Search

Traumatic brain injury (TBI) is a major public health concern that can result in long-term neurological impairments. Calpain is a calcium-dependent cysteine protease that is activated within minutes after TBI, and sustained calpain activation is known to contribute to neurodegeneration and blood-brain barrier dysregulation. Based on its role in disease progression, calpain inhibition has been identified as a promising therapeutic target. Efforts to develop therapeutics for calpain inhibition would benefit from the ability to measure calpain activity with spatial precision within the injured tissue. In this work, we designed an activity-based nanotheranostic (ABNT) that can both sense and inhibit calpain activity in TBI. To sense calpain activity, we incorporated a peptide substrate of calpain flanked by a fluorophore/quencher pair. To inhibit calpain activity, we incorporated calpastatin peptide, an endogenous inhibitor of calpain. Both sensor and inhibitor peptides were scaffolded onto a polymeric nanoscaffold to create our ABNT. We show that in the presence of recombinant calpain, our ABNT construct is able to sense and inhibit calpain activity. In a mouse model of TBI, systemically administered ABNT can access perilesional brain tissue through passive accumulation and inhibit calpain activity in the cortex and hippocampus. In an analysis of cellular calpain activity, we observe the ABNT-mediated inhibition of calpain activity in neurons, endothelial cells, and microglia of the cortex. In a comparison of neuronal calpain activity by brain structure, we observe greater ABNT-mediated inhibition of calpain activity in cortical neurons compared to that in hippocampal neurons. Furthermore, we found that apoptosis was dependent on both calpain inhibition and brain structure. We present a theranostic platform that can be used to understand the regional and cell-specific therapeutic inhibition of calpain activity to help inform drug design for TBI.

PLACK syndrome (OMIM 616295) is a rare genodermatosis associated with peeling skin, leukonychia, acral punctate keratosis, cheilitis, and knuckle pads and is caused by loss‐of‐function mutations in the CAST gene, which encodes calpastatin, a calcium‐dependent protease. This case report highlights a case of PLACK syndrome presenting with the unique findings of striate hyperkeratosis on the palms as well as life‐threatening cardiomyopathy. We review why CAST mutations might impact cardiac function and raise awareness of the potential association between PLACK syndrome and cardiac manifestations. [ABSTRACT FROM AUTHOR]

This research aimed to examine the effects of haplotype groups observed in three different loci on the Calpastatin gene on ultrasonographic MLD measurements and seasonal live weight in five different types of sheep (GBK, HM, K, KM, and R). In the CAST intron 1, intron 5, and intron 12 regions, 15 SNPs were found. The HWE p-value for SNP2, SNP3, SNP5, SNP6, SNP7, SNP8, and SNP14 is less than 0.05, and except SNP9 and SNP10, all SNPs have a MAF of more than 0.01. SNP1, SNP2, and SNP7 made up one haplotype block. The haploblock has 3 haplogroups. The most common haplotype group was H1 (-AGG-), which had a frequency of 0.52; H2 (-TGG-) and H3 (-TAA-) had rates of 0.35 and 0.13, respectively. Based on ultrasonographic MLD readings and live weights, there were no statistically significant differences between haplotype H1 and H3, but there were statistically significant differences between haplotype H2 lambs. The effect of the H2 haplotype on 90-day MLD depth revealed a statistically significant difference between the HM and KM and K and KM breeds. This distinction persisted until the 180th day of life before disappearing into adulthood. Similarly, the effect of H2 haplotype on the skin thickness at day 90 was significant between K and KM and between K and R, whereas the effect of H2 haplotype on fat thickness demonstrated a substantial difference between HM and KM at one year of age.

The calpastatin (CAST) gene is one of the genes associated with meat quality. Studies have shown that this gene is associated with quality traits such as body weight gain, carcass yield, meat tenderness and fat content in meat. In this study, the polymorphism of the CAST gene was determined using the PCR-RFLP method in Holstein, Simmental and Brown Swiss cattle. As a result of genotyping, three genotypes (MM, MN and NN) and two alleles (M and N) were determined. It was found that the MM genotype and the M allele had the highest frequency in all breeds examined, while the NN genotype had the lowest frequency. The NN genotype was only found in the Brown Swiss breed. All Simmental animals used in the study belonged to the MM genotype. [ABSTRACT FROM AUTHOR]

Marker-assisted selection for improving reindeer meat production is at the early stages of development, which requires analysis of variation within the candidate genes for meat production. The calpastatin and androgen receptor genes were chosen as such genes to study their variability in reindeer. In different domesticated animal species, polymorphisms and indels in the androgen receptor gene were associated with growth and weight characteristics. Based on the results from many studies, sequence variation in the calpastatin CAST gene region was associated with meat quality and meat production of livestock. Principal component analysis of CAST variability grouped wild and domestic deer from Yakutia, as well as wild and domestic deer from Amur oblast, which implies gene flow between local breeds of domesticated deer and wild populations. At the same time, in the case of three microsatellite loci found in the present study in the intron of the androgen receptor gene, principal component analysis separated wild and domestic deer. [ABSTRACT FROM AUTHOR]

