• Fisheries and Aquatic Sciences
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• 제19권4호
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• pp.21.1-21.11
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• 2016

Background: The evaluation of suitable reference genes as normalization controls is a prerequisite requirement for launching quantitative reverse transcription-PCR (RT-qPCR)-based expression study. In order to select the stable reference genes in abalone Haliotis discus hannai tissues (gill and hepatopancreas) under heavy metal exposure conditions (Cu, Zn, and Cd), 12 potential candidate housekeeping genes were subjected to expression stability based on the comprehensive ranking while integrating four different statistical algorithms (geNorm, NormFinder, BestKeeper, and ${\Delta}CT$ method). Results: Expression stability in the gill subset was determined as RPL7 > RPL8 > ACTB > RPL3 > PPIB > RPL7A > EF1A > RPL4 > GAPDH > RPL5 > UBE2 > B-TU. On the other hand, the ranking in the subset for hepatopancreas was RPL7 > RPL3 > RPL8 > ACTB > RPL4 > EF1A > RPL5 > RPL7A > B-TU > UBE2 > PPIB > GAPDH. The pairwise variation assessed by the geNorm program indicates that two reference genes could be sufficient for accurate normalization in both gill and hepatopancreas subsets. Overall, both gill and hepatopancreas subsets recommended ribosomal protein genes (particularly RPL7) as stable references, whereas traditional housekeepers such as ${\beta}-tubulin$ (B-TU) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were ranked as unstable genes. The validation of reference gene selection was confirmed with the quantitative assay of MT transcripts. Conclusions: The present analysis showed the importance of validating reference genes with multiple algorithmic approaches to select genes that are truly stable. Our results indicate that expression stability of a given reference gene could not always have consensus across tissue types. The data from this study could be a good guide for the future design of RT-qPCR studies with respect to metal regulation/detoxification and other related physiologies in this abalone species.

• The Korean Journal of Physiology and Pharmacology
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• 제20권6호
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• pp.621-628
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• 2016

Hypertension can be caused by various factors while the predominant causes include increase in body fluid volume and resistance in the circulatory system that elevate the blood pressure. Consumption of probiotics has been proven to attenuate hypertension; however, the effect is much strain-dependent. In this study, a newly isolated Lactobacillus casei (Lb. casei ) strain C1 was investigated for its antihypertensive properties in spontaneously hypertensive rats (SHR). Lactic acid bacteria (LAB) suspension of 11 log colony-forming unit (CFU) was given to SHR (SHR+LAB, n=8), and phosphate buffer saline (PBS) was given as a control in SHR (SHR, n=8) and in Wistar rats as sham (WIS, n=8). The treatment was given via oral gavage for 8 weeks. The results showed that the weekly systolic blood pressure (SBP), mean arterial pressure (MAP), diastolic blood pressure (DBP) and aortic reactivity function were remarkably improved after 8 weeks of bacterial administration in SHR+LAB. These effects were mostly attributed by restoration of wall tension and tensile stress following the bacterial treatment. Although not statistically significant, the level of malondialdehye (MDA) in SHR+LAB serum was found declining. Increased levels of glutathione (GSH) and nitric oxide (NO) in SHR+LAB serum suggested that the bacterium exerted vascular protection through antioxidative functions and relatively high NO level that induced vasodilation. Collectively, Lb. casei strain C1 is a promising alternative for hypertension improvement.

• Nutritional Sciences
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• 제7권3호
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• pp.138-143
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• 2004

Conjugated linoleic acid (CLA) has been shown to have a range of biological activities, including anti-carcinogenic, anti-atherosclerotic, anti-adipogenic and anti-diabetogenic effects. Recent reports also showed that CLA has free radical scavenging capacity, which may have health benefits for human beings. The current study was performed to investigate the effect of structured lipid (SL)-containing CLA on plasma lipids and hepatic antioxidant enzyme activity. Sprague-Dawley mts were fed 5% and 10% SL-containing normal diet for 6 wks and these groups were compared to rats fed 5% and 10% corn oil. In plasma lipids, total-cholesterol was not affected by fat source or dietary fat level while triglyceride level decreased significantly in groups fed 10% fat diet compared to the other groups. Plasma thiobarbituric acid reactive substances (TBARS) level decreased significantly in the S5 and S10 groups compared to the C5 and C10 groups, although hepatic TBARS level was not altered by fat source. On the other hand, in terms of hepatic antioxidant enzyme activity, superoxide dismutase activity increased in the S10 group, whereas catalase activity decreased in the S10 group. Glutathione peroxidase activity decreased significantly in the SL groups compared to the C5 group. Glutathione reductase activity increased and glucose-6-phosphate dehydrogenase activity decreased in the C10 group compared to the C5 and C5 groups. In conclusion, the free radical scavenging activity of CLA seemed to suppress oxidative stress, which reduced lipid peroxidation resulting in lower hepatic antioxidant enzyme activity.

