Muscle Samples Research Articles

Islet-derived exosomal miR-204 accelerates insulin resistance in skeletal muscle by suppressing sirtuin 1: An in vivo study in a mouse model of high-fat diet-induced obesity.

The interaction between pancreatic islets and skeletal muscle plays a pivotal role in the development of insulin resistance. The present study aimed to elucidate the impact of non-hormonal molecules from islets on the insulin sensitivity of skeletal muscle cells. We developed a mouse model of obesity through a high-fat diet, assessing glucose tolerance and conducting miRNA sequencing on skeletal muscle samples. An in vitro model was established by treating cells with palmitic acid, and exosomes in the supernatant were characterized using scanning electron microscopy and CD63 expression analysis. Intracellular miR-204-5p levels were quantified by RT-PCR. Our in vivo model demonstrated a robust correlation between miR-204-5p level alterations and obesity-induced insulin resistance. Elevated fatty acid levels were observed to increase miR-204-5p in both skeletal muscle and islets. In cellular studies, palmitic acid increased miR-204-5p in MIN-6 islet β-cells but not in C2C12 skeletal muscle cells. Exosomes containing miR-204-5p, secreted by palmitic acid-treated MIN6 cells, were identified through morphological examination, immunoblotting for the exosomal marker CD63, and intraexosomal miR-204-5p level measurement. C2C12 cells were shown to uptake islet-derived miR-204-5p exosomes, as evidenced by the uptake of Exo-Red labeled exosomes. TargetScan analysis identified a highly conserved binding site for miR-204-5p in the 3' UTR of Sirt mRNA. Functional studies indicated that miR-204-5p overexpression reduced glucose consumption and uptake in C2C12 cells, decreased Sirt expression, and impaired insulin signaling, as evidenced by reduced Akt phosphorylation and membrane Glut4 levels. Our findings reveal that miR-204-5p contributes to the development of insulin resistance in obesity and acts as a signaling molecule in the crosstalk between pancreatic islets and skeletal muscle.

Occurrence of Trichinella spiralis in farmed wild boars (Sus scrofa): an underrated risk in China.

Natural infection by Trichinella sp. has been reported in humans and more than 150 species of animals, especially carnivorous and omnivorous mammals. Although the presence of Trichinella sp. infection in wild boars (Sus scrofa) has been documented worldwide, limited information is known about Trichinella circulation in farmed wild boars in China. This study intends to investigate the prevalence of Trichinella sp. in farmed wild boars in China. Seven hundred and sixty-one (761) muscle samples from farmed wild boars were collected in Jilin Province of China from 2017 to 2020. The diaphragm muscles were examined by artificial digestion method. The overall prevalence of Trichinella in farmed wild boars was 0.53% [95% confidence interval (CI): 0.51–0.55]. The average parasite loading was 0.076 ± 0.025 larvae per gram (lpg), and the highest burden was 0.21 lpg in a wild boar from Fusong city. Trichinella spiralis was the only species identified by multiplex polymerase chain reaction. The 5S rDNA inter-genic spacer region of Trichinella was amplified and sequenced. The results showed that the obtained sequence (GenBank accession number: OQ725583) shared 100% identity with the T. spiralis HLJ isolate (GenBank accession number: MH289505). Since the consumption of farmed wild boars is expected to increase in the future, these findings highlight the significance of developing exclusive guidelines for the processing of slaughtered farmed wild boar meat in China.

Mitochondrial quality control measures, systemic inflammation, and lower-limb muscle power in older adults: a PROMPT secondary analysis

