• Title/Summary/Keyword: Nuclear localization

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Gene Structure and Function of fkhE, a Forkhead Gene in a Filamentous Fungus Aspergillus nidulans (Aspergillus nidulans forkhead 유전자 fkhE의 구조와 기능 분석)

  • Park, Mi-Hye;Kim, Hyoun-Young;Kim, Jong-Hwa;Han, Kap-Hoon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.160-166
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    • 2010
  • A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.

Dexamethasone Facilitates NF-κB Signal Pathway in TNF-α Stimulated Rotator Cuff Tenocytes

  • Ji, Jong-Hun;Kim, Young-Yul;Patel, Kaushal;Cho, Namjoon;Park, Sang-Eun;Ko, Myung-Sup;Park, Suk-Jae;Kim, Jong Ok
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.297-303
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    • 2019
  • Corticosteroids are commonly used for pain control in rotator cuff tear. Deregulated $NF-{\kappa}B$ activation is a hallmark of chronic inflammatory diseases and has been responsible for the pathogenesis of rotator cuff tear. The Dexamethasone(DEXA) is a synthetic corticosteroid. The purpose of this study was to examine the exact effect of dexamethasone on $NF-{\kappa}B$ signaling in rotator cuff tear. We measured $NF-{\kappa}B$ expression in four groups: control, $TNF-{\alpha}$-treated, DEXA-treated, and combined treatment with $TNF-{\alpha}$ and DEXA. Tenocytes were isolated from patients with rotator cuff tears and pre-incubated with $TNF-{\alpha}$ (10 ng/ml), DEXA ($1{\mu}M$), or both of them for 10 min, 1 h, and 2 h. Expression of p65, p50, and p52 in the nuclei and cytosol was analyzed by western blotting and immunofluorescence imaging using confocal microscopy. We also evaluated nucleus/cytosol (N/C) ratios of p65, p50, and p52. In our study, the combined treatment with DEXA and $TNF-{\alpha}$ showed increased N/C ratios of p65, p50, and p52 compared with those in the $TNF-{\alpha}$ group at all time points. Additionally, in the DEXA group, N/C ratios of p65, p50, and p52 gradually increased from 10 min to 2 h. In conclusion, DEXA promoted the nuclear localization of p65, p50, and p52, but was not effective in inhibiting the inflammatory response of $TNF-{\alpha}$-stimulated rotator cuff tear.

The TGFβ→TAK1→LATS→YAP1 Pathway Regulates the Spatiotemporal Dynamics of YAP1

  • Min-Kyu Kim;Sang-Hyun Han;Tae-Geun Park;Soo-Hyun Song;Ja-Youl Lee;You-Soub Lee;Seo-Yeong Yoo;Xin-Zi Chi;Eung-Gook Kim;Ju-Won Jang;Dae Sik Lim;Andre J. van Wijnen;Jung-Won Lee;Suk-Chul Bae
    • Molecules and Cells
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    • v.46 no.10
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    • pp.592-610
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    • 2023
  • The Hippo kinase cascade functions as a central hub that relays input from the "outside world" of the cell and translates it into specific cellular responses by regulating the activity of Yes-associated protein 1 (YAP1). How Hippo translates input from the extracellular signals into specific intracellular responses remains unclear. Here, we show that transforming growth factor β (TGFβ)-activated TAK1 activates LATS1/2, which then phosphorylates YAP1. Phosphorylated YAP1 (p-YAP1) associates with RUNX3, but not with TEAD4, to form a TGFβ-stimulated restriction (R)-point-associated complex which activates target chromatin loci in the nucleus. Soon after, p-YAP1 is exported to the cytoplasm. Attenuation of TGFβ signaling results in re-localization of unphosphorylated YAP1 to the nucleus, where it forms a YAP1/TEAD4/SMAD3/AP1/p300 complex. The TGFβ-stimulated spatiotemporal dynamics of YAP1 are abrogated in many cancer cells. These results identify a new pathway that integrates TGFβ signals and the Hippo pathway (TGFβ→TAK1→LATS1/2→YAP1 cascade) with a novel dynamic nuclear role for p-YAP1.

The Evaluation for Attenuation Map using Low Dose in PET/CT System (PET/CT 시스템에서 감쇠지도를 만들기 위한 저선량 CT 평가)

