Muscle Fragments Research Articles

7962 Transformation of Poorly Differentiated Papillary Thyroid Carcinoma to Anaplastic Thyroid Carcinoma, an Atypical Course!

Abstract Disclosure: T. Altaweel: None. S. Makadsi: None. M. Hussein: None. J. Samantray: None. Introduction: Anaplastic thyroid cancers are undifferentiated, rare and aggressive. The transformation of papillary thyroid cancer to anaplastic thyroid cancer is documented in literature, however, rare. In this report, we present a case of Anaplastic thyroid carcinoma likely arising from a co-existing poorly differentiated Papillary carcinoma. Case Presentation: A 74-year-old male with past medical history of tobacco smoking who presented with the onset of voice hoarseness for 6 months. He has also noticed a left neck mass that is in the mid supraclavicular region. A Neck CT neck was performed at the time of presentation and was significant for a 4.6x3.4x5.9 cm left thyroid mass with displacement of the trachea. FNA biopsy showed papillary carcinoma with focal features of Tall Cell Variant. Poorly differentiated components with fragments of stroma and muscle were seen and suggested invasion. These findings raised the possibility of anaplastic thyroid carcinoma in the setting of papillary carcinoma and poorly differentiated carcinoma. BRAF V600E pathogenic variant mutation along with TERT mutation were found. Patient underwent direct laryngoscopy, bronchoscopy with biopsies, and tracheostomy with surgical pathology confirming papillary thyroid cancer with muscles invasion. Combination dabrafenib and trametinib were started. No surgical resection of thyroid tissue was planned. A follow up CT showed a significant decrease in the low-density soft tissue involving the medial thyroid gland bilaterally, consistent with interim positive response to therapy. Three months later, CT neck showed near complete resolution of low-density soft tissue involving the medial thyroid and parathyroid glands and subglottic trachea consistent with further interim positive response to therapy. Six months post initiation of therapy, CT Neck showed focal low density in the mid and inferior right thyroid gland which may represent residual and/or recurrent thyroid carcinoma. That density was further demonstrated on a repeat CT neck as a 1.8 cm hypodense nodule in the inferior right thyroid gland and concerning for recurrent thyroid tumor with possible infiltration of the trachea. Conclusion: ATC prognosis is affected by many factors such as metastasis, age, treatment strategies, socioeconomic status and the presence of BRAF V600E and TERT mutations. Diagnosis can be challenging. Our patient presented months after his symptoms started and this is an atypical course of Anaplastic Thyroid Carcinoma as it usually progresses aggressively and rapidly. Given the pathology reports and the more slow presentation of this patient’s carcinoma, it can be suggested that this is a case of papillary carcinoma with a later transformation to ATC. In these cases, surgical resection Presentation: 6/2/2024

Derivation and long-term maintenance of porcine skeletal muscle progenitor cells

Culture of muscle cells from livestock species has typically involved laborious enzyme-based approaches that yield heterogeneous populations with limited proliferative and myogenic differentiation capacity, thus limiting their use in physiologically-meaningful studies. This study reports the use of a simple explant culture technique to derive progenitor cell populations from porcine muscle that could be maintained and differentiated long-term in culture. Fragments of semitendinosus muscle from 4 to 8 week-old piglets (n = 4) were seeded on matrigel coated culture dishes to stimulate migration of muscle-derived progenitor cells (MDPCs). Cell outgrowths appeared within a few days and were serially passaged and characterised using RT-qPCR, immunostaining and flow cytometry. MDPCs had an initial mean doubling time of 1.4 days which increased to 2.5 days by passage 14. MDPC populations displayed steady levels of the lineage-specific markers, PAX7 and MYOD, up until at least passage 2 (positive immunostaining in about 40% cells for each gene), after which the expression of myogenic markers decreased gradually. Remarkably, MDPCs were able to readily generate myotubes in culture up until passage 8. Moreover, a decrease in myogenic capacity during serial passaging was concomitant with a gradual increase in the expression of the pre-adipocyte markers, CD105 and PDGFRA, and an increase in the ability of MDPCs to differentiate into adipocytes. In conclusion, explant culture provided a simple and efficient method to harvest enriched myogenic progenitors from pig skeletal muscle which could be maintained long-term and differentiated in vitro, thus providing a suitable system for studies on porcine muscle biology and applications in the expanding field of cultured meat.

