Muscle Biopsy Research Articles

The effects of whey, pea, and collagen protein supplementation beyond the recommended dietary allowance on integrated myofibrillar protein synthetic rates in older males: a randomized controlled trial.

Skeletal muscle mass is determined predominantly by feeding- and activity-induced fluctuations in muscle protein synthesis (MPS). Older individuals display a diminished MPS response to protein ingestion, referred to as age-related anabolic resistance, that contributes to the progression of age-related muscle loss - sarcopenia. We aimed to determine the impact of consuming higher- versus lower-quality protein supplements above the recommended dietary allowance (RDA) on integrated MPS rates. We hypothesized that increasing total protein intake above the RDA, regardless of the source, would support higher integrated rates of myofibrillar protein synthesis. Thirty-one healthy older males (72 ± 4 y) underwent a controlled diet with protein intake set at the RDA - control phase (CON; days 1-7). In a double-blind, randomized controlled fashion, participants were assigned to consume an additional 50g (2x25g) of Whey (WHEY, n=10), Pea (PEA, n=11), or collagen (COLL, n=10) protein each day (25 g at breakfast and lunch) during the supplemental phase (SUPP; days 8-15). Deuterated water ingestion and muscle biopsies assessed integrated MPS and acute anabolic signaling. Postprandial blood samples were collected to determine feeding-induced aminoacidemia. Integrated-MPS was increased during SUPP with WHEY (1.59 ± 0.11 %/d, p<0.001) and PEA (1.59 ± 0.14 %/d, p<0.001) when compared with RDA (1.46 ± 0.09 %/d WHEY; 1.46 ± 0.10 %/d PEA); however, it remained unchanged with COLL. Supplemental protein was sufficient to overcome anabolic signaling deficits (mTORC1 and rpS6), corroborating the greater postprandial aminoacidemia. Our findings demonstrate that supplemental protein provided at breakfast and lunch over the current RDA enhanced anabolic signaling and integrated MPS in older males; however, the source of additional protein may be an important consideration in overcoming age-related anabolic resistance. Clinical Trial Registry number and website where it was obtained This trial (NCT04026607) was registered clinicaltrials.gov.

Senolytic effects of exercise in human muscles require acute inflammation.

Higher intensity exercise, despite causing more tissue damage, improved aging conditions. We previously observed decreased p16INK4a mRNA in human skeletal muscle after high-intensity interval exercise (HIIE), with no change following equivalent work in moderate-intensity continuous exercise. This raises the question of whether the observed senolytic effect of exercise is mediated by inflammation, an immune response induced by muscle damage. In this study, inflammation was blocked using a multiple dose of ibuprofen (total dose: 1200 mg), a commonly consumed nonsteroidal anti-inflammatory drug (NSAID), in a placebo-controlled, counterbalanced crossover trial. Twelve men aged 20-26 consumed ibuprofen or placebo before and after HIIE at 120% maximum aerobic power. Multiple muscle biopsies were taken for tissue analysis before and after HIIE. p16INK4a+ cells were located surrounding myofibers in muscle tissues. The maximum decrease in p16INK4a mRNA levels within muscle tissues occurred at 3 h post-exercise (-82%, p < 0.01), gradually recovering over the next 3-24 h. A concurrent reduction pattern in CD11b mRNA (-87%, p < 0.01) was also found within the same time frame. Ibuprofen treatment attenuated the post-exercise reduction in both p16INK4a mRNA and CD11b mRNA. The strong correlation (r = 0.88, p < 0.01) between p16INK4a mRNA and CD11b mRNA in muscle tissues suggests a connection between the markers of tissue aging and pro-inflammatory myeloid differentiation. In conclusion, our results suggest that the senolytic effect of high-intensity exercise on human skeletal muscle is mediated by acute inflammation.

