• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.335-345
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• 1996

A critical role of oxygen-derived free radicals has been implicated in ischemia/reperfusion (I/R)-induced brain damage. In this study, we have produced experimental I/R to the brains of Mongolian gerbil (Meriones unguiculatus) by a transient occlusion and release of the common carotid arteries. We have attempted to determine whether the oxidative stress is generated upon I/R and whether this oxidative stress is linked to the cell damage. Since hippocampus has been suggested as one of the most vulnerable regions of the brain to the oxidative stress, we analyzed samples from hippocampus in comparison with those from cortex. In addition, we have examined the expression of heat shock protein 70kD species (HSP70) in these regions in order to evaluate a possible role of this protein in I/R-induced brain damage. To determine whether the oxidative stress is produced upon I/R, we measured the glutathione oxidation, GSSG/ (GSH + 2xGSSG), as an index of oxidative stress. We found an increase of the glutathione oxidation primarily in hippocampus upon I/R. To determine whether this oxidative stress is linked to the cell damage, we measured the degree of lipid peroxidation upon I/R. We found an increase of lipid peroxidation in both regions. However, the magnitude of increases was greater in hippocampus than in cortex. In addition, we found that changes in both the magnitude and the temporal patterns of glutathione oxidation closely correlated with those of lipid peroxidation. Our study provides biochemical evidences that the oxidative stress is generated upon I/R and this oxidative stress is linked to the oxidative cell damage. Our study also provides evidences that the degree of oxidative stress as well as oxidative cell damage is greater in hippocampus than in cortex. We could not find difference in the basal level of HSP70 expression between hippocampus and cortex, indicating that the intrinsic vulnerability of hippocampus cannot be explained by the lower level of HSP70 expression. We did find, however, that the induction of HSP70 expression upon I/R was impaired in the hippocampus. This impairment appeared to be at the transcriptional level. These results suggest that the measurement of HSP70 induction may be employed as a useful predictor of differential cellular susceptibilities to the I/R-induced brain damage.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.5
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• pp.657-662
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• 2008

To develop transgenic orchardgrass (Dactylis glomerata L.) resistant to high temperature, the recombinant DgHSP17.2 gene was introduced into orchardgrass plants using the Agrobacterium-mediated transformation method and expressed constitutively under the control of the CaMV 35S promoter. The results of genomic DNA PCR and Southern analysis showed a DNA band and hybridization signal on agarose gel and X-ray film in transgenic orchardgrass plants harboring the recombinant DgHSP17.2 gene, but a DNA band and hybridization signal were not observed in the wild type and empty vector control plants. The same result was also obtained in RT-PCR and Southern blot analysis, and these transgenic orchardgrass plants did not show any morphological aberration both in the culture bottle and soil mixture. When leaf discs cut from transgenic orchardgrass plants with recombinant DgHsp17.2 gene were exposed to lethal temperature (heat treatment at $60^{\circ}C$ for 50 min), 60-80% of the leaf discs showed only damage symptoms, but non-transgenic leaf discs showed a lethal condition. These results indicate that the DgHsp17.2 gene may act as a protector from heat stress in plants.

• The Korean Journal of Zoology
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• v.37 no.4
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• pp.554-564
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• 1994

