• Title/Summary/Keyword: Desmin

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Desmin Binding Property of Nebulin Isoforms

  • Jeon Eun-Hee;Lee Yeong-Mi;Lee Min-A;Kim Ji-Hee;Choi Jae-Kyong;Park Eun-Ran;Kim Hyun-Suk;Ahn Seung-Ju;Min Byung-In;Joo Young-Mi;Kim Chong-Rak
    • Biomedical Science Letters
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    • v.12 no.2
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    • pp.73-79
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    • 2006
  • Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.

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The Diagnostic Utility of Mesothelial Markers in Distinguishing between Reactive Mesothelial Cell and Adenocarcinoma Cells in Serous Effusions with Cytospin Preparation (장액성 삼출액의 도말 표본에서 반응성 중피세포와 샘암종 세포의 감별에 중피세포 표지자의 유용성)

  • Kang, Mi-Sun;Choi, Soo-Im
    • The Korean Journal of Cytopathology
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    • v.17 no.2
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    • pp.108-115
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    • 2006
  • Evaluation of serous effusions can include immunocytochemical stains that differentiate reactive mesothelial cell from adenocarcinoma cell. Among several positive mesothelial cell markers, we used desmin, CK5/6, WT1 and calretinin all known to have high sensitivity and specificity as selective mesothelial cell markers. We studied smears obtained with cytospin from 15 malignant and eight benign effusions. The mesothelial cells were positively stained by desmin, CK5/6, WT1 and calretinin in 60.9%, 29.1%, 26.7% and 56.5%, respectively among 8 benign and 15 malignant effusions; the adenocarcinoma cells were positively stained 6.7%, 13.3%, 1.0% and 0.0%, respectively among 15 malignant effusions. The percentage of positively stained mesothelial cells were somewhat lower for all antibodies compared to the results of previous studies. This was likely due to the differences in preparation methods and fixatives among studies. In conclusion, the use of desmin and calretinin were more valuable than CK5/6 and WT1 for distinguishing between reactive mesothelial cell and adenocarcinoma cells in serous effusion; however, choice of the proper preparation methods and fixatives are also important

Proteomic Analysis of Differentially Expressed Proteins in Bovine Endometrium with Endometritis

  • Choe, Chang-Yong;Park, Jeong-Won;Kim, Eun-Suk;Lee, Sung-Gyu;Park, Sun-Young;Lee, Jeong-Soon;Cho, Myung-Je;Kang, Kee-Ryeon;Han, Jae-Hee;Kang, Da-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.4
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    • pp.205-212
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    • 2010
  • Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, $\alpha$-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin $\beta$ subunit, and potassium channel tetramerisation domaincontaining 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, $\alpha$-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and $\alpha$-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and $\alpha$-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis.

Effect of Exercise on Muscle Recovery of Sciatic Nerve Injured Rats (궁둥신경 손상을 유발한 흰쥐에서 운동이 근육재생에 미치는 영향)

  • Ro, Hyo-Lyun
    • Proceedings of the KAIS Fall Conference
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    • 2010.05b
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    • pp.855-858
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    • 2010
  • 이 연구의 목적은 흰쥐의 궁둥신경의 손상을 준 뒤 달리기운동을 시켜 운동이 근육재생에 효과가 있는지를 밝힐 목적으로 시행하였다. 수컷 Sprague-Dawley계 흰쥐의 궁둥신경을 압궤손상을 준후, 하루 60분 씩 주 5회 운동을 시켰다. 궁둥신경 압궤손상 후 12일(5일간 운동), 19일(10일간 운동), 26일(15일간 운동), 33일(20일간 운동 및 61일(40일간 운동)에 흰쥐를 희생시켜 장딴지근육을 절취하여 냉동절편을 만들었으며 중간세사인 desmin과 vimentin에 대한 면역조직화학적 염색방법을 시행하였고 NADH-TR을 이용한 효소조직화학적 방법으로 뼈대근육의 변성 등을 관찰하였다. 중간세사단백 중 desmin은 근육섬유의 변성 및 재생과정 모두에서 발현되며 vimentin은 재생과정에서만 발현되었다. 대조군보다 실험군에서 desmin과 vimentin의 면역 반응성이 높았고, 실험군의 근육섬유들은 손상 후 61일째에 횡단면에서 각을 갖고 있어 정상 근육섬유임을 입증해 주었으나 대조군의 근육섬유는 정상으로 회복되지 않은 둥근 근육섬유가 관찰되었다. 19일째의 NADH-TR반응에서 대조군에서는 근육 섬유형 군집(fiber type grouping)으로 인해 diformazan 과립이 뭉쳐져 있음을 볼 수 있으나 실험군에서는 점점 diformazan 과립이 고르게 분포됨을 볼 수 있었다. 재신경지배가 일어 날 때 나타나는 표적근육섬유(target fiber)는 NADH-TR반응에서 26일 실험군의 일부 근육섬유에서 처음으로 관찰되었다. 61일째 NADH-TR반응에서 대조군은 아직도 diformazan 과립이 근육섬유성 군집을 보였으나 실험군에서는 정상군과 다름없는 염색성이 관찰되었다. 이상의 결과로 흰쥐 궁둥신경 압궤손상 후 트레드밀 달리기 운동이 흰쥐 다리 뼈대근육의 중간세사발현에 효과가 있다고 생각된다.