Different roles of circulating and intramuscular GDF15 as markers of skeletal muscle health

IntroductionGrowth Differentiation Factor 15 (GDF15) is a mitokine expressed in response to various stresses whose circulating levels increase with age and are associated with numerous pathological conditions, including muscle wasting and sarcopenia. However, the use of circulating GDF15 (c-GDF15) as a biomarker of sarcopenia is still debated. Moreover, the role of GDF15 intracellular precursor, pro-GDF15, in human skeletal muscle (SM-GDF15) is not totally understood. In order to clarify these points, the association of both forms of GDF15 with parameters of muscle strength, body composition, metabolism and inflammation was investigated.Methodsthe levels of c-GDF15 and SM-GDF15 were evaluated in plasma and muscle biopsies, respectively, of healthy subjects (HS) and patients with lower limb mobility impairment (LLMI), either young (&amp;lt;40 years-old) or old (&amp;gt;70 years-old). Other parameters included in the analysis were Isometric Quadriceps Strength (IQS), BMI, lean and fat mass percentage, Vastus lateralis thickness, as well as circulating levels of Adiponectin, Leptin, Resistin, IGF-1, Insulin, IL6, IL15 and c-PLIN2. Principal Component Analysis (PCA), Canonical Discriminant Analysis (CDA) and Receiving Operating Characteristics (ROC) analysis were performed.Resultsc-GDF15 but not SM-GDF15 levels resulted associated with decreased IQS and IGF-1 levels in both HS and LLMI, while only in LLMI associated with increased levels of Resistin. Moreover, in LLMI both c-GDF15 and SM-GDF15 levels were associated with IL-6 levels, but interestingly SM-GDF15 is lower in LLMI with respect to HS. Furthermore, a discrimination of the four groups of subjects based on these parameters was possible with PCA and CDA. In particular HS, LLMI over 70 years or under 40 years of age were discriminated based on SM-GDF15, c-GDF15 and Insulin levels, respectively.Conclusionour data support the idea that c-GDF15 level could be used as a biomarker of decreased muscle mass and strength. Moreover, it is suggested that c-GDF15 has a different diagnostic significance with respect to SM-GDF15, which is likely linked to a healthy and active state.

Lysosomal dysfunction and overload of nucleosides in thymidine phosphorylase deficiency of MNGIE

Inherited deficiency of thymidine phosphorylase (TP), encoded by TYMP, leads to a rare disease with multiple mitochondrial DNA (mtDNA) abnormalities, mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). However, the impact of TP deficiency on lysosomes remains unclear, which are important for mitochondrial quality control and nucleic acid metabolism. Muscle biopsy tissue and skin fibroblasts from MNGIE patients, patients with m.3243 A > G mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS) and healthy controls (HC) were collected to perform mitochondrial and lysosomal functional analyses. In addition to mtDNA abnormalities, compared to controls distinctively reduced expression of LAMP1 and increased mitochondrial content were detected in the muscle tissue of MNGIE patients. Skin fibroblasts from MNGIE patients showed decreased expression of LAMP2, lowered lysosomal acidity, reduced enzyme activity and impaired protein degradation ability. TYMP knockout or TP inhibition in cells can also induce the similar lysosomal dysfunction. Using lysosome immunoprecipitation (Lyso- IP), increased mitochondrial proteins, decreased vesicular proteins and V-ATPase enzymes, and accumulation of various nucleosides were detected in lysosomes with TP deficiency. Treatment of cells with high concentrations of dThd and dUrd also triggers lysosomal dysfunction and disruption of mitochondrial homeostasis. Therefore, the results provided evidence that TP deficiency leads to nucleoside accumulation in lysosomes and lysosomal dysfunction, revealing the widespread disruption of organelles underlying MNGIE.Graphical

Atypical skin conditions of the neck and back as a dermal manifestation of anti-HMGCR antibody-positive myopathy