Supernumerary teats (SNT) are a prevalent and intricate polygenic trait that is frequently observed at birth in sheep. These additional teats have been linked to a heritable trait associated with maternal capability, especially in ewes with high fertility. Investigating the genomic basis of these characteristics is an ongoing endeavor and poses an intriguing research inquiry with implications for both biological understanding and practical applications. In order to delve into this scientific enigma, a specific cohort of 20 local fat-tailed Ghezel ewes, each possessing four functional teats, was carefully chosen to form the case group. In parallel, a control group comprising 25 healthy ewes with the standard two normal teats was established for comparative analysis. Blood samples were meticulously collected from both groups of Ghezel ewes demonstrating distinct phenotypic expressions, specifically those with either two or four functional teats. Following the sample collection, the genomic DNA extraction process was carried out using a commercial DNA purification kit provided by Qiagen company, known for its high-quality DNA extraction products. We conducted an examination of four PCR-RFLP markers on four candidate genes (IGF1, Leptin, Calpastatin, and FBP30). Through our research, we unveiled the allele distribution and genotype frequency of these potential genes in both the case and control groups of sheep. Our findings demonstrated notable disparities in the genotype and gene frequencies between the case and control groups for each candidate gene. Furthermore, we calculated the odds ratio (OR) and 95 percent confidence intervals (CIs) under Hardy-Weinberg disequilibrium using various genetic models. The χ2 test was also utilized to compare the genotype and allele distribution of these candidate gene polymorphisms between the case and control groups. The results of our study are particularly intriguing as they indicate that individuals with an AA genotype for the leptin gene are significantly more likely to have four additional functional teats (OR=7.7143, 95 percent CI=1.6025, Pvalue= 0.0108). Additionally, the results in a dominant model (AA vs. GA+GG) revealed an OR of 0.1296 and 95 percent CI of 0.02669-0.6240, with a p-value of 0.0108, further highlighting the association between leptin genotypes and supernumerary teats (SNT). Moreover, the risk of having more than four functional teats is heightened by the GG genotype in the FBP3 gene (P<0.13). However, our analysis did not uncover any additional significant correlation between the polymorphism of another investigated candidate gene and supernumerary teats (SNT). Overall, this topic presents an intriguing area that warrants further research to gain a comprehensive understanding of the biological mechanisms governing the development of four functional teats in ewes with large litter sizes. [ABSTRACT FROM AUTHOR]

Diabetes mellitus (DM) is a major risk factor for atrial structural remodeling and atrial fibrillation (AF). Calpain activity is hypothesized to promote atrial remodeling and AF. The purpose of this study was to investigate the role of calpain in diabetes-associated AF, fibrosis, and calcium handling dysfunction. DM-associated AF was induced in wild-type (WT) mice and in mice overexpressing the calpain inhibitor calpastatin (CAST-OE) using high-fat diet feeding followed by low-dose streptozotocin injection (75 mg/kg). DM and AF outcomes were assessed by measuring blood glucose levels, fibrosis, and AF susceptibility during transesophageal atrial pacing. Intracellular Ca2+ transients, spontaneous Ca2+ release events, and intracellular T-tubule membranes were measured by in situ confocal microscopy. WT mice with DM had significant hyperglycemia, atrial fibrosis, and AF susceptibility with increased atrial myocyte calpain activity and Ca2+ handling dysfunction relative to control treated animals. CAST-OE mice with DM had a similar level of hyperglycemia as diabetic WT littermates but lacked significant atrial fibrosis and AF susceptibility. DM-induced atrial calpain activity and downregulation of the calpain substrate junctophilin-2 were prevented by CAST-OE. Atrial myocytes of diabetic CAST-OE mice exhibited improved T-tubule membrane organization, Ca2+ handling, and reduced spontaneous Ca2+ release events compared to littermate controls. This study confirmed that DM promotes calpain activation, atrial fibrosis, and AF in mice. CAST-OE effectively inhibits DM-induced calpain activation and reduces atrial remodeling and AF incidence through improved intracellular Ca2+ homeostasis. Our results support calpain inhibition as a potential therapy for preventing and treating AF in DM patients. [ABSTRACT FROM AUTHOR]

