• BMB Reports
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• 제32권3호
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• pp.273-278
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• 1999

Cytolysin produced by Vibrio vulnificus has been known to be lethal to mice by increasing vascular permeability and neutrophil sequestration in the lung. In the present study, a cytotoxic mechanism of V. vulnificus cytolysin on the neutrophil was investigated. Cytolysin rapidly bound to neutrophils and induced cell death, as determined by the trypan blue exclusion test. V. vulnificus cytolysin caused the depletion of cellular ATP without the release of ATP or lactate dehydrogenase. Formation of transmembrane pores was evidenced by the rapid efflux of potassium and 2-deoxy-D-[$^3H$]glucose from cytolysin-treated neutrophils. It was further confirmed by the rapid flow of monovalent ions in the patch clamp of cytolysin-treated neutrophil membrane. The pore formation was accompanied by the oligomerization of cytolysin monomers on the neutrophil membrane as demonstrated by immunoblot, which exhibited a 210 kDa band corresponding to a tetramer of the native cytolysin of $M_r$ 51,000. These findings indicate that V. vulnificus cytolysin rapidly binds to the neutrophil membrane and oligomerizes to form small transmembrane pores, which induce the efflux of potassium and the depletion of cellular ATP leading to cell death without cytolysis.

• Animal Bioscience
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• 제34권6호
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• pp.957-966
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• 2021

Objective: Ovarian follicular development, which dependent on the proliferation and differentiation of granulosa cells (GCs), is a complex biological process in which miRNA plays an important role. Our previous study showed that miR-458b-5p is associated with ovarian follicular development in chicken. The detailed function and molecular mechanism of miR-458b-5p in GCs is unclear. Methods: The luciferase reporter assay was used to verify the targeting relationship between miR-458b-5p and catenin beta-1 (CTNNB1), which is an important transcriptional regulatory factor of the Wnt/β-catenin pathway. The cell counting kit-8 (CCK-8) assay, flow cytometry with propidium iodide (PI) and annexin V-fluorescein isothiocyanate (FITC) labeling were applied to explore the effect of miR-458b-5p on proliferation, cell cycle and apoptosis of chicken GCs. Quantitative real-time polymerase chain reaction and Western blot were used to detect the mRNA and protein expression levels. Results: We demonstrated that the expression of miR-458b-5p and CTNNB1 showed the opposite relationship in GCs and theca cells of hierarchical follicles. The luciferase reporter assay confirmed that CTNNB1 is the direct target of miR-458b-5p. Using CCK-8 assay and flow cytometry with PI and Annexin V-FITC labeling, we observed that transfection with the miR-458b-5p mimics significantly reduced proliferation and has no effects on apoptosis of chicken GCs. In addition, miR-458b-5p decreased the mRNA and protein expression of CD44 molecule and matrix metallopeptidase 7, which are the downstream effectors of CTNNB1 in Wnt/β-Catenin pathway and play functional roles in cell proliferation. Conclusion: Taken together, the data indicate that miR-458b-5p regulates ovarian GCs proliferation through Wnt/β-catenin signaling pathway by targeting CTNNB1, suggesting that miR-458b-5p and its target gene CTNNB1 may potentially play a role in chicken ovarian follicular development.

• 한국화재소방학회논문지
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• 제27권6호
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• pp.50-56
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• 2013

