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Regional Differences of Proteins Expressing in Adipose Depots Isolated from Cows, Steers and Bulls as Identified by a Proteomic Approach

  • Cho, Jin Hyoung;Jeong, Jin Young;Lee, Ra Ham;Park, Mi Na;Kim, Seok-Ho;Park, Seon-Min;Shin, Jae-Cheon;Jeon, Young-Joo;Shim, Jung-Hyun;Choi, Nag-Jin;Seo, Kang Seok;Cho, Young Sik;Kim, MinSeok S.;Ko, Sungho;Seo, Jae-Min;Lee, Seung-Youp;Chae, Jung-Il;Lee, Hyun-Jeong
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1197-1206
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    • 2016
  • Adipose tissue in the loin muscle area of beef cattle as a marbling factor is directly associated with beef quality. To elucidate whether properties of proteins involved in depot specific adipose tissue were sex-dependent, we analyzed protein expression of intramuscular adipose tissue (IMAT) and omental adipose tissue (OMAT) from Hanwoo cows, steers, and bulls of Korean native beef cattle by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis, quantitative polymerase chain reaction (PCR) and western blot analysis. Two different adipose depots (i.e. intramuscular and omental) were collected from cows (n = 7), steers (n = 7), or bulls (n = 7). LC-MS/MS revealed a total of 55 and 35 proteins in IMAT and OMAT, respectively. Of the 55 proteins identified, 44, 40, and 42 proteins were confirmed to be differentially expressed in IMAT of cows, steers, and bulls, respectively. In OMAT of cows, steers, and bulls, 33, 33, and 22 were confirmed to be differentially expressed, respectively. Tropomyosin (TPM) 1, TPM 2, and TPM3 were subjected to verification by quantitative PCR and western blot analysis in IMAT and OMAT of Hanwoo cows, steers, and bulls as key factors closely associated with muscle development. Both mRNA levels and protein levels of TPM1, TPM2, and TPM3 in IMAT were lower in bulls compared to in cows or steers suggesting that they were positively correlated with marbling score and quality grade. Our results may aid the regulation of marbling development and improvement of meat quality grades in beef cattle.

Association of Six Susceptibility Loci with Prostate Cancer in Northern Chinese Men

  • Zhang, Yu-Rong;Xu, Yong;Yang, Kuo;Liu, Ming;Wei, Dong;Zhang, Yao-Guang;Shi, Xiao-Hong;Wang, Jian-Ye;Yang, Fan;Wang, Xin;Liang, Si-Ying;Zhao, Cheng-Xiao;Wang, Fei;Chen, Xin;Sun, Liang;Zhu, Xiao-Quan;Zhu, Ling;Yang, Yi-Ge;Tang, Lei;Jiao, Hai-Yan;Huo, Zheng-Hao;Yang, Ze
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6273-6276
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    • 2012
  • Background/Aim: Six prostate cancer (PCa) susceptibility loci were identified in a genome-wide association study (GWAS) in populations of European decent. However, the associations of these 6 single-nucleotide polymorphisms (SNPs) with PCa has remained tobe clarified in men in Northern China. This study aimed to explore the loci associated with PCa risk in a Northern Chinese population. Methods: Blood samples and clinical information of 289 PCa patients and 288 controls from Beijing and Tianjin were collected. All risk SNPs were genotyped using polymerase chain reaction (PCR)-high resolution melting curve technology and gene sequencing. Associations between PCa and clinical covariates (age at diagnosis, prostate-specific antigen [PSA], Gleason score, tumor stage, and level of aggressiveness) and frequencies of alleles and genotypes of these SNPs were analyzed using genetic statistics. Results: Among the candidate SNPs, 11p15 (rs7127900, A) was associated with PCa risk (P = 0.02, odds ratio [OR] = 1.64, 95% confidence interval [CI] = 1.09-2.46). Genotypes showed differences between cases and controls on 11p15 (rs7127900, A), 11q13 (rs7931342, T), and HNF1B (rs4430796, A) (P = 0.03, P = 0.01, and P = 0.04, respectively). The genotype TG on 11q13 (rs7931342, T) was positively associated with an increased Gleason score (P = 0.04, OR = 2.15, 95% CI = 1.02-4.55). Patients carrying TG on 17q24 (rs1859962, G) were negatively associated with an increased body mass index (BMI) (P = 0.03, OR = 0.44, 95% CI = 0.21-0.92) while those with AG on HNF1B (rs4430796, A) were more likely to have PSA increase (P = 0.002). Conclusion: Our study suggests that 11p15 (rs7127900, A) could be a susceptibility locus associated with PCa in Northern Chinese. Genotype TG on 11q13 (rs7931342, T) could be related to an increased Gleason score, AG on HNF1B (rs4430796, A) could be associated with PSA increase, and TG on 17q24 (rs1859962, G) could be negatively associated with an increased BMI in Chinese men with PCa.