Context: Selection for growth rate has received considerable attention in beef cattle but the evidence for an improvement in the efficiency of feed conversion is equivocal. Aim: To examine whether feed efficiency by beef cattle finished in a feedlot had been changed in response to divergence selection for growth rate. Methods: The Angus cattle used came from three lines of cattle selected for over five generations for fast growth rate to yearling age (High-line), slow growth (Low-line), or from an unselected Control-line. Over sequential years, a cohort of steers, then of heifers and then of steers, representative of the lines, were measured for feedlot performance, and carcase- and meat-quality traits. The animals were fed a high-energy feedlot ration and after an adjustment period they underwent a performance test of at least 70 days of duration. After slaughter, muscle samples were taken for subsequent measurement of the components of the endogenous calpain proteolytic enzyme system. Their carcasses underwent a standard chiller assessment and meat samples were taken after 1 day and 14 days (steers) or 17 days (heifers) for objective measurement of tenderness. Key results: Cattle from the High-line grew 48% faster (P < 0.05), and ate 48% more feed (P < 0.05) than did those from the Low-line, but had similar (P > 0.05) feed conversion ratio and residual feed intake. There were no differences between the High-line and Low-line in the visual meat-quality attributes of meat colour, fat colour and marbling, and no differences in the objective measurements of tenderness and connective-tissue toughness. There was no evidence of a selection response in the circulating concentrations of the metabolites and hormones measured, nor in the endogenous calpain proteolytic enzyme system in muscle. Conclusions: The superior growth demonstrated by the High-line cattle over the feedlot test was accompanied by a higher feed intake, with no evidence for an improvement in feed efficiency. Implications: Selection for growth rate is a powerful tool to alter animal performance but the beef industry needs to be cognisant of the proportional increase in feed requirement from breeding bigger animals. Selection for growth rate is a powerful tool to alter beef cattle performance but the evidence for an improvement in feed efficiency is equivocal. This experiment found that selection for faster growth in Angus cattle resulted in heavier cattle that ate more in the feedlot, with no evidence of improvement in feed efficiency. Breeding programs that place emphasis on growth rate need to be cognisant of the proportional increase in feed requirement by bigger animals. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) is the most malignant brain tumor, characterized by cell heterogeneity comprising stem cells (GSCs) responsible for aggressiveness. The calpain/calpastatin (calp/cast) proteolytic system is involved in critical physiological processes and cancer progression. In this work we showed the expression profile of hcast 3-25 (a Type III calpastatin variant devoid of inhibitory units) and the members of the system in several patient-derived GSCs exploring the relationship between hcast 3-25 and activation/activity of calpains. Each GSC shows a peculiar calp/cast mRNA and protein expression pattern, and hcast 3-25 is the least expressed. Differentiation promotes upregulation of all the calp/cast system components except hcast 3-25 mRNA, which increased or decreased depending on individual GSC culture. Transfection of hcast 3-25-V5 into two selected GSCs indicated that hcast 3-25 effectively associates with calpains, supporting the digestion of selected calpain targets. Hcast 3-25 possibly affects the stem state promoting a differentiated, less aggressive phenotype.

Background: This study aims to identify circulating biomarkers by using proteomic analysis associated with sac shrinkage or expansion in patients undergoing endovascular aneurysm repair (EVAR) for abdominal aortic aneurysms (AAAs). Methods: Plasma samples were analysed from 32 patients treated with EVAR between 10/2009 and 10/2020. Patients were divided into two groups based on postoperative sac behaviour: sac shrinkage (≥5 mm reduction) and no shrinkage (stabilisation or expansion). Proteomic analysis was performed using high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS), with abundant protein depletion to enhance the detection of low-abundant proteins. Results: Of the 32 patients, 20 exhibited sac shrinkage, and 12 showed no shrinkage. Proteomic analysis identified 632 proteins, with significant differential abundance observed after adjusting for relevant clinical parameters. Notably, neurogranin (NRGN) levels were significantly associated with hypertension and smoking, while casein alpha S1 (CSN1S1) levels varied with statin use. Differentially abundant proteins related to aortic diameter included calpastatin, SCUBE3, and ubiquitin-conjugating enzyme E2, among others. Conclusions: Proteomic profiling revealed distinct biomarker patterns associated with sac behaviour in EVAR-treated AAA patients. These findings suggest potential therapeutic targets for enhancing EVAR outcomes and underscore the need for further investigation into the biological mechanisms underlying aneurysm sac shrinkage and stability.

