Actin and some actin binding proteins such as tropomyosin, -actinin and troponin T were localized by simultaneous double immunogold labeling in several developmental stages of Cryptosporidium parvum. All of the observed developmental stages have many particles of tropomyosin and actin around pellicle and cytoplasm. Tropomyosin was labeled much more than the actin when these two proteins were labeled simultaneously. And alpha-actinin was labeled mostly in the pellicle, but troponin T labeling was very rarely observed. From this study, it was suggested that tropomyosin seems to be one of the major proteins of C. parvum, so it must be playing important roles in C. parvum.
parasitology-protozoa ; actin ; tropomyosin ; alpha-actinin ; troponin T ; Cryptosporidium parvum

Cardiomyopathy, Dilated ; Cardiomyopathy, Restrictive/genetics* ; Humans ; Mutation ; Pedigree ; Tropomyosin/genetics*

Cap myopathy is pathologically characterized by cap structures comprising well-demarcated areas under the sarcolemma and containing deranged myofibrils and scattered Z-disks. Clinically it presents with slowly progressive muscle weakness, myopathic face, and frequent respiratory insufficiency. Four genes have been reported to be associated with the disease: TPM2, TPM3, ACTA1, and NEB. Here we describe that a patient presenting with mild limb weakness with facial affection showed cap structures on muscle pathology and carried a heterozygous TPM3 mutation.
Extremities ; Humans ; Muscle Weakness ; Muscular Diseases* ; Mutation, Missense* ; Myofibrils ; Pathology ; Respiratory Insufficiency ; Sarcolemma ; Tropomyosin

BACKGROUND: Neurotrophin-3 (NT-3), a member of the NT family, has only been considered an ancillary compound that provides anti-apoptotic benefits by inactivating tropomyosin receptor kinase C (TrkC)-induced apoptotic signals. However, little is known about the clinical relevance of NT-3 expression itself in neuroblastoma. The purpose of this study was to assess NT-3 expression in patients with neuroblastoma and its relevance to clinicopathologic findings and treatment outcomes. METHODS: In this study, expression of NT-3 and TrkC was analyzed using immunohistochemistry in 240 patients with newly diagnosed neuroblastoma. RESULTS: The results of the study revealed that NT-3 expression was associated with older age at diagnosis, localized tumors, and more differentiated tumors but was not associated with early treatment response (degree of residual tumor volume after three cycles of chemotherapy) and progression-free survival (PFS). However, when analysis was confined to patients with MYCN amplified tumors, NT-3 expression was associated with better early treatment response with borderline significance (P = 0.092) and higher PFS (86.9% vs. 58.2%; P = 0.044). In multivariate analysis in patients with MYCN amplified tumors, NT-3 was independent prognostic factor (hazard ratio, 0.246; 95% confidence interval, 0.061–0.997; P = 0.050). In another subgroup analysis, the early treatment response was better if NT-3 was expressed in patients without TrkC expression (P = 0.053) while it was poorer in patients with TrkC expression (P = 0.023). CONCLUSION: This study suggests that NT-3 expression in neuroblastoma has its own clinical significance independent of TrkC expression, and its prognostic significance differs depending on the status of MYCN amplification and/or TrkC expression.
Diagnosis ; Disease-Free Survival ; Humans ; Immunohistochemistry ; Multivariate Analysis ; Neoplasm, Residual ; Neuroblastoma ; Phosphotransferases ; Tropomyosin

Objective: To investigate the expression pattern of tropomyosin 2(TPM2) in aorta of patients with aortic dissection and explore its clinical implication. Methods: Thirteen cases with acute type A aortic dissection(TAAD) diagnosed by transabdominal aortic angiography from 2015 in Tongji Hospital were included. During the operation, the aortic wall tissues of these patients were collected. Ten patients with heart transplantation were selected as control group, and normal aortic wall tissues were taken. The hematoxylin-eosin (HE) and Verhoeff's Van Gieson (EVG) staining were performed to observe the morphological changes of aorta. The mRNA expression level of TPM2 was measured by real-time fluorescent quantitative-PCR, and the protein levels of TPM2 were detected by Western blot and immunohistochemical staining. Image The J software was used to collect the optical density values of each point on the image, obtain the integrated optical density(IOD) value, and calculate the average density(%, IOD/area of the target distribution area). Results: HE and EVG staining revealed medial degeneration and broken elastic fiber in aorta of TAAD patients. The mRNA expression levels of TPM2 were significantly upregulated in aorta of TAAD patients as compared to the control group (P<0.05), so as the TPM2 protein expression levels ((9.73±1.20)% vs. (0.11±0.04)%, P<0.05). And TPM2 was mainly expressed in cytoplasm. Conclusion: The increased expression of TPM2 in TAAD patients hints that TPM2 might be involved in the pathogenesis of aortic dissection.
Aneurysm, Dissecting/genetics* ; Aorta ; Aortic Aneurysm, Thoracic/genetics* ; Gene Expression ; Humans ; RNA, Messenger ; Tropomyosin/metabolism*

OBJECTIVETo investigate the dynamic expression of TPM1 in rat model of hepatic fibrosis and hepatic stellate cells induced by TGFβ1.