Decellularized Bovine Skeletal Muscle Scaffolds: Structural Characterization and Preliminary Cytocompatibility Evaluation.

Skeletal muscle degeneration is responsible for major mobility complications, and this muscle type has little regenerative capacity. Several biomaterials have been proposed to induce muscle regeneration and function restoration. Decellularized scaffolds present biological properties that allow efficient cell culture, providing a suitable microenvironment for artificial construct development and being an alternative for in vitro muscle culture. For translational purposes, biomaterials derived from large animals are an interesting and unexplored source for muscle scaffold production. Therefore, this study aimed to produce and characterize bovine muscle scaffolds to be applied to muscle cell 3D cultures. Bovine muscle fragments were immersed in decellularizing solutions for 7 days. Decellularization efficiency, structure, composition, and three-dimensionality were evaluated. Bovine fetal myoblasts were cultured on the scaffolds for 10 days to attest cytocompatibility. Decellularization was confirmed by DAPI staining and DNA quantification. Histological and immunohistochemical analysis attested to the preservation of main ECM components. SEM analysis demonstrated that the 3D structure was maintained. In addition, after 10 days, fetal myoblasts were able to adhere and proliferate on the scaffolds, attesting to their cytocompatibility. These data, even preliminary, infer that generated bovine muscular scaffolds were well structured, with preserved composition and allowed cell culture. This study demonstrated that biomaterials derived from bovine muscle could be used in tissue engineering.

Development of Technology for the Production of Meat Products of the "Halal" Category for Functional Nutrition

The article is devoted to the development of technology for meat products of the "Halal" category. The authors consider the main problems of the meat industry in the field of the development of meat products of the "Halal" category in accordance with the National Program "Development of meat farming for 2018-2027", as well as the peculiarities of the production of meat products of the "Halal" category in Kazakhstan. A technology for the production of sausages for frying of the "Halal" category based on minced turkey and beef meat for functional nutrition has been developed, organoleptic, physico-chemical parameters of the new product have been studied. The mass fraction of protein is 8 %, the mass fraction of fat is 7 %. The authors of the article have developed a technological scheme for the production of a new product of the "Halal" category, which includes: acceptance of raw materials, cutting, deboning, veining, grinding to 2-3 mm of meat raw materials. At the next stage, meat is cooked at t 2-4 0C for 12 hours, minced meat is prepared (adding jerusalem artichoke 3 %, adding vitamin premix "Rovifarin" in an amount of 1.0 %, as well as spices and garlic according to the developed recipe). At the next stage, the minced meat is stuffed into natural shells with a diameter of (32-44) mm, cooled at t (4 + 2) 0C, stored at t (4 + 2) 0C for 36 hours. During the microstructural study of a sample of boiled and smoked sausage produced using traditional technology, it was found that the mass of the sample was formed from large fragments of muscle, fat and connective tissue (0.7-1.4) microns. The layout of the structural elements of the minced meat is dense, vacuoles and microcapillaries are rounded in shape, sometimes without clear boundaries, merged, with a size of (60-300) microns.

Pesticide residues detected in Colossoma macropomum by the modified QuEChERS and GC-MS/MS methods