• 한국결정성장학회지
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• 제11권3호
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• pp.115-119
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• 2001

수산화아피타이트와 이트리아 부분 안정화된 지르코니아의 바를 달리하는 3층 구조를 가즌ㄴ 생체 재료용 경사기능재료(FGMs)를 spark plasma sintering(SPS)과 hot pressing(HP) 장비로 제조하였다. HAp 원료에 대한 전처리를 실시할 경우 HAp의 소결성이 개선되었다. 전처리하여 얻은 FGM 복합체의 최고밀도는 전처리하지 않은 FGM 보다 낮은 온도에서 얻을 수 있었다. SPS로 FGM 소결체를 제조할 경우 10 MPa의 가압조건에서 8분 동안 소결 할 경우, $1200^{\circ}C$의 온도에서도 HAp의 TCP로의 분해 반응은 일어나지 않았으나. 이 온도에서 FGm의 지르코니아는 정방정에서 압방정으로와 상변화가 일어났다. 지르코니아 첨가에 따라. 즉 지르코니아의 응력 유기 상전이에 의해, HAp-ZrO$_2$ FGm 소결체의 기계적 물성이 증진된 것으로 예산된다. 치밀하고 고강도의 FGM을 제조하는 방법으로서 SPS가 HP공정에 비해 우수한 것으로 나타났다.

• 식물조직배양학회지
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• 제27권5호
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• pp.415-421
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• 2000

The disaccharide trehalose ($\alpha$-D-glucopyranosyl-$\alpha$-D-glucopyranoside) is found in variety of organ-isms that are able to withstand almost complete desiccation. In order to identify the function of trehalose in plants, we isolated Arabidopsis trehalase (AtTRE) gene that encodes the enzyme able to hydrolyze trehalose to glucose, and trehalose-6-phosphate synthase isolog, TPS3 gene by RT-PCR. The AtTRE had the substrate specificity to hydrolyze only trehalose, and a broad pH range of enzyme activity. The AtTRE promoter/GUS reporter gene was expressed in cotyledons, mature leaf tissues including guard cells, and developing siliques. The GUS expression driven by AtTPS3 promoter was significant in root tissues, and the level of GUS activity was much higher than that of the pBll 21 control seedlings. The knockout of AtTPS3 gene in Arabidopsis resulted in the retarded root development, whereas the overexpression of AtTPS3 increased the root elongation in the presence of sucrose in MS medium. Possible functions of AtTRE and AtTPS3 in plant will be discussed. In addition, ectopic expression of yeast TPS1 driven by the inducible promoters in tobacco and potato conferred the plants on the drought and freezing tolerances.

• Journal of Microbiology and Biotechnology
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• 제28권7호
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• pp.1156-1167
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• 2018

The aim of this study was to isolate and characterize lead (Pb)-solubilizing bacteria from heavy metal-contaminated mine soils and to evaluate their inoculation effects on the growth and Pb absorption of Brassica juncea. The isolates were also evaluated for their plant growth-promoting characteristics as well as heavy metal and salt tolerance. A total of 171 Pb-tolerant isolates were identified, of which only 15 bacterial strains were able to produce clear haloes in solid medium containing PbO or $PbCO_3$, indicating Pb solubilization. All of these 15 strains were also able to dissolve the Pb minerals in a liquid medium, which was accompanied by significant decreases in pH values of the medium. Based on 16S rRNA gene sequence analysis, the Pb-solubilizing strains belonged to genera Bacillus, Paenibacillus, Brevibacterium, and Staphylococcus. A majority of the Pb-solubilizing strains were able to produce indole acetic acid and siderophores to different extents. Two of the Pb-solubilizing isolates were able to solubilize inorganic phosphate as well. Some of the strains displayed tolerance to different heavy metals and to salt stress and were able to grow in a wide pH range. Inoculation with two selected Pb-solubilizing and plant growth-promoting strains, (i.e., Brevibacterium frigoritolerans YSP40 and Bacillus paralicheniformis YSP151) and their consortium enhanced the growth and Pb uptake of B. juncea plants grown in a metal-contaminated soil. The bacterial strains isolated in this study are promising candidates to develop novel microbe-assisted phytoremediation strategies for metal-contaminated soils.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1345-1358
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• 2017

The impact of overproduction of a heterologous protein on the metabolic system of host Lactococcus lactis was investigated. The protein expression profiles of L. lactis IL1403 containing two near-identical plasmids that expressed high- and low-level of the green fluorescent protein (GFP) were examined via shotgun proteomics. Analysis of the two strains via high-throughput LC-MS/MS proteomics identified the expression of 294 proteins. The relative amount of each protein in the proteome of both strains was determined by label-free quantification using the spectral counting method. Although expression level of most proteins were similar, several significant alterations in metabolic network were identified in the high GFP-producing strain. These changes include alterations in the pyruvate fermentation pathway, oxidative pentose phosphate pathway, and de novo synthesis pathway for pyrimidine RNA. Expression of enzymes for the synthesis of dTDP-rhamnose and N-acetylglucosamine from glucose was suppressed in the high GFP strain. In addition, enzymes involved in the amino acid synthesis or interconversion pathway were downregulated. The most noticeable changes in the high GFP-producing strain were a 3.4-fold increase in the expression of stress response and chaperone proteins and increase of caseinolytic peptidase family proteins. Characterization of these host expression changes witnessed during overexpression of GFP was might suggested the metabolic requirements and networks that may limit protein expression, and will aid in the future development of lactococcal hosts to produce more heterologous protein.

