• 생명과학회지
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• 제20권7호
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• pp.1005-1011
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• 2010

$\gamma$-aminobutyric acid (GABA)는 중추신경계에서 억제성으로 작용하는 주요한 신경전달물질이다. GABA 수송체(GAT)는 연접간격에 존재하는 GABA를 세포 내로 재 흡수하여 GABA의 농도를 조절한다. 그런데 GABA 수송체가 어떻게 조절되는지는 아직 밝혀지지 않았다. 본 연구에서는 효모 two-hybrid system을 사용하여 뇌의 주요 GABA 수송체인 GAT1의 C-말단과 특이적으로 결합하는 ubiquitin-specific protease 14 (Usp14)를 분리하였다. Usp14는 GABA 수송체 GAT1및 GAT2와는 결합하지만, 다른 GAT isoform과는 결합하지 않았다. GAT1과의 결합에는 Usp14의 C-말단부위가 필수적으로 관여함을 확인하였다. 또한 이 단백질간의 결합을 GST pull-down assay로 확인하였으며, 생쥐 뇌 균질액의 co-immunoprecipitation을 통하여 in vivo에서도 GAT1과 Usp14가 결합함을 확인하였다. 이러한 결과들은 Usp14가 GAT1과 결합하여 세포막에 존재하는 GAT1의 수를 조절하는 역할을 할 가능성을 시사한다.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.14-40
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• 2002

15-Deoxy-${\Delta}^{12, 14}$-prostaglandin $J_2$ (15-deoxy-$PGJ_2$), a naturally occurring ligand activates the peroxisome proliferator-activated $receptor-{\gamma}(PPAR-{\gamma}$). Activation of $PPAR-{\gamma}$ has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-$PGJ_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-$PGJ_2$ (0.2 to 1.6 ${\mu}M$) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/ml). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-$PGJ_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-$PGJ_2$(0.8 ${\mu}M$) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-$PGJ_2$ on the expression of p38 MAP kinase and activation of AP-1, The promoting ability of 15-deoxy-$PGJ_2$ did not occur through $PPAR-{\gamma}$, as synthetic PPAR-${\gamma}$ agonist andantagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), $PPAR-{\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and $PGE_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of f 5-deoxy-$PGJ_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-$PGJ_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 single pathway may be important in the promoting activity of 15-deoxy-$PGJ_2$ cells.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.14-40
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• 2002

15-Deoxy- Δ$\^$12,14/-prostaglandin J$_2$ (15-deoxy-PGJ$_2$), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$). Activation of PPAR-y has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-PGJ$_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-PGJ$_2$ (0.2 to 1.6 ${\mu}$M) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/$m\ell$). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-PGJ$_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-PGJ$_2$ (0.8 ${\mu}$M) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-PGJ$_2$ on the expression of p38 MAP kinase and activation of AP-1. The promoting ability of 15-deoxy-PGJ$_2$ did not occur through PPAR-${\gamma}$, as synthetic PPAR-${\gamma}$ agonist and antagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), PPAR-${\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and PGE$_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of 15-deoxy-PGJ$_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-PGJ$_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 signal pathway may be important in the promoting activity of 15-deoxy-PGJ$_2$ on the differentiation of PC12 cells.

• Tuberculosis and Respiratory Diseases
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• 제75권3호
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• pp.104-110
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• 2013

Background: Osteopontin (OPN) and carbonic anhydrase IX (CAIX), which are expressed on the surface of tumor cells, are associated with hypoxia during tumor development and progression. However, the roles of these proteins in the plasma of patients with non-small cell lung cancer (NSCLC) are poorly understood. Herein, we hypothesized that plasma OPN and CAIX levels could be used as diagnostic and prognostic tumor markers in patients with NSCLC. Methods: Fifty-three patients with NSCLC and 50 healthy control subjects were enrolled. We selected controls without malignancy and matched them with NSCLC patient cases according to age and gender. Blood samples were collected at the time of diagnosis; the plasma levels of OPN and CAIX were measured by enzyme-linked immunosorbent assays. Results: The plasma levels of OPN in the patients with NSCLC were significantly elevated as compared to those in the controls (p=0.016). However, there was no difference in the plasma level of CAIX between the NSCLC patients and controls. NSCLC patients with a distant metastasis had a remarkable increase in plasma OPN compared with patients without metastasis (p=0.026), but no such correlation was found for CAIX. There was no difference in overall survival rates according to the plasma level of OPN between the two groups (by Kaplan-Meier survival analysis). Conclusion: Plasma OPN levels were elevated in patients with NSCLC as compared with the controls, with greater elevation of OPN levels in the advanced stages of disease. Therefore, plasma OPN may have utility as a diagnostic, but not prognostic, biomarker of advanced NSCLC.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.224-228
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• 2010

