solubility, (c) sarcoplasmic protein binding to myofilaments and myofibrils, (d) integrity of the cytoskeleton and cell adhesion proteins, (e) sarcomere integrity and myofibrillar proteins, and (f) myosin denaturation and rigor bond modification. New data are presented to support the proposed role of structural elements in muscle causing achromatic light scattering and their contribution to the surface color of meat. In addition, the relationships between lightness and water holding capacity and pH are explored and the economic impact of dark meat for the meat industry is discussed.

Doxorubicin is an anthracycline-based chemotherapeutic that causes myotoxicity with symptoms persisting beyond treatment. Patients experience muscle pain, weakness, fatigue, and atrophy, but the underlying mechanisms are poorly understood. Studies investigating doxorubicin-induced myotoxicity have reported disrupted mitochondrial function. Mitochondria are responsible for regulating both cellular energy status and Ca(2+) handling, both of which impact contractile function. Moreover, loss of mitochondrial integrity may initiate muscle atrophy. Skeletal muscle mitochondrial dysregulation may therefore contribute to an overall loss of skeletal muscle quality and performance that may be mitigated by appropriately targeted mitochondrial therapies. We therefore assessed the impact of doxorubicin on muscle performance and applied a multiplexed assay platform to diagnose alterations in mitochondrial respiratory control. Mice received a clinically relevant dose of doxorubicin delivered systemically and were euthanized 72 h later. We measured extensor digitorum longus and soleus muscle forces, fatigue, and contractile kinetics in vitro, along with Ca(2+) uptake in isolated sarcoplasmic reticulum. Isolated skeletal muscle mitochondria were used for real-time respirometry or frozen for protein content and activity assays. Doxorubicin impaired muscle performance, which was indicated by reduced force production, fatigue resistance, and sarcoplasmic reticulum–Ca(2+) uptake, which were associated with a substrate-independent reduction in respiration and membrane potential but no changes in the NAD(P)H/NAD(P)(+) redox state. Protein content and dehydrogenase activity results corroborated these findings, indicating that doxorubicin-induced mitochondrial impairments are located upstream of ATP synthase within the electron transport system. Collectively, doxorubicin-induced lesions likely span mitochondrial complexes I–IV, providing potential targets for alleviating doxorubicin myotoxicity.

Calcium ion (Ca2+) cycle plays a crucial role in the contraction and relaxation of cardiomyocytes. The sarcoplasmic reticulum (SR) acts as an organelle for storing Ca2+, which mediated the release and re-uptake of Ca2+ during contraction and relaxation. Disorders of SR function lead to the dysfunction of Ca2+ cycle and myocardial cell function. The sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 2a (SERCA2a) acts as a subtype of SERCA expressed in the heart, which mediates the contraction of cardiomyocytes and Ca2+ in the cytoplasm to re-enter into the SR. The rate of uptake of Ca2+ by the SR determines the rate of myocardial relaxation. The regulation of SERCA2a activity controls the contractility and relaxation of the heart, affecting cardiac function. The expression and activity of SERCA2a are reduced in failing hearts. Gene therapy by increasing the expression of SERCA2a in the heart has been proven effective. In addition, SERCA2a is regulated by a variety of factors, including transmembrane micropeptides, protein kinases, and post-translational modifications (PTMs). In this review, we discuss the regulatory factors of SERCA2a and provide new insights into future treatments and the direction of heart failure research. In addition, gene therapy for SERCA2a has recently emerged as therapeutic option and hence will be discussed in this review.

Heat stress causes oxidative damage and quality reduction in poultry. Here, a tandem mass tag proteomic approach was applied to investigate the proteomic differences in duck meat from birds exposed to heat stress. Altogether 212 differential proteins were identified, including 178 down-regulated and 34 up-regulated proteins, compared to the control. Malondialdehyde and carbonyl content and cooking loss of the chest muscle significantly increased under heat stress. The proteomic analysis indicated that heat stress suppressed mitochondrial functions and respiratory chains, which might be responsible for the higher oxidation level. The results also revealed potential protective proteins involved in the defensive mechanisms against heat stress in duck muscles, such as sarcoplasmic/endoplasmic reticulum calcium ATPases, Mn-superoxide dismutase, heat shock protein family B member 7, methyltransferase like 21C, myosin-binding protein C, and carbonic anhydrase 3. These results provide potential targets for the research and identification of oxidative meat products due to heat stress.

Among the physiological functions declining with aging, decreased secretion of gastric fluid, achlorhydria, is commonly observed. We evaluated the digestion of meat (chicken, beef, and pork) and soy proteins using in vitro conditions mimicking gastrointestinal (GI) digestion in adults (control, C) and elderly individuals with achlorhydria (EA). Changes in degrees of hydrolysis (DH), SDS-PAGE profiles, peptide concentrations, and proteomic profiles during digestion were investigated. Digestion under the EA conditions markedly decreased DH, especially for soy proteins. SDS-PAGE profiling and proteomics showed that myofibrillar/sarcoplasmic proteins from meat and glycinin/beta-conglycinin from soy were most affected by digestion conditions. Our results indicated that differences in the digestibility of meat protein between EA and control conditions gradually narrowed from the gastric to the intestinal phase for meat protein, while a pronounced difference persisted in the intestinal phase for soy protein. Our work provides new insight of value for future dietary recommendations for elderly individuals.