METHODThirty male SD rats were divided into control group (n = 6) and model group (n = 24). The rat model of hepatic fibrosis was established by intraperitoneal injection of dimethylnitrosamine(DMN). The sera were collected from portal vein and liver tissues were taken from animals 2, 4, 6, 8 weeks HSC-T6 cells were cultured and induced 48 hours by 5 ng/ml TGF-β1. The pathological changes of liver were observed by Hematoxylin-Eosin and Masson Staining. Reverse Transcription-polymerase chain reaction (RT-PCR), immunohistochemistry and Western-blotting were used to determine the mRNA and protein expressions of TPM1, TGFβ1 and α-SMA in rat models and HSC-T6 cells and the localization of TPM1 in rat models.

RESULTSRat models of hepatic fibrosis were successfully established. TPM1 was lowly stained in the wall of blood vessels in portal areas in normal livers, in fibrotic livers TPM1 was mainly stained along the fibrotic septum. The mRNA and protein expressions of TPM1 and α-SMA in rat models of hepatic fibrosis increased at the week 2 and peaked at week 6, which was statistical significance compared to control group, P < 0.05; TGF-β1 increased at week 2 and it was higher than the levels in other groups at week 4, which was statistical significance compared to control group P < 0.05; Correlation analysis showed that TPM1 positively correlated with α-SMA and TGF-β1, rs = 0.688, rs = 0.692, P < 0.01. In HSC-T6, the mRNA expressions of TPM1 and α-SMA increased after being induced by TGF -beta1. compare with control group, the differences were significant, P less than 0.05.

CONCLUSIONTPM1 may be playing an important role in the occurrence and development of liver fibrosis. Maybe it could become a potential therapeutic target for hepatic fibrosis.

Animals ; Hepatic Stellate Cells ; metabolism ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Tropomyosin ; metabolism

Crustacean shellfish allergy is an important cause of food allergy and anaphylaxis in Asia. The major allergen in shellfish allergy is tropomyosin, a pan-allergen that is also found in house dust mites and cockroaches. Tropomyosins from house dust mites (HDMs) have a high sequence homology to shellfish tropomyosins, and cross-reactivity between HDM and shrimp tropomyosins has been demonstrated. Exposure to inhaled tropomyosins from house dust mites has been postulated to be the primary sensitizer for shellfish allergy, in a reaction analogous to the oral allergy (inhalant-food) syndrome. This notion is supported by indirect data from the effects of HDM immunotherapy on shellfish allergy, and strong correlations of shellfish and HDM sensitization. HDM immunotherapy has been reported to induce both shrimp allergy in non-allergic patients and shrimp tolerance in shrimp-allergic patients. Epidemiological surveys have also demonstrated a strong correlation between shellfish and HDM sensitization in both hospital-based and community-based studies. Unexposed populations have also been shown to develop sensitization-shellfish sensitization in orthodox Jews with no history of shellfish consumption was associated with HDM sensitization. Reciprocally, HDM sensitization in an Icelandic population living in a HDM-free environment was associated with shrimp sensitization. In vitro IgE inhibition studies on sera in shrimp-allergic Spanish patients indicate that mites are the primary sensitizer in shrimp-allergic patients living in humid and warm climates. Current data supports the hypothesis that tropomyosin is the link between HDM and shellfish allergies. The role of tropomyosin in HDM and shellfish allergies is a fertile field for investigation as it may provide novel immunotherapeutic strategies for shellfish allergy.
Anaphylaxis ; Asia ; Climate ; Cockroaches ; Dust* ; Food Hypersensitivity ; Humans ; Hypersensitivity ; Iceland ; Immunoglobulin E ; Immunotherapy ; Jews ; Mites ; Pyroglyphidae* ; Sequence Homology ; Shellfish* ; Tropomyosin*

OBJECTIVE: We previously described that Diva is highly expressed in matured metaphase II (MII) oocytes compared to immature germinal vesicle (GV) oocytes in mouse.1 We report here that the expression of Diva transcript as well as protein is oocyte-specific. To elucidate its physiological role in oocyte, the binding partner(s) of Diva has been identified by using immunoprecipitation (IP) followed by Mass Spectrometry. METHODS: NIH/3T3 cells were transiently transfected for 24 h with either empty vector for control or FLAG-tagged mouse Diva construct, and IP was performed with anti-FLAG antibody. The immuno-isolated complexes were resolved by SDS-PAGE on a 12% gel followed by Coomassie Blue staining. For in-gel digestion, 15 bands of interest were excised manually and digested with trypsin. All mass spectra were acquired at a positive reflector mode by a 4700 Proteomics Analyzer (Applied Biosystems, Framingham, MA). Proteins were identified by searching the NCBI nonredundant database using MASCOT Peptide Mass Fingerprint software (Matrixscience, London). RESULTS: Diva-associated complexes were formed in FLAG-tagged mouse Diva-overexpressed NIH/3T3 cells via IP using anti-FLAG-conjugated beads. Among the excised 15 bands, actin and actin-binding proteins such as tropomyosin, tropomodulin 3, and alpha-actinin were identified. Binding between Diva and actin or tropomyosin was confirmed by IP followed by Western blot analysis. Both bindings were also detected endogenously in mouse ovaries, indicating that Diva works with actin and tropomyosin. CONCLUSIONS: This is the first report that immuno-isolated Diva-associated complexes are related to actin filament of the cytoskeletal system. When we consider the association of Diva with actin and tropomyosin, oocyte-specific Diva may play a role in modulating the cytoskeletal system during oocyte maturation.
Actin Cytoskeleton ; Actinin ; Actins ; Animals ; Blotting, Western ; Dermatoglyphics ; Digestion ; Electrophoresis, Polyacrylamide Gel ; Female ; Immunoprecipitation ; Mass Spectrometry* ; Metaphase ; Mice ; Microfilament Proteins ; Oocytes ; Ovary ; Proteomics ; Tropomodulin ; Tropomyosin ; Trypsin