BackgroundImmune-mediated necrotizing myopathy (IMNM) is an idiopathic inflammatory myopathy (IIM). Though patients with IMNM were not considered to show skin rash, several reports have showed atypical skin conditions in patients with anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) antibody-positive IMNM (HMGCR-IMNM). The incidence and phenotype of skin conditions in patients with HMGCR-IMNM are not fully known.ResultsAmong the 100 IIM patients diagnosed from April 2015 through August 2022, 34 (34%) presented some form of skin condition, with 27 having typical skin rashes; this included 13 patients with dermatomyositis (DM), 8 with anti-synthetase syndrome (ASS), and 6 with IMNM. Meanwhile, 8 of 19 patients with HMGCR-IMNM (42%) presented atypical skin lesions, but no patients with other IIMs did (p < 0.001). Skin eruption with ash-like scales was observed in four HMGCR-IMNM patients, and non-scaly red patches and lumps in the other four patients; accordingly, their skin manifestations were considered as other dermal diseases except for IIM. However, skin and muscle biopsies revealed the atypical skin conditions of patients with HMGCR-IMNM to have the same pathological background, formed by Bcl-2-positive lymphocyte infiltrations.ConclusionsHMGCR-IMNM patients frequently have atypical skin conditions of the neck and back. Skin biopsy specimens from these lesions showed the same Bcl-2-positive lymphocytic infiltrations as muscle biopsy specimens regardless of the different gross dermal findings. Thus, such atypical skin conditions may be suggestive for HMGCR-IMNM.

Clinical features and genetic analysis of a child with Central core disease due to compound heterozygous variants of RYR1 gene

To explore the clinical features and genetic etiology of a child with Central core disease (CCD). A child with CCD who was treated at the Children's Hematology Department of the First Affiliated Hospital of Zhengzhou University in February 2022 was selected as the study subject. Muscle biopsy was performed. Peripheral blood samples were collected from the child and his parents for the extraction of genomic DNA. The child was subjected to whole exome sequencing (WES), and candidate variant was verified by Sanger sequencing. The child, a 12-year-old boy, had manifested motor retardation, facial weakness, ptosis, pectus carinatum, scoliosis, etc. Muscle biopsy showed that the central nucleus muscle fibers and atrophic muscle fibers were mainly type I. WES revealed that the child has harbored c.10561G>A (p.G3521S) and c.3448T>C (p.C1150R) compound heterozygous variants of the RYR1 gene. Sanger sequencing confirmed that they were inherited from his mother and father, respectively. Based on the guidelines from the American College of Medical Genetics and Genomics, both variants were considered as likely pathogenic (PS4+PM1+PM2_Supporting+PP3；PM1+PM2_Supporting+PM3+PP3). By combining his clinical manifestation and results of muscle pathology and genetic testing, the child was diagnosed with CCD, which may be attributed to the c.10561G>A (p.G3521S) and c.3448T>C (p.C1150R) compound heterozygous variants of the RYR1 gene.

Alterations in Skeletal Muscle Insulin Signaling DNA Methylation: A Pilot Randomized Controlled Trial of Olanzapine in Healthy Volunteers

Antipsychotics are associated with severe metabolic side effects including insulin resistance; however, the mechanisms underlying this side effect are not fully understood. The skeletal muscle plays a critical role in insulin-stimulated glucose uptake, and changes in skeletal muscle DNA methylation by antipsychotics may play a role in the development of insulin resistance. A double-blind, placebo-controlled trial of olanzapine was performed in healthy volunteers. Twelve healthy volunteers were randomized to receive 10 mg/day of olanzapine for 7 days. Participants underwent skeletal muscle biopsies to analyze DNA methylation changes using a candidate gene approach for the insulin signaling pathway. Ninety-seven methylation sites were statistically significant (false discovery rate &lt; 0.05 and beta difference between the groups of ≥10%). Fifty-five sites had increased methylation in the skeletal muscle of olanzapine-treated participants while 42 were decreased. The largest methylation change occurred at a site in the Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-Alpha (PPARGC1A) gene, which had 52% lower methylation in the olanzapine group. Antipsychotic treatment in healthy volunteers causes significant changes in skeletal muscle DNA methylation in the insulin signaling pathway. Future work will need to expand on these findings with expression analyses.