β-catenin is an important regulator of malignant progression. 17β-Estradiol (E2), an important sex hormone in women, promotes the growth and metastasis of triple-negative breast cancer (TNBC). However, whether β-catenin is involved in E2-induced metastasis of TNBC remains unknown. In this study, we show that E2 induces the proliferation, migration, invasion, and metastasis of TNBC cells. E2 induces β-catenin protein expression and nuclear translocation, thereby regulating the expression of target genes such as Cyclin D1 and MMP-9. The inhibition of β-catenin reversed the E2-induced cell malignant behaviors. Additionally, E2 activated Calpain by increasing intracellular Ca2+ levels and reducing calpastatin levels. When Calpain was inhibited, E2 did not induce the proliferation, migration, invasion, or metastasis of TNBC cells. In addition, E2 promoted translocation of YAP into the nucleus by inhibiting its phosphorylation. Calpain inhibition reversed the E2-induced YAP dephosphorylation. Inhibition of YAP transcriptional activity reversed the effects of E2 on the proliferation, migration, invasion, and β-catenin of TNBC cells. In conclusion, we demonstrated that E2 induced metastasis-related behaviors in TNBC cells and this effect was mediated through the Calpain/YAP/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Hypoxic-ischemic encephalopathy (HIE) is a complex pathophysiological process with multiple links and factors. It involves the interaction of inflammation, oxidative stress, and glucose metabolism, and results in acute and even long-term brain damage and impairment of brain function. Calpain is a family of Ca2+-dependent cysteine proteases that regulate cellular function. Calpain activation is involved in cerebral ischemic injury, and this involvement is achieved by the interaction among Ca2+, substrates, organelles, and multiple proteases in the neuronal necrosis and apoptosis pathways after cerebral ischemia. Many calpain inhibitors have been developed and tested in the biochemical and biomedical fields. This study reviewed the potential role of calpain in the treatment of HIE and related mechanism, providing new insights for future research on HIE. [ABSTRACT FROM AUTHOR]

Copyright of Revista Colombiana de Ciencia Animal - RECIA is the property of Revista Colombiana de Ciencia Animal - RECIA and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Bibliometric analysis is commonly used to guide decisions regarding research funding and the creation of research policies and provides scientists with helpful information about research trends, patterns, and impact. Thus, researchers can track collaborators in this subject and find prospective scientific alliances. Additionally, researchers can develop new research themes by constantly monitoring the most recent trend study topics in this area. Therefore, we performed a comprehensive bibliometric analysis of 4085 documents scanned in the Web of Science (WoS) database on some major effect genes associated with meat yield traits in livestock between 1981 and 2023. The analysis shows that interest in this topic has recently grown. The fact that numerous scholars participated in the investigations, which major research groups conducted, demonstrates the growth of this field's collaborative working culture. The publication of studies in this field in high-impact journals such as Meat Science, Journal of Animal Science, and Animal Genetics reveals the scientific impact of this field. Keywords used in studies in this field are generally related to investigating the genetic factors affecting livestock's growth, muscle development, and meat quality characteristics. In country-based studies, China and the United States have the most studies in this field. The citation records of articles reveal the significant impact of this field in literature. The scientific community has recently been particularly interested in investigating some major effect genes associated with meat yield traits in livestock. [ABSTRACT FROM AUTHOR]

In connection with the increasing interest in the production of young mutton and lamb, priority is given to the study of genes that control meat production. Alleles of genes calpastatin (CAST) and somatotropin (GH) may act as potential markers of sheep meat productivity. However, until now there is no information on the influence of these genes on the indicators of meat productivity of sheep of Russian breeds. Based on this, the purpose of this research was to study the polymorphism of the CAST and GH genes in meat and wool sheep of the ½ Poll Dorset x ½ North Caucasian meat - wool genotype bred in the Stavropol Territory (Russia) and their influence on the traits of meat productivity. Genotyping of sheep for somatotropin and calpastatin genes was carried out by polymerase chain reaction (PCR) with further study of restriction fragment length polymorphism (RFLP). Three genotypes were identified for the GH gene (AA, AB, and BB) and two for CAST (MM and MN). The highest frequency of occurrence for the GH gene was characterized by the heterozygous AB genotype (42.8%), for the CAST gene - the homozygous MM genotype (87.9%). These genotypes were correlated with quantitative and qualitative parameters of meat productivity. The best indicators of meat productivity were in the bright AB, BB, and MN genotypes of the growth hormon and calpastatin genes. The slaughter weight of individuals of the AB, BB genotype of the GH gene and the MN genotype of the CAST gene is higher by 6.3, 7.3, and 5.2%, respectively. According to the point assessment of the "marbling" of meat, animals with the indicated genotypes outnumbered their peers by 1.8; 2.1 and 3.7 points. [ABSTRACT FROM AUTHOR]