Induction of Apoptosis by Ethanol Extract of Lythrum anceps (Koehne) Makino in Human Leukemia U937 Cells (인체백혈병 U937 세포에서 부처꽃 에탄올추출물에 의한 apoptosis 유도)

  • Jeong, Jin-Woo;Kim, Chul Hwan;Lee, Young-Kyung;Hwang, Yong;Lee, Ki Won;Choi, Kyung-Min;Kim, Jung Il
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.279-286
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    • 2020
  • Purple loosestrife-Lythrum anceps (Koehne) Makino is a herbaceous perennial plant belonging to the Lythraceae family. It has been used for centuries in Korea and other Asian traditional medicine. It has been showed pharmacological effects, including anti-oxidant and anti-microbial effects. However, the mechanisms underlying its anti-cancer effect are not yet understood. In this study, we investigated the mechanism of apoptosis signaling pathways by ethanol extract of Lythrum anceps (Koehne) Makino (ELM) in human leukemia U937 cells. Treatment with ELM significantly inhibited cell growth in a dose-dependent manner by inducing apoptosis, as evidenced by the formation of apoptotic bodies (ApoBDs), DNA fragmentation and increased populations of sub-G1 ratio. Induction of apoptosis by ELM was connected with up-regulation of death receptor (DR) 4 and DR5, pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 protein, and inhibitor of apoptosis protein (IAP) family proteins, depending on dosage. This induction was associated with Bid truncation, mitochondrial dysfunction, proteolytic activation of caspases (-3, -8 and -9) and cleavage of poly(ADP-ribose) polymerase protein. Therefore, our data indicate that ELM suppresses U937 cell growth by activating the intrinsic and extrinsic apoptosis pathways, and thus may have applications as a potential source for an anti-leukemic chemotherapeutic agent.

Anti-aging Effect of Inula britannica var. chinensis Flower Extract According to the Extraction Temperature (추출 온도에 따른 금불초 꽃 추출물의 항노화 효능)

  • Jeon, Ji Min;Yoo, Dae Sung;Cheon, Jong Woo;Kwon, Soon Sik;Jeon, So Ha;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.1
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    • pp.109-120
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    • 2014
  • In this study, the extracts of Inula britannica var. chinensis (I. britannica) flower were extracted at three different temperatures (room temperature, $45^{\circ}C$, and $65^{\circ}C$) and their anti-aging effects were studied. Before investigating anti-aging effects of the extracts, their cytotoxicity was tested on B16F10, Hs683, and HaCaT cells. All extracts showed no cytotoxicity at the concentration less than 0.1% (v/v). Melanin synthesis inhibitory activities in B16F10 cells and reverse transcriptase polymerase chain reaction (RT-PCR) in Hs683 and HaCaT cells were used to see their anti-aging effects. The room temperature extract at 0.1% showed 24.5% melanin synthesis inhibition, which was better than the $45^{\circ}C$ and $65^{\circ}C$ extracts. In addition, expression rates of the room temperature extract at 0.1% on HAS-1, HAS-2, and HAS-3 related to hyaluronan synthase genes were 123.3%, 137.8%, 133.2%, respectively. which were higher than reference material of L-ascorbic acid. Expression rates of the $45^{\circ}C$ extract at 0.1% on TNF-${\alpha}$, COX-2, and IL-$1{\alpha}$, which are inflammatory related genes, was suppressed to 30.3%, 12.8%, 25.7%, respectively. It was better in anti-in flammatory effect than the room temperature and $65^{\circ}C$ extracts. As results, we showed that I. britannica var. chinensis flower extarcts decreased melanin production and expression of inflammatory related genes and increased the expression rate of hyaluronan synthase genes. Thus, it is believed that the extracts affect anti-aging effects of skin through whitening, moisturizing, and anti-inflammatory processes and could be applicable to cosmetics as a functional cosmetic ingredient.