Inflammation-associated intramyocellular lipid alterations in human pancreatic cancer cachexia.

Cancer cachexia is a multifactorial metabolic syndrome characterized by systemic inflammation and ongoing skeletal muscle loss resulting in weakness, poor quality of life, and decreased survival. Whereas lipid accumulation in skeletal muscle is associated with cancer cachexia as well as the prognosis of cancer patients, surprisingly little is known about the nature of the lipids that accumulate in the muscle during cachexia, and whether this is related to inflammation. We aimed to identify the types and distributions of intramyocellular lipids in patients with and without cancer cachexia. Rectus abdominis muscle biopsies were collected during surgery of patients with pancreatic ductal adenocarcinoma (n=10 without cachexia, n=20 cachectic without inflammation (CRP<10mg/L), n=10 cachectic with inflammation (CRP≥10mg/L). L3-CT scans were analysed to assess body composition based on validated thresholds in Hounsfield units (HU). Muscle sections were stained with Oil-Red O and H&E to assess general lipid accumulation and atrophy. Untargeted lipidomic analyses were performed on laser-microdissected myotubes using LC-MS/MS. The spatial distribution of intramyocellular lipids with differential abundance between groups was visualized by mass-spectrometry imaging. Genes coding for inflammation markers and enzymes involved in de novo ceramide synthesis were studied by qPCR. Muscle radiation attenuation was lower in cachectic patients with inflammation (median 24.3 [18.6-30.8] HU) as compared with those without inflammation (34.2 [29.3-38.7] HU, P=0.033) or no cachexia (37.4 [33.9-42.9] HU, P=0.012). Accordingly, intramyocellular lipid content was lower in non-cachectic patients (1.9 [1.6-2.1]%) as compared with those with cachexia with inflammation (5.5 [4.5-7.3]%, P=0.002) or without inflammation (4.8 [2.6-6.0]%, P=0.017). Intramyocellular lipid accumulation was associated with both local IL-6 mRNA levels (rs=0.57, P=0.015) and systemic CRP levels (rs=0.49, P=0.024). Compared with non-cachectic subjects, cachectic patients had a higher relative abundance of intramyocellular glycerophospholipids and a lower relative abundance of glycerolipids. Furthermore, increases in several intramyocellular lipids such as SM(d36:1), PC(34:1), and TG(48:1) were found in cachectic patients with inflammation and correlated with specific cachexia features. Altered intramyocellular lipid species such as PC(34:1), LPC(18:2), and TG(48:1) showed an uneven distribution in muscle sections of cachectic and non-cachectic patients, with areas featuring abundance of these lipids next to areas almost devoid of them. Intramyocellular lipid accumulation in patients with cachexia is associated with both local and systemic inflammation, and characterized by changes in defined lipid species such as glycerolipids and glycerophospholipids.

Clinical and genetic characteristics of myotonia congenita in Chinese population

ABSTRACT Myotonia congenita (MC) is a rare hereditary muscle disease caused by variants in the CLCN1 gene. Currently, the correlation of phenotype-genotype is still uncertain between dominant-type Thomsen (TMC) and recessive-type Becker (BMC). The clinical data and auxiliary examinations of MC patients in our clinic were retrospectively collected. Electromyography was performed in 11 patients and available family members. Whole exome sequencing was conducted in all patients. The clinical and laboratory data of Chinese MC patients reported from June 2004 to December 2022 were reviewed. A total of 11 MC patients were included in the study, with a mean onset age of 12.64 ± 2.73 years. The main symptom was muscle stiffness of limbs. Warm-up phenomenon and percussion myotonia were found in all patients. Electromyogram revealed significant myotonic charges in all patients and two asymptomatic carriers, while muscle MRI and biopsy showed normal or nonspecific changes. Fourteen genetic variants including 6 novel variants were found in CLCN1. Ninety-eight Chinese patients were re-analyzed and re-summarized in this study. There were no significant differences in the demographic data, clinical characteristics, and laboratory findings between 52 TMC and 46 BMC patients. Among the 145 variants in CLCN1, some variants, including the most common variant c.892 G>A, could cause TMC in some families and BMC in others. This study expanded the clinical and genetic spectrum of Chinese patients with MC. It was difficult to distinguish between TMC and BMC only based on the clinical, laboratory, and genetic characteristics.

