Fiber Predominance Research Articles

Inter-Individual Variability in Muscle Fiber-Type Distribution Affects Running Economy but Not Running Gait at Submaximal Running Speeds.

Running economy is an important determinant of endurance running performance, yet insights into characteristics contributing to its inter-individual variability remain limited. Although slow-twitch muscle fibers are more energy-efficient than fast-twitch fibers during the (near-)isometric contractions common during submaximal running, current literature lacks a consensus on whether a relationship between muscle fiber-type distribution and running economy exists. This study aims to resolve the ongoing debate by addressing potential confounding factors often overlooked in prior research, such as the effect of different running speeds, the homogeneity of investigated groups, and the potential impact of the adopted running gait. We selected two groups with predetermined distinct muscle fiber-type distribution in their triceps surae muscle by measurement of carnosine via 1H-MRS, one predominantly slow (ST; n = 11; carnosine z-score = -1.31) and the other predominantly fast (FT; n = 10; z-score = 0.83). Across a range of running speeds (2-4 m/s), we measured running economy (W/kg) through indirect calorimetry, along with running kinematics, kinetics and muscle activity of the lower limb. The ST-group exhibited, on average, 7.8% better running economy than the FT-group (p = 0.01) and this difference was consistent across speeds. Both groups demonstrated almost identical kinematics, kinetics, and muscle activity patterns across submaximal running speeds. Overall, our findings indicate that distinct muscle fiber-type distribution explains some of the observed variability in running economy, for which a predominance of energy-efficient slow-twitch fibers appear beneficial. In contrast, muscle fiber-type distribution does not affect running gait substantially.

Three-dimensional virtual histology of the rat uterus musculature using micro-computed tomography.

Contractions of the uterus play an important role in menstruation and fertility, and contractile dysfunction can lead to chronic diseases such as endometriosis. However, the structure and function of the uterus are difficult to interrogate in humans, and thus animal studies are often employed to understand its function. In rats, anatomical studies of the uterus have typically been based on histological assessment, have been limited to small segments of the uterine structure, and have been time-consuming to reconstruct at the organ scale. This study used micro-computed tomography imaging to visualise the muscle structures in the entire non-pregnant rat uterus and assess its use for 3D virtual histology. An assessment of the rodent uterus is presented to (i) quantify muscle thickness variations along the horns, (ii) identify predominant fibre orientations of the muscles and (iii) demonstrate how the anatomy of the uterus can be mapped to 3D volumetric meshes via virtual histology. Micro-computed tomography measurements were validated against measurements from histological sections. The average thickness of the myometrium was found to be 0.33 ± 0.11 mm and 0.31 ± 0.09 mm in the left and right horns, respectively. The micro-computed tomography and histology thickness calculations were found to correlate strongly at different locations in the uterus: at the cervix, r = 0.87, and along the horn from the cervical end to the ovarian end, respectively, r = 0.77, r = 0.89 and r = 0.54, with p < 0.001 in every location. This study shows that micro-computed tomography can be used to quantify the musculature in the whole non-pregnant uterus and can be used for 3D virtual histology.

Muscle ultrastructure and histopathological findings in a Brazilian single-centre series of genetically classified telethoninopathy patients

BackgroundTelethoninopathy or TCAP-gene related Limb Girdle Muscular Dystrophy is a rare genetic disease that was first described in Brazil. There are around 100 families reported worldwide. Due to its rarity, detailed information on muscle biopsy light and electron microscopic features are lacking.Cases presentationRetrospective study of consecutive muscle biopsies performed in patients from a Neuromuscular Outpatient Clinic between 2011 and 2023. Inclusion criteria: telethoninopathy diagnosed by both immunohistochemistry and molecular studies. Seven patients (0.7% or 7/953) were found: five male and two female, admitted from 6 to 54 years old. Detailed light and electron microscopy findings are illustrated. Muscle imaging is presented. A dystrophic pattern on muscle biopsy was found in 57% (4/7) of the patients. Other 43% (3/7) presented myopathic features such as variation in fibre calibre, nuclear internalization, rimmed vacuoles, and oxidative irregularities. Morphometry disclosed type 1 lobulated fibres that were 34%, 52%, and 57% smaller than type 2 fibres, respectively, in three patients, without type 1 fibre predominance. Electron microscopy demonstrated nuclear pseudoinclusions, pyknosis, multifocal loss of the sarcolemma, and 17 nm intrasarcoplasmic filamentous inclusions. All patients presented: (1) complete absence of the immunohistochemical expression of telethonin, and (2) the homozygous c.157C > T, p.(Gln53*) pathogenic variant in exon 2 of the TCAP gene.ConclusionAnti-telethonin immunohistochemistry may be helpful in unsolved cases with nonspecific myopathic abnormalities, specially with small type 1 lobulated fibres. Appropriate diagnosis is important for adequate genetic counselling.