The encapsulation of bacteria may be used to harness them for longer periods of time in order to make them viable, whereas antibiotic treatment would result in controlled release of therapeutic molecules. Encapsulated Escherichia coli GFP (green fluorescent protein) (E. coli GFP) was used here as a model for therapeutic substance - GFP fragments release (model of bioactive substances). Our aim was to evaluate the performance of bacteria encapsulated in hollow fibers (HFs) treated with antibiotic for induction of cell death. The polypropylene-surface-modified HFs were applied for E. coli encapsulation. The encapsulated bacteria were treated with tetracycline in vitro or in vivo during subcutaneous implantation into mice. The HF content was evaluated in a flow cytometer, to assess the bacteria cell membrane permeability changes induced by tetracycline treatment. It was observed that the applied membranes prevented release of bacteria through the HF wall. The E. coli GFP culture encapsulated in HF in vitro proved the tetracycline impact on bacteria viability and allows the recognition of the sequence of events within the process of bacteria death. Treatment of the SCID mice with tetracycline for 8 h proved the tetracycline impact on bacteria viability in vivo, raising the necrotic bacteria-releasing GFP fragments. It was concluded that the bacteria may be safely enclosed within the HF at the site of implantation, and when the animal is treated with antibiotic, bacteria may act as a local source of fragments of proteins expressed in the bacteria, a hypothetical bioactive factor for the host eukaryotic organism.

• Fisheries and Aquatic Sciences
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• 제18권1호
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• pp.51-56
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• 2015

A 8-week trial was conducted to evaluate the effects of photoperiod manipulation on the growth performance and hematological parameters of juvenile Caspian roach, Rutilus rutilus caspicus (average weight $1.46{\pm}0.12g$ mean${\pm}$SD) reared under five photoperiods (24 h Light, 24L; 18 h Light & 6 h Dark, 18L:6D; 12 h Light & 12 h Dark, 12L:12D; 6 h Light & 18 h Dark, 6L:18D; 24 h Dark, 24D) with constant light intensity 1,500 lx on the water surface. Triplicate of 20 fish were allocated into each of 15 fiberglass tanks of 50 L capacity and they were fed three times per day with the commercial feed (SFK, Co., Sari - Iran) contains 50.0% protein and 10.5% lipid. At the end of experimental period, final body weight, weight gain and specific growth rates of fish exposed to 24L were significantly higher than those of fish exposed to 12L:12D, 6L:18D and 24D (P < 0.05). Red blood cell and hemoglobin of fish exposed to 24L were significantly higher than those of fish exposed to 24D. No significant difference observed in hematocrit, white blood cell and plasma glucose among the different treatments groups. Therefore, these results demonstrated that the growth performance of juvenile Caspian roach can be significantly stimulated by using 24L and 18L:6D photoperiods without any measurable significant stress response such as plasma glucose concentration.

• Biomolecules & Therapeutics
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• 제22권5호
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• pp.414-419
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• 2014

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that regulate cell-matrix composition and are also involved in processing various bioactive molecules such as cell-surface receptors, chemokines, and cytokines. Our group recently reported that MMP-3, -8, and -9 are upregulated during microglial activation and play a role as proinflammatory mediators (Lee et al., 2010, 2014). In particular, we demonstrated that MMP-8 has tumor necrosis factor alpha (TNF-${\alpha}$)-converting enzyme (TACE) activity by cleaving the prodomain of TNF-${\alpha}$ and that inhibition of MMP-8 inhibits TACE activity. The present study was undertaken to compare the effect of MMP-8 inhibitor (M8I) with those of inhibitors of other MMPs, such as MMP-3 (NNGH) or MMP-9 (M9I), in their regulation of TNF-${\alpha}$ activity. We found that the MMP inhibitors suppressed TNF-${\alpha}$ secretion from lipopolysaccharide (LPS)-stimulated BV2 microglial cells in an order of efficacy: M8I>NNGH>M9I. In addition, MMP inhibitors suppressed the activity of recombinant TACE protein in the same efficacy order as that of TNF-${\alpha}$ inhibition (M8I>NNGH>M9I), proving a direct correlation between TACE activity and TNF-${\alpha}$ secretion. A subsequent pro-TNF-${\alpha}$ cleavage assay revealed that both MMP-3 and MMP-9 cleave a prodomain of TNF-${\alpha}$, suggesting that MMP-3 and MMP-9 also have TACE activity. However, the number and position of cleavage sites varied between MMP-3, -8, and -9. Collectively, the concurrent inhibition of MMP and TACE by NNGH, M8I, or M9I may contribute to their strong anti-inflammatory and neuroprotective effects.