A healthcare claims analysis to identify and characterize patients with suspected X-Linked Myotubular Myopathy (XLMTM) in the Brazilian Healthcare System

BackgroundX-linked myotubular myopathy (XLMTM) is a rare, life-threatening congenital disease, which is not well-defined. To our knowledge, no studies characterizing the XLMTM disease burden have been conducted in Brazil. We identified and described patients with suspected XLMTM using administrative claims data from the Brazilian public healthcare system.MethodsData from 2015 to 2019 were extracted from the DATASUS database. As no XLMTM-specific ICD-10 code was available, a stepwise algorithm was applied to identify patients with suspected XLMTM by selecting male patients with a congenital myopathies code (G71.2), aged < 18 years at index date (first claim of G71.2), with an associated diagnostic procedure (muscle biopsy/genetic test) and without spinal muscular atrophy or Duchenne muscular dystrophy. We attempted to identify patients with suspected severe XLMTM based on use of both respiratory and feeding support, which are nearly universal in the care of XLMTM patients. Analyses were performed for the overall cohort and stratified by age at index date < 5 years old and ≥ 5 years old.ResultsOf 173 patients with suspected XLMTM identified, 39% were < 5 years old at index date. Nearly all (N = 166) patients (96%) were diagnosed by muscle biopsy (91% of patients < 5 years old and 99% of patients ≥ 5 years old), six (3.5%) were diagnosed by clinical evaluation (8% of patients < 5 years old and 1% of patients ≥ 5 years old), and one was diagnosed by a genetic test. Most patients lived in Brasilia (n = 55), São Paulo (n = 33) and Minas Gerais (n = 27). More than 85% of patients < 5 years old and approximately 75% of patients ≥ 5 years old had physiotherapy at the index date. In both age groups, nearly 50% of patients required hospitalization at some point and 25% required mobility support. Respiratory and feeding support were required for 3% and 12% of patients, respectively, suggesting that between 5 and 21 patients may have had severe XLMTM.ConclusionIn this real-world study, genetic testing for XLMTM appears to be underutilized in Brazil and may contribute to underdiagnosis of the disease. Access to diagnosis and care is limited outside of specific regions with specialized clinics and hospitals. Substantial use of healthcare resources included hospitalization, physiotherapy, mobility support, and, to a lesser extent, feeding support and respiratory support.

104 Effects of providing vitamin and mineral supplementation throughout gestation on subsequent F1 replacement heifer liver and muscle oxygen consumption and mitochondrial function throughout pregnancy