ObjectivesThe study was conducted to explore associations between markers of mitochondrial quality control (MQC) from vastus lateralis muscle biopsies, serum inflammatory markers, and measures of muscle power assessed by two different tools in a sample of older adults. DesignSecondary analysis of data collected in the PeppeR develOpMental ProjecT (PROMPT) at the University of Florida (Gainesville, FL, USA). MethodsForty-three older adults (n = 20 women) were included in the study. Muscle volume of the calf and thigh was quantified by three-dimensional magnetic resonance imaging. Lower-limb muscle power was estimated using 5-time sit-to-stand (5STS) muscle power equations and isokinetic test. Protein markers of MQC were measured in muscle samples by Western immoblotting (n = 12–23), while type I and II fiber cross-sectional area (CSA) and their proportion were quantified using immunohistochemistry (n = 12). Cytochrome C oxidase enzyme activity was measured spectrophotometrically. Finally, inflammatory markers were quantified in the serum using a multiplex immunoassay (n = 39). ResultsMean age of participants was 78.1 ± 5.5 years, and the average body mass index was 26.2 ± 4.5 kg/m2. Markers of mitochondrial biogenesis (i.e., PGC-1α), mitochondrial import proteins (i.e., cHsp70 and mtHsp70), and type I fiber CSA were significantly associated with muscle power estimated via both 5STS muscle power equations and isokinetic test (p < 0.05). Specific associations were also found according to the muscle power assessment method. 5STS muscle power measures were negatively correlated with ClvCasp3, P-AMPK, T-AMPK, P-p38, GM-CSF, INF-γ, IL1b, IL6, IL8, and TNF-α, whereas positive associations were found with BAX (p < 0.05). In contrast, isokinetic measures were significantly and positively correlated with RIP140, Hsp60, and type II muscle fiber CSA (p < 0.05). ConclusionsMarkers of mitochondrial biogenesis (PGC-1α), mitochondrial import proteins (cHsp70 and mtHsp70), and type I muscle fiber CSA were significantly linked to lower-limb muscle power in older adults. These results suggest that muscle power is influenced by mitochondrial signaling. We also found that the relationship between mitochondrial mediators, inflammatory markers, and muscle power varied according to the assessment tool used.

Carbohydrate storage in cells: a laboratory activity for the assessment of glycogen stores in biological tissues.

Carbohydrates and fats constitute our primary energy sources. The importance of each of these energy substrates varies across cell types and physiological conditions. For example, the brain normally relies almost exclusively on glucose oxidation, whereas skeletal muscle shifts from lipids toward higher carbohydrate oxidation rates as exercise intensity increases. Understanding how carbohydrates are stored in our cells and which tissues contain significant carbohydrate stores is crucial for health professionals, especially given the role of carbohydrate metabolism in various pathophysiological conditions. This laboratory activity uses a simple and low-cost iodine binding method to quantify glycogen in mouse skeletal muscle and liver samples. By integrating the results of this activity with literature data, students can determine overall glycogen storage in the human body. The primary goal of the activity is to enhance students' understanding of the importance and limitations of glycogen stores in energy metabolism.NEW & NOTEWORTHY Carbohydrates are one of the primary energy sources utilized by our cells. Liver and skeletal muscle glycogen, which are the main carbohydrate reserves in the body, play a central role in energy metabolism, especially during periods of fasting and exercise. In this laboratory activity, students measure glycogen levels in tissues to gain insights into how carbohydrates are stored in our cells and understand the role and limitations of liver and muscle carbohydrate stores.

Mitochondrial pathways and sarcopenia in the geroscience era

Sarcopenia is associated with structural, ultrastructural, and molecular abnormalities of skeletal muscle. Mitochondrial dysfunction is a pivotal factor involved in muscle aging and sarcopenia. Mitochondrial bioenergetics are significantly reduced in muscles of older adults which is associated with whole-body aerobic capacity, muscle strength, and physical performance. Transcriptional profiling of muscle samples from older adults also revealed inverse correlations between gene expression patterns of autophagy and mitophagy and muscle volume and physical performance. This is in line with the proposition that mitochondrial quality control (MQC) processes are key to organellar and tissue health. MQC encompasses mitochondrial biogenesis, dynamics, and mitophagy. The latter has recently been included among the hallmarks of aging and alterations in MQC have been associated with chronic sterile inflammation as well as muscle atrophy and dysfunction. Several biomarkers spanning MQC, inflammation, metabolism, intercellular communication, and gut microbiota have been linked to sarcopenia. Findings from these initial studies hold promise to inform geroscience-based research in the field of sarcopenia by offering a plausible biological framework for developing gerotherapeutics and monitoring their effects.

