• Asian-Australasian Journal of Animal Sciences
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• 제19권8호
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• pp.1065-1070
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• 2006

The avian uncoupling protein (avUCP) is a member of the mitochondrial transporter superfamily that uncouples proton entry in the mitochondrial matrix from ATP synthesis. The sequencing analysis method was used to identify nucleotide polymorphisms within the avUCP gene in Korean native chicken (KNC). This study identified ten single nucleotide polymorphisms (SNPs) in the avUCP gene. We analyzed the SNPs of the avUCP gene to investigate whether polymorphism in the gene might be responsible for quantitative variations in economic traits in KNC. Three significant polymorphic sites for economic traits were avUCP C+282T (mean body weight, p<0.05), avUCP C+433T (daily percent lay, p<0.05), and avUCP T+1316C (daily percent lay, p<0.05). The frequency of each SNP was 0.125 (C+282T in avUCP gene exon 1 region), 0.150 (C+433T in avUCP gene intron 1 region), and 0.15 (T+1316C in avUCP gene exon 3 region), respectively. Among the identified SNPs, one pair of SNPs (genotype CC, C+282T and TT, avUCP C+433T) showed the highest daily percent lay (p<0.05) and mean body weight (p<0.05) and the frequency was 0.067. This study of the avUCP gene could be useful for genetic studies of this gene and selection on economic traits for KNC.

• 한국해양생명과학회지
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• 제1권2호
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• pp.79-87
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• 2016

카드뮴과 같은 중금속은 독성이 높아 수서 생물과 인간에게 해로운 영향을 미친다고 알려져 있다. 본 연구에서는 해양 섬모충 Euplotes crassus에서 카드뮴이 해독 기전에 관여하는 ABC transporters (ABCs)와 glutathione S-transferase (GST)의 유전자 발현에 미치는 영향을 조사하였다. 총 7개의 ABCs 유전자와 1개의 GST 유전자 일부를 클로닝하여 유전자 분석을 실시하였고, 카드뮴(0.1~1 mg/l) 노출에 따른 이들 유전자의 발현 양상을 quantitative real time RT-PCR (qRT-PCR)을 이용하여 분석하였다. 염기서열 분석과 계통 분석 결과 이들 ABCs 유전자가 ABC transporter의 특징을 가지며, ABC-B/C family에 속하는 것을 확인하였고, GST 유전자는 theta isoform과 유사한 것으로 나타났다. 카드뮴에 8시간 노출시킨 결과 ABC transporter 유전자의 경우 ABCB21 유전자를 제외하고는 대부분 농도 의존적으로 유전자 발현이 유의하게 증가하였다. GST 유전자는 0.5 mg/l에서 가장 높은 유전자 발현 양상을 보였으며, 1 mg/l에서는 발현량이 대조군 수준으로 감소되었다. 본 연구 결과는 E. crassus의 ABC transporter와 GST 유전자가 카드뮴에 의해 유도되는 독성에 대한 방어 기전에 참여하는 것을 의미한다.

• Journal of Gastric Cancer
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• 제15권4호
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• pp.223-230
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• 2015

Purpose: The purpose of this pilot study was to evaluate the association between adenosine triphosphate-based chemotherapy response assays (ATP-CRAs) and subsets of tumor infiltrating lymphocytes (TILs) in gastric cancer. Materials and Methods: In total, 15 gastric cancer tissue samples were obtained from gastrectomies performed between February 2007 and January 2011. Chemotherapy response assays were performed on tumor cells from these samples using 11 chemotherapeutic agents, including etoposide, doxorubicin, epirubicin, mitomycin, 5-fluorouracil (5-FU), oxaliplatin, irinotecan, docetaxel, paclitaxel, methotrexate, and cisplatin. TILs in the tissue samples were evaluated using antibodies specific for CD3, CD4, CD8, Foxp3, and Granzyme B. Results: The highest cancer cell death rates were induced by etoposide (44.8%), 5-FU (43.1%), and mitomycin (39.9%). Samples from 10 patients who were treated with 5-FU were divided into 5-FU-sensitive and -insensitive groups according to median cell death rate. No difference was observed in survival between the two groups (P=0.216). Only two patients were treated with a chemotherapeutic agent determined by an ATP-CRA and there was no significant difference in overall survival compared with that of patients treated with their physician's choice of chemotherapeutic agent (P=0.105). However, a high number of CD3 TILs was a favorable prognostic factor (P=0.008). Pearson's correlation analyses showed no association between cancer cell death rates in response to chemotherapeutic agents and subsets of TILs. Conclusions: Cancer cell death rates in response to specific chemotherapeutic agents were not significantly associated with the distribution of TIL subsets.