BACKGROUND AND OBJECTIVES: Left ventricle burdened by longstanding volume-overload, undergoes various structural and functional alterations. Accordingly, the expressions of multiple classes of genes are likely to be altered. However, the profile of gene expressions, specifically in a volume-overloaded left ventricle in humans, has not been explored. SUBJECTS AND METHODS: The pattern of gene expression was studied, using a cDNA microarray, in myocardium from 4 normal subjects and 5 patients with chronic regurgitant valvular heart disease whose end-diastolic left ventricular dimension measures 65 mm or more, but whose systolic function remained preserved. RESULTS: We identified 58 differentially expressed genes that were functionally classifiable in the volume-overloaded myocardium. Those genes involved in cell cycle/growth (up/down-regulation: 9/1), signal transduction (4/1) were mostly overexpressed in the volume-overloaded myocardium. The distributions of the gene expressions were variable for those involved in transcription/translation (up/down-regulation: 6/7) and apoptosis (2/2). The genes related to the myocyte structure (troponin T3, tropomyosin, etc)(up/down-regulation: 1/10), as well as those related to metabolism (2/5), were underexpressed. The gene expression patterns from RT-PCR and Western blot, with randomly selected genes, were similar to those from the cDNA microarray. CONCLUSION: Altered expression was identified in multiple genes in the volume-overloaded human left ventricle prior to the development of heart failure. The genes related to cell growth and signal transduction were mostly overexpressed, while those related to cellular structure and metabolism appeared to be underexpressed. These results might help in the elucidation of cellular mechanisms for the remodeling process associated with chronic volume-overloading.
Apoptosis ; Blotting, Western ; Cellular Structures ; Gene Expression* ; Heart Failure* ; Heart Valve Diseases ; Heart Ventricles ; Heart* ; Humans* ; Metabolism ; Muscle Cells ; Myocardium* ; Oligonucleotide Array Sequence Analysis ; Signal Transduction ; Transcriptome* ; Tropomyosin

BACKGROUND: Shrimp is one of the major causative crustacean food allergen. An investigation has been reported that tropomyosin belonged to muscle protein is a major allergen within shrimp. But there have been a few investigations on shrimp allergen in Korea. The aim of this study is to evaluate skin reactivity and specific IgE sensitization to Metapenaeus joyneri which is one of the major shrimp in this country, and to identitify IgE binding components and evaluate allergenic relationship with other species. METHODS: We performed skin prick test with M. joyneri extract in 1,738 patients. ELISA was performed for detection of serum specific IgE antibody. To evaluate the cross allergenecity between M. joyneri and other crustaceans (crab, lobster, crayfish), Dermatophagoides pteronyssinus (Dpt), triton shell, abalone and buckwheat. ELISA inhibition tests were performed with each four patient's sera showing high specific IgE antibody. To identify IgE binding components, SDS-PAGE followed by IgE-Immunoblot were applied. RESULTS: 211 patients (12.2%) showed positive responses (A/H >or=2+) on skin prick test. Serum specific IgE antibodies were detected in 61 patients (37.2%) of 164 sensitzed patients. ELISA inhibition test using four patient's sera showed significant inhibitions by M. joyneri. and other crustaceans including lobster, crab and crayfish, partial inhibitions were noted by Dpt, triton shell, buckwheat and abalone. SDS-PAGE and IgE-imunoblot with patients' individual sera sensitized to M. joyneri showed 12 IgE binding components (31, 32, 38-44, 57, 70, 81 kDa) and two (31, 32 kDa) were bound to IgE in more than 50% of sera tested. Five (43, 44, 57, 70 and 81 kDa) were bound to IgE in more than 25% of sera tested. CONCLUSION: Specific IgE was detected in 37.2% of allergy patients sensitized to M. joyneri. Twelve IgE binding components and two (31, 32 kDa) major allergens were indentified. Cross allergenecity was noted with other crustaceans.
Allergens ; Antibodies ; Astacoidea ; Dermatophagoides pteronyssinus ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Fagopyrum ; Humans ; Hypersensitivity ; Immunoglobulin E* ; Korea ; Muscle Proteins ; Neptune ; Penaeidae ; Skin ; Tropomyosin