  • Nam, So-Ra;Cho, Hyo-Min;Jung, Ji-Young;Lee, Chang-Lae;Lim, Han-Sang;Park, Hoon-Hee;Kim, Hee-Joung
    • Progress in Medical Physics
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    • v.18 no.3
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    • pp.134-138
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    • 2007
  • The current PET/CT system with high quality CT images not only increases diagnostic value by providing anatomic localization, but also shortens the acquisition time for attenuation correction than primary PET system. All commercially available PET/CT system uses the CT scan for attenuation correction instead of the transmission scan using radioactive source such as $^{137}Cs,\;^{68}Ge$. However the CT scan may substantially increase the patient dose. The purpose of this study was to evaluate quality of PET images reconstructed by CT attenuation map using various tube currents. in this study, images were acquired for 3D Hoffman brain phantom and cylindrical phantom using GE DSTe PET/CT system. The emission data were acquired for 10 min using phantoms after injecting 44.03 MBq of $^{18}F-FDG$. The CT images for attenuation map were acquired by changing tube current from 10 mA to 95 mA with fixed exposure time of 8 sec and fixed tube voltage of 140 kVp. The PET images were reconstructed using these CT attenuation maps. Image quality of CT images was evaluated by measuring SD (standard deviation) of cylindrical phantom which was filled with water and $^{18}F-FDG$ solution. The PET images were evaluated by measuring the activity ratio between gray matter and white matter in Hoffman phantom images. SDs of CT images decrease by increasing tube current. When PET images were reconstructed using CT attenuation maps with various tube currents, the activity ratios between gray matter and white matter of PET images were almost same. These results indicated that the quality of the PET images using low dose CT data were comparable to the PET images using general dose CT data. Therefore, the use of low dose CT is recommended than the use of general dose CT, when the diagnostic high quality CT is not required. Further studies may need to be performed for other system, since this study is limited to the GE DSTe system used in this study.

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Renal Expression of TonEBP and Urea Transporter in the Water-deprived Mongolian Gerbil(Meriones unguiculatus) (절수시 Mongolian Gerbil 콩팥에서 TonEBP와 Urea transporter의 발현 변화)

  • Park, Yong-Deok;Kim, Sung-Joong;Jung, Ju-Young
    • Applied Microscopy
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    • v.37 no.4
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    • pp.271-280
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    • 2007
  • Tonicity-responsive enhancer binding protein(TonEBP) is a transcriptional factor essential in the function and development of the renal medulla. TonEBP plays a critical role in protecting renal medullary cells from the deleterious effect of hypertonicity. TonEBP is a key regulator of urinary concentration via stimulation of transcription of urea transporter(UT) in a manner independent of vasopressin. UT in the renal inner medulla is important for the conservation of body water due to its role in the urine concentrating mechanism. Mongolian gerbil(Meriones unguiculatus) has been as an model animal for studying the neurological disease such as stroke and epilepsy because of the congenital incomplete in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animal. In this study, we divide 3 groups of which each group include the 5 animals. In the study of 7 or 14 days water restricted condition, we investigated the TonEBP and UT-A by using a immunohistochemistry in the kidney. In the normal kidney, the distribution of TonEBP is generally localized on nuclei of inner medullary cells. Nuclear distribution of TonEBP is generally increased throughout the medulla in 7 and 14 days dehydrated group compared with control group. Increased nuclear localization was particularly dramatic in thin limbs. In control groups, UT-A was expressed in inner stripe of outer medulla(ISOM) and inner medulla(IM). UT-A was present in the terminal part of the short-loop of descending thin limbs (DTL) in ISOM and also present in the inner medullary collecting duct(IMCD), where the intensity of it gradually increased toward the papillary tip. In the dehydrated kidney, UT-A immunoreactivity was increased in the short-loop of DTL in ISOM and in the long-loop of DTL in the initial part of IM, where was expressed moderate positive reaction in the normal kidney. Also it was up regulated in the IMCD in initial & middle part of IM. However UT-A down regulated in the IMCD, where the intensity of it gradually decreased toward the papillary tip. These findings suggest that increased levels of TonEBP in medulla and UT-A in shot-loop of DTL and IMCD play a important role for maintain fluid balance in the water-deprived mongolian gerbil kidney.

Evaluation of Usefulness of Radio-iodine SPECT/CT in Differentiated Thyroid Cancer (분화성갑상선암에서 방사성요오드 SPECT/CT의 유용성 평가)

  • Lee, Jeong-Won;Lee, Ho-Young;Oh, So-Won;Kim, Seok-Ki;Jeong, Ki-Wook;Kim, Seon-Wook;Kang, Keon-Wook
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.5
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    • pp.350-358
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    • 2007
  • Purpose: Localizing and differentiating a metastatic lesion of differentiated thyroid cancer (DTC) by using radio iodine whole body scan could be difficult because a whole body scan (WBS) lacks anatomic information. This study was performed to evaluate the usefulness of radio-iodine SPECT/CT for differentiating equivocal lesions. Materials & Methods: Among 253 patients with DTC who had undergone radio-iodine scan between February and July 2006, 26 patients were enrolled (M:F = 8:18, Age $50.7{\pm}12.5$ years) in this study. The patients had abnormal uptakes in the WBSs that necessitated precise anatomical localization for differentiating between a metastatic lesion and a false-positive lesion. SPECT/CT was performed for the region with abnormal uptake in the WBS. WBS and SPECT/CT were evaluated visually. Metastases were diagnosed based on the results of the radio-iodine scan along with the results of other radiological examinations and serological tests. Results: Based on the WBS images, 13 were suspected with cervical lymph node (LN) metastases in 16 patients with abnormal neck uptake, and in the 11 patients with abnormal extra-cervical uptakes, extra-cervical metastases were doubtful in all. After SPECT/CT was performed, the diagnostic results were altered for 16 patients (62%). SPECT/CT revealed that only 5 patients had cervical LN metastases, while 3 patients had extra-cervical (mediastinal) LN metastases. Overall, there was a 58% (15/26) change in diagnoses and plans for treatment due to SPECT/CT. Among 8 patients suspected with metastases on SPECT/CT, 6 patients underwent another radio-iodine therapy. In 96% (24/25) of the patients, the results of SPECT/CT corresponded with those of further radiological examinations and with other clinical information. Conclusion: Radio-iodine SPECT/CT images permitted the differentiation of abnormal radio-iodine uptake and improved anatomical interpretation in DTC.