세균이 세포내 공생하는 xD strain과 모 세포주인 tD strain Amoeba proteus의 열충격 대응의 차이를 알아 보기 위하여 방사선 동위원소로 표지된 아미노산을 Ca2+_less Chalkley's 용액에서 음작용 경로를 통하여 90분 동안 흡수하게 하고, 저온 및 고온 스트레스에 대하여 새로 합성되는 스트레스 대응 단백질의 양상을 1, 2차원 전기영동 및 자기방사 사진법에 의해서 비교하였다 저온(10"C) 충격에 대응하여 아메바는 두 strain 모두 56.0 kDa, pl 6.0 단백질을 강하게 발현하였으며, xD strain에서는 tD strain과 달리 저온 충격 초기에 66 0 kDa, pl 5.5 단백질의 발현이 중단되었다. 한편 고온(33"C) 열충격에 대하여 두 strain 아메바에서 모두 10여종의 단백질이 새합성되는 것으로 확인되었으며, tD 아메바에는 이들 단백질의 새합성이 완만하게 이루어지는데 비하여 xD 아메바에서는 그중 66.0 kDa 단백질이 고온 대응 단백질로서 신속하게 새합성되는 것으로 나타났다. 이외에도 2차원 전기 영동 분석을 통하여 열충격에 의해서 발현이 촉진되는 다수의 단백질들을 탐지하였다 탐지된 아메바의 열충격 단백질은 분자량에 따라 hsp100군 2종, hsp90군, 3종, hsp70군 및 hsp60군 각 1종, 그리고 small csp군 4종으로 분류해 볼 수 있었다 두 분석의 결과를 종합해 보면 tD 아메바에는 저온 및 고온 충격에 대하여 열충격 단백질의 합성이 완만하게 상승하는 데 비하여 xD strain에서는 신속하게 이루어졌다. 이상의 결과로 보아 아메바의 세포내 공생 세균은 숙주의 열충격 대응기작에 변화를 야기한 것으로 판단된다한 것으로 판단된다. 10mg과 20mg의 estrogen 처리구 사이에 유두 직경, 길이 그리고 용적의 증가량에 있어서는 차이가 없었다. 10mg 및 20mg의 estrogen 처리는 초발정일령을 각각 20일 및 124일 단축시켰다. 전체적으로 이러한 결과는 송아지에 estradiol의 삽입은 성장과 유선 발달을 촉진시키고 초발정일령을 단축시킬수 있다는 것을 강력하게 지적한다. 일치하지 않으므로 더욱 정밀한 조사를 실시하여 분류학상의 위치를 정확히 밝혀 볼 필요가 있을 것으로 생각되었다.연한 도구이자 정신활동으로 보게함으로써, 주제 및 연구방법에서 획일성보다 다양성과 창조성이 강조되고 있다. 그리고 연구에 있어서 주제 의 다양성을 통해 보다 현실생활에 밀접하게 연결되어야 할 필요성은 학문이나 과학의 사회 성에 대한 새로운 인식을 가져다 주고 있다. 이러한 지리교육과정의 좌표의 변화된 측면들 을 고려하여, 지리교육과정의 새로운 방향은 다음의 세가지로 모색될 수 있다. 첫째, 爭點中 心 地理敎育課程이다. 사회쟁점에 대한 접근은 쟁점의 이해와 문제해결에의 지리적 관점의 활용을 통해 학습내용의 시사성과 사실성을 높힐 수 있다. 이때 문제해결능력을 통해 현대 시민의 자질 및 능력을 기를 수 있음은 물론, 다른 한편으로 실제세계 즉 학생의 실생활, 사 회, 국가, 세계에서 일어나는 일들과의 관련성을 갖게 함으로써, 내적 동기화와 외적인 자극 을 강력하게 결합할 수 있을 것이다. 이는 개인적 유관적합성과 사회적 유관적합성을 동시 에 확보하는 데 유리할 것이다. 둘째, 思考中心 地理敎育課程이다. 지리교육은 학생들을 지 식 및 기능의 숙달자가 되도록 할 것이 아니라 기본적 문장해독력의 수준을 넘어 능력있는 사고자로 길러내는 것을 목표로 하여야 한다.

• Journal of Animal Science and Technology
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• v.57 no.12
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• pp.44.1-44.9
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• 2015

Background: Heat shock proteins play an important role in protection from stress stimuli and metabolic insults in almost all organisms. Methods: In this study, computational tools were used to deeply analyse the physicochemical characteristics and, using homology modelling, reliably predict the tertiary structure of the blunt snout bream (Ma-) Hsp70 and Hsc70 proteins. Derived three-dimensional models were then used to predict the function of the proteins. Results: Previously published predictions regarding the protein length, molecular weight, theoretical isoelectric point and total number of positive and negative residues were corroborated. Among the new findings are: the extinction coefficient (33725/33350 and 35090/34840 - Ma-Hsp70/ Ma-Hsc70, respectively), instability index (33.68/35.56 - both stable), aliphatic index (83.44/80.23 - both very stable), half-life estimates (both relatively stable), grand average of hydropathicity (-0.431/-0.473 - both hydrophilic) and amino acid composition (alanine-lysine-glycine/glycine-lysine-aspartic acid were the most abundant, no disulphide bonds, the N-terminal of both proteins was methionine). Homology modelling was performed by SWISS-MODEL program and the proposed model was evaluated as highly reliable based on PROCHECK's Ramachandran plot, ERRAT, PROVE, Verify 3D, ProQ and ProSA analyses. Conclusions: The research revealed a high structural similarity to Hsp70 and Hsc70 proteins from several taxonomically distant animal species, corroborating a remarkably high level of evolutionary conservation among the members of this protein family. Functional annotation based on structural similarity provides a reliable additional indirect evidence for a high level of functional conservation of these two genes/proteins in blunt snout bream, but it is not sensitive enough to functionally distinguish the two isoforms.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.110-122
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• 2022