대향류 비예혼합 연료-공기 유동장에서 고온연료의 점화특성과 형성된 화염의 소화특성에 미치는 복사효과에 대해 수치계산을 통해 검토하였다. 화학반응의 계산을 위해 GRI-v3.0의 상세화학반응기구를 사용하였으며, 단열계산과 광학적으로 얇은 복사모델을 적용하여 계산을 수행하였다. 대향류 유동장의 점화와 소화점을 정확히 찾기 위하여 화염제어 연속계산법을 적용하였다. 결과를 통해 스트레인율 변화에 대해 최고 온도보다는 최고 H 라디칼 농도가 점화와 소화거동을 이해하는데 더 적합하다는 것을 확인하였다. 최고 H 라디칼 농도변화 거동을 통해 기존에 알려진 S-곡선, C-곡선 및 O-곡선 등을 확인하였다. 복사열손실 분율($f_r$)과 공간에 대해 적분된 열발생률(IHRR)을 통해 $f_r$이 가장 큰 점에서 복사효과에 의한 소화가 발생하였으며, 화염신장 소화점에서는 IHRR이 가장 높지만 화염에서의 전도에 의한 열손실로 인해 소화가 되는 것을 확인하였다. 복사는 화염신장 소화점에는 거의 영향이 없지만 복사 소화점과 점화점에는 큰 영향을 주는 것을 알 수 있었다. 또한 연료의 온도가 높아질수록 복사에 의한 소화점의 스트레인율과 화염신장에 의한 스트레인율 사이의 영역이 넓어지게 되어 화염 안정성이 향상되고 있음을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제31권10호
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• pp.1331-1342
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• 2021

In this study, we evaluated the mechanism of long non-coding RNA MIR22 host gene (LncRNA MIR22HG) in osteosarcoma cells. Forty-eight paired osteosarcoma and adjacent tissues samples were collected and the bioinformatic analyses were performed. Target genes and potential binding sites of MIR22HG, microRNA (miR)-629-5p and tet methylcytosine dioxygenase 3 (TET3) were predicted by Starbase and TargetScan V7.2 and confirmed by dual-luciferase reporter assay. Cell Counting Kit-8, colony formation and flow cytometry assays were utilized to determine the viability, proliferation and apoptosis of transfected osteosarcoma cells. Pearson's analysis was introduced for the correlation analysis between MIR22HG and miR-629-5p in osteosarcoma tissue. Relative expressions of MIR22HG, miR-629-5p and TET3 were measured by quantitative real-time polymerase chain reaction or Western blot. MiR-629-5p could competitively bind with and was negatively correlated with MIR22HG, the latter of which was evidenced by the high expression of miR-629-5p and low expression of MIR22HG in osteosarcoma tissues. Overexpressed MIR22HG repressed the viability and proliferation but enhanced apoptosis of osteosarcoma cells, which was reversed by miR-629-5p upregulation. TET3 was the target gene of miR-629-5p, and the promotive effects of upregulated miR-629-5p on the viability and proliferation as well as its repressive effect on apoptosis were abrogated via overexpressed TET3. To sum up, overexpressed MIR22HG inhibits the viability and proliferation of osteosarcoma cells, which was achieved via regulation of the miR-629-5p/TET3 axis.

• Applied Biological Chemistry
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• 제39권1호
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• pp.70-76
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• 1996

일련의 pyridylsulfonyl urea들을 합성하고 25%(v/v) 디옥산 수용액의 넓은 pH범위에서 가수분해 반응속도 상수를 측정하였다. pH-효과, 용매 효과($m{\ll}1,\;n{\ll}3$ 및 ${\mid}m{\mid}{\ll}{\mid}{\ell}{\mid}$), 일반 염기-효과, 산-해리상수(pKa, 3: 4.9 및 5: lit.4.6), 열역학적 활성화 파라미터(${\Delta}H^{\neq}=0.025\;Kcal.mol.^{-1}$ 및 ${\Delta}S^{\neq}=0.54{\sim}\;-2.19\;e.u.$) 및 생성물 분석 결과로부터 반응속도식을 유도하고 가수분해 반응 메카니즘을 제안하였다. 즉, 비(H)치환체, 1-(4,6-dimethoxypyrimidine-2-yl)-3-(2-pyridylsulfonyl) urea, 3은 산성용액에서 A-2형(또는 $A_{AC}2$)반응 그리고 염기성 용액 에서는 $(E_1)_{anion}$ 메카니즘으로 가수분해 반응이 일어난다. 반면에 trifluoromethyl-치환체, 1-(4,6-dimethoxypyrimidine-2-yl)-3-(3-trifluorornethyl-2-pyridylsulfonyl) urea, 5(Flazasulfuron)는 산성 용액중에서 $A-S_N2Ar$형의 반응으로 생성된 conjugate acid($5H^+$), 그리고 pH 9.0 이상에서는 $(E_1)_{anion}$ 및 $(E_1CB)_R$ 반응으로 생성된 conjugate base(CB)를 거쳐 산성 및 염기성 용액중에서 모두 5원자 고리 중간체를 경유하는 Smile 자리옮김 반응으로 산성에서는 3-trifluoromethyl-2-pyridylpyrimidinyl urea(PPU) 그리고 염기성에서는 3-trifluoromethyl-2-pyridyl-4,6-dimethoxy-pyrimidinyl amine(PPA)을 생성하는 가수분해 반응이 일어남을 알았으며 5는 3보다 약 3.5배 빠른 반응속도를 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제19권12호
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• pp.1635-1643
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• 2009

