There have been numerous hypotheses about the pathogenesis of idiopathic scoliosis, but it is still unclear. There are some reports that abnormalities of contractile proteins may play a role in the pathogenesis of idiopathic scoliosis. The purpose of this report is to study the quantitative abnormalities of contractile proteins in muscles and nonactivated and activated platelets, and to determine whether or not the abnormalities in contractile proteins may play a role in the pathogenesis of idiopathic scoliosis. The materials were 21 idiopathic scoliosis patients aged from 13 years to 28 years(average 19.2 years) and 20 persons aged from 17 years to 25 years(average 20.1 years) as a control group. The electrophoretic analysis(SDS-PAGE method) was done on platelets both unstimulated and stimulated with thrombin and also on proteins of paraspinal muscles and gluteus maximus of idiopathic scoliosis patient and paraspinal muscles of control group. The results are as follows. 1. The myosin/actin ratios of triton-insoluble fractions to paraspinal muscles in convex sides of main curvatures of scoliosis patients(1.69±0.81) were significantly decreased compared to those of concave sides(2.55±1.28), gluteus maximus muscles(2.56±1.70) and control group(2.61±1.01). 2. There were no significant differences between scoliosis group and control group in the actin/myosin ratios of triton-insoluble fractions of the platelets both nonactivated and activated by thrombin. In conclusion, abnormalities of contractile protein in paraspinal muscles of convex side may play a role in the pathogenesis of idiopathic scoliosis, rather than abnormalities of systemic contractile protein.
Actins ; Blood Platelets ; Contractile Proteins ; Humans ; Muscles ; Myosins ; Paraspinal Muscles ; Scoliosis ; Thrombin

Diabetes mellitus is known to be associated with a specific cardiomyopathy. This is evident from the clinical-pathological work and the epidemiologic data. An investigation was made in this study to determine whether diabetic cardiomyopathy in rats is associated with an alteration of biochemical characteristics of cardiac contractile proteins. Rats were made diabetic with intravenous injection of streptozotocin and hearts removed 8 weeks later for the isolation of myofibrils. The basal ATPase activity of myofibrils from diabetic hearts was significantly lower than that of the controls, suggesting the presence of some subtle structural and conformational changes in diabetic myofibrils. The activating effect of Mg ions on the myofibrillar actomyosin system of rat heart muscle was also demonstrated. Sodium dodecylsulfate gel electrophoresis showed the presence of myosin heavy chain, light chain 1 and 2, actin and troponin but failed to reveal differences in the patterns of these contractile proteins of light subunits between diabetics and controls. The deficiency in utilization of energy rich phosphates by the myofibrillar protein may be one of of the main mechanisms of cardiodepression observed in diabetic hearts. The cardiac myofibrillar ATPase activity may be one of useful measurements in evaluating pathophysiological states of cardiac contractile proteins.
Actins ; Actomyosin ; Adenosine Triphosphatases* ; Animals ; Cardiomyopathies ; Contractile Proteins ; Diabetes Mellitus ; Diabetic Cardiomyopathies ; Electrophoresis ; Heart ; Injections, Intravenous ; Ions ; Myocardium ; Myofibrils ; Myosin Heavy Chains ; Phosphates ; Rats* ; Sodium ; Streptozocin ; Troponin

The main task of skeletal muscle is contraction and relaxation for body movement and posture maintenance. During contraction and relaxation, Ca²⁺ in the cytosol has a critical role in activating and deactivating a series of contractile proteins. In skeletal muscle, the cytosolic Ca²⁺ level is mainly determined by Ca²⁺ movements between the cytosol and the sarcoplasmic reticulum. The importance of Ca²⁺ entry from extracellular spaces to the cytosol has gained significant attention over the past decade. Store-operated Ca²⁺ entry with a low amplitude and relatively slow kinetics is a main extracellular Ca²⁺ entryway into skeletal muscle. Herein, recent studies on extracellular Ca²⁺ entry into skeletal muscle are reviewed along with descriptions of the proteins that are related to extracellular Ca²⁺ entry and their influences on skeletal muscle function and disease.
Contractile Proteins ; Cytosol ; Extracellular Space ; Kinetics ; Muscle, Skeletal* ; Posture ; Relaxation ; Sarcoplasmic Reticulum