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Ultrastructural Changes of Fat-storing Cells in Experimental Hepatic Fibrosis (실험적 간섬유화에서 Fat-storing Cell의 미세구조의 변화에 대한 연구)

  • Kim, Mi-Jin;Choi, Won-Hee;Lee, Tae-Sook
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.224-238
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    • 1992
  • Hepatic fibrosis was induced in Sprague-Dawley rats to evaluate the ultrastructural changes of fat-storing cells(Ito cells). For experimental induction of liver fibrosis, the rats were administered intraperitoneally with 0.5ml of 50% $Ccl_4$ solution per Kg body weight, twice weekly for 12 weeks. The rats were sacrified every week. The liver tissues were examined under light and eletron microscopes. And the immunohistochemical study of desmin was also performed. The results were summarized as follows : Light microscopic findings : The cellular infiltrations with inflammatory cells and Kupffer cells developed from 1 week after $Ccl_4$ injection, and were the most severe in 4 weeks. The strong immunoreactivity for desmin was also evident in 4 weeks. The centrilobular necrosis and fibrosis developed from 2 weeks after injection, and the necrosis persisted until 8 weeks. The progress of fibrosis was accompanied by decreases in cellular infiltration and reactivity for desmin, and increased gradual nodular formation was also observed. The cirrhosis was developed after 10 weeks. Electron microscopic findings : An increase in number of fat-storing cells was observed from 1 week after injection. Transitional cells characterized by a depletion of lipid droplets and a hypertrophy of the rER appeared after 2 weeks. The number of transitional cells with abundant collagen fibers in the extracellular spaces increased in 4 weeks. With progression of fibrosis the number of fat-storing cells decreased and proliferating fibroblasts with dilated rER were observed. According to these results it was revealed that there was an apparent transition from fat-storing cells to transitional cells and to fibroblasts. These cells had a few similar characteristics and may belong to the same cell population. Thus it was suggested that fat-storing cells might play an important role in hepatic fibrosis.

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Differentiation of mouse embryonic stem cell into smooth muscle cells by DBcAMP and retinoic acid (DBcAMP와 retinoic acid를 이용한 마우스 배아줄기의 평활근세포 분화)

  • Park, Sung-Soo;Kang, Ju-Won
    • Korean Journal of Veterinary Service
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    • v.31 no.4
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    • pp.449-456
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    • 2008
  • The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

Postmortem Proteolysis of Breast and Leg Muscles from Taiwan Colored Chickens and Silkie Bantams

  • Tsai, Shih-Fen;Lin, Chia-Ying;Lu, Jin-Jenn;Chou, Rong-Ghi R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.739-743
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    • 2006
  • Postmortem proteolysis of breast (BM) and leg (LM) muscles from Taiwan colored chickens (TCC) and silkie bantams (SB) at $5^{\circ}C$ were compared. Myofibrils were prepared from BM and LM samples that were randomly taken from the carcasses of SB and TCC after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. pH of samples was determined, and degradation of myofibrillar proteins was analyzed by the SDS-PAGE and western blots. The results showed that pH was lower in BM than in LM samples from both avian strains. Appearance of 30 kDa components and disappearance of titin and nebulin were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data might suggest that postmortem proteolysis occurred more rapidly in BM than in LM from both TCC and SB.