Variation in microplastic characteristics among amphibian larvae: a comparative study across different species and the influence of human activity

Microplastic pollution is a significant global environmental issue, and impacts span from individual organisms to the entire ecosystems. This study investigated the properties of microplastics in amphibian larvae, shedding light on their environmental interactions and potential ecological consequences. We examined microplastics extracted from amphibian larvae of 10 taxa, sampled from sites experiencing different levels of human impact. Our findings revealed a predominance of blue microplastics and fibres, each comprising 53% of the total microplastics in amphibian larvae. Microplastic fibres were also notably longer than other morphological types of microplastics. Furthermore, we observed variations in the surface area of microplastics among different amphibian families. An interesting observation from our research is the apparent positive relationship between the size of amphibian larvae and the length of granular and flake-shaped microplastics. Conversely, we observed a negative relationship between the length of these microplastics and human environmental impact. These insights significantly contribute to the understanding of microplastic pollution in freshwater environments, highlighting its complexities beyond marine ecosystems. Our research emphasises the intricate relationships between microplastics and freshwater organisms, underscoring the need for comprehensive strategies to mitigate microplastic pollution.

Influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis in Wistar rats.

The aim of this study was to investigate the influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis (AP) in Wistar rats. Fifty-six rats were submitted to pulp exposure of the lower left first molar for the induction of AP. On the same day, intraperitoneal antibiotic therapy was administered once a day, for 15days, until euthanasia. The groups were formed according to the different treatments (n = 8): C-control; GEN-treated with gentamicin (10mg/Kg); AC-treated with amoxicillin (100mg/Kg); MZ-treated with metronidazole (40mg/Kg); AMP-treated with ampicillin (100mg/Kg); AMC group-treated with amoxicillin + clavulanic acid (100mg/kg); CLI-treated with clindamycin (60mg/kg). After euthanasia, the jaws were collected and processed for (1) histological and histometric analysis using hematoxylin and eosin staining, (2) analysis of collagen fibers using Picrosirius Red staining and (3) bacteriological analysis using Brown-Brenn staining. The data were analyzed statistically (p < 0.05). AP induction was confirmed in all groups. The AMC group had the lower intensity of inflammatory infiltrate (p = 0.028) and less periapical bone resorption compared to control (p = 0.006). Regarding collagen maturation, PSR staining revealed a predominance of mature collagen fibers in all groups. The AC and AMC groups had the lower amount of mature fibers and the highest amount of immature fibers, compared to all other groups (p < 0.001). All groups showed bacterial contamination; however, the AC and AMC groups showed a lower extent of bacterial contamination compared to the control (p < 0.001). It can be concluded that systemic antibiotic therapy influences the development and progression of induced AP.

Recurring homozygous ACTN2 variant (p.Arg506Gly) causes a recessive myopathy.

ACTN2, encoding alpha-actinin-2, is essential for cardiac and skeletal muscle sarcomeric function. ACTN2 variants are a known cause of cardiomyopathy without skeletal muscle involvement. Recently, specific dominant monoallelic variants were reported as a rare cause of core myopathy of variable clinical onset, although the pathomechanism remains to be elucidated. The possibility of a recessively inherited ACTN2-myopathy has also been proposed in a single series. We provide clinical, imaging, and histological characterization of a series of patients with a novel biallelic ACTN2 variant. We report seven patients from five families with a recurring biallelic variant in ACTN2: c.1516A>G (p.Arg506Gly), all manifesting with a consistent phenotype of asymmetric, progressive, proximal, and distal lower extremity predominant muscle weakness. None of the patients have cardiomyopathy or respiratory insufficiency. Notably, all patients report Palestinian ethnicity, suggesting a possible founder ACTN2 variant, which was confirmed through haplotype analysis in two families. Muscle biopsies reveal an underlying myopathic process with disruption of the intermyofibrillar architecture, Type I fiber predominance and atrophy. MRI of the lower extremities demonstrate a distinct pattern of asymmetric muscle involvement with selective involvement of the hamstrings and adductors in the thigh, and anterior tibial group and soleus in the lower leg. Using an in vitro splicing assay, we show that c.1516A>G ACTN2 does not impair normal splicing. This series further establishes ACTN2 as a muscle disease gene, now also including variants with a recessive inheritance mode, and expands the clinical spectrum of actinopathies to adult-onset progressive muscle disease.