The aim of this study was to provide new insight into the mechanism whereby the housekeeping enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) locates to cell walls of Lactobacillus plantarum 299v. After purification, cytosolic and cell wall GAPDH (cw-GAPDH) forms were characterized and shown to be identical homotetrameric active enzymes. GAPDH concentration on cell walls was growth-time dependent. Free GAPDH was not observed on the culture supernatant at any time during growth, and provoked cell lysis was not concomitant with any reassociation of GAPDH onto the cell surface. Hence, with the possibility of cw-GAPDH resulting from autolysis being unlikely, entrapment of intracellular GAPDH on the cell wall after a passive efflux through altered plasma membrane was investigated. Flow cytometry was used to assess L. plantarum 299v membrane permeabilization after labeling with propidium iodide (PI). By combining PI uptake and cw-GAPDH activity measurements, we demonstrate here that the increase in cw-GAPDH concentration from the early exponential phase to the late stationary phase is closely related to an increase in plasma membrane permeability during growth. Moreover, we observed that increases in both plasma membrane permeability and cw-GAPDH activity were delayed when glucose was added during L. plantarum 299v growth. Using a double labeling of L. plantarum 299v cells with anti-GAPDH antibodies and propidium iodide, we established unambiguously that cells with impaired membrane manifest five times more cw-GAPDH than unaltered cells. Our results show that plasma membrane permeability appears to be closely related to the efflux of GAPDH on the bacterial cell surface, offering new insight into the understanding of the cell wall location of this enzyme.

• 복식문화연구
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• 제21권6호
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• pp.967-973
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• 2013

During high speed sewing, the needle thread is exposed to dynamic loading, short strike loading, inertia forces, friction, rubbing, force of check spring, bending, pressure, friction, impact, shock and thermal influence. The dynamic thread loading/tension alters throughout the stitch formation cycle and along its passage through the machine. The greatest tensile force occurs at the moment of stitch stretching, when the take up lever pulls for required thread length through the tension regulator. These stresses act on the thread repeatedly and the thread passes 50-80 times through the fabric, the needle eye and the bobbin case mechanism, before getting incorporated into the seam, which result in upto 40% loss in tensile strength of the sewing thread. This damage in the sewing thread adversely affects its processing and functional performance. In this paper, the contribution of dynamic loading, passage through needle and fabric, and bobbin thread interaction in the loss in tensile properties has been studied. It is observed that the loss in tensile properties occurs mainly due to the bobbin thread interaction. Dynamic loading due to the action of take up lever also causes substantial loss in tenacity and breaking elongation of cotton threads.

• 한국자원식물학회지
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• 제1권1호
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• pp.8-22
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• 1988