Made available in DSpace on 2022-04-28T17:23:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-11-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) In the present study, we describe a co-infection of two microsporidians within the abdominal muscle of the Amazon River prawnMacrobrachium amazonicum from Brazil, detected through light and transmission electron microscopy and histopathological analysis. Two spore types and respective early developmental stages (meronts, sporonts, and sporoblasts) appeared grouped in numerous adjacent clusters among the muscle fibers of the host. Spores were initially divided into two morphotypes, Sp. 1 and Sp. 2, based on differences in shape, size, and internal organization. The Sp. 1 spores are pyriform to ovoid (4.3 +/- 0.3 x 3.7 +/- 0.4 mu m in size) and resided in groups of eight within a sporophorous vesicle. Sp. 1 spores had bilayered walls and long tape-like external filaments with irregular morphology and size, forming a complex contiguous membranous system attached to the spore wall. The umbrella-like anchoring disc of the spores was in continuity with an anisofilar polar tube arranged in 7-8 (rarely 9) coils. The Sp. 2 spores were rod-like in shape (3.1 +/- 0.5 x 0.8 +/- 0.1 mu m in diameter) and the polar tube had 7-8 coils. Their merogonic and sporogonic stages occurred within the sarcoplasm of the muscle cells in close contact with the myofibrils. Based on ultrastructural organization of the sporogonic stages, the Sp. 1 and Sp. 2 morphotypes probably belong to the genera Inodosporus and Myospora, respectively. Univ Estadual Paulista UNESP, Programa Posgrad Ciencias Biol Zool, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil FIT MG, Fac Inovacao & Tecnol Minas Gerais, BR-37900060 Passos, MG, Brazil Univ Estadual Paulista UNESP, Lab Morfol Invertebrados LMI, Dept Biol, Fac Ciencias Agr & Ve Jaboticabal, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista UNESP, Ctr Aquicultura CAUNESP, Fac Ciencias Agr & Ve Jaboticabal, BR-14884900 Jaboticabal, SP, Brazil Univ Porto, Inst Invest & Inovacao Saude I3S, P-4200135 Porto, Portugal Univ Porto ICBAS UP, Lab Cell Biol, Inst Biomed Sci, P-4050313 Porto, Portugal Univ Inst Hlth Sci TOXRUN, Toxicol Res Unit, CESPU, P-4585116 Gandra, Portugal Univ Porto, Lab Anim Pathol, Interdisciplinary Ctr Marine & Environm Res CIIMA, P-4450208 Matosinhos, Portugal Univ Estadual Paulista UNESP, Programa Posgrad Ciencias Biol Zool, Inst Biociencias, BR-13506900 Rio Claro, SP, Brazil Univ Estadual Paulista UNESP, Lab Morfol Invertebrados LMI, Dept Biol, Fac Ciencias Agr & Ve Jaboticabal, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista UNESP, Ctr Aquicultura CAUNESP, Fac Ciencias Agr & Ve Jaboticabal, BR-14884900 Jaboticabal, SP, Brazil CAPES: 001 CAPES: 1989/2014 CAPES: 23038.004309/2014-5 FAPESP: JP 2005#04707-5 FAPESP: 2018/13685-5 CNPq: 309298/2020-2

Mag-Fluo-4 has revealed differences in the kinetics of the Ca2+ transients of mammalian fiber types (I, IIA, IIX, and IIB). We simulated the changes in [Ca2+] through the sarcomere of these four fiber types, considering classical (troponin –Tn–, parvalbumin –Pv–, adenosine triphosphate –ATP–, sarcoplasmic reticulum Ca2+ pump –SERCA–, and dye) and new (mitochondria –MITO–, Na+/Ca2+ exchanger –NCX–, and store-operated calcium entry –SOCE–) Ca2+ binding sites, during single and tetanic stimulation. We found that during a single twitch, the sarcoplasmic peak [Ca2+] for fibers type IIB and IIX was around 16 µM, and for fibers type I and IIA reached 10–13 µM. The release rate in fibers type I, IIA, IIX, and IIB was 64.8, 153.6, 238.8, and 244.5 µM ms−1, respectively. Both the pattern of change and the peak concentrations of the Ca2+-bound species in the sarcoplasm (Tn, PV, ATP, and dye), the sarcolemma (NCX, SOCE), and the SR (SERCA) showed the order IIB ≥ IIX &gt, IIA &gt, I. The capacity of the NCX was 2.5, 1.3, 0.9, and 0.8% of the capacity of SERCA, for fibers type I, IIA, IIX, and IIB, respectively. MITO peak [Ca2+] ranged from 0.93 to 0.23 µM, in fibers type I and IIB, respectively, while intermediate values were obtained in fibers IIA and IIX. The latter numbers doubled during tetanic stimulation. In conclusion, we presented a comprehensive mathematical model of the excitation–contraction coupling that integrated most classical and novel Ca2+ handling mechanisms, overcoming the limitations of the fast- vs. slow-fibers dichotomy and the use of slow dyes.

The sodium/calcium exchangers (NCX) use the extracellular sodium concentration to facilitate the extrusion of calcium out of the cell. Alongside the plasma membrane Ca2+-ATPase (PMCA) and sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA), as well as the sodium/potassium/calcium exchangers (NKCX, SLC24 family), NCX allow recovery of intracellular calcium back to basal levels after cellular stimulation. When intracellular sodium ion levels rise, for example, following depolarisation, these transporters can operate in the reverse direction to allow calcium influx and sodium efflux, as an electrogenic mechanism. Structural modelling suggests the presence of 9 TM segments, with a large intracellular loop between the fifth and sixth TM segments [1].