Abstract The study aimed to assess the impact of maternal vitamin/mineral supplementation during gestation on the energy utilization in liver and muscle tissues of F1 heifer offspring. We hypothesized that VTM supplementation during gestation would mitigate mitochondrial energy loss and uncoupling of liver and muscle tissue in pregnant offspring. At breeding, crossbred Angus heifers [n = 31; initial body weight (BW) = 412.5 ± 53.68 kg] were randomly assigned to either a basal diet targeting BW gains of 0.45 kg•heifer-1•d-1(CON; n = 14) or the basal diet plus 113 g•heifer-1•d-1of a vitamin/mineral supplement (VTM; n = 17). Heifers remained on their respective dietary treatments until calving. After calving, all F1 heifer calves were raised in a single management group with their dams until weaning (BW = 209 ± 4.82 kg; VTM, n = 17; CON, n = 12) and fed a common diet throughout postnatal development. At breeding and throughout gestation, F1 heifers (n = 9, VTM; n = 7, CON) were allotted daily feed intakes of 1.5% of BW on a dry-matter basis as a total mixed ration and used to evaluate cellular energy metabolism throughout pregnancy. Liver and Longissimus dorsi muscle biopsies were collected on d 179 and d 247 +/- 3 of gestation and mitochondrial function was assessed via high resolution respirometry (Oroboros Instruments, Innsbruck, Austria). The following respiratory states were evaluated: LEAK respiration (L), OXPHOS capacity (P), NADH-linked OXPHOS (PI), and electron transfer capacity (E). Data were analyzed using the MIXED procedure of SAS with repeated measures where appropriate. No treatment × trimester interaction (P ≥ 0.13) was observed for any of the respiratory states in liver and muscle. Hepatic oxygen consumption was not influenced by maternal gestational VTM supplementation (P ≥ 0.68), or stage of gestation (P ≥ 0.27). In muscle, L respiration was the only state affected by maternal dietary treatments, being greater (P = 0.05) in CON than VTM heifers. Additionally, L respiration was greater (P = 0.02) during the third trimester compared with the second trimester. This observation in CON heifers is important because increased L respiration (proton leak) results in decreased ATP production. Protons that escape the intermembrane space, are dissipated as heat rather than be used for ATP synthesis; thus, oxygen consumption increases to compensate for the reduction of protons entering ATP turnover. Results show increased muscle mitochondria inefficiency in offspring with dams lacking supplementation during pregnancy. The increased incidence of proton leak results in potentially less efficient cattle as protons escape the electron transport chain rather than be transformed for energy utilization need of the animal, thus VTM supplementation throughout pregnancy impacts fetal development with potential effects in live offspring 18 to 22 mo post-parturition.

297 Effects of plasma zinc concentration on longissimus thoracis metabolic status of beef steers

Abstract The objective of this study was to examine the role of Zn status on muscle glucose and other metabolites. Angus steers [n = 144; body weight (BW) = 525 ± 30 kg] with varying plasma Zn concentration and implant status served as the initial pool for this secondary experiment. Steers were assigned to one of two implant (IMP) treatments: no implant (NO) or Component TE-200 (TE-200; Elanco, Greenfield, IN) on d 0. Zinc was supplemented as ZnSO4 at 0 mg Zn/kg dry matter (DM; analyzed 54 mg Zn/kg DM), 30 mg Zn/kg DM, or 100 mg Zn/kg DM, starting 60 d before implant day. Steers were fed via GrowSafe bunks (GrowSafe Systems Ltd., Airdie, AB, Canada), and steer was experimental unit. Steers were fed in two blocks with a 14-d stagger for sample logistics. Blood and muscle biopsies of the longissimus thoracis were collected on d 40 after implant, a minimum of 2 h post-feed delivery. Plasma Zn was quantified via ICP-OES and 144 samples were stratified into quantiles by plasma Zn concentration. Samples (n = 12 low and 12 high from NO and TE-200 groups) were identified and designated as low plasma Zn (LOW, 1.1 mg Zn/L average) and high plasma Zn (HI, 1.6 mg Zn/L average) treatments (TRT). Muscle samples from these 48 steers were homogenized and derivatized for subsequent analysis via gas chromatography-mass spectrometry (GCMS) for non-targeted metabolomic analysis. Data were analyzed as a complete randomized design using the MIXED procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC) with fixed effects of TRT, IMP, BLOCK, and TRT×IMP. A TRT×IMP effect was observed for muscle glycine (P = 0.02) where LOW/IMP was similar to HI/NO but had greater glycine concentrations than LOW/NO and HI/IMP. There was a tendency for a TRT×IMP (P = 0.07) effect where LOW/IMP and HI/NO tended to have greater concentrations of hydroxyisovaleric acid. Given the role of glycine in protein synthesis and hydroxyisovaleric acid in leucine metabolism these data suggest differential protein responses to implants in cattle of lesser vs. greater plasma Zn concentrations. Hydroxybutyric acid, a ketone body which can be utilized for energy, tended to be greater in muscle of steers receiving an IMP (P = 0.09). Contrary to our hypothesis that greater plasma Zn would support differential muscle glucose metabolism, no effects of plasma Zn TRT or IMP were noted for glucose, lactate, and other detected metabolites. Both LOW and HI plasma Zn concentrations were still considered adequate, and future research should explore the impact of distinctly deficient and adequate plasma Zn concentration on glucose metabolism pathways in muscle.