The sheep breeeding for meat is of great interst at present due to rising meat intake among people. Over the recent decade, selective breeding with molecular markers has provided the opportunity to improve the sheep muscular system and the carcass weight and yield with decreasing the fat accumulation. Recognition of genes affecting the characteristics of farm animal products has allowed the researchers to identify single nucleotide polymorphisms (SNPs) associated with phenotypic traits improving the productive performance of livestock. Alleles of genes calpastatin (CAST) and somatropin, the synthetic form of growth hormone (GH) can represent the potential markers for sheep meat productivity. The objective of the survey is to study polymorphic variants of the CAST and GH genes in meat and wool sheep of the genotype of cross 1/2 Poll Dorset ×1/2 North-Caucasian meat-and-wool breed, reared in the Stavropol krai and their relationships with the traits important in meat production. Sheep genotyping for the somatotropin and calpastatin genes was performed with the method of PCR restriction fragment length polymorphism (PCR-RFLP). The evaluated genes were polymorphic. Three genotypes (AA, AB, and BB) of gene GH and two genotypes (MM and MN) of gene CAST have been identified. Genotypes AB and MM were characterised by the highest frequency of occurrence (42.8 and 87.9%, respectively). The highest values for meat production were recorded in the ewes with the AB, BB, and MN genotypes. The slaughter weight of specimens with genotype MN of the CAST gene was greater than that in the sheep with homozygous genotype (MM) by 5.9% and in the animals with genotypes AB and BB of gene GH, compared to the ewes with genotype AA, by 6.8 and 7.5%, respectively. The animals with genotypes indicated above were more efficient than their peers were in the muscle-tissue protein contents by 0.6, 3.5, and 1.5%. [ABSTRACT FROM AUTHOR]

Reperfusion after ischemia would cause massive myocardial injury, which leads to oxidative stress (OS). Calcium homeostasis imbalance plays an essential role in myocardial OS injury. CaV1.2 calcium channel mediates calcium influx into cardiomyocytes, and its activity is modulated by a region of calpastatin (CAST) domain L, CSL54-64. In this study, the effect of Ahf-caltide, derived from CSL54-64, on myocardial OS injury was investigated. Ahf-caltide decreased the levels of LDH, MDA and ROS and increased heart rate, coronary flow, cell survival and SOD activity during OS. In addition, Ahf-caltide permeated into H9c2 cells and increased CaV1.2, CaVβ2 and CAST levels by inhibiting protein degradation. At different Ca2+ concentrations (25 nM, 10 μM, 1 mM), the binding of CSL to the IQ motif in the C terminus of the CaV1.2 channel was increased in a H2O2 concentration-dependent manner. CSL54-64 was predicted to be responsible for the binding of CSL to CaV1.2. In conclusion, Ahf-caltide exerted a cardioprotective effect on myocardial OS injury by stabilizing CaV1.2 protein expression. Our study, for the first time, proposed that restoring calcium homeostasis by targeting the CaV1.2 calcium channel and its regulating factor CAST could be a novel treatment for myocardial OS injury. [ABSTRACT FROM AUTHOR]

The single-nucleotide polymorphism (SNP) form of genes is a valuable source of information regarding their suitability for use as specific markers of desirable traits in beef cattle breeding. For several decades, breeding work focused on improving production efficiency through optimizing the feed conversion ratio and improving daily gains and meat quality. Many research teams previously undertook research work on single-nucleotide polymorphism in myostatin (MSTN), thyroglobulin (TG), calpain (CAPN), and calpastatin (CAST) proteins. The literature review focuses on the most frequently addressed issues concerning these genes in beef cattle production and points to a number of relevant studies on the genes’ polymorphic forms. The four genes presented are worth considering during breeding work as a set of genes that can positively influence productivity and production quality.

Abstract Feeding is a determining factor in the various characteristics of sheep meat and animal performance, the objectives were to evaluate the effect of supplementation of ewe lambs finished in different nutritional planes on the gene expression of CASP3, CAPN1, CAPN2 and CAST and its possible association with meat quality. Samples of the Longissimus lumborum muscle of 24 ewe lambs were used, distributed in 3 groups (n=8): P (pasture), PS (pasture and supplement) and F (feedlot). Physicochemical analyses were performed for centesimal analysis, pH, lipid oxidation, Warner-Bratzler shear force and RT-qPCR for the analysis of relative gene expression of the following genes: CASP3, CAPN1, CAPN2 and CAST. There is an increase in daily weight gain and ethereal extract values in the meat of confined animals, due to the greater energy intake in the nutrition of these animals. Animals kept only on pasture have lower lipid oxidation in meat than other treatments because of the lower percentage of lipids. The Warner-Bratzler shear force is considerably higher in the meat of animals kept only on pasture but is still considered tender. The different nutritional systems do not interfere with the gene expression of CASP3, CAPN1, CAPN2 and CAST in ewe lambs.