Dietary oleic acid intake increases the proportion of type 1 and 2X muscle fibers in mice

Skeletal muscle is one of the largest metabolic tissues in mammals and is composed of four different types of muscle fibers (types 1, 2A, 2X, and 2B); however, type 2B is absent in humans. Given that slow-twitch fibers are superior to fast-twitch fibers in terms of oxidative metabolism and are rich in mitochondria, shift of muscle fiber types in direction towards slower fiber types improves metabolic disorders and endurance capacity. We previously had reported that oleic acid supplementation increases type 1 fiber formation in C2C12 myotubes; however, its function still remains unclear. This study aimed to determine the effect of oleic acid on the muscle fiber types and endurance capacity. An in vivo mouse model was used, and mice were fed a 10% oleic acid diet for 4 weeks. Two different skeletal muscles, slow soleus muscle with the predominance of slow-twitch fibers and fast extensor digitorum longus (EDL) muscle with the predominance of fast-twitch fibers, were used. We found that dietary oleic acid intake improved running endurance and altered fiber type composition of muscles, the proportion of type 1 and 2X fibers increased in the soleus muscle and type 2X increased in the EDL muscle. The fiber type shift in the EDL muscle was accompanied by an increased muscle TAG content. In addition, blood triacylglycerol (TAG) and non-esterified fatty acid levels decreased during exercise. These changes suggested that lipid utilization as an energy substrate was enhanced by oleic acid. Increased proliferator-activated receptor γ coactivator-1β protein levels were observed in the EDL muscle, which potentially enhanced the fiber type transitions towards type 2X and muscle TAG content. In conclusion, dietary oleic acid intake improved running endurance with the changes of muscle fiber type shares in mice. This study elucidated a novel functionality of oleic acid in skeletal muscle fiber types. Further studies are required to elucidate the underlying mechanisms. Our findings have the potential to contribute to the field of health and sports science through nutritional approaches, such as the development of supplements aimed at improving muscle function.

The diagnosis of asbestosis in the 21st century: a clinicopathological correlation of 102 cases

ABSTRACT Asbestosis, defined as diffuse pulmonary fibrosis caused by inhalation of asbestos fibers, occurs after heavy exposures to asbestos dust over several decades. Because workplace exposures have been significantly curtailed since the banning of asbestos in insulation products, we were interested in examining the clinicopathological characteristics of cases diagnosed in the 21st century. The consultation files of one of the authors (VLR) were reviewed for cases of asbestosis diagnosed since 1/1/2001. 102 cases were identified, with a median age of 75 years (range: 45–89). There were 100 men and 2 women. The women were from Turkey and Brazil (none from the United States). Malignancies were present in 78 cases, including 38 lung cancers, 29 pleural mesotheliomas, and 8 peritoneal mesotheliomas. The grade of asbestosis was available in 88 cases (median severity of 2; scale: 1–4). Pleural plaque was present in 94% of cases. The most common exposure categories were insulators (39), shipyard workers (16), asbestos manufacturing (9), boiler workers (8) and pipefitter/welders (6). The median duration of exposure was 33 years (range: 2–49 years). Lung fiber burden analysis was performed in 34 cases, with amosite being the predominant fiber type. Results were compared with similar information for 475 cases diagnosed prior to 1/1/2001.