Expression and Possible Role of Phospholipase C $\beta1$ and $\gamma1$ in Mouse Oocyte Maturation and Preimplantation Embryo Development (생쥐 난자의 성숙과 착상전 배발생에서의 Phospholipase C $\beta1$$\gamma1$의 발현 및 기능)

  • Lee, Young-Hyun;Geum, Dong-Ho;Shim, Chan-Seob;Suh, Phan-Gil;Kim, Kyung-Jin
    • Development and Reproduction
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    • v.2 no.1
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    • pp.9-20
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    • 1998
  • It has been wel known that phospholipase C(PLC) plays an important role in the intracellular signaling in a variety of cell types. However, involvement of PLC in mouse oocyte maturation and preimplantation embryo development remains unknown. The present study examined the expression patterns of the mouse PLC \beta 1 and \gamma 1 during oocyte maturatio and preimplantation embryo development study examined the expression patterns of the mouse PLC \beta 1 and \gamma 1 during oocyte maturation and preimplantation embryo development by the competitive reverse transcription-polymerase chain reaction (RT-PCR method). PLC \gamma 1 mRNA (0.1 fg) was readily detected in germinal vesicle (GV)-stage oocyte and its level was reduced as meiotic resumption proceeded. PLC-\beta 1 mRNA (<0.1 fg) as detected at low level at GV-stage oocytes and scarcely detected at germinal vescle breakdown (GVBD)-stage oocytes. After fertilization, both PLC \beta 1 and \gamma 1 mRNA levels began to increase at morula-stage embryos (0.2 fg) and were more prominent in blastocyst-stage embryos(1 fg). to elucidate the possible involvement of PLC via protein kinase C(PKC) pathway during oocyte maturation and preimplantation embryo development , the effects of sphingosine (PKC inhibitor), sn-$diC_{8}$(PKC activator) anc U73122 (PLC ingibitor) were examined. Treatment of GV-stage oocytes with sphingosine (20 \mu M) facilitated the meiotic resuption by 10-20 over the control within 1 h as judged by GVBD, whereas U73122 failed to show any significant effect. U73122 (10 \mu M) effectively blocked the compaction of morula, while sn-$diC_{8}$(50 \mu M). In summary, the present study shows that the mouse PLC \beta 1 and \gamma 1 are expressed in a developmental stage-specific manner and PLC-PKC pathway may be involved in early preimplantation embryo development.

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Effect of Growth Hormone and Androgen on Vitellogenin and Estrogen Receptor Gene Expression in the Japanese eel, Anguilla japonica (뱀장어 Vitellogenin과 Estrogen 수용체 유전자 발현에 대한 성장호르몬 및 웅성호르몬의 영향)

  • Kwon, Hyuk-Chu;Choi, Seong-Hee;Kim, Eun-Hee;Kwon, Joon-Yeong
    • Development and Reproduction
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    • v.10 no.2
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    • pp.97-103
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    • 2006
  • Vitellogenin(Vg) is a sex specific serum protein present in sexually maturing female blood of oviparous vertebrates. Estrogen($E_2$) is a main inducer of hepatic Vg synthesis. We investigated the effects of androgen and growth hormone(GH) on regulation of Vg and estrogen receptor(ER) genes in Japanese eel. Immature eels($200{\sim}250\;g$) were given a single injection of $E_2(5{\sim}5,000\;{\mu}g/kg\;bw)$ alone, or in combination with eel recombinant GH(eGH, $1{\sim}10\;{\mu}g/kg$) or methyltestosterone(MT, $1{\sim}5\;mg/kg$) and sacrificed 10 days after the hormone treatments. Expression levels of ER and Vg genes from the liver were determined by means of reverse transcription and polymerase chain reaction(RT-PCR). Administration of $E_2$ stimulated Vg gene expression in a dose dependent manner. Levels of Vg mRNA after the injection of $E_2(500\;{\mu}g/kg)$ with MT(5mg/kg) or eGH($10\;{\mu}g/kg$) were much higher than in that of $E_2$ alone($500\;{\mu}g/kg$). Whereas, injection of either vehicle, eGH ($10\;{\mu}g/kg$) or MT(5mg/kg) alone did not induce the expression of Vg gene in the liver. ER mRNA was detected from the fish treated with vehicle alone. $E_2$ injection($5{\sim}500\;{\mu}g/kg\;bw$) increased this ER expression but dose dependent response was not clear. Addition of MT(5mg/kg) or eGH($10\;{\mu}g/kg$) did not affect $E_2-stimulated$ ER mRNA expression. This study confirms the necessity of $E_2$ as the primary factor for Vg gene expression and requirement of additional hormones such as MT or GH for the full expression of Vg mRNA, and suggests that the additive effect of MT or GH on Vg gene expression would be mediated by some unknown factors other than ER.