Single-Nucleus RNA Sequencing Unravels Early Mechanisms of Human Becker Muscular Dystrophy.

The transcriptional heterogeneity at a single-nucleus level in human Becker muscular dystrophy (BMD) dystrophic muscle has not been explored. Here, we aimed to understand the transcriptional heterogeneity associated with myonuclei, as well as other mononucleated cell types that underly BMD pathogenesis by performing single-nucleus RNA sequencing. We profiled single-nucleus transcriptional profiles of skeletal muscle samples from 7 BMD patients and 3 normal controls. A total of 17,216 nuclei (12,879 from BMD patients and 4,337 from controls) were classified into 13 known cell types, including 9 myogenic lineages and 4 non-myogenic lineages, and 1 unclassified nuclear type according to their cell identities. Among them, type IIx myonuclei were the first to degenerate in response to dystrophin reduction. Differential expression analysis revealed that the fibro-adipogenic progenitors (FAPs) population had the largest transcriptional changes among all cell types. Sub-clustering analysis identified a significantly compositional increase in the activated FAPs (aFAPs) subpopulation in BMD muscles. Pseudotime analysis, regulon inference, and deconvolution analysis of bulk RNA-sequencing data derived from 29 BMD patients revealed that the aFAPs subpopulation, a distinctive and previously unrecognized mononuclear subtype, was profibrogenic and expanded in BMD patients. Muscle quantitative real-time polymerase chain reaction and immunofluorescence analysis confirmed that the mRNA and protein levels of the aFAPs markers including LUM, DCN, and COL1A1 in BMD patients were significantly higher than those in controls, respectively. Our results provide insights into the transcriptional diversity of human BMD muscle at a single-nucleus resolution and new potential targets for anti-fibrosis therapies in BMD. ANN NEUROL 2024;96:1070-1085.

Real-time monitoring of cattle (Bos taurus) tissue using a novel point-of-care (POC) polymerase spiral reaction (PSR) colorimetric assay

Adulteration of cattle meat with other species raises critical public health concerns, as existing detection methods lack sensitivity, specificity and practicality. We address this gap by introducing a novel Point-of-Care (POC) polymerase spiral reaction (PSR) assay. This rapid, on-site tool enables highly specific and sensitive identification of cattle tissue, even in resource-limited settings. This innovation paves the way for improved food safety and consumer protection. The cattle-specific PSR assay employs COX3 gene-targeting primers and a chromogenic indicator triad (HNB, Malachite Green and Phenol Red) for rapid visual detection. Cross-amplification absence with other meats (buffalo, sheep, goat and pig) and concordance with SYBR Green I and agarose gel electrophoresis confirm specificity. PSR at 62 °C for 60minutes exhibits a 100 fg detection limit and identifies 0.1% meat admixing. Efficacy is demonstrated across a diverse range of cattle meat samples, blind samples and processed cattle muscle samples. Compatibility with both conventional (snap chill) and commercial DNA extraction kits enables on-site analysis within 90minutes, facilitating rapid field sample analysis.

Training Status Influences Regulation of Muscle and PBMC TLR4 Expression and Systemic Cytokine Responses to Vigorous Endurance Exercise.

A bout of vigorous endurance exercise transiently activates Toll-like receptor 4 (TLR4) and reduces TLR4 protein expressed on peripheral blood mononuclear cells (PBMCs). Endurance training, on the other hand, reduces TLR4-mediated signaling and minimizes the physiological stress imposed by exercise. Less is known about what occurs in skeletal muscle regarding TLR4 regulation and signaling. Therefore, this study aimed to investigate the regulation of TLR4 expressed in different tissue types (PBMCs and skeletal muscle samples) between endurance-trained and untrained men following vigorous endurance exercise and determine the effect of training status on cytokine responses associated with TLR4 activation. Endurance-trained (n = 7) and untrained (n = 5) men cycled for 1-hr at their respiratory compensation point, with blood and skeletal muscle samples collected pre- and 3-hours post-exercise. In response to vigorous exercise, untrained men experienced a decrease in inhibitor of κBα (IκBα) protein (suggesting IκB degradation and the activation of TLR4-associated transcription factor NF-κB) and TLR4 protein levels, along with a simultaneous increase in TLR4 mRNA expression in both skeletal muscle and PBMCs. Moreover, this exercise session led to elevated levels of circulating interleukin-6, tumor necrosis factor-α, and interleukin-1β. Collectively, these results suggest a heightened TLR4-mediated signaling pathway in untrained men. However, no changes in these targets were observed in endurance-trained men, possibly indicating a potential mechanism by which regular endurance training blunts systemic inflammation. These findings highlight the potential of endurance training to mitigate TLR4-mediated signaling, such as systemic inflammation, and shed light on the effects of exercise on TLR4 expression in PBMCs and skeletal muscle.