The reduction of milk yield caused by heat stress in summer is the main condition restricting the economic benefits of dairy farms. To examine the impact of hyperthermia on bovine mammary epithelial (MAC-T) cells, we incubated the MAC-T cells at thermal-neutral (37℃, CON group) and hyperthermic (42℃, HS group) temperatures for 6 h. Subsequently, the cell viability and apoptotic rate of MAC-T cells, apoptosis-related genes expression, casein and amino acid transporter genes, and the expression of the apoptosis-related proteins were examined. Compared with the CON group, hyperthermia significantly decreased the cell viability (p < 0.05) and elevated the apoptotic rate (p < 0.05) of MAC-T cells. Moreover, the expression of heat shock protein (HSP)70, HSP90B1, Bcl-2-associated X protein (BAX), Caspase-9, and Caspase-3 genes was upregulated (p < 0.05). The expression of HSP70 and BAX (pro-apoptotic) proteins was upregulated (p < 0.05) while that of B-cell lymphoma (BCL)2 (antiapoptotic) protein was downregulated (p < 0.05) by hyperthermia. Decreased mRNA expression of mechanistic target of rapamycin (mTOR) signaling pathway-related genes, amino acid transporter genes (SLC7A5, SLC38A3, SLC38A2, and SLC38A9), and casein genes (CSNS1, CSN2, and CSN3) was found in the heat stress (HS) group (p < 0.05) in contrast with the CON group. These findings illustrated that hyperthermia promoted cell apoptosis and reduced the transport of amino acids into cells, which inhibited the milk proteins synthesis in MAC-T cells.

• Journal of Periodontal and Implant Science
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• v.53 no.5
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• pp.347-361
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• 2023

Purpose: Exosomes are membrane vesicles that are present in body fluids and contain proteins, lipids, and microRNA (miRNA). Periodontal tissue examinations assess the degree of periodontal tissue destruction according to the probing depth (PD), clinical attachment loss (CAL), bleeding on probing, and X-ray examinations. However, the accurate evaluation of the prognosis of periodontitis is limited. In this study, we collected saliva from patients before and after initial periodontal therapy (IPT) and compared changes in the clinical parameters of periodontitis with changes in the components of salivary exosomes. Methods: Saliva was collected from patients with stage III and IV periodontitis at the first visit and post-IPT. Exosomes were purified from the saliva, and total protein and RNA were extracted. Changes in expression levels of C6, CD81, TSG101, HSP70, and 6 kinds of miRNA were analyzed by western blots and real-time polymerase chain reaction. Results: Patients with increased C6 expression after IPT had significantly higher levels of periodontal inflamed surface area (PISA), miR-142, and miR-144 before and after IPT than patients with decreased C6 expression after IPT. Patients with decreased and unchanged CD81 expression after IPT showed significantly higher PD, CAL, and PISA before IPT than after IPT. Patients with decreased and unchanged TSG101 expression after IPT had significantly higher PD before IPT than after IPT. Patients with increased HSP70 expression after IPT had significantly higher PD and PISA before and after IPT than patients with unchanged HSP70 after IPT. The expression levels of miR-142, miR-144, miR-200b, and miR-223 changed with changes in the levels of C6, CD81, TSG101, and HSP70 in the salivary exosomes of periodontitis patients before and after IPT. Conclusions: The expression levels of proteins and miRNAs in salivary exosomes significantly changed after IPT in periodontitis patients, suggesting that the components of exosomes could serve as biomarkers for periodontitis.