This study aimed to detect pesticides in the muscle tissue of farmed tambaqui (Colossoma macropomum), in relation to good management practices (GMP) and prophylaxis and biosecurity measures. There were 54 fish farms randomly selected from the 138 found in the Microregion of Zona da Mata, RO - Brazil, for visits and collection of epidemiological data. There were extracted 24 muscle fragments were extracted from dorsolateral portion of the tail of five specimens tambaqui per fish farming. The methods of detection and quantification of pesticide residues were modified QuEChERS and GC-MS/MS. Most fish farms are small businesses and their production areas are smaller than a rural module. These ventures are an income alternative for rural producers, this information is confirmed in percentage of 88.89% (48/54) of the rural properties visited have livestock and agriculture as their main productive activity. Water monitoring was carried out in 70.37% of fish farms. However, with frequency of two water analyzes per year. Preventive and prophylactic measures taken were performed in 30% (17/54). In addition, 7% (4/54) of fish farmers reported using sodium chloride and/or using formalin as a secondary preventive measure. It is also important to mention that there was a report of administration of potassium permanganate in a fish farm, 2% (1/54). There were 12.49% (3/54) positive for at least one pesticide. The chemical compounds found Azosxystrobin (<LOQ), Epoxiconazole (<LOQ) and Chlorpyrifos (<LOQ). The presence of Azosxystrobin, Epoxiconazol and Chlorpyrifos in tambaqui muscle tissue samples was detected above acceptable limits by the PNCRC.

Mechanism of textural properties changes of cooked chicken in early postmortem: Effect of protein degradation induced by calpain on heating shrinkage.

Although the relationship between myofibrillar protein status and cooked meat quality is well documented, its underlying mechanism still need to be clarified. In this study, the effect of calpain-induced myofibrillar degradation on the cooked chicken quality was discussed by comparing the difference in muscle fiber's heat shrinkage state. In early postmortem, the protein around Z-line was degraded, which would cause the unstable Z-line and released into the sarcoplasm, according to WB results. This phenomenon will aggravate the lateral contraction of muscle fragments during the heating process. Then along comes a higher cooking loss and lower texture properties of meat. Above findings indicate that the Z-line dissociation caused by calpain in the early postmortem period is an essential reason for the quality difference of mature chicken. This study provided a fresh light on the mechanism underlying the impact of myofibril degradation in the early postmortem on the quality of cooked chicken.

Caracterização microbiológica de duas espécies de peixes neotropicais oriundas de área quilombola maranhense, Brasil

The aim of study is to feature the microbiological quality of two neotropical fish species from the quilombola area of Maranhão State, Brazil. In order to do so, 21 samples of Hoplerythrinus unitaeniatus and 21 samples of Cichlasoma bimaculatum were captured in flooded environment. Collected fish were euthanized in laboratory environment; muscle fragments were removed for microbiological analyses focused on enumerating molds and yeasts, viable strict and facultative mesophilic microorganisms and coagulase-positive staphylococci; on counting total and thermotolerant coliforms; and on investigating Escherichia coli and Salmonella sp. Microbiological results were compared to the Brazilian legislation, which establishes the list of microbiological standards for food products. Among the assessed fish, 9.52% were classified as non-acceptable for human consumption, based on the Salmonella parameter. Enumerated coagulase-positive staphylococci ranged from < 10 to 3.9 x 104 CFU/g; 9.52% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. E. coli counting ranged from 3.6 to > 1,100MPN/g; 4.76% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. Total and thermotolerant coliforms’ counting and the enumeration of viable strict and facultative aerobic microorganisms, as well as of molds and yeasts, have evidenced high microbial population rates; this finding suggests poor hygienic conditions at capture site, contaminated raw material and risk of incidence of enteropathogens. This finding has evidenced imbalance in the investigated environment, as well as compromised aquatic biodiversity.

Brazilian Horses from Bahia State Are Highly Infected with Sarcocystis bertrami.

The protozoan Sarcocystis bertrami (syn. Sarcocystis fayeri) infects horses and has dogs as definitive hosts. Herein we aimed to detect S. bertrami in Brazilian horses destined for human consumption and to determine the frequency of infection in the examined animals. Muscle fragments from 51 horses were collected in a slaughterhouse in Bahia State during three different seasons of the year. Samples from six tissues from each animal were prepared for macroscopic and microscopic evaluation, using tissue grinding, squash and histology. Sarcocystis sp. was observed in 100% of the examined horses. Selected samples were processed for transmission electron microscopy (TEM). Species identification was confirmed using a PCR targeted to the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Histological examination revealed sarcocysts with variable sizes and shapes, and dispersed within the muscle fibers. When observed by TEM, the sarcocyst wall was wavy and covered by an electrodense layer. The villar protrusions were digitiform and bent. To our knowledge, this study is the first morphological and molecular confirmation of S. bertrami in horses in Brazil and South America.