Identification of differentially expressed genes and polymorphisms related to intramuscular oleic-to-stearic fatty acid ratio in pigs.

The intramuscular oleic-to-stearic fatty acid ratio (C18:1n-9/C18:0) is an important indicator of the biosynthesis and desaturation of fatty acids in muscle. By using an RNA-Seq approach in muscle samples from 32 BC1_DU (25% Iberian and 75% Duroc) pigs with divergent values (high: H and low: L) of C18:1n-9/C18:0 fatty acids ratio, a total of 81 differentially expressed genes (DEGs) were identified. Functional analyses of DEGs indicate that mainly peroxisome proliferator-activated receptor signaling pathway (associated genes: PPARG, SCD, PLIN1, and FABP3) was overrepresented. Notably, SCD is directly involved in the conversion of C18:0 to C18:1n-9, and PPARG is a transcription factor regulating lipid metabolism genes, including SCD. However, other DEGs (e.g., ACADVL, FADS3, EPHB2, HGFAC, NGFR, NR0B2, MDH1, MMAA, PPP1R1B, SFRP5, RAB30, and TRARG1) are plausible candidate genes to explain the phenotypic differences of the C18:1n-9/C18:0 ratio. Interestingly, seven genetic variants within the SCD (including the well-known AY487830:g.2228T>C SNP and other novel genotyped polymorphisms) are associated with two haplotypes. Although the haplotypes are segregating at different frequencies in the H and L groups, they do not fully explain the desaturation ratios or the SCD expression levels. A more complex model, including polyunsaturated fatty acids such as C18:2n-6, C20:4n-6, and C18:3n-3, is suggested to explain the regulation of the C18:1n-9/C18:0 desaturation ratio in porcine muscle.

Novel goose parvovirus in naturally infected ducks suffering from locomotor disorders: molecular detection, histopathological examination, immunohistochemical signals, and full genome sequencing

ABSTRACT In this study, we investigated the pathological effects of novel goose parvovirus (NGPV) infection on the skeletal muscle, brain, and intestine of naturally affected ducks suffering from locomotor dysfunction as a new approach for a deeper understanding of this clinical form. For this purpose, a total of 97 diseased ducks, representing 24 flocks of different duck breeds (14–75 days old), were clinically examined. In total, 72 tissue pools of intestine, brain, and skeletal muscle samples were submitted for molecular identification. Typical clinical signs among the examined ducks suggested parvovirus infection. Regarding postmortem examination, all examined ducks showed muscle emaciation (100%) either accompanied by congestion (34%) or paleness (66%). Slight congestion, either in the brain (82.5%) or intestine (75.25%), was predominantly detected. Based on molecular identification, the intestine had the highest percentage of positive detection (91.7%), followed by the skeletal muscle (70.8%), and the brain (20.8%). The main histopathological alterations were myofibre atrophy and degeneration, marked enteritis accompanied by lymphocytic infiltration in the lamina propria and submucosa, while the affected brains showed vasculitis, diffuse gliosis, and Purkinje cell degeneration in the cerebellum. Next-generation sequencing further confirmed the presence of a variant strain of goose parvovirus (vGPV) that is globally known as NGPV and closely related to Chinese NGPV isolates. Using immunohistochemistry, the NGPV antigen was positively detected in the muscle fibres, enterocytes, and Purkinje cells in the cerebellum. These findings provided proof of the involvement of virus replication in the locomotor disorders linked to NGPV infection in ducks.