The first evidence of microplastic presence in the River Nile in Khartoum, Sudan: Using Nile Tilapia fish as a bio-indicator

The extent of microplastics in African freshwater systems remains less investigated. In Sudan, there is no single study reporting microplastics in water bodies. This scoping study aimed to investigate the presence and characteristics of microplastics (MPs) in Nile Tilapia fish from the River Nile in Khartoum, Sudan. The digestive tracts of the fish were digested using 10% potassium hydroxide, and microplastic particles were extracted by density separation using sodium iodide. 567 particles of different sizes (0.04–4.94 mm), shapes (fibers, fragments, films, foams, and pellets), and colours (mostly green, black, blue, and grey) were identified as microplastics. The average abundance of microplastics was 72.02 ± 62.06 particles/kg, and the average intensity was 18.90 ± 9.17 MPs/fish. Small-sized (<1 mm), fibrous-shaped, and coloured microplastics were most abundant in all samples, representing 56%, 85%, and 84%, respectively. Surface examination by SEM showed signs of fragmentation such as cracks, pits, and pores. Two polymer types (high-density polyethylene and polypropylene) were identified by Raman spectroscopy. The predominance of fibers and fragments (94.5%) over pellets (0.35%) and the apparent signs of fragmentation may indicate that MPs are mostly secondary MPs. Wastewater effluent, domestic discharge, and recreational activities are the potential sources. This scoping investigation provided the first data on microplastic presence in the River Nile in Khartoum, and it could be used to guide future studies to fill research gaps in the region.

Characteristic of Microplastic on Coral Reef Sediment and Sea Urchin (Diadema sp.) in Tidung Island, Jakarta Bay, Indonesia

Microplastics are recognized as common contaminants of coral ecosystem in Tidung Island, affecting both sediment and sea urchins residing in this environment. Therefore, this study aimed to determine the characteristics of microplastics found in sediment, the mouth, and the digestive system of sea urchins (Diadema sp.), assessing the relationship between coral cover percentage and microplastic type and size, and the transfer of microplastics from sediment to sea urchins. Sampling was conducted twice, namely in October 2021 and October 2022. Microplastics in sediments were extracted using ZnCl2. The destruction process of sea urchins used 30% H2O2 and FeSO4.7H2O, while the microplastics were identified with Attenuated Total Reflection Fourier Transform Infrared Spectroscopy (ATR-FTIR). The results showed that the microplastics found in the sediments and sea urchins were similar in terms of shape, color, size, and plastic polymers. The forms of microplastics found in this study were fibre, fragment, and foam with fibre predominance, based on the results obtained. The size of microplastics found in sediments and sea urchins was dominated by sizes &gt;1000 µm. Fibres were found in hard coral (HC) and dead coral (DC) conditions while fragments and foams were present in turf algae (TA), sponge (SP), and soft coral (SC) conditions. The microplastics found in sea urchins originate in part from sediments because they have similar characteristics. Sea urchins ingested microplastics from sediment, particularly those grown by algae as a food source. The increase in the number of microplastics found in sediment could potentially result in higher abundance in the biota.

Microplastics in inland and offshore sediments in the Apulo-Lucanian region (Southern Italy)

Inland and offshore sediments from Southern Italy were studied in order to evaluate the occurrence and nature of microplastics (MPs). Inland sediments were collected in the Bradano and Basento rivers (Apulo-Lucanian region, Southern Italy), while offshore sediments were collected on the continental shelf near Bari (Adriatic Sea) and Metaponto (Ionian Sea). MPs were detected and characterized using optical microscopy, micro-Fourier-Transform Infrared spectroscopy (μ-FTIR) and micro-Raman analyses. The number of MPs present varied between 144 and 1246 kg−1 of dry sediment (468.8 ± 410,7 MPs kg−1) with a predominance of black fibers; no correlation emerged between MPs and sediment grain size. In river sediments, the occurrence of MPs is associated with local pollution, whereas the offshore occurrence of MPs depends on seasonal river flow and submarine canyons. Compositional analyses suggest that the main source of MPs in the studied sediments is sewage discharge from residential areas.

Microplastics in the sediments along the eastern Arabian Sea shelf: Distribution, governing factors and risk assessment