Laboratory Experience with the Microscopic Method for the Detection of Insects in Poultry Feeds.

The use of insects and their processed animal proteins (PAPs) for animal nutrition creates the need for research into methods useful for routine surveillance for their presence. The aim of this study was to evaluate a modified microscopic method for the detection of particles of insects in poultry feed. A total of 90 samples including PAP of insects (Hermetia illucens and Tenebrio molitor), poultry feeds produced with different levels (0-27%) of insect PAP content, and other poultry feeds spiked with insect PAP at 1% were investigated using a modified microscopic method with a double sedimentation protocol. Characteristic features of insects including cuticulae, muscles, bristles and tracheoles were determined in the microscopic images obtained. In all spiked samples, characteristic fragments of insects were detected. The fragments of muscle and tracheoles only indicated the presence of material from members of the insect class but could not facilitate identification of organisms to species level. The results obtained with this double sedimentation protocol for the isolation of insect PAP from feed for poultry have shown that the method can be used in routine analysis.

Histopathological Evidence of Canalicular Agenesis in Patients With Punctal Agenesis: Punctum Update Study 2.

The purpose of this study was to histologically assess the presence and extent of the canalicular tissue in cases of punctal agenesis. A prospective, interventional study was performed on 10 consecutive cases of punctal agenesis over a study period of 6 months. Exclusion criteria included cases with punctal dysgenesis or incomplete punctal canalization, secondary/acquired cases of punctal effacement, or any prior interventions. Following the exploration, the entire specimen of the tissue (length = 8 mm and depth = 3 mm) was sent for histopathological analysis to ascertain the presence and extent of the canalicular tissue. The mean age of the patients at the time of exploration was 7.5 years (range 6-10 years). Histopathological analysis showed tissue surfaces lined with stratified squamous epithelium with occasional interspersed goblets cells (conjunctival epithelium), a few chronic inflammatory cells, and muscle fragments. Epithelium suggestive of proximal lacrimal drainage system was absent. Canalicular tissues were found to be absent in all cases of punctal agenesis. The present series did not find any histologic evidence of canalicular tissues in cases of punctal agenesis. The findings of this study do not support retrograde approaches in cases of punctal agenesis.

Pathohistological characteristics of muscles in patients with chronic lower limb ischemia

To assess the signs of damage (the extent of necrosis and apoptosis, capillary density (CD)) and regeneration (the cross-sectional muscle fiber area (CSMFA), the proportion of centrinucleated muscle fibers (CNMFs), and that of connective tissue), by using the gastrocnemius medial head biopsy specimens obtained from patients with heterogeneous forms of chronic lower limb obliterating diseases (CLLODs). The investigation included the biopsy specimens obtained from 44 men under 65 years of age (their mean age was 54±9.8 years) with Stage IIB-IV chronic limb ischemia (according to the A.V. Pokrovsky-Fontaine classification) with its history of at least six months. The nosological entities were atherosclerotic lesion in 33 patients (distal atherosclerosis n=13), multistage lesion (n=8), and Leriche's syndrome (n=12); autoimmune-mediated vascular injury in 11 patients (Buerger's disease (n=7) and nonspecific aortoarteritis (n=4)). The similar muscle fragments obtained during autopsy from the deceased without obvious signs of cardiovascular system diseases were examined as a control. It was found that there was a statistically significant difference between the nosological entities, as compared to the control in terms of CD and CSMFA (a decrease), the proportion of CNMFs and that of connective tissue (an increase). No substantial differences were found in the studied parameters between the nosological entities. The findings may suggest the universal mechanism for damage to striated muscle tissue because of circulatory hypoxia, regardless of its etiology and the common character of tissue compensatory-adaptive responses (regeneration).