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Effect of Di(2-ethyl hexyl)phthalate(DEHP) on the Onset of Puberty in Female Rat (암컷 흰쥐의 사춘기 개시에 미치는 di(2-ethyl hexyl)phthalate(DEHP)의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.2
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    • pp.147-154
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    • 2006
  • Phthalates such as di(2-ethyl hexyl)phthalate(DEHP) are industrial chemicals with wide-ranging human exposures because of their use in plastics and other common consumer products. Consequently, their adverse effects as endocrine disruptor in the reproductive physiology of both laboratory rodents and human have been studied extensively. The present study was undertaken to examine whether prepubertal exposure to DEHP affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. DEHP(100mg/kg/day) was administered daily from postnatal day 25(PND 25) through the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day. Gross anatomy and weight of reproductive tissues were compared to test the DEHP's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. Specific radioimmunoassay was carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed VO was shown in the DEHP group(PND $37.3{\pm}0.7$) compared to the control group(PND $35.3{\pm}0.7$; p<0.05). DEHP treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.05). Interestingly, elevation of serum LH levels was shown in the DEHP group(p<0.05). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals. Numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from DEHP-treated animals. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the DEHP-treated group. These effects were probably due to the inhibitory effects of DEHP on the synthesis and secretion of estrogen from granulosa cells. In the semiquantitative RT-PCR studies, the transcriptional activities of PR in both ovary(p<0.05) and uterus(p<0.01) from DEHP-treated animals were significantly lower than those from the control animals. The present studies demonstrated that the acute exposure to DEHP during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.

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Amended Soil with Biopolymer Positively Affects the Growth of Camelina sativa L. Under Drought Stress (가뭄 조건 하에서 바이오폴리머 혼합 토양이 Camelina sativa L.의 생장에 미치는 긍정적 영향)

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Lee, Hyeon-Sook;Sin, Jung-Ho;Kim, Eun-Suk;Woo, Hyo-Seop;Ahn, Sung-Ju
    • Ecology and Resilient Infrastructure
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    • v.5 no.3
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    • pp.163-173
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    • 2018
  • The biopolymer (BP) used in this study is mainly composed of xanthan gum and ${\beta}$-glucan derived from microorganism and has been introduced as a novel material for soil stabilization. However, the broad applicability of BP has been suggested in the field of geotechnical engineering while little information is available about the effects of BP on the vegetation. The goal of this study is to find the BP effects on the growth of Camelina sativa L. (Camelina) under drought condition. For more thorough evaluation of BP effects on the plant growth, we examined not only morphological but also physiological traits and gene expression patterns. After 25 days of drought treatment from germination in the soil amended with 0, 0.25, 0.5, and 1% BP, we observed that the BP concentration was strongly correlated the growth of Camelina. When plants were grown under drought stress, Camelina in 0.5% BP mixture showed better physiological parameters of the leaf stomatal conductance, electrolyte leakage and relative water content compared to those in control soil without BP. Plant recovery rate after re-watering was higher and the development of lateral root was lower in BP amended soil. RNA expression of Camelina leaf treated with/without drought for 7 and 10 days showed that aquaporin genes transporting solutes at bio-membrane, CsPIP1;4, 2;1, 2;6 and TIP1;2, 2;1, were induced more in