A Guide to Conducting a Pathological Examination: Muscle Biopsy

Muscle biopsy for inflammatory myopathies facilitates the direct observation of muscle pathology at the site of inflammation by obtaining a small amount of tissue for pathological diagnosis. In clinical settings, muscle pathology derived from a muscle biopsy provides information limited to the tissue sampled. Thus, clinical information must supplement this to ascertain whether the obtained muscle pathology aligns with patient clinical presentation of myositis. This paper discusses considerations for effectively utilizing muscle biopsy in diagnosing inflammatory muscle diseases in clinical practice. Additionally, we briefly explore interpreting the pathological findings of myositis obtained from biopsy samples.

Treatment resistance in inclusion body myositis: the role of mast cells

Inclusion body myositis is the commonest acquired myopathy in those over 50 years of age. Although it is classified as an idiopathic inflammatory myopathy and the most frequent finding on muscle biopsy in inclusion body myositis is an endomysial inflammatory infiltrate, it is clinically distinct from other myositis, including a lack of response to immunosuppressive medication. Neurogenic changes are commonly reported in inclusion body myositis and inflammatory changes are observed in muscle following neurogenic injury. The objective of our study was to explore whether neurogenic inflammation plays a role in the pathogenesis of inclusion body myositis, possibly explaining its resistance to immunosuppression. The number of mast cells and presence of neuropeptides, substance P and calcitonin gene-related peptide, were assessed in 48 cases of inclusion body myositis, 11 cases of steroid responsive myositis, two cases of focal myositis associated with neurogenic injury, and ten normal controls. The number of mast cells in inclusion body myositis focal and myositis associated to neurogenic injury were significantly greater than that observed in steroid responsive myositis. Our findings suggest that neurogenic inflammation mediated through mast cells may play a role in the pathogenesis of inclusion body myositis, and focal myositis associated to neurogenic injury, and thus, explain in some part its lack of response to immunosuppressive treatments.

Transcriptomic gene signatures measure satellite cell activity in muscular dystrophies

Transcriptomic gene signatures measure satellite cell activity in muscular dystrophies

Deep learning-based classification of the capillary ultrastructure in human skeletal muscles

Deep learning-based classification of the capillary ultrastructure in human skeletal muscles

Sporadic Inclusion Body Myositis

Sporadic inclusion body myositis (sIBM) is an intractable and progressive skeletal muscle disease of unknown etiology. Muscle biopsy typically reveals endomysial inflammation, invasion of mononuclear cells into non-necrotic fibers, and rimmed vacuoles, suggesting that inflammation and degeneration co-exist in the pathomechanism. According to a nationwide survey conducted by a research team of the Ministry of Health, Labor, and Welfare, the number of patients is increasing in Japan as well. The clinical progression shows a slow and chronic deterioration. sIBM is usually diagnosed five years after onset. Muscle weakness and atrophy in the quadriceps, wrist flexors, and finger flexors are typical neurological findings of sIBM. Dysphagia and asymmetric weakness are often found. Serum creatine kinase is usually below 2,000 IU/L. sIBM is generally refractory to current therapy, such as steroids or immunosuppressants. Understanding the pathomechanism of sIBM is crucial for developing effective therapeutic strategies.

A Laing distal myopathy-associated proline substitution in the β-myosin rod perturbs myosin cross-bridging activity.