Study protocol: understanding the pathophysiologic mechanisms underlying delirium in older people undergoing hip fracture surgery

BackgroundPostoperative delirium (POD) is a common complication of older people undergoing hip fracture surgery, which negatively affects clinical- and healthcare-related outcomes. Unfortunately, POD pathophysiology is still largely unknown, despite previous studies showing that neuroinflammation, neuroendocrine dysfunction, increased reactive oxidative stress (ROS), and endothelial dysfunctions may be involved. There is also evidence that many of the pathophysiological mechanisms which are involved in delirium are involved in sarcopenia too.This article describes the protocol of a pilot study to evaluate the feasibility of a larger one that will explore the pathophysiological mechanisms correlating POD with sarcopenia. We will analyse whether various biomarkers reflecting neuroinflammation, ROS, neuroendocrine disorders, and microvasculature lesions will be simultaneously expressed in in the blood, cerebrospinal fluid (CSF), and muscles of patients developing POD.MethodsTwo centres will be involved in this study, each recruiting a convenient sample of ten older patients with hip fracture. All of them will undergo a baseline Comprehensive Geriatric Assessment, which will be used to construct a Rockwood-based Frailty Index (FI). Blood samples will be collected for each patient on the day of surgery and 1 day before. Additionally, CSF and muscle fragments will be taken and given to a biologist for subsequent analyses. The presence of POD will be assessed in each patient every morning until hospital discharge using the 4AT. Delirium subtypes and severity will be assessed using the Delirium Motor Subtype Scale-4 and the Delirium-O-Meter, respectively. We will also evaluate the patient’s functional status at discharge, using the Cumulated Ambulation Score.DiscussionThis study will be the first to correlate biomarkers of blood, CSF, and muscle in older patients with hip fracture.

Adaptation of the two-dimensional electrophoresis method for canned meat

Studies of the qualitative indicators of canned meat in accordance with regulatory documents are carried out on average samples of specimens, but when studying by proteomic methods, such sampling does not allow high-quality separation of protein components due to the high fat content in the product. When two-dimensional electrophoresis was carried out on an average sample, fragments of the main muscle and connective tissue proteins of beef were found in small quantities, but the electrophoretogram was not very informative. A significantly better separation was achieved after removing the fat fraction from the product. When studying broth from canned meat, the largest amount of intensely coloured high-molecular-weight protein fractions with a mass of more than 50 kDa was revealed. The electrophoretogram of the meat pieces showed a wide range of proteins across the entire molecular weight range of the polyacrylamide gel, including major muscle proteins. The study of broth together with meat pieces but after fat removal is optimal for the primary screening of the protein component of canned meat.

S3196 Diagnosis and Management of Phlegmonous Gastritis in a Young Female Without Predisposing Risk Factors