• Korean Journal of Poultry Science
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• v.44 no.1
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• pp.1-9
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• 2017

The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

• Food Science and Preservation
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• v.8 no.2
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• pp.131-139
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• 2001

Kiyomi tangor(Citus unshiu x sinensis) was stored at 3$\^{C}$ and 85% relative humidity, and the changes in firmness, pectin degrading enzymes activity and other physicochemical properties of citrus fruits during storage were investigated. Decay ratio and weight loss during 180 days’ storage were increased gradually to 13.0% and 12.9%, respectively. Firmness of fruits with 2 mm probe was decreased gradually from 808.7 g-force to 406.4 g-force, and moisture of peel and flesh were decreased from 76.5% to from 89.6% to 87.6% during storage, respectively. Exo-polygalacturonase activity of peel after 150 days’ storage were increased gradually to 558.09 units/100g. Pectin methylesterase activity of peel and flesh were increased from 14.7 units/g to 2.3 units/g, and from 9.4 units/ml to 2.7 units/ml at 150days’ storage, respectively. Endo-polygalacturonase activities were not changed notably during storage. Alcohol-insoluble solid(AIS) of peel was not changed notably. During storage of the fruits water soluble pectin(WSP) of peel and flesh were increased from 474.49 mg/100g to 614.29mg/100g, and from 66.91mg/100g to 92.74mg/100g as wet basis, respectively. Hexameta-phosphate soluble pectin(HMP) of peel were decreased from 405.5mg/100g to 270.43mg/100g, hydochloric acid soluble pectin(HSP) of peel was also decreased from 544.02mg/100g to 412.64mg/100g during storage. Total pectin substance(TPS) of peel and flesh were decreased from 1,424.01mg/100g to 1,297.36mg/100g, and from 165.51mg/100g to 171.54mg/100g, respectively. Composition ratio of pectin was in order of WSP > HSP > HMP.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.11
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• pp.1598-1608
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• 2013

An optimum dietary carbohydrate content is important for maximum fish growth. In this study, we fed Wuchang bream (Megalobrama amblycephala) with either control diet (30.42%) or high carbohydrate diet (52.92%) for 90 d. Fish were fed to apparent satiation three times daily in an aquarium with automatic temperature control and circulated water. Growth performance, plasma biochemical parameters, hepatic morphology and enzyme activities were determined. It was shown that compared to fish fed control diet, fish fed high carbohydrate diet had higher plasma triglyceride and cortisol levels for d 90, and lower alkaline phosphatase level for d 45, lower hepatic superoxide dismutase and total antioxidative capacity for d 90, higher malondialdehyde for d 45 and glycogen content for d 45 and 90 (p<0.05). Histological and transmission electron microscopy studies showed that hepatocytes of fish fed high carbohydrate diet contained large lipid droplets, causing displacement of cellular organelles to periphery of hepatocytes. The relative level of hepatic heat shock protein 70 (HSP70) mRNA of Wuchang bream fed high carbohydrate diet was significantly higher than that of fish fed the control diet for 90 d (p<0.05). These changes led to decreased specific growth rate and increased feed conversion ratio (p<0.05). Upon hypoxia challenge, fish fed high carbohydrate diet had higher cumulative mortality than those fed the control diet (p<0.05). These results suggested that high dietary carbohydrate (52.92%) was detrimental to the growth performance and health of Wuchang bream.

• Journal of Plant Biotechnology
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• v.43 no.3
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• pp.302-310
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• 2016

A study aimed at identifying putative drought responsive genes that confer tolerance to water stress deficit in tea plants was conducted in a 'rain-out shelter' using potted plants. Eighteen months old drought tolerant and susceptible tea cultivars were each separately exposed to water stress or control conditions of 18 or 34% soil moisture content, respectively, for three months. After the treatment period, leaves were harvested from each treatment for isolation of RNA and cDNA synthesis. The cDNA libraries were sequenced on Roche 454 high-throughput pyrosequencing platform to produce 232,853 reads. After quality control, the reads were assembled into 460 long transcripts (contigs). The annotated contigs showed similarity with proteins in the Arabidopsis thaliana proteome. Heat shock proteins (HSP70), superoxide dismutase (SOD), catalase (cat), peroxidase (PoX), calmodulinelike protein (Cam7) and galactinol synthase (Gols4) droughtrelated genes were shown to be regulated differently in tea plants exposed to water stress. HSP70 and SOD were highly expressed in the drought tolerant cultivar relative to the susceptible cultivar under drought conditions. The genes and pathways identified suggest efficient regulation leading to active adaptation as a basal defense response against water stress deficit by tea. The knowledge generated can be further utilized to better understand molecular mechanisms underlying stress tolerance in tea.