Comparison of Postmortem Proteolysis between Breast and Leg Muscles in Chiayi Native Chickens

  • Cha, Shih-Ting;Tseng, Tsai-Fuh;Ho, Sy-Shyan;Chou, Rong-Ghi R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.5
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    • pp.721-724
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    • 2002
  • Postmortem Proteolysis of breast (BM) and leg (LM) muscles in Chiayi native chickens at $5^{\circ}C$ were compared. Myofibrils were purified from BM and LM samples that were randomly taken from carcasses after 0, 1, 3, 7 and 14 days of storage at $5^{\circ}C$. Fragmentation of myofibrils were determined, and degradation of myofibrillar proteins were analyzed by the SDS-PAGE and western blots. The results showed that myofibril fragmentation index (MFI) was significantly (p<0.05) higher in BM than in LM samples. Disappearance of titin and nebulin and appearance of the 30 kDa component were more rapidly as seen on SDS-PAGE in BM than in LM samples. Western blots labeled with a monoclonal antibody to desmin also demonstrated that desmin degraded more quickly in BM samples. Our data suggested that postmortem proteolysis occurred more rapidly in breast muscles in Chiayi native chickens.

Immunohistochemical and Electron Microscopic Studies on Intermediate Filament of Epithelial Cell and Non-Epithelial Cells (상피 및 비상피세포들의 Intermediate Filament에 대한 면역조직화학적 및 전자현미경적 연구)

  • Kim, Il;Kim, Hyo-Sung;Rho, Young-Bok
    • Applied Microscopy
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    • v.21 no.1
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    • pp.46-62
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    • 1991
  • The intermediate filament is one of the most important constituents of the intracytoplasmic cytoskeleton microtubule, actin, myosin and intermediate filament. It is composed of keratin, desmin, vimentin, neurofilament and glial filament, and has important role as a cellular marker, epithelial or mesenchymal origin. So it will be important to differentiated from some poorly or undifferentiated neoplasm to provide adequate therapeutic modalities. This study was performed by using immunohistochemical staining and electron microscopic observation to find out intermediate filaments of epithelial and non-epithelial tumor cells evaluate the degree of differentiation in tumors and therefore to provide some diagnostic and therapeutic modalities. The materials consisted of 83 epithelial and non-epithelial elements bearing 23 normal control, 28 epithelial tumors, and 32 non-epithelial tumors, that are resected for definite treatment at Chosun University Hospital from June, 1988 to June, 1990. Immunohistochemical stain for keratin, desmin and vimentin, and electron microscopic study were performed in all cases. The results obtained were as follows. 1. Immunohistochemical stain for intermediate filament were very useful diagnostic aid for differentiated epithelial tumor to non-epithelial tumor in diagnostic neoplasia. 2. In the electron microscopic finding, the size of intermediate filaments were possible differentiated to cell components of epithelial tumor and non-epithelial tumors.

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Primary Leiomyosarcoma of The Lung -One case report- (폐의 원발성 평활근육종)

  • 김형수;지현근;이원용;김응중;홍기우;남은숙
    • Journal of Chest Surgery
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    • v.31 no.9
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    • pp.907-910
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    • 1998
  • Primary leiomyosarcoma is an extremely rare tumor. We exprienced a case of 46-year-old man with a mass located in the left upper lobe bronchus which was discovered on a chest CT. Cytology of the sputum and bronchoscopic biopses did not reveal any malignant cells. The patient underwent a left sleeve upper lobectomy. The tumor was growing from the left upper lobe bronchus and had partially destroyed the lung parenchyme. The pathologic feature of the tumor was composed of fascicular arrayed cellular spindle cells with blunt-ended nuclei revealed mild to moderate pleomorphism and frequent mitoses (15/10HPF). The immunohistochemical staining was revealed positive reaction for antibody to smooth muscle actin and desmin. We conclude that the tumor is leiomyosarcoma of the lung and then report with a review of the literatures.

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