Introduction: Phlegmonous gastritis (PG) is a rare, but serious, condition characterized by bacterial infection of the gastric wall. Since 1862, less than 500 cases of PG have been reported in the literature, limiting knowledge of its management. We discuss a case of PG in a previously healthy female who was successfully treated with a combination of medical and surgical therapies. Case Description/Methods: A 32-year-old female presented to the emergency department with 10 days of worsening epigastric pain, fever, chills, and nausea. Symptoms onset after consuming a seafood platter containing shrimp, scallops, crab, and crawfish from a local restaurant. She had no significant past medical history. CT demonstrated heterogeneously enhancing wall thickening of the distal stomach and the first portion of the duodenum with adjacent fat stranding. Lab work was significant for leukocytosis and elevated CRP. Following admission, the patient was empirically started on piperacillin-tazobactam with general surgery and gastroenterology consulted. On hospital day 2, an EGD was performed which showed a raised lesion in the antrum with central ulceration and an overlying yellow exudate. Biopsies of the lesion were consistent with active gastritis with cryptitis. No organisms, including H pylori, were visualized on routine stains. MRI revealed a 7.5 x 4.3 cm mass in the gastric antrum with central necrosis and multiple septations. EGD with EUS on hospital day 6 noted a mass in the antrum arising from the muscular layer with areas of necrosis. FNA was performed, showing gastric columnar cells, smooth muscle fragments, and a cluster of bland-appearing spindle cells. The following day, she underwent an open distal gastrectomy with Billroth I reconstruction. Significant adhesions and a desmoid reaction involving the distal stomach to the falciform ligament was present. Tissue pathology showed a perforated ulcer with perigastric abscess and acute serositis. No malignant cells were identified. The post-operative course was uncomplicated, and the patient was discharged home on hospital day 11. Discussion: This case highlights the need to consider PG in the differential diagnosis of persistent abdominal pain, although patients may lack classic risk factors for this condition, such as immunosuppression or recent intervention. Discussion should be prompted regarding the optimal treatment regimen for PG, as there are currently no guidelines regarding standardized therapy.Figure 1.: Fig 1A: An abdominal X-ray showing the faint outline of the trichobezoar impacted within the stomach and leading to the convoluted small intestinal tract. Fig 1B: An abdominal CT scan showing the trichobezoar compacted within the stomach and the small intestines. Fig 1C: Post-operative picture of the removed trichobezoar that shows it took on the form of the stomach shape along with the tapered, elongated end of the form of the small intestine.

Abstract 2656: Muscle wasting in head and neck carcinomas: Investigations on the underlying transcriptional reprogrammation

Abstract Introduction: Cancer cachexia is a multifactorial syndrome associated with a skeletal muscle atrophy that affects at least 55% of patients bearing Head and Neck Squamous Cell Carcinomas (HNSCCs). To our knowledge, molecular mechanisms of muscle alterations have never been investigated on clinical muscle specimens from HNSCC patients. The aim of this study was to perform transcriptome profiling by bulk RNAseq on muscle fragments from HNSCC patients with and without cachexia and from nude mice bearing human HNSCC xenografts. Methods: Our study was made in three parts: 1) In Vitro investigations on human myoblasts co-cultivated with HNSCC cells. 2) In Situ investigations on muscles collected from nude mice xenografted with HNSSCs cells (FADU). 3) Exploration of clinical muscle samples collected from HNSCC patients. RNA sequencing was used to assess gene expression changes and gene function enrichment related to cachexia, in myoblasts co-cultivated with malignant cells as well as skeletal muscle samples collected from mice xenografted with HNSCC cells (FADU) or HNSCC patients. Results: In vitro experimentations showed significant impact of malignant cells on myoblast proliferation and differentiation. In nude mice, FADU xenografts resulted in distant loss of muscle mass at various anatomic locations. RNA sequencing of muscle samples from cachectic patients showed significant differences in gene expression compared with control muscles samples from tumor-free donors. Commonly modified pathways included immune and inflammatory response, mitochondrial metabolism and striated muscle differentiation. The most remarkable finding was the up-regulation of the BIRC3 transcript which encodes the anti-apoptotic protein c-IAP2. This alteration was observed both in human cachectic muscles and murine muscle fragments from tumor-bearing mice. Conclusions: Although the mechanism of BIRC3 up-regulation in cachectic muscle cells is not well understood, this finding is likely to be important for patient management in a context of promising therapeutic results obtained using Smac mimetics. Because these compounds are inhibitors of the c-IAP protein family, it will be important in the future to monitor their positive or negative effects on muscle alterations in cachectic HNSCC patients. Citation Format: Héla Hachicha, Iryna Pirozhkova, Aurore Gelin, François Bidault, Valérie Rouffiac, Thibault Dayris, Thierry Ragot, Ingrid Breuskin, Caroline Even, Philippe Gorphe, Pierre Busson. Muscle wasting in head and neck carcinomas: Investigations on the underlying transcriptional reprogrammation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2656.

Acetyl-CoA-driven respiration in frozen muscle contributes to the diagnosis of mitochondrial disease.