The intercalated disk is a cardiac specific structure composed of three main protein complexes—adherens junctions, desmosomes, and gap junctions—that work in concert to provide mechanical stability and electrical synchronization to the heart. Each substructure is regulated through a variety of mechanisms including proteolysis. Calpain proteases, a class of cysteine proteases dependent on calcium for activation, have recently emerged as important regulators of individual intercalated disk components. In this review, we will examine how calcium homeostasis regulates normal calpain function. We will also explore how calpains modulate gap junctions, desmosomes, and adherens junctions activity by targeting specific proteins, and describe the molecular mechanisms of how calpain dysregulation leads to structural and signaling defects within the heart. We will then examine how changes in calpain activity affects cardiomyocytes, and how such changes underlie various heart diseases. [ABSTRACT FROM AUTHOR]

Background: The family with sequence similarity 20-member C (FAM20C) kinase, a Golgi casein kinase, which is responsible for phosphorylating the majority of the extracellular phosphoproteins within S-x-E/pS motifs, and is fundamentally associated with multiple biological processes to maintain cell proliferation, biomineralization, migration, adhesion, and phosphate homeostasis. In dissecting how FAM20C regulates downstream molecules and potential mechanisms, however, there are multiple target molecules of FAM20C, particularly many phenomena remain elusive, such as changes in cell-autonomous behaviors, incompatibility in genotypes and phenotypes, and others. Methods: Here, assay for transposase-accessible chromatin using sequencing (ATAC-seq), RNA sequencing (RNA-seq), proteomics, and phosphoproteomics were performed in Fam20c-dificient osteoblasts and to facilitate an integrated analysis and determine the impact of chromatin accessibility, genomic expression, protein alterations, signaling pathway, and post translational modifcations. Results: By combining ATAC-seq and RNA-seq, we identified TCF4 and Wnt signaling pathway as the key regulators in Fam20c-dificient cells. Further, we showed Calpastatin/Calpain proteolysis system as a novel target axis for FAM20C to regulate cell migration and F-actin cytoskeleton by integrated analysis of proteomics and phosphoproteomics. Furthermore, Calpastatin/Calpain proteolysis system could negatively regulate the Wnt signaling pathway. Conclusion: These observations implied that Fam20c knockout osteoblasts would cause cell homeostatic imbalance, involving changes in multiple signaling pathways in the conduction system. [ABSTRACT FROM AUTHOR]

Tenderness is an important characteristic of meat quality. Calpastatin and calpain play important roles in meat tenderization. However, it is not clear how phosphorylation affects the regulation of calpastatin on μ-calpain and, consequently, meat tenderness. Calpastatin with high and low phosphorylation levels were obtained in vitro corresponding to the treatments by protein kinase A (PKA) and alkaline phosphatase. Then, calpain was incubated with calpastatin with different phosphorylation levels, and the effect of calpastatin on calpain activity under different phosphorylation levels was analyzed. The results showed that PKA promoted the phosphorylation of calpastatin, and a high phosphorylation level was maintained during incubation. The degradation rate of μ-calpain in AP group was higher than that in the other groups, meaning there was lower inhibition of calpastatin on calpain activity. The degradation of calpastatin was lower and its structure was more stable after phosphorylation. One more serine 133 site of calpastatin was identified in PKA group compared with the other groups. Phosphorylation at serine 133 of calpastatin enhanced its inhibition on calpain activity by maintaining its structural stability, thus inhibiting the tenderization of meat. [ABSTRACT FROM AUTHOR]