Assessment of the Synovial Inflammation in Rheumatoid Arthritis with $^{99m}Tc$-labelled Polyclonal Human IgG(HIG): Prospective Comparison with Gadolinium Enhanced MRI ($^{99m}Tc$-labelled HIG 스캔을 이용한 류마티스 관절염 환자에서 활막염증의 평가 : 조영증강 자기공명영상과의 전향적인 비교)

  • Ryu, Young-Hoon;Lee, Jong-Doo;Suh, Jin-Suck;Park, Chang-Yun;Jeon, Pyoung;Na, Jae-Beom;Lee, Soo-Kon
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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    • pp.84-91
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    • 1995
  • Many clinical and laboratory tests have been employed to evaluate disease activity in rheumatioid arthritis. $^{99m}Tc$-labelled polyclonal IgG(HIG) has been demonstrated to accumulate in focal sites of infection or inflammation in both animals and human subjects. The purpose of this study was to distinguish arthritis with active inflammation from those without active inflammation and to correlate relative intensities of $^{99m}Tc$-labelled HIG uptake of the rheumatoid arthritis with clinical and MR indices of the joint inflammation. This study included twelve patients with active rheumatoid arthritis, two with ankylosing spondylitis and one with degenerative osteoarthritis without active inflammation. A Whole-body and spot images were obtained 4 hours after intravenous injection of 20mCi of $^{99m}Tc$-labelled HIG. Scintigrams were assessed visually by 3 experienced radiologists, and graded as normal or mildly and markedly increased uptake within the joints, and the degree of uptake was compared with clinical and radiologic severity of synovial inflammation. MRI studies were done on the involved joints consisted of wrist(n = 11), knee(n = 2) and hip joint(n= 2). Active synovitis was defined when marked elevation of ESR and gadolinium enhancement of synovium on MRI were demonstrated. Markedly increased radiotracer uptake was seen in 10 of 11 rheumatoid arthritic patients with active synovitis whereas normal or mildly increased uptakes were noted in others, including rheumatoid arthritic patient(n=1) and non-rheumatoid patients(n = 3) without active synovitis. This study showed that the localization of involved joints in rheumatoid arthritis could be detected with $^{99m}Tc$-labelled HIG and that the degree of uptake correlated well with the degree and activity of inflammation. In conclusion, $^{99m}Tc$-labelled HIG scan is a useful method in the evaluation of active inflammation in rheumatoid arthritis.

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Expression of Yippee-Like 5 (YPEL5) Gene During Activation of Human Peripheral T Lymphocytes by Immobilized Anti-CD3 (인체 말초혈액의 활성화 과정 중 yippee-like 5 (YPEL5) 유전자의 발현 양상)

  • Jun, Do-Youn;Park, Hye-Won;Kim, Young-Ho
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1641-1648
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    • 2007
  • Yippee-like proteins, which have been identified as the homolog of Drosophila yippee protein containing a zinc-finger domain, are known to be highly conserved among eukaryotes. However, their functional roles are still poorly understood. Recently we initiated ordered differential display (ODD)-polymerase chain reaction (PCR) to isolate genes of which expressions are altered following activation of human T cells. On the ODD-PCR image, one PCR-product detected in unstimulated T cells was not detectable at the time when the activated T cells traversed near $G_1/S$ boundary following activation by immobilized anti-CD3. Cloning and nucleotide sequence analysis revealed that the PCR-product was yippee-like 5 (YPEL5) gene, which was known as a human homolog of the Drosophila yippee gene. Northern blot analysis confirmed the amount of ${\sim}2.2$ kb YPEL5 mRNA expression detectable in unstimulated T cells was sustained until 1.5 hr after activation and then rapidly declined to undetectable level by 5 hr. Ectopic expression of YPEL5 gene in human cervix epitheloid carcinoma HeLa cells caused a significant reduction in cell proliferation to the level of 47% of the control. Expression of GFP-YPEL5 fusion protein in HeLa cells showed its nuclear localization. These results demonstrated that the expression level of human YPEL5 mRNA was negatively regulated in the early stage of T cell activation, and suggested that YPEL5 might exert an inhibitory effect on the cell proliferation as a nuclear protein.