Freezing human biopsies is common in clinical practice for storage. However, this technique disrupts mitochondrial membranes, hampering further analyses of respiratory function. To contribute to laboratorial diagnosis of mitochondrial diseases, this study sought to develop a respirometry approach using O2k (Oroboros Ins.) to measure the whole electron transport chain (ETC) activity in homogenates of frozen skeletal muscle biopsies. We enrolled 16 patients submitted to muscle biopsy in the process of routine diagnostic investigation: four with mitochondrial disease and severe mitochondrial dysfunction; seven with exercise intolerance and multiple deletions of mitochondrial DNA, presenting mild to moderate mitochondrial dysfunction; five without mitochondrial disease, as controls. Whole homogenates of muscle fragments were prepared using grinder-type equipment. O2 consumption rates were normalized using citrate synthase activity. Transmission electron microscopy confirmed mitochondrial membrane discontinuation, indicating increased permeability of mitochondrial membranes in homogenates from frozen biopsies. O2 consumption rates in the presence of acetyl-CoA lead to maximum respiratory rates sensitive to rotenone, malonate and antimycin. This protocol of acetyl-CoA-driven respiration (ACoAR), applied in whole homogenates of frozen muscle, was sensitive enough to identify ETC abnormality, even in patients with mild to moderate mitochondrial dysfunction. We demonstrated adequate repeatability of ACoAR and found significant correlation between O2 consumption rates and enzyme activity assays of individual ETC complexes. We present preliminary data on a simple, low cost and reliable procedure to measure respiratory function in whole homogenates of frozen skeletal muscle biopsies, contributing to diagnosis of mitochondrial diseases in humans.

Twenty-one days of low-intensity eccentric training improve morphological characteristics and function of soleus muscles of mdx mice

Duchene muscular dystrophy (DMD) is caused by the absence of the protein dystrophin, which leads to muscle weakness, progressive degeneration, and eventually death due to respiratory failure. Low-intensity eccentric training (LIET) has been used as a rehabilitation method in skeletal muscles after disuse. Recently, LIET has also been used for rehabilitating dystrophic muscles, but its effects are still unclear. The purpose of this study was to investigate the effects of 21 days of LIET in dystrophic soleus muscle. Thirty-six male mdx mice were randomized into six groups (n = 6/each): mdx sedentary group; mdx training group-3 days; mdx training group-21 days; wild-type sedentary group; wild-type training group-3 days and wild-type training group-21 days. After the training sessions, animals were euthanized, and fragments of soleus muscles were removed for immunofluorescence and histological analyses, and measurements of active force and Ca2+ sensitivity of the contractile apparatus. Muscles of the mdx training group-21 days showed an improvement in morphological characteristics and an increase of active force when compared to the sedentary mdx group. The results show that LIET can improve the functionality of dystrophic soleus muscle in mice.

Влияние величины деформации в грудном отделе позвоночника на морфологическую картину параспинальных мышц у больных с идиопатическим сколиозом тяжелой степени

Introduction Morphological, biochemical and histopathological characteristics of paraspinal muscles have been well described in patients with idiopathic scoliosis. However, there is a paucity of literature on morphological picture of paraspinal muscles in patients with severe idiopathic scoliosis. Objective To explore how the magnitude of thoracic spine deformity affects morphological characteristics of paraspinal muscles in patients with severe idiopathic scoliosis. Material and methods A total 21 patients with idiopathic scoliosis were reviewed. The patients were assigned to two groups depending on magnitude of preoperative thoracic spine deformity. A major group consisted of patients with scoliosis of ≥ 60º (n = 11) and control group included patients with a curve < 60º (n=10). Biopsy samples were taken from muscle fragments (multifidus muscle) at the apex of the curve (major arc) at the Th6-Th10 level on the convex side while approaching to the posterior spinal structures during surgical correction of kyphoscoliosis using light microscope. The specimens were examined histologically with light microscope. Results Patients with severe idiopathic scoliosis (≥ 60º) showed evident muscle fiber atrophy and dystrophy of different extent. There were nucleus free areas and homogenized muscle fragments observed with connective tissue layers being swollen and frayed with multiple fibroblasts seen in nearly all the samples. Arterial walls were normally fibrotized. Structural changes ranged from minimal with a curve of 60º to degrading muscles with the fibers replaced by fat and connective tissues with a curve angle of 145º. Conclusion Morphological characteristics of paraspinal muscles have been shown to be severely affected by the magnitude of thoracic spine deformity in patients with severe idiopathic scoliosis. The findings should be considered in surgical correction of kyphoscoliosis with special focus on postoperative vascular and neurotropic therapy to allow wound healing and lower complication rate.