Alternative polyadenylation landscape of longissimus dorsi muscle with high and low intramuscular fat content in cattle.

Intramuscular fat content is one of the most important factors affecting beef quality. However, the role of alternate polyadenylation (APA) in intramuscular fat deposition remains unclear. We compared APA events in muscle samples from high and low intramuscular fat (IMF) cattle, based on RNA-seq data. A total of 363 significant APAs were identified. Notably, the number of shortened 3'UTR events exceeded the number of lengthened 3'UTR events, and genes associated with shortened 3'UTR events were enriched in fatty acid metabolism-related pathways. Most APA events had alternative 3'UTR (aUTR) lengths of 200-300bp. As the 3'UTR lengthened, the aUTR also lengthened (R2 = 0.79). These findings indicate that genes with longer 3'UTRs are more likely to be regulated by APA in the muscle of cattle with high IMF. To determine whether the identified APA events drove alterations in the expression of fat deposition-related genes, we analyzed the relationship between APA events and differentially expressed genes and identified several genes critical for fat deposition (e.g., PFKL and SLC1A5). Since miRNAs usually bind to the 3'UTR region of protein-coding genes and affect gene expression, we constructed an miRNA-APA network to detect several key miRNAs that may regulate fat deposition. We identified 10 important miRNAs that affect changes in IMF content, which may be gained (gained miRNA-binding sites) or lost (lost miRNA-binding sites) owing to 187 differential APA events. Our study characterized the APA profiles of cattle with high and low intramuscular fat content and provided further insights into the relationship between APA, miRNA, and fat deposition.

Anabolic Sensitivity in Healthy, Lean, Older Men Is Associated With Higher Expression of Amino Acid Sensors and mTORC1 Activators Compared to Young.

Sarcopenia is thought to be underlined by age-associated anabolic resistance and dysregulation of intracellular signalling pathways. However, it is unclear whether these phenomena are driven by ageing per se or other confounding factors. Lean and healthy young (n = 10, 22 ± 3 years, BMI; 23.4 ± 0.8 kg/m2) and old men (n = 10, 70 ± 3 years, BMI; 22.7 ± 1.3 kg/m2) performed unilateral resistance exercise followed by intake of essential amino acids (EAA). Muscle biopsies were collected from the rested and the exercised leg before, immediately after and 60 and 180 min after EAA intake. Muscle samples were analysed for amino acid concentrations, muscle protein synthesis (MPS) and associated anabolic signalling. Following exercise, peak plasma levels of EAA and leucine were similar between groups, but the area under the curve was ~11% and ~28% lower in Young (p < 0.01). Absolute levels of muscle EAA and leucine peaked 60 min after exercise, with ~15 and ~21% higher concentrations in the exercising leg (p < 0.01) but with no difference between groups. MPS increased in both the resting (~0.035%·h-1 to 0.056%·h-1, p < 0.05) and exercising leg (~0.035%·h-1 to 0.083%·h-1, p < 0.05) with no difference between groups. Phosphorylation of S6K1Thr389 increased to a similar extent in the exercising leg in both groups but was 2.8-fold higher in the resting leg of Old at the 60 min timepoint (p < 0.001). Phosphorylation of 4E-BP1Ser65 increased following EAA intake and exercise, but differences between legs were statistically different only at 180 min (p < 0.001). However, phosphorylation of this site was on average 78% greater across all timepoints in Old (p < 0.01). Phosphorylation of eEF2Thr56 was reduced (~66% and 39%) in the exercising leg at both timepoints after EAA intake and exercise, with no group differences (p < 0.05). However, phosphorylation at this site was reduced by ~27% also in the resting leg at 60 min, an effect that was only seen in Old (p < 0.01). Total levels of Rheb (~45%), LAT1 (~31%) and Rag B (~31%) were higher in Old (p < 0.001). Lean and healthy old men do not manifest AR as evidenced by potent increases in MPS and mTORC1 signalling following EAA intake and exercise. Maintained anabolic sensitivity with age appears to be a function of a compensatory increase in basal levels of proteins involved in anabolic signalling. Therefore, our results suggest that age per se does not appear to cause AR in human skeletal muscle.