Vincristine and vinblastine isolated from Vinco spp. , and podophyllotoxine derivatives isolated from Podophyllum spp. are usefulas anticancerous components obtaned from higher plants. More thanten antineoplastic compounds are now following them as anticancerousagents from higher plants. In my laboratory, Sarcoma 180A has beenused as the first screening test. By this method, I have found outsome kinds of antineoplastic constituents from active plants extracts .For instance, bisaborane type compounds were isolated from Curcumaxanthorrhiza, one of Indonesian plants; a morphinane type compoundfromCocculus trilobus; cyclic hexapeptides from Rubia akane and R.cordiorta. Seven components having antineoplastic actirity wereisolated from Rubia spp. except. R. tinctoria. Their structures wereelucidated except RA-Vl by chemical reaction and variovs instrumentalanalysis as shown in Fig. Among of them, RA-Vll showed strong activityagainst P388 Lymphocytic leukemia, L2O, B16 melanoma, Lewis lungcarcinoma, colon 38 and Ehrlich carcinoma. RA-V revealed excellentactivity against MM2 mammary carcinoma. The· value of acute LD5O ofRA-ViI were 10. Omg/kg( iP) and 16.5mg/kg( po ) respectiveIy . Therapruticratio was 400, compared with 10 of mitomycin C. QSAR was also appliedto these compounds by elongation of ether and ester side chains atR'. Mechanism of action of RA-Vll was also investigated and wasassumed to be inhibition of protein biosynthesis .

• Molecules and Cells
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• 제45권12호
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• pp.911-922
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• 2022

A structural protein of SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), nucleocapsid (N) protein is phosphorylated by glycogen synthase kinase (GSK)-3 on the serine/arginine (SR) rich motif located in disordered regions. Although phosphorylation by GSK-3β constitutes a critical event for viral replication, the molecular mechanism underlying N phosphorylation is not well understood. In this study, we found the putative alpha-helix L/FxxxL/AxxRL motif known as the GSK-3 interacting domain (GID), found in many endogenous GSK-3β binding proteins, such as Axins, FRATs, WWOX, and GSKIP. Indeed, N interacts with GSK-3β similarly to Axin, and Leu to Glu substitution of the GID abolished the interaction, with loss of N phosphorylation. The N phosphorylation is also required for its structural loading in a virus-like particle (VLP). Compared to other coronaviruses, N of Sarbecovirus lineage including bat RaTG13 harbors a CDK1-primed phosphorylation site and Gly-rich linker for enhanced phosphorylation by GSK-3β. Furthermore, we found that the S202R mutant found in Delta and R203K/G204R mutant found in the Omicron variant allow increased abundance and hyper-phosphorylation of N. Our observations suggest that GID and mutations for increased phosphorylation in N may have contributed to the evolution of variants.

• Journal of Ecology and Environment
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• 제47권3호
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• pp.85-102
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• 2023

Background: Ecophysiological characteristics of Rosa rugosa were analyzed under different environmental factors from May to October 2022. Photosynthesis, chlorophyll fluorescence, chlorophyll content, leaf water content (LWC), osmolality, carbohydrate content, and total ion content were measured to compare the physiological characteristics of R. rugosa at two study sites (i.e., in large pots and in the Goraebul coastal sand dune area). Results: When R. rugosa was exposed to high temperatures, photosynthetic parameters including net photosynthetic rate (P_N) and stomatal conductance (g_s) in both experiment areas declined. In addition, severe photoinhibition occurs when R. rugosa is continuously exposed to high photosynthetically active radiation (PAR), and because of this, relatively low Y(II) (i.e., the quantum yield of photochemical energy conversion in photosystem II [PSII]) and high Y(NO) (i.e., the quantum yield of non-regulated, non-photochemical energy loss in PSII) in the R. rugosa of the pot were observed. As the high Y(NPQ) (i.e., the quantum yield of regulated non-photochemical energy loss in PSII) of R. rugosa in the coastal sand dune, they dissipated the excessed photon energy through the non-photochemical quenching (NPQ) mechanism when they were exposed to relatively low PAR and low temperature. Rosa rugosa in the coastal sand dune has higher chlorophyll a and carotenoid content. The high chlorophyll a + b and low chlorophyll a/b ratios seemed to optimize light absorption in response to low PAR. High carotenoid content played an important role in NPQ. As a part of the osmotic regulation in response to low LWCs, R. rugosa exposed to high temperatures and continuously high PAR used soluble carbohydrates and ions to maintain high osmolality. Conclusions: We found that F_v/F_m was lower in the potted plants than in the coastal sand dune plants, indicating the vulnerability of R. rugosa to high temperatures and PAR levels. We expect that the suitable habitat range for R. rugosa will shrink and move to north under climate change conditions.