Despite the omnipresence of microplastics (MPs), the studies around the western continental shelf of Indian Ocean (Eastern Arabian Sea-EAS) are uncovered and understudied. Thus, the present study was focused to understand the spatial distribution, characterization and risk assessment of MPs in sediment across seven coastal transects (10 to 50 m) all along the EAS shelf. The highest MPs concentration (MPs/kg d.w.) was detected in the northern EAS (NEAS; 2260 ± 1050) followed by central (CEAS; 1550 ± 1012) and southern (SEAS; 1300 ± 513) shelves. Among all distinct locations, the highest concentration of MPs (2500 ± 1042) was detected in the north coastal sediments off Mumbai, followed by off Mangalore (1480 ± 1169) in the center and off Kochi (1350 ± 212) in the south. MPs were found in the form of fibres, fragments and films with a predominance of fibres (~70–80 %). Approximately 74.6 % of the total MPs were in the size range of 300 μm to 5 mm. The surface of detected MPs was rough, irregular, and mechanical weathering features such as pits, grooves also observed and spotted with bacterial community structures. Polypropylene (PP; 34 %), polyisoprene (PIP; 19 %), butyl rubber (18 %), and low-density polyethylene (LDPE; 13 %) were dominant polymers. The pollution load index highlighted minor risk while the polymer hazard index exhibited a hazard level of V. Litter discharge, fishing activities, and active marine navigation are among the many high-risk sources of plastic contamination in this region. Due to the prevailing winds, currents, low sea surface height, and high precipitation, the conditions in the EAS are favorable for the accumulation of both sea-based and land-based particles. Hence, this study provides novel insights into the potential risks posed by MP to the IO rim and associated marine ecosystem which will enhance our knowledge of the ecological implications and consequences of MP pollution, ultimately aiding in developing effective management and mitigation strategies.

Monitoring of anthropogenic microplastic pollution in antarctic fish (emerald rockcod) from the Terranova Bay after a quarter of century

Monitoring the occurrence of microplastic contamination in the Antarctic area is the key to implement policy measures for waste regulations in the research stations. Antarctic fish Trematomus bernachii is a suitable species for establishing microplastic contamination and for investigating changes over time in the concentration and type of microplastics in the Antarctic region. In this paper a total of 78 fish, caught during the 37th Italian Antarctic expedition (2021–2022) in the Ross Sea (Antarctica) were analysed. Different microfibers and dyes were identified by Raman spectroscopy and the results were compared with those obtained for fish sampled in 1998. Differences in polymer type emerged with a predominance of synthetic fibers with respect to natural ones. These changes appear to be related to the increased human activities in the Antarctica over the last twenty years and highlights the need to improve the environmental sustainability of the numerous research stations operating throughout that area.

Comparing microplastics ingested by the bogue Boops boops (Teleostei: Sparidae) to those available in its feeding areas: A case study on the Portuguese west coast

Understanding the exposure of wild marine fish to microplastics (MPs) has been frequently assessed through the quantification and characterization of MPs found in stomachal contents. However, whether such particles mirror the MPs available in the environment or suggest a selectivity ability in the species, it has been scarcely analyzed. Here, in addition to investigating temporal patterns in the abundance and diversity of MPs ingested by Boops boops (Linnaeus, 1758) caught in coastal waters between Setúbal and Sesimbra (Portugal), we also aimed at comparing them with the patterns of MPs occurring locally, namely where the bogue feeds: at both surface water and seabed sediments. A total of 200 fish, 50 per month, were supplied at the local fish first sale concurrently with the water and sediments sampling (from October 2018 to January 2019). Fibers accounted for 93% of all MPs ingested and were also the main type of MPs extracted from sediments. Both food intake (inferred by fish gastrointestinal tract) and abundance of MPs per fish (1.80 ± 1.26; mean ± SD, n = 102) significantly decreased during the 4 month-period analyzed. Such decreasing tendency is potentially related to this species reproduction season. Despite the distinct fluctuations observed in the abundance of MPs found in both feeding areas of the bogue, it was possible to detect similarities between fish and sediments regarding the predominance of fibers, namely those belonging to the smaller size class. Yet, the prevailing colors differed, suggesting a selection capability of the bogue. Further studies taking place in spring/summer months (out of the reproduction season) are thus needed to clarify how MPs ingestion will vary in this species and consequently interfere with its ability to mirror the MP pollution patterns in this coastal region.