• Journal of Microbiology and Biotechnology
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• 제28권4호
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• pp.542-550
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• 2018

Heat shock protein 90 (Hsp90) is treated as a molecular therapeutic target for the prevention and treatment of cancer. Geldanamycin (GA) was the first identified natural Hsp90 inhibitor, but hepatotoxicity has limited its clinical application. Nevertheless, a new GA analog (WK-88-1) with the non-benzoquinone skeleton, obtained from genetically engineered Streptomyces hygroscopicus, was found to have anticancer activity against two human breast cancer cell lines. WK-88-1 produced concentration-dependent inhibition of cell proliferation, cell cycle arrest, and apoptosis in estrogen receptor (ER)-positive MCF-7 and ER-negative MDA-MB-231 cell lines. Detailed analysis showed that WK-88-1 downregulated some key cell cycle molecules (CDK1 and cyclin B1) and lead to $G_2/M$ cell cycle arrest. Further studies also showed that WK-88-1 could induce human breast cancer cell apoptosis by downregulating Hsp90 client proteins (Akt, p-Akt, IKK, c-Raf, and Bcl-2), decreasing the ATP level, increasing reactive oxygen species production, and lowering the mitochondrial membrane potential. Meanwhile, we discovered that WK-88-1 significantly decreased the levels of Her-2 and $ER-{\alpha}$ in MCF-7 cells but not in MDA-MB-231 cells. In addition, WK-88-1 significantly increased caspase-3, -8, and -9 activities and the cleavage of PARP in a concentration-dependent manner (with the exception of caspase-3 and PARP in MCF-7 cells). Taken together, our preliminary results suggest that WK-88-1 has the potential to play a role in breast cancer therapy.

• 한국수정란이식학회지
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• 제25권4호
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• pp.281-286
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• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• International Journal of Oral Biology
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• 제31권2호
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• pp.53-65
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• 2006

Calcium concentration in the bone resorption lacunae is high and is in the mM concentration range. Both osteoblast and osteoclast have calcium sensing receptor in the cell surface, suggesting the regulatory role of high extracellular calcium in bone metabolism. In vitro, high extracellular calcium stimulated osteoclastogenesis in coculture of mouse osteoblasts and bone marrow cells. Therefore we examined the genes that were commonly regulated by both high extracellular calcium and $1,25(OH)_2vitaminD_3(VD3)$ by using mouse oligo 11 K gene chip. In the presence of 10 mM $[Ca^{2+}]e$ or 10 nM VD3, mouse calvarial osteoblasts and bone marrow cells were co-cultured for 4 days when tartrate resistant acid phosphatase-positive multinucleated cells start to appear. Of 11,000 genes examined, the genes commonly regulated both by high extracellular calcium and by VD3 were as follows; 1) the expression of genes which were osteoclast differentiation markers or were associated with osteoclastogenesis were up-regulated both by high extracellular calcium and by VD3; trap, mmp9, car2, ctsk, ckb, atp6b2, tm7sf4, rab7, 2) several chemokine and chemokine receptor genes such as sdf1, scya2, scyb5, scya6, scya8, scya9, and ccr1 were up-regulated both by high extracellular calcium and by VD3, 3) the genes such as mmp1b, mmp3 and c3 which possibly stimulate bone resorption by osteoclast, were commonly up-regulated, 4) the gene such as c1q and msr2 which were related with macrophage function, were commonly down-regulated, 5) the genes which possibly stimulate osteoblast differentiation and/or mineralization of extracellular matrix, were commonly down-regulated; slc8a1, admr, plod2, lox, fosb, 6) the genes which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were commonly up-regulated; s100a4, npr3, mme, 7) the genes such as calponin 1 and tgfbi which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were up-regulated by high extracellular calcium but were down-regulated by VD3. These results suggest that in coculture condition, both high extracellular calcium and VD3 commonly induce osteoclastogenesis but suppress osteoblast differentiation/mineralization by regulating the expression of related genes.

• 한국작물학회지
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• 제65권2호
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• pp.113-123
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• 2020

본 논문은 과산화수소 엽면 처리를 통해 수수에서 한발 스트레스 완화 효과가 있는지 알아보기 위하여 과산화수소 처리에 따른 생육 및 생리적 특성 변화와 단백질 변화를 분석하였다. 1. 포장 실험에서 한발 스트레스는 수수 생장을 감소시켰으나, 과산화수소 처리구에서는 과산화수소 무처리구보다 생육이 우수하였다. 또한 한발 조건에서 과산화수소 처리는 무처리에 비해 수량 관련 형질들의저하를 막는 효과를 보였다. 2. 엽 녹색도(SPAD) 및 엽록소 형광(Fv/Fm), 광합성 형질 조사에서 과산화수소 처리가 과산화수소 무처리보다 높았고, 포장 실험에서 엽내 과산화수소 함량은 적습+H₂O₂ 처리가 적습+H₂O 처리보다 과산화수소 함량이 227.8 µmol·g^-1 높았고, 한발 조건에서 엽내 과산화수소 함량은 한발+H₂O₂ 처리가 한발+H₂O 처리보다 16.7 µmol·g^-1으로 낮았다. 3. 한발 조건에서 과산화수소 처리는 여러 단백질의 발현을 변화시켰으며, 특히 광합성 관련 단백질인 ATP synthase deltal chain, cytochrome b6-f complex iron-sulfur subunit, ATP synthase subunit gamma, putative uncharacterized protein Sb02g002690, Sb07g027500와 Superoxide 라디칼을 제거해주는 superoxide dismutase가 증가한 것을 확인하였다. 그리고 단백질 보호와 복원에 관련된 heat shock protein의 발현 증가도 확인되었다. 종합적으로 과산화수소 엽면 처리가 한발 하에서 광합성 관련 단백질의 발현을 증가와 기공 개도를 높여 광합성 능력을 향상시켰다. 특히 엽내에 축적될 수 있는 활성산소종을 제거하는 항산화 능력이 높아져 한발 스트레스 대한 내성을 높여 수수의 생육과 수량성 저하가 억제되는 것을 확인할 수 있었다.