Human levator veli palatini muscle: a novel source of mesenchymal stromal cells for use in the rehabilitation of patients with congenital craniofacial malformations

BackgroundBone reconstruction in congenital craniofacial differences, which affect about 2–3% of newborns, has long been the focus of intensive research in the field of bone tissue engineering. The possibility of using mesenchymal stromal cells in regenerative medicine protocols has opened a new field of investigation aimed at finding optimal sources of multipotent cells that can be isolated via non-invasive procedures. In this study, we analyzed whether levator veli palatini muscle fragments, which can be readily obtained in non-invasive manner during palatoplasty in cleft palate patients, represent a novel source of MSCs with osteogenic potential.MethodsWe obtained levator veli palatini muscle fragments (3–5 mm3), during surgical repair of cleft palate in 5 unrelated patients. Mesenchymal stromal cells were isolated from the muscle using a pre-plating technique and other standard practices. The multipotent nature of the isolated stromal cells was demonstrated via flow cytometry analysis and by induction along osteogenic, adipogenic, and chondrogenic differentiation pathways. To demonstrate the osteogenic potential of these cells in vivo, they were used to reconstruct a critical-sized full-thickness calvarial defect model in immunocompetent rats.ResultsFlow cytometry analysis showed that the isolated stromal cells were positive for mesenchymal stem cell antigens (CD29, CD44, CD73, CD90, and CD105) and negative for hematopoietic (CD34 and CD45) or endothelial cell markers (CD31). The cells successfully underwent osteogenic, chondrogenic, and adipogenic cell differentiation under appropriate cell culture conditions. Calvarial defects treated with CellCeram™ scaffolds seeded with the isolated levator veli palatini muscle cells showed greater bone healing compared to defects treated with acellular scaffolds.ConclusionCells derived from levator veli palatini muscle have phenotypic characteristics similar to other mesenchymal stromal cells, both in vitro and in vivo. Our findings suggest that these cells may have clinical relevance in the surgical rehabilitation of patients with cleft palate and other craniofacial anomalies characterized by significant bone deficit.

Blackleg in inadequately immunized calves and their recovery following antibiotic therapy.

There is consensus regarding the importance of blackleg vaccination as a preventive measure, and proper immunization protocols are available. However, few studies have evaluated the effectiveness of vaccine protection against Clostridium chauvoei and the treatment of the disease in calves exhibiting early or advanced clinical courses. This study describes twelve blackleg cases in unvaccinated calves and in calves that received a single dose of the vaccine. It also reports the recovery of some calves after antibiotic therapy. Two necropsies of cattle dead from blackleg were performed. Fragments of skeletal muscle from these two cattle were immersed in paraffin for multiplex polymerase chain reaction (PCR) analysis. Twelve calves up to nine months of age developed signs of blackleg and eight died. Ten of those 9-month-old calves had received only the first dose of a blackleg vaccine at 4 months of age, but no booster. The last two affected calves belonged to a herd that had never been vaccinated. Four out of five calves treated with penicillin for 6-7 days recovered from the disease. The diagnosis of blackleg was based on necropsy, histopathological findings and detection of C. chauvoei in skeletal muscle samples of two necropsied calves using PCR. The occurrence of cases only in calves that did not receive a booster dose or were not vaccinated indicated that the vaccine used was effective when performed as recommended by the manufacturer. However, neglecting the booster resulted in casualties due to blackleg.