Effects of inhibition of 5-lipoxygenase and 12-lipoxygenase pathways on skeletal muscle fiber regeneration

We investigated the effect of inhibition of 5-lipoxigenase (LOX) and 12-LOX pathways on the regeneration of skeletal muscle fibers after injury induced by a myotoxin (MTX) phospholipase A2 from snake venom in an in vivo experimental model. Gastrocnemius muscles of mice injected with MTX presented an increase in 5-LOX protein expression, while 12-LOX was found to be a constitutive protein of skeletal muscle. Animals that received oral treatments with 5-LOX inhibitor MK886 or 12-LOX inhibitor baicalein 30 min and 48 h after MTX-induced muscle injury showed a reduction in the inflammatory process characterized by a significant decrease of cell influx and injured fibers in the degenerative phase (6 and 24 h after injury). In the beginning of the regeneration process (3 days), mice that received MK886 showed fewer new basophilic fibers, suggesting fewer proliferative events and myogenic cell fusion. Furthermore, in the progression of tissue regeneration (14–21 days), the mice treated with 5-LOX inhibitor presented a lower quantity of central nucleus fibers and small-caliber fibers, culminating in a muscle that is more resistant to the stimulus of fatigue during muscle regeneration with a predominance of slow fibers. In contrast, animals early treated with the 12-LOX inhibitor presented functional fibers with higher diameters, less resistant to fatigue and predominance of fast heavy-chain myosin fibers as observed in control animals. These effects were accompanied by an earlier expression of myogenic factor MyoD. Our results suggest that both 5-LOX and 12-LOX pathways represent potential therapeutic targets for muscle regeneration. It appears that inhibition of the 5-LOX pathway represses only the degenerative process by reducing tissue inflammation levels. Meanwhile, inhibition of the 12-LOX pathway also favors the anticipation of maturation and earlier recovery of muscle fiber activity function after injury.

Development of a biomimetic arch-like 3D bioprinted construct for cartilage regeneration using gelatin methacryloyl and silk fibroin-gelatin bioinks

In recent years, engineering biomimetic cellular microenvironments have been a top priority for regenerative medicine. Collagen II, which is arranged in arches, forms the predominant fiber network in articular cartilage. Due to the shortage of suitable microfabrication techniques capable of producing 3D fibrous structures, in vitro replication of the arch-like cartilaginous tissue constitutes one of the major challenges. Hence, in the present study, we report a 3D bioprinting approach for fabricating arch-like constructs using two types of bioinks, gelatin methacryloyl (GelMa) and silk fibroin-gelatin (SF-G). The bioprinted SF-G constructs displayed increased proliferation of the encapsulated human bone marrow-derived mesenchymal stem cells compared to the GelMA constructs. Biochemical assays, gene, and protein expression exhibited the superior role of SF-G in forming the fibrous collagen network and chondrogenesis. Protein-protein interaction study using Metascape evaluated the function of the proteins involved. Further GeneMANIA and STRING analysis using Col 2A1, SOX 9, ACAN, and the genes upregulated on day 21 in RT-PCR, i.e. β-catenin, TGFβR1, Col 1A1 in SF-G and PRG4, Col 10A1, MMP 13 in GelMA validated our in vitro results. These findings emphasized the role of SF-G in regulating the Wnt/β-catenin and TGF-β signaling pathways. Hence, the 3D bioprinted arch-like constructs possess a substantial potential for cartilage regeneration.

A homozygous loss of function variant in POPDC3: From invalidating exercise intolerance to a limb-girdle muscular dystrophy phenotype

Recessive pathogenic variants in POPDC3 have recently been associated with the rare limb-girdle muscular dystrophy (LGMD) subtype LGMDR26. We studied three siblings and a distantly related individual with a skeletal muscle disorder, harboring the c.486–6T>A splice site variant in POPDC3 in homozygosity. Immunohistochemistry, western blot, and mRNA experiments on patients’ skeletal muscle tissue as well as on patients’ myoblasts were performed to study the pathogenicity of the predicted loss of function mechanism of the variant. Patients mainly presented with invalidating myalgia and exercise intolerance and limited to no segmentary muscle weakness. CK levels were markedly elevated in all patients. A loss of function mechanism at the RNA level was shown (r.485_486insauag, p.Ile163*). Muscle biopsies performed in three out of four patients showed non-specific myopathic features with a marked type 2 fiber predominance and the presence of a large number of severely atrophic fibers with pyknotic nuclear clumps. We show that skeletal muscle symptoms in LGMDR26 may range from an overt late juvenile to young adult-onset limb-girdle muscular dystrophy phenotype to severe exercise intolerance and myalgia, with consistently highly elevated CK levels. We further prove a clear LOF mechanism of POPDC3 in this rare disorder.