Calcium/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) regulates bone remodeling through its effects on osteoblasts and osteoclasts. However, its role in osteocytes, the most abundant bone cell type and the master regulator of bone remodeling, remains unknown. Here we report that the conditional deletion of CaMKK2 from osteocytes using Dentine matrix protein 1 (Dmp1)-8kb-Cre mice led to enhanced bone mass only in female mice owing to a suppression of osteoclasts. Conditioned media isolated from female CaMKK2-deficient osteocytes inhibited osteoclast formation and function in in vitro assays, indicating a role for osteocyte-secreted factors. Proteomics analysis revealed significantly higher levels of extracellular calpastatin, a specific inhibitor of calcium-dependent cysteine proteases calpains, in female CaMKK2 null osteocyte conditioned media, compared to media from female control osteocytes. Further, exogenously added non-cell permeable recombinant calpastatin domain I elicited a marked, dose-dependent inhibition of female wild-type osteoclasts and depletion of calpastatin from female CaMKK2-deficient osteocyte conditioned media reversed the inhibition of matrix resorption by osteoclasts. Our findings reveal a novel role for extracellular calpastatin in regulating female osteoclast function and unravel a novel CaMKK2-mediated paracrine mechanism of osteoclast regulation by female osteocytes. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare the influence of beef production systems using additive combinations of growth-promotant technologies on meat quality. Steer calves (n  = 120) were assigned to 1 of 4 treatments: 1) no technology (NT; control), 2) antibiotic treated (ANT; NT plus therapeutic antibiotics, monensin, and tylosin), 3) implant treated (IMP; ANT plus a series of three implants), and 4) beta-agonist treated (BA; IMP plus ractopamine-HCl). Muscle biopsy samples from the longissimus lumborum were extracted from a subset (n  = 4 per treatment) of steers to evaluate expression of calpain-1, calpain-2, and calpastatin using real-time RT-PCR. Following carcass chilling, objective color (L *, a *, and b *) was evaluated. The right strip loin was removed from each carcass, portioned into 2.54-cm steaks, and designated to 7, 14, or 21 d postmortem aging periods for analysis of cook loss and Warner–Bratzler shear force (WBSF). The anterior face of each strip loin was used for analysis of crude fat and moisture. Treatment influenced (P  < 0.001) L *, a *, and b *. The NT and IMP treatments had greater (P  < 0.01) L * values, ANT was intermediate, and BA had the lowest (P  < 0.01) L * values. The NT and IMP treatments had higher (P  < 0.01) a * and b * values compared with ANT, which were higher (P  < 0.01) than BA. Steaks from implanted steers (IMP and BA) tended (P ≤ 0.067) to exhibit higher a * and b * than steaks from nonimplanted steers. Cattle in the NT and ANT treatments produced steaks with increased (P  < 0.01) crude fat percentage compared with the IMP and BA treatments, which were similar (P  > 0.05). Percent moisture of NT steaks was lower (P  < 0.01) than all other treatments, ANT was intermediate, and IMP and BA were similar (P  > 0.05) and had the highest (P  < 0.01) moisture content. Cook loss tended to be greater (P  = 0.088) for implanted steers (IMP and BA) compared to nonimplanted steers (NT and ANT). Steaks from NT and ANT treatments were more tender (P  < 0.05) than IMP and BA, which were similar (P  > 0.05). Thus, WBSF was lower (P  < 0.001) in nonimplanted than implanted steaks. Expression of calpastatin was increased (P ≤ 0.025) in ANT and BA treatments, and there was a tendency for expression of calpain-2 to be increased (P  = 0.081) in ANT compared to NT. These results suggest that production systems with limited use of growth promoting technology produced strip loins with more crude fat, less moisture and cook loss, and improved tenderness. [ABSTRACT FROM AUTHOR]

This article, titled "[Retracted] Calpastatin peptide attenuates early brain injury following experimental subarachnoid hemorrhage," was published in the journal Experimental & Therapeutic Medicine in 2020. However, it has since been retracted due to concerns raised by a reader. The reader pointed out that an image in the article was very similar to an image in another article published in a different journal. After discussing the issue with the authors, the editor decided to retract the paper. The authors accepted this decision, and the editor apologizes for any inconvenience caused. [Extracted from the article]

Objective The aim of this study was to evaluate the effect of ageing and feeding strategies on the calpain protease system and meat quality traits in Braford steers. Methods Thirty Braford steers were employed; 15 animals were supplemented with corn silage during finishing and 15 were kept only on pasture. Meat quality traits and calpain system protein activity were evaluated in longissimus thoracis et lumborum (LTL) steaks aged for 2, 7, 14, and 21 days. Results Aged meat showed higher pH and calcium content, while Warner Bratzler shear force (WBSF) decreased to day 21. No interaction between ageing and diet was seen for quality traits. Steers finished with corn silage showed higher values of water holding capacity, WBSF and free calcium, and lower values of pH and cooking loss. Calpain and calpastatin activities decreased with ageing. Finishing steers on pasture produced higher values of calpains and lower values of calpastatin activities. The higher values of calpain 1 activity were observed in muscles aged 2 days from pasture finished animals, and the lower activity of the inhibitor in the 21 days aged samples of the same group. Conclusion These results suggest a diet by ageing interaction in calpains and calpastatin and this interaction impact in Warner Bratzler Shear Force in Braford LTL muscle.

Myelin forming around axons provides electrical insulation and ensures rapid and efficient transmission of electrical impulses. Disruptions to myelinated nerves often result in nerve conduction failure along with neurological symptoms and long-term disability. In the central nervous system, calpains, a family of calcium dependent cysteine proteases, have been shown to have a role in developmental myelination and in demyelinating diseases. The roles of calpains in myelination and demyelination in the peripheral nervous system remain unclear. Here, we show a transient increase of activated CAPN1, a major calpain isoform, in postnatal rat sciatic nerves when myelin is actively formed. Expression of the endogenous calpain inhibitor, calpastatin, showed a steady decrease throughout the period of peripheral nerve development. In the sciatic nerves of Trembler-J mice characterized by dysmyelination, expression levels of CAPN1 and calpastatin and calpain activity were significantly increased. In lysolecithin-induced acute demyelination in adult rat sciatic nerves, we show an increase of CAPN1 and decrease of calpastatin expression. These changes in the calpain-calpastatin system are distinct from those during central nervous system development or in acute axonal degeneration in peripheral nerves. Our results suggest that the calpain-calpastatin system has putative roles in myelination and demyelinating diseases of peripheral nerves. [ABSTRACT FROM AUTHOR]