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that alpha-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and beta-actin, actin-capping protein(beta subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.
Actins ; Carrier Proteins ; Collagen ; Collagen Type I ; Collagen Type V ; Connective Tissue ; Cytoskeletal Proteins ; DNA, Complementary* ; Epidermal Growth Factor ; Fibroblasts* ; Gene Expression Profiling ; Gene Expression* ; Humans ; Insulin-Like Growth Factor II ; Muscle, Smooth ; Myosin Heavy Chains ; Myosin Light Chains ; Myosins ; Nerve Growth Factor ; Oligonucleotide Array Sequence Analysis* ; Osteoblasts ; Periodontal Ligament* ; Receptors, Fibroblast Growth Factor ; Receptors, Platelet-Derived Growth Factor ; Regeneration ; RNA ; RNA, Messenger

BACKGROUND AND OBJECTIVES: Most studies in cardiac regeneration have explored bone marrow mesenchymal stem cells (BM-MSC) with variable therapeutic effects. Amniotic fluid MSC (AF-MSC) having extended self-renewal and multi-potent properties may be superior to bone marrow MSC (BM-MSC). However, a comparison of their cardiomyogenic potency has not been studied yet.METHODS: The 5-azacytidine (5-aza) treated AF-MSC and BM-MSC were evaluated for the expression of GATA-4, Nkx2.5 and ISL-1 transcripts and proteins by quantitative RT-PCR and Western blotting, respectively as well as for the expression of cardiomyogenic differentiation markers cardiac troponin-T (cTNT), beta myosin heavy chain (βMHC) and alpha sarcomeric actinin (ASA) by immunocytochemistry.RESULTS: The AF-MSC as compared to BM-MSC had significantly higher expression of GATA-4 (183.06±29.85 vs. 9.80±0.05; p<0.01), Nkx2.5 (8.3±1.4 vs. 1.82±0.32; p<0.05), and ISL-1 (39.59±4.05 vs. 4.36±0.39; p<0.01) genes as well as GATA-4 (2.01±0.5 vs. 0.6±0.1; p<0.05), NKx2.5 (1.9±0.14 vs. 0.8±0.2; p<0.01) and ISL-1 (1.7±0.3 vs. 0.9±0.1; p<0.05) proteins. The AF-MSC also had significantly elevated expression of cTNT (5.0×10⁴±0.6×10⁴ vs. 3.5 ×10⁴±0.8×10⁴; p<0.01), β-MHC (15.7×10⁴±0.9×10⁴ vs. 8.2×10⁴±0.6×10⁴; p<0.01) and ASA (18.6×10⁴±4.9×10⁴ vs. 13.1×10⁴±3.0×10⁴; p<0.05) than BM-MSC.CONCLUSIONS: Our data suggest that AF-MSC have greater cardiomyogenic potency than BM-MSC, and thus may be a better source of MSC for therapeutic applications in cardiac regenerative medicine.
Actinin ; Amniotic Fluid ; Antigens, Differentiation ; Azacitidine ; Blotting, Western ; Bone Marrow ; Female ; Humans ; Immunohistochemistry ; Mesenchymal Stromal Cells ; Regeneration ; Regenerative Medicine ; Therapeutic Uses ; Troponin T ; Ventricular Myosins