Exploring differential miRNA expression profiles in muscular and visceral adipose tissue of patients with severe obesity.

This study aims to investigate the differential miRNA expression profile between the visceral white adipose tissue and the skeletal muscle of people with obesity undergoing bariatric surgery. Skeletal muscle and visceral adipose tissue samples of 10 controls and 38 people with obesity (50% also with type 2 diabetes) undergoing bariatric surgery were collected. miRNA expression profiles were analyzed using Next-Generation Sequencing and subsequently validated using RT-PCR. Approximately 69% of miRNAs showed similar expression in both tissues, however, 55 miRNAs were preferentially expressed in visceral adipose tissue and 53 in skeletal muscle. miR-122b-5p was uniquely identified in skeletal muscle, while miR-1-3p and miR-206 were upregulated in skeletal muscle. Conversely, miR-224-5p and miR-335-3p exhibited upregulation in visceral adipose tissue. Notably, distinctions related to the presence of type 2 diabetes were observed solely in the expression of miR-1-3p and miR-206 in visceral adipose tissue. This is the first study unveiling distinct miRNA expression profiles in paired samples of visceral adipose tissue and skeletal muscle in humans. The identification of obesity-specific miRNAs in these tissues opens up promising avenues for research into potential biomarkers for obesity diagnosis and treatment.

Immunofluorescent Localization of Ca²⁺-Sensor Proteins in The Somatic Motor Muscles of The Earthworm &lt;i&gt;Lumbricus terrestris&lt;/i&gt;

The method of immunofluorescent staining of earthworm somatic muscle samples showed the presence of calmodulin, Ca²⁺-calmodulin-dependent protein kinases type 1 and type 2, synaptotagmin type 2 and type 7 and calcineurin A. These proteins are detected in both synaptic and extrasynaptic regions of the motor muscle. However, for synaptotagmin type 2 and type 7, calcineurin A, their predominant localization in the area of neuromuscular synapses has been established. Besides, synaptic localization for synaptotagmin 7 and calcineurin A is most clearly expressed.

Beef Carcasses Aged at Mild Temperature to Improve Sustainability of Meat Production

Beef carcass aging, which enhances tenderness and flavor through proteolysis, is traditionally costly and slow, requiring long-term storage at temperatures near 0 °C. To reduce energy consumption, a new technique using moderate cooling room temperatures was tested. Six carcasses of Holstein bulls were used. From each carcass, two shoulders were processed in different ways: one was refrigerated at 8 °C (W), and after spraying with a solution with calcium chloride and sodium chloride, was coated with sodium alginate. The other shoulder was stored at 2 ± 1 °C as a cold control (C). After five days of aging, the shoulders were dissected, and two muscles (Caput longum triceps brachii and Supraspinatus) were subjected to physico-chemical analysis, microbiological safety assessment, and sensory testing. The remaining samples of both muscles were stored in domestic conditions for an additional 5 days at various temperatures (2, 4, 8 °C), where the same physic-chemical and sensory tests were conducted. The results showed that moderate aging temperature improved meat quality, significantly reducing the shear force (p = 0.001) and increasing sarcomere length, the myofibrillar fragmentation index, and sensory tenderness (p = 0.042, p = 0.039, and p = 0.027, respectively). However, domestic storage post-dissection should not exceed 4 °C to prevent rapid lipid oxidation, as observed at 8 °C for both muscles (p &lt; 0.001). Mild aging temperature maintained legal safety standards, enhanced certain meat qualities, and promoted enzymatic activity similar to traditional dry aging while reducing high energy consumption.