Identification and separation of endogenous proteases are important to find out their role in changes of biochemical process in post‐mortem muscle. Diverse studies were conducted to find out the activity of these enzymes but only a very few studies were carried out on avian species. In this study, a simple casein zymography method was explained for detection of calpain‐1 and calpain‐2 with the standardization of extraction buffer and that were quantified biochemically after one‐step separation and purification on anion exchange chromatography. DEAE Sephacel matrix was used to purify calpain‐1, calpain‐2, and calpastatin. SDS‐PAGE analysis showed native and autolyzed forms of calpains. Biochemical test indicated concentrations of calpain‐1, calpain‐2, and calpastatin being higher in breast than thigh muscles across all four breeds of chicken. A simple, effective, and low‐cost extraction, purification, and separation technique was developed for accurate identification of calpain‐1and calpain‐2 on casein zymography method and quantification by biochemical test. Novelty Impact Statement: Identification and isolation of endogenous proteases are necessary to determine their significance in the PM aging of chicken meat from various breeds. In this study, a straightforward casein zymography approach for in‐vitro detection of calpain‐1 and calpain‐2 activity is devised. This will aid in the optimization of PM aging duration for meat tenderization. [ABSTRACT FROM AUTHOR]

Na V 1.5 channel is an integral membrane protein involved in the initiation and conduction of action potentials. IQ motif is located in the C-terminal domain of Na V 1.5 sodium channel, which is highly conserved in human sodium channel subtypes. IQ motif is involved in the Ca2+-dependent regulation through interaction with the regulatory proteins such as calpastatin domain L (CS L). Mutations in SCN5A, the gene encoding Na V 1.5 channel, have been linked to many cardiac arrhythmias, such as Long QT syndrome type 3 (LQT3) and Brugada syndrome (BRS). LQT3-associated mutations in Na V 1.5 IQ motif, IQ Q1909R and IQ R1913H , have been reported to affect the late I Na. A BRS-associated mutation in Na V 1.5 IQ motif, IQ A1924T, has been reported to affect the peak I Na. But the detailed pathogenic mechanisms of LQT3 and BRS remains unclear. To explore the binding properties of CS L to IQ motif and its muants associated with LQT3/BRS, molecular docking and GST pull down assay were performed in this study. As a result, S58 and E59 in CS L activating channel effect region L54-64 were involved in the conformation of the CS L /IQ WT complex by protein-protein docking. IQ motif could bind to CS L in a [CS L ]-dependent and [Ca2+]-dependent manner by pull down assay. However, the binding affinities of IQ Q1909R and IQ R1913H to CS L were decreased and its reaction rates with CS L were slower. The binding characteristics of IQ A1924T to CS L was opposite in a [Ca2+]-dependent manner and its binding efficacy became smaller. The changes of the binding characteristics of IQ mutants to CS L would affect the regulation of Na V 1.5 channel, which may be related to LQT3 and BRS. • CS L could bind to Na V 1.5 channel IQ motif in a [CS L ]-dependent and [Ca2+]-dependent manner. • The binding sites of CS L to IQ motif were within the region of CS L that reprime the activity of Ca V 1.2 channel, L54-64. • LQTs-associated mutants, IQ Q1909R and IQ R1913H weakened the binding activity to CS L and the reaction rates were slower. • The binding characteristics of BRS-associated mutant, IQ A1924T , to CS L was reversed in the [Ca2+]-dependent manner. [ABSTRACT FROM AUTHOR]

Background: This study aims to identify circulating biomarkers by using proteomic analysis associated with sac shrinkage or expansion in patients undergoing endovascular aneurysm repair (EVAR) for abdominal aortic aneurysms (AAAs)., Methods: Plasma samples were analysed from 32 patients treated with EVAR between 10/2009 and 10/2020. Patients were divided into two groups based on postoperative sac behaviour: sac shrinkage (≥5 mm reduction) and no shrinkage (stabilisation or expansion). Proteomic analysis was performed using high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS), with abundant protein depletion to enhance the detection of low-abundant proteins., Results: Of the 32 patients, 20 exhibited sac shrinkage, and 12 showed no shrinkage. Proteomic analysis identified 632 proteins, with significant differential abundance observed after adjusting for relevant clinical parameters. Notably, neurogranin (NRGN) levels were significantly associated with hypertension and smoking, while casein alpha S1 (CSN1S1) levels varied with statin use. Differentially abundant proteins related to aortic diameter included calpastatin, SCUBE3, and ubiquitin-conjugating enzyme E2, among others., Conclusions: Proteomic profiling revealed distinct biomarker patterns associated with sac behaviour in EVAR-treated AAA patients. These findings suggest potential therapeutic targets for enhancing EVAR outcomes and underscore the need for further investigation into the biological mechanisms underlying aneurysm sac shrinkage and stability.