Proline substitutions within the coiled-coil rod region of the β-myosin gene (MYH7) are the predominant mutations causing Laing distal myopathy (MPD1), an autosomal dominant disorder characterized by progressive weakness of distal/proximal muscles. We report that the MDP1 mutation R1500P, studied in what we believe to be the first mouse model for the disease, adversely affected myosin motor activity despite being in the structural rod domain that directs thick filament assembly. Contractility experiments carried out on isolated mutant muscles, myofibrils, and myofibers identified muscle fatigue and weakness phenotypes, an increased rate of actin-myosin detachment, and a conformational shift of the myosin heads toward the more reactive disordered relaxed (DRX) state, causing hypercontractility and greater ATP consumption. Similarly, molecular analysis of muscle biopsies from patients with MPD1 revealed a significant increase in sarcomeric DRX content, as observed in a subset of myosin motor domain mutations causing hypertrophic cardiomyopathy. Finally, oral administration of MYK-581, a small molecule that decreases the population of heads in the DRX configuration, significantly improved the limited running capacity of the R1500P-transgenic mice and corrected the increased DRX state of the myofibrils from patients. These studies provide evidence of the molecular pathogenesis of proline rod mutations and lay the groundwork for the therapeutic advancement of myosin modulators.

The Influence of a Genetic Variant in CCDC78 on LMNA-Associated Skeletal Muscle Disease.

Mutations in the LMNA gene-encoding A-type lamins can cause Limb-Girdle muscular dystrophy Type 1B (LGMD1B). This disease presents with weakness and wasting of the proximal skeletal muscles and has a variable age of onset and disease severity. This variability has been attributed to genetic background differences among individuals; however, such variants have not been well characterized. To identify such variants, we investigated a multigeneration family in which affected individuals are diagnosed with LGMD1B. The primary genetic cause of LGMD1B in this family is a dominant mutation that activates a cryptic splice site, leading to a five-nucleotide deletion in the mature mRNA. This results in a frame shift and a premature stop in translation. Skeletal muscle biopsies from the family members showed dystrophic features of variable severity, with the muscle fibers of some family members possessing cores, regions of sarcomeric disruption, and a paucity of mitochondria, not commonly associated with LGMD1B. Using whole genome sequencing (WGS), we identified 21 DNA sequence variants that segregate with the family members possessing more profound dystrophic features and muscle cores. These include a relatively common variant in coiled-coil domain containing protein 78 (CCDC78). This variant was given priority because another mutation in CCDC78 causes autosomal dominant centronuclear myopathy-4, which causes cores in addition to centrally positioned nuclei. Therefore, we analyzed muscle biopsies from family members and discovered that those with both the LMNA mutation and the CCDC78 variant contain muscle cores that accumulated both CCDC78 and RyR1. Muscle cores containing mislocalized CCDC78 and RyR1 were absent in the less profoundly affected family members possessing only the LMNA mutation. Taken together, our findings suggest that a relatively common variant in CCDC78 can impart profound muscle pathology in combination with a LMNA mutation and accounts for variability in skeletal muscle disease phenotypes.

Pelizaeus disease Merzbacher like: About an observation

Pelizaeus-Merzbacher disease (PM) is an X-linked leukodystrophy causing developmental delay, nystagmus, hypotonia, spasticity and variable intellectual deficit. Three forms are described according to age of onset and severity: neonatal forms (+ severe), classical form (moderate, first 2 months of life) and transient form (the least serious 2-3 years). Prevalence is estimated at 1/400,000. We report the sighting of a 12-year-old and 8-month-old girl, with no significant history, from a non-indigenous marriage. Presents since the age of 5 months a bilateral horizontal nystagmus, myoclonia and a psychomotor retardation and whose examination revealed a pyramidal syndrome and a discreet hepatomegaly. The biological balance objectivéhyperlactatemie, hyperammoniémieet Chromatography of Amino Acids in urine showed an increase in glycine, alanine and tyrosine with Amino Acid chromatography in the blood and chromatography of Organic Acids normal. MRI brain objectified a demyelinating aspect of the white substance and atrophy of the corpus callosum. PEV pathological retina-cortical conduction, PEA altered left, Muscle biopsy absence of shredded red fibers and genetic study confirms PelizaeusMerzbacher disease like.