• Journal of Chest Surgery
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• 제50권2호
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• pp.126-129
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• 2017

The identification of circulating tumor cells (CTCs) is clinically important for diagnosing cancer. We have previously developed a size-based filtration platform followed by epithelial cell adhesion molecule immunofluorescence staining for detecting CTCs. To characterize CTCs independently of cell surface protein expression, we incorporated a chromosomal fluorescence in situ hybridization (FISH) assay to detect abnormal copy numbers of chromosomes in cells collected from peripheral blood samples by the size-based filtration platform. Aneuploid cells were detected in the peripheral blood of patients with lung cancer. Unexpectedly, aneuploid cells were also detected in the control group, which consisted of peripheral blood samples from patients with benign lung diseases, such as empyema necessitatis and non-tuberculous mycobacterial lung disease. These findings suggest that chromosomal abnormalities are observed not only in tumor cells, but also in benign infectious diseases. Thus, our findings present new considerations and bring into light the possibility of false positives when using FISH for cancer diagnosis.

• Applied Microscopy
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• 제12권2호
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• pp.35-44
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• 1982

The fat bodies of cabbage worm (Pieris rapae) and silk worm (Bomyx mori) during metamorphosis was comparatively studied by electron microscope. 1. Cell oranelles: Golgi apparatus were not observed in both species. It is observed that RER of cabbage worms initiate to degenerate in prepupa stage with complete degeneration at adult stage, while that of silk worms shows similar degenerative pattern. However, mitochondria of cabbage worms are transformed into autophagic vacuole from prepupa stage until adult stage whereas those of silk worm shows a decrease in number in prepupa stage but maintains a certain level until adult stage. 2. Storage substance in cell: Lipid droplets in cabbage worms were observed to increase in numbers during larval stage but afterward decrease in number with an enlargement in size. However immediately after their pupal stage, they almost disappear. On the contrary lipid droplets in silk worms show rather increase in number until adult stage. Protein storage granules in bothspecies were arised from autophagic vacuoles(lysosome) . Fat cells of cabbage worm in adult stage turn out to be residual bodies which last until final stage, but those of silk worm rapidly decrease. Glycogen particles in both species reach maximum at last larval instar and thee gradually decrease thereafter. 3. Fat body sheath: The average width of fat body sheath was measured to be $0.2{\mu}m$ and $0.6{\mu}m$ and surface of fat cells adjacent to fat body sheath in silk worm is heavily infolded.

• 한국동물학회지
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• 제35권2호
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• pp.149-160
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• 1992

Using the protein A-gold complex, the mvoabrillogenesis and actin localization of cultured myoblast were invastisated. In the superstructural changes of mvogenic cell during differentiation, pectoral myoblasts contained large nucleus and numerous ribosomes but no myofibrils during the first 24 hr of cultures. Mvoblast initiated to differentiate at 3-day of culture contained the primitive myofibrillar structure. At 96 hr of culture, the mvofibrillar structure showed reletively discernable Z band but pools defined A, H and M bands. The feature of sarcomeric structure showed more defined form at cultur 5 day. In the aspect of actin localization, actin wvas diffusely detected throughout the cytoplasm of myogenic cell and nucleus during the proliferating stage. At 72 hr of culture, with the appearantc oi primitive mvofibrils, gold particles were observed in surrounding of myofibrils but still presented in overall of cytoplasm, especially in the surface and lumen of endoplasmic reticulum. With the gradual increase of culture time, local distribution of actin was readily detected within cytoplasm. In the 5-day specimen of cultures, gold particles precisely indicate the sites of actin localifation within the sarcomere. These results indicate the time of onset of myofibrill appearance and the biosynthetic and incorporation pathway of actin molecules into sarcomeric structure during myofibrillogenesis. Thus, in the present study, the first mvoabrillar structure was detected at culture 3 day, and the initiation of assembly into a typical sarcmeric structure was observed at culture 5 day. It seems, however, that the course of events on myofibrillogenesis of cultured myoblasts can be changed with great dependence of culture conditions including the number and groluth rate of mononucleated mvoblasts after seeding although the fundamental process shows identical appearances.