Clinical and functional characterization of a long survivor congenital titinopathy patient with a novel metatranscript-only titin variant

Congenital titinopathies are an emerging group of a potentially severe form of congenital myopathies caused by biallelic mutations in titin, encoding the largest existing human protein involved in the formation and stability of sarcomeres. In this study we describe a patient with a congenital myopathy characterized by multiple contractures, a rigid spine, non progressive muscular weakness, and a novel homozygous TTN pathogenic variant in a metatranscript-only exon: the c.36400A > T, p.Lys12134*. Muscle biopsies showed increased internalized nuclei, variability in fiber size, mild fibrosis, type 1 fiber predominance, and a slight increase in the number of satellite cells. RNA studies revealed the retention of intron 170 and 171 in the open reading frame, and immunoflourescence and western blot studies, a normal titin content. Single fiber functional studies showed a slight decrease in absolute maximal force and a cross-sectional area with no decreases in tension, suggesting that weakness is not sarcomere-based but due to hypotrophy. Passive properties of single fibers were not affected, but the observed increased calcium sensitivity of force generation might contribute to the contractural phenotype and rigid spine of the patient. Our findings provide evidence for a pathogenic, causative role of a metatranscript-only titin variant in a long survivor congenital titinopathy patient with distal arthrogryposis and rigid spine.

Fiber type and metabolic characteristics of skeletal muscle in 16 breeds of domestic dogs.

The domestic dog (Canis lupus familiaris) species comprises hundreds of breeds, each differing in physical characteristics, behavior, strength, and running capability. Very little is known about the skeletal muscle composition and metabolism between the different breeds, which may explain disease susceptibility. Muscle samples from the triceps brachii (TB) and vastus lateralis (VL) were collected post mortem from 35 adult dogs, encompassing 16 breeds of varying ages and sex. Samples were analyzed for fiber type composition, fiber size, oxidative, and glycolytic metabolic capacity (citrate synthase [CS], 3-hydroxyacetyl-coA dehydrogenase [3HAD], creatine kinase [CK], and lactate dehydrogenase [LDH] enzyme activities). There was no significant difference between the TB and VL in any of the measurements. However, there were large intra species variation, with some variables confirming the physical attributes of a specific breed. Collectively, type IIA was the predominant fiber type followed by type I and type IIX. The cross-sectional areas (CSA) of the fibers were all smaller when compared to humans and similar to other wild animals. There was no difference in the CSA between the fiber types and muscle groups. Metabolically, the muscle of the dog displayed high oxidative capacity with high activities for CS and 3HAD. Lower CK and higher LDH activities than humans indicate a lower and higher flux through the high energy phosphate and glycolytic pathways, respectively. The high variability found across the different breeds may be attributed to genetics, function or lifestyle which have largely been driven through human intervention. This data may provide a foundation for future research into the role of these parameters in disease susceptibility, such as insulin resistance and diabetes, across breeds.

Diversity and Differential Expression of MicroRNAs in the Human Skeletal Muscle with Distinct Fiber Type Composition.

The ratio of fast- and slow-twitch fibers in human skeletal muscle is variable and largely determined by genetic factors. In this study, we investigated the contribution of microRNA (miRNA) in skeletal muscle fiber type composition. The study involved biopsy samples of the vastus lateralis muscle from 24 male participants with distinct fiber type ratios. The miRNA study included samples from five endurance athletes and five power athletes with the predominance of slow-twitch (61.6-72.8%) and fast-twitch (69.3-80.7%) fibers, respectively. Total and small RNA were extracted from tissue samples. Total RNA sequencing (N = 24) revealed 352 differentially expressed genes between the groups with the predominance of fast- and slow-twitch muscle fibers. Small RNA sequencing showed upregulation of miR-206, miR-501-3p and miR-185-5p, and downregulation of miR-499a-5p and miR-208-5p in the group of power athletes with fast-twitch fiber predominance. Two miRtronic miRNAs, miR-208b-3p and miR-499a-5p, had strong correlations in expression with their host genes (MYH7 and MYH7B, respectively). Correlations between the expression of miRNAs and their experimentally validated messenger RNA (mRNA) targets were calculated, and 11 miRNA-mRNA interactions with strong negative correlations were identified. Two of them belonged to miR-208b-3p and miR-499a-5p, indicating their regulatory links with the expression of CDKN1A and FOXO4, respectively.