Using an enzyme preparation and vitamin C to increase the meat productivity of broiler chickens

Purpose: to study the effect of vitamin C and MEK Feedbest®VGPro when introduced into the compound feed of broiler chickens with excessive background of heavy metals on the indices of meat productivity, food and biological value, ecological safety of their meat.Materials and methods. The objects of the study were meat chickens of the ROSS-508 cross. At the age of one day, 4 groups of 100 birds each were formed from them in accordance with the principle of analog groups. In the composition of the used recipes of complete compound feed in all three age periods of growing, the main grain and protein ingredients were represented by locally produced feed: corn, wheat, peas and rapeseed meal. Growing of experimental broilers for meat was completed at the age of 42 days and their control slaughter was carried out, for which 5 birds were selected from each group. The concentration of salts of heavy metals was determined in the samples of feed and meat of the experimental birds.Results. The experiment substantiated the feasibility of combined introduction of MEC Feedbest®VGPro at a rate of 100 g/t of feed and vitamin C at a rate of 500 g/t of feed into diets with excessive content of heavy metal salts. Compared to the control analogues, broilers of the 3rd experimental group showed a reliable (p&lt;0,05) decrease in fat by 0,26 % and a simultaneous increase in dry matter, protein and the value of the protein-quality indicator in the pectoral muscle samples. Combined additions of the tested preparations contributed to a decrease in the concentration of zinc by 63,17 % (p&lt;0,05), cadmium by 61,11 % (p&lt;0,05) and lead by 62,34 % (p&lt;0,05) in poultry meat samples from the 3rd experimental group, compared to the control. Moreover, in the meat samples of broilers of the 3rd experimental group, the maximum permissible concentrations of none of these elements were exceeded.

Trophic ecology, based on stable isotope values, of long-finned pilot whales Globicephala melas stranded on the Icelandic coast

In Iceland, occurrences of long-finned pilot whales Globicephala melas and strandings in coastal waters are irregular and thought to be related to prey movements; however, little is known about the population’s trophic ecology, making it difficult to assess its role in the regional ecosystem. To investigate the trophic niche of the species, we measured δ13C and δ15N values from skin and muscle samples collected from 16 strandings (12 single and 4 mass strandings) spanning 33 yr (1988-2021). We tested the effects of sex, age class and body length on δ13C and δ15N values within the species. We found that δ13C values did not differ between sexes or age classes, nor did they vary with body length. Values of δ15N did not vary with sex or body length, but varied slightly between adults and juveniles, albeit not sufficiently to support a difference in trophic levels. Whales from different mass stranding events showed similar stable isotope niche widths (approximately 0.20‰2) and overlapping isotope niches (generally &gt;45% of the standard ellipse area corrected for small sample size). Overall, pilot whales in this region showed a narrow isotopic niche width and low intraspecific variation in isotopic niche. Future analyses using complementary chemical tracers and stomach contents will be important to identify the factors driving these patterns. The first measurements of the stable isotope niche of pilot whales presented here provide a baseline for future studies to determine niche overlap between cetaceans in Iceland, contributing to their management and conservation.

The sodium/ascorbic acid co-transporter SVCT2 distributes in a striated membrane-enriched domain at the M-band level in slow-twitch skeletal muscle fibers

BackgroundVitamin C plays key roles in cellular homeostasis, functioning as a potent antioxidant and a positive regulator of cell differentiation. In skeletal muscle, the vitamin C/sodium co-transporter SVCT2 is preferentially expressed in oxidative slow fibers. SVCT2 is up-regulated during the early fusion of primary myoblasts and decreases during initial myotube growth, indicating the relevance of vitamin C uptake via SVCT2 for early skeletal muscle differentiation and fiber-type definition. However, our understanding of SVCT2 expression and function in adult skeletal muscles is still limited.ResultsIn this study, we demonstrate that SVCT2 exhibits an intracellular distribution in chicken slow skeletal muscles, following a highly organized striated pattern. A similar distribution was observed in human muscle samples, chicken cultured myotubes, and isolated mouse myofibers. Immunohistochemical analyses, combined with biochemical cell fractionation experiments, reveal a strong co-localization of SVCT2 with intracellular detergent-soluble membrane fractions at the central sarcomeric M-band, where it co-solubilizes with sarcoplasmic reticulum proteins. Remarkably, electrical stimulation of cultured myofibers induces the redistribution of SVCT2 into a vesicular pattern.ConclusionsOur results provide novel insights into the dynamic roles of SVCT2 in different intracellular compartments in response to functional demands.

Pathological Changes and CYP1A1 Expression as Biomarkers of Pollution in Sarpa Salpa and Diplodus Sargus.