BACKGROUND AND OBJECTIVES: We investigated the effects of different concentrations of serum, 5-azacytidine, and culture time on the cardiomyogenic differentiation of P19 embryonal carcinoma stem cells in the course of developing an efficient protocol for generating the cardiomyogenic lineage. MATERIALS AND METHODS: P19 cells were plated at a density of 1x10(6) cells on 10-cm bacterial dishes for 96 hours in the presence of 1% dimethyl sulfoxide to form embryoid bodies. The embryoid bodies were cultured in medium with 2% or 10% fetal bovine serum for an additional 10 or 15 consecutive days in the presence of 0, 1, or 3 microM 5-azacytidine. RESULTS: Quantitative real-time polymerase chain reaction (PCR) analysis showed that the messenger ribonucleic acid (mRNA) expression of cardiac muscle-specific genes, such as GATA4, alpha-actin, alpha-myosin heavy chain, and cardiac troponin T, were significantly higher in the 15-day culture groups than in the 10-day culture groups. Furthermore, the cardiac muscle-specific genes were expressed more in the high-serum groups compared to the low-serum groups regardless of the culture time. Cardiomyogenic differentiation of the P19 cells was most effective in 1 microM 5-azacytidine regardless of the serum concentrations. In addition, the stimulation effects of 5-azacytidine on cardiomyogenic differentiation were more significant under low-serum culture conditions compared to high-serum culture conditions. Cardiomyogenic differentiation of P19 cells was further confirmed by immunostaining with cardiac muscle-specific antibodies. CONCLUSION:Taken together, these results demonstrated that cardiomyogenic differentiation of P19 cells was enhanced by a combination of different experimental factors.
Actins ; Antibodies ; Azacitidine ; Carcinoma, Embryonal ; Cell Differentiation ; Dimethyl Sulfoxide ; Embryoid Bodies ; Embryonal Carcinoma Stem Cells ; Myocytes, Cardiac ; Real-Time Polymerase Chain Reaction ; RNA ; Safrole ; Troponin T ; Ventricular Myosins

PURPOSE: Aspirin exacerbated respiratory disease (AERD) results in a severe asthma attack after aspirin ingestion in asthmatics. The filamin A interacting protein 1 (FILIP1) may play a crucial role in AERD pathogenesis by mediating T cell activation and membrane rearrangement. We investigated the association of FILIP1 variations with AERD and the fall rate of forced expiratory volume in one second (FEV1). METHODS: A total of 34 common FILIP1 single nucleotide polymorphisms (SNPs) were genotyped in 592 Korean asthmatic subjects that included 163 AERD patients and 429 aspirin-tolerant asthma (ATA) controls. RESULTS: This study found that 5 SNPs (P=0.006-0.01) and 2 haplotypes (P=0.01-0.03) of FILIP1 showed nominal signals; however, corrections for the multiple testing revealed no significant associations with the development of AERD (P corr>0.05). In addition, association analysis of the genetic variants with the fall rate of FEV1, an important diagnostic marker of AERD, revealed no significant evidence (P corr>0.05). CONCLUSIONS: Although further replications and functional evaluations are needed, our preliminary findings suggest that genetic variants of FILIP1 might be not associated with the onset of AERD.
Aspirin ; Asthma ; Contractile Proteins ; Eating ; Forced Expiratory Volume ; Haplotypes ; Humans ; Hypersensitivity ; Membranes ; Microfilament Proteins ; Negotiating ; Polymorphism, Single Nucleotide

OBJECTIVE: The aims of this study were to examine whether a passive stretch stimulus by means of a functional appliance induces changes in the fiber composition of masticatory muscles and whether these changes are similar to the changes in stretched limb muscle fibers by using RT-PCR, western blot, and immunohistochemical assays. METHODS: Five male New Zealand White rabbits were fitted with a prefabricated inclined plane on the maxillary central incisors to force the mandible forward (- 2 mm) and downward (- 4 mm). Further, 1 hind limb was extended and constrained with a cast so that the extensor digitorum longus (EDL) was stretched when the animal used the limb. The animals were sacrificed after 1 week and the masseter, lateral pterygoid, and EDL were processed and compared with those from control animals (n = 3). RESULTS: The stretched EDL had a significantly higher percentage of slow fibers, whereas the stretched masticatory muscles did not show changes in the composition of the major contractile proteins after 7 days. CONCLUSIONS: The transition of fiber phenotypes in response to a stretch stimulus may take longer in the masticatory muscles than in the limb muscles.
Animals ; Blotting, Western ; Contractile Proteins ; Extremities ; Humans ; Incisor ; Male ; Mandible ; Masticatory Muscles ; Muscles ; Myosin Heavy Chains ; Phenotype ; Rabbits