• 농업과학연구
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• 제45권3호
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• pp.455-461
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• 2018

Deoxynivalenol (DON), a Fusarium mycotoxin, causes health hazards for both humans and livestock. Therefore, the aim of this study was to investigate the metabolic profiles of the liver, serum, and urine of piglets fed DON using proton nuclear magnetic resonance ($^1H-NMR$) spectroscopy. The $^1H-NMR$ spectra of the liver, serum, and urine samples of the piglets provided with feed containing 8 mg DON/kg for 4 weeks were aligned and identified using the icoshift algorithm of MATLAB $R^2013b$. The data were analyzed by multivariate analysis and by MetaboAnalyst 4.0. The DON-treated groups exhibited discriminating metabolites in the three different sample types. Metabolic profiling by $^1H-NMR$ spectroscopy revealed potential metabolites including lactate, glucose, taurine, alanine, glycine, glutamate, creatine, and glutamine upon mycotoxin exposure (variable importance in the projection, VIP > 1). Forty-six metabolites selected from the principal component analysis (PCA) helped to predict sixty-five pathways in the DON-treated piglets using metabolite sets containing at least two compounds. The DON treatment catalyzed the citrate synthase reactions which led to an increase in the acetate and a decrease in the glucose concentrations. Therefore, our findings suggest that glyceraldehyde-3-phosphate dehydrogenase, citrate synthase, ATP synthase, and pyruvate carboxylase should be considered important in piglets fed DON contaminated feed. Metabolomics analysis could be a powerful method for the discovery of novel indicators underlying mycotoxin treatments.

• 한국임상약학회지
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• 제8권1호
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• pp.29-34
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• 1998

Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

• Asian-Australasian Journal of Animal Sciences
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• 제19권4호
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• pp.554-562
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• 2006

Two lines of mice, M16 selected for rapid growth and a randomly selected control ICR as well as their reciprocal crosses were used to study the effects of genotype on whole-body energetics and intestinal responses to monensin. Six mice, eight weeks of age, from each line or reciprocal cross were assigned to one of two treatments, 1) drinking water containing 20 mmol/L monensin dissolved in 0.5% V/V ethanol, and 2) drinking water containing 0.5% V/V ethanol (control) for two weeks. After 11 days (age of 9 weeks and 4 days), whole-body $O_2$ consumption was measured. At the end of two weeks, jejunal $O_2$ consumption, intestinal tissue composition and histomorphometrics as well as the rate and efficiency of glucose absorption were estimated. In comparison with the control, monensin administration in drinking water resulted in less daily water intake (13.4 vs. 15.5 ml/mouse, p<0.01), less protein to DNA ratio of jejunal mucosa (5.41 vs. 6.01 mg/mg, p<0.05), lower villus width (88 vs. $100{\mu}m$, p<0.05), and less jejunal tissue $O_2$ consumption enhancement by alcohol (7.2 vs. 10.5%, p<0.01) in mice. Other than those changes, monensin had little (p>0.05) effect on variables measured in either line of mice or their reciprocal cross. In contrast, the M16 line, selected for rapid growth, as compared to the ICR controls or the reciprocal crosses, had less initial (pre-monensin treatment) whole-body $O_2$ consumption per gram of body weight (1.68 vs. $2.11-2.34{\mu}mol/min{\cdot}g$ BW, p<0.01) as compared to the ICR and reciprocal crosses. In addition, the M16 mice exhibited greater growth (412 vs. 137-210 mg/d, p<0.05), better feed efficiency (41.7 vs. 19.9-29.3 mg gain/g feed, p<0.05), shorter small intestines adjusted for fasted body weight (1.00 vs. 1.22-1.44 cm/g FBW, p<0.05), wider villi (109 vs. $87-93{\mu}m$, p<0.05), more mature height of enterocytes (28.8 vs. $24.4-25.1{\mu}m$, p<0.05) and a lower rate (91 vs. $133-145{\eta}mol\;glucose/min{\cdot}g$ jejunum, p<0.05) and less energetic efficiency (95 vs. $59-72{\eta}mol$ ATP expended/${\eta}mol$ glucose uptake, p<0.05) of glucose absorption compared to the ICR line and the reciprocal cross. Monensin had little (p>0.05) effect on whole-body $O_2$ consumption and jejunal function, whilst selection for rapid growth resulted in an apparent down-regulation of intestinal function. These data suggest that genetic selection for increased growth does not result in concomitant changes in intestinal function. This asynchrony in the selection for production traits and intestinal function may hinder full phenotypic expression of genotypic growth potential.