• Journal of Animal Science and Technology
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• 제56권7호
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• pp.23.1-23.7
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• 2014

Background: Scanning of the genome for selection signatures between breeds may play important role in understanding the underlie causes for observable phenotypic variations. The discovery of high density single nucleotide polymorphisms (SNPs) provide a useful starting point to perform genome-wide scan in pig populations in order to identify loci/candidate genes underlie phenotypic variation in pig breeds and facilitate genetic improvement programs. However, prior to this study genomic region under selection in commercially selected Berkshire and Korean native pig breeds has never been detected using high density SNP markers. To this end, we have genotyped 45 animals using Porcine SNP60 chip to detect selection signatures in the genome of the two breeds by using the $F_{ST}$ approach. Results: In the comparison of Berkshire and KNP breeds using the FDIST approach, a total of 1108 outlier loci (3.48%) were significantly different from zero at 99% confidence level with 870 of the outlier SNPs displaying high level of genetic differentiation ($F_{ST}{\geq}0.490$). The identified candidate genes were involved in a wide array of biological processes and molecular functions. Results revealed that 19 candidate genes were enriched in phosphate metabolism (GO: 0006796; ADCK1, ACYP1, CAMK2D, CDK13, CDK13, ERN1, GALK2, INPP1; MAK, MAP2K5, MAP3K1, MAPK14, P14KB, PIK3C3, PRKC1, PTPRK, RNASEL, THBS1, BRAF, VRK1). We have identified a set of candidate genes under selection and have known to be involved in growth, size and pork quality (CART, AGL, CF7L2, MAP2K5, DLK1, GLI3, CA3 and MC3R), ear morphology and size (HMGA2 and SOX5) stress response (ATF2, MSRB3, TMTC3 and SCAF8) and immune response (HCST and RYR1). Conclusions: Some of the genes may be used to facilitate genetic improvement programs. Our results also provide insights for better understanding of the process and influence of breed development on the pattern of genetic variations.

• 미생물학회지
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• 제55권1호
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• pp.17-24
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• 2019

극지에 서식하는 세균은 강한 빛과 자외선을 받는다. 이 연구에서 우리는 북극에 서식하는 지의류 Cetraria sp.에서 분리한 호냉성 세균 Sphingomonas sp. PAMC 26621의 생장에 빛이 미치는 영향을 조사하였다. 이 세균은 암조건에서 명조건에서 보다 생장이 느렸다. 놀랍게도, 이 세균은 M9 최소배지에 티아민 혹은 아스코브르산을 첨가하면 명조건에서 생장이 증가하였지만, N-acetylcysteine을 첨가한 배지에서는 생장의 변화가 없었다. 첨가한 티아민과 아스코브르산은 포도당-6 인산 탈수소효소와 항산화 효소의 활성을 증가시켰다. 이 연구의 결과는 지의류와의 공생에서 제공된 티아민이 Sphingomonas sp. PAMC26621의 빛에 의한 산화적 스트레스를 완화시키는 항산화제 역할을 함을 의미한다. 이 연구는 강한 빛과 자외선이 만연한 북극에 서식하는 세균에 대한 생리적, 생화학적 관점에서 고찰할 점을 제시한다.

• Toxicological Research
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• 제35권3호
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• pp.249-255
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• 2019

Cadmium is a strong toxic heavy metal which presents in paints and liquid wastes and causes oxidative stress in fish. On the other hand, lead is widely used for different purposes, e.g. lead pipes, it targets vital organs such as liver and kidney causing biochemical alterations. The present study evaluates the effects of 60 days exposure to Cd and Pb either single or combined together in African catfish. Sixty-four fishes were divided into 3 groups and exposed to $CdCl_2$ (7.02 mg/L) or $PbCl_2$ (69.3 mg/L) or a combination of them along with control group. Activities of acid phosphatase (ACP), lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G-6-PDH) were estimated. Moreover, gill, liver and kidney were assayed for activities of superoxide dismutase (SOD), catalase (CAT) and levels of glutathione (GSH) and malondialdehyde (MDA). Individual exposure showed that both Cd and Pb significantly decreased LDH activity and SOD activity in the kidney. Pb significantly increased G-6-PDH activity and decreased GSH level in the gill. CAT activity in liver and kidney elevated significantly on Cd exposure while lead caused a significant depletion in the liver and significant elevation in the kidney. Both Cd and Pb significantly increased MDA levels in liver and kidney while Pb increased its level in gills. The combined exposure resulted in normalization of LDH, G-6-PDH activity, and CAT activity in liver and kidney as well as GSH level in both tissues and MDA in gill and kidney. The combination increased SOD activity and MDA level in liver and decreased SOD activity in kidney and GSH level in gills. In conclusion, the antioxidant system of African catfish was adversely affected by prolonged exposure to Cd and Pb. The combined exposure caused less damage than individual exposure and returned most parameters to those of controls.