PURPOSE: To elucidate the molecular basis of muscle atrophy in cellular adaptation point of view, gene expression profiling in rat muscle atrophy model was performed. The functions changed by muscle atrophy were analyzed. MATERIAL AND METHODS: Sciatic nerve and femoral nerve were resected in right leg to make muscle atrophy model in rat. The left leg was considered as a compensatory hypertrophy model. The suppression subtractive hybridization (SSH) was done to identify the profile of differential gene expression during muscle atrophy followed by nerve injury in rat. The DNA fragments obtained in SSH were labeled with biotin and used as cDNA tags for isolation of full-length cDNA from cDNA library. Differentially expressed genes were confirmed by reverse dot blot hybridization. RESULTS: Down regulation of genes were much more predominant than up regulation. The profile of down regulated genes were composed of genes coding muscle contractile proteins, enzymes involving carbohydrate metabolism including glycolysis and glycogenolysis, enzymes in oxidative phoshorylation, and proteins related with calcium release. The target genes were isolated by enrichment using cDNA tags from cDNA library for further functional studies. We identified some novel genes related with muscle atrophy by nerve injury. CONCLUSION: During the process of muscle atrophy, genes coding muscle contractile proteins, enzymes in carbohydrate metabolism, enzymes in oxidative phosphorylation, and proteins related with calcium release were down regulated.
Animals ; Atrophy ; Biotin ; Calcium ; Carbohydrate Metabolism ; Chimera ; Clinical Coding ; Contractile Proteins ; DNA ; DNA, Complementary ; Down-Regulation ; Femoral Nerve ; Gene Expression ; Gene Expression Profiling ; Gene Library ; Glycogenolysis ; Glycolysis ; Hypertrophy ; Leg ; Muscle, Skeletal ; Muscles ; Muscular Atrophy ; Oxidative Phosphorylation ; Proteins ; Rats ; Sciatic Nerve ; Up-Regulation

OBJECTIVETo explore the genetic basis and phenotypic correlation with disease severity in a large cohort of Chinese patients with hypertrophic cardiomyopathy (HCM).

METHODSA total of 179 unrelated Chinese HCM patients admitted to our department from 2002 to 2011 were enrolled in this study. Direct gene sequencing of β-myosin heavy chain (MYH7), myosin binding protein-C ( MYBPC3), and cardiac troponin T (TNNT2) were performed and clinical data were obtained in these patients.

RESULTSA total of 34 mutations were identified in 40 patients (22.3%), 79.4% (27/34) mutations occurred only once and a possible hot spot, A26 in MYH7, was found. Distribution of mutations was 52.9% (18/34) (MYBPC3), 35.3% (12/34) ( MYH7) and 11.8% (4/34) (TNNT2) respectively. Double mutations were identified in 2.2% (4/179) patients. Genotype-positive patients were associated with an earlier symptom onset, severer left ventricular hypertrophy, a higher incidence of syncope, and were more likely to have positive family history of HCM or sudden cardiac death (SCD) , and were more likely to progress into heart failure (24.2% vs. 5.0%, P = 0.002) and at a higher risk of SCD (9.1% vs. 0, P = 0.009) during the 6.5-year follow-up. No statistical difference in any clinical parameters and outcomes was found between patients carrying MYBPC3 and MYH7 mutations. Double mutations were associated with malignant clinical progression in this cohort. Different phenotype severity could be seen in HCM patients with same genotype (e. g. MYH7-1736T, TNNT2-R92W).

CONCLUSIONMYBPC3 is the most predominant gene mutation in this HCM cohort. The presence of a sarcomere mutation in patients with HCM is associated with poor clinical outcome, although no specific genes or mutations can exactly predict the severity of clinical phenotypes.

Asian Continental Ancestry Group ; Cardiomyopathy, Hypertrophic ; Carrier Proteins ; Death, Sudden, Cardiac ; Disease Progression ; Genotype ; Humans ; Hypertrophy, Left Ventricular ; Mutation ; Phenotype ; Sarcomeres ; Troponin T ; Ventricular Myosins