In a marine ecosystem, the most sensitive organisms to environmental changes, mainly to anthropic pressures, are fishes and invertebrates. Therefore, they are considered the ideal targets to indirectly evaluate the health of an entire ecosystem. Teleost fishes, particularly those that occupy the highest trophic levels, can accumulate toxic substances through their diet. In this study, we used two fish species with sedentary behavior and trophic habits, Diplodus sargus and Sarpa salpa, caught in two areas at different anthropic pressures divided into the Gulf of Naples (Na) and the Gulf of Salerno (Sa). This study aimed to correlate the pathological alterations in target organs in both species with known concentrations of polychlorinated biphenyls (PCBs) and heavy metals (lead and cadmium) to the expression of CYP1A1. Histological examination took into consideration circulatory disorders, increase in melanomacrophages (MMs) number, inflammation in kidney and hepatopancreas and gonadal stage maturation. Next, the pathological and morphological changes found were compared to immunohistochemical expression of CYP1A1 in the same samples. Chemical analysis of PCBs, based on 28, 52, 101, 138, 153, and 180 congeners, and heavy metals, were performed on hepatopancreas and muscle samples. Higher median values of PCBs concentration were detected in both species in the Salerno area (8.1 ng/g in Diplodus sargus muscles and 51.1 ng/g in Sarpa salpa hepatopancreas, respectively), although the values were consistently below the legal limits. No critical values were found for lead and cadmium. Therefore, we hypothesized that CYP1A1 and pathological alterations were more expressed in fish from Salerno area. The pathological changes showed a statistically significant difference in inflammation of the kidneys (p < 0.0001) between S. salpa of both Gulfs. In addition, we found a statistically significant difference in the assessment of the increase in MMs/MMCs (p = 0.0384) and circulation disorders (p = 0.0325) of hepatopancreas in D. sargus of both Gulfs. As not all the variables considered showed statistical significance, the analysis of the results does not fully support the correlation between the highest levels of contaminants found in the Salerno area and the expression of CYP1A1. Our data could be a starting point for future studies to better correlate the role of CYP1A1 to pollutants, considering this is the first study involving two of the most common species in the Mediterranean Sea. Thus, future studies could include other species to improve and increase records.

Temperature alters antioxidant status and induces cell damage in the Amazonian fish tambaqui

Since Amazonian fish live close to their maximum thermal limits, this makes them vulnerable to the effects of global warming. The aim of this study was to evaluate the oxidative stress and antioxidant enzymatic and biochemical responses of the plasma, liver and muscle of tambaqui (Colossoma macropomum) exposed to a rising gradient of water temperature. One hundred and twenty (N=120) juvenile tambaqui were exposed to four temperature levels, these being: the environmental temperature of the season (Tenv – 25.7-30 ºC), 31 ºC, 34 ºC and 37 ºC, following a completely randomized design with three replicates for a period of 60 days. Liver and muscle samples were used to determine the levels of the enzymes superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (GPx) and lipid peroxidation (LPO). Plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured. A histopathological damage assessment (HAI) was performed using liver samples and the results showed an increase in lipid peroxidation in the muscle and liver of animals kept at 37 ºC in relation to other temperatures. Enzyme responses were tissue-specific in the liver and muscle. In the liver, the reduction of CAT, SOD and GPx levels of the animals was observed at 37 ºC compared to those maintained at Tenv and SOD and GPx in relation to animals maintained at 31 and 34 ºC. The GPx enzyme showed higher activity at 34 and 37 ºC compared to the other evaluated temperatures. At 37 ºC, plasma levels of ALT and AST were higher than the other temperatures evaluated, as well as an increase in histopathological damage. In this way, in a scenario of warming of the waters of the Amazon or even of the systems used for rearing of the species, the tambaqui will be able to cope with temperatures of up to 34 ºC, without affecting its antioxidant capacity. However, at 37 ºC, oxidative stress levels and increased liver damage suggest a reduction in antioxidant capacity due to tissue impairment of the organ and general loss of animal performance as it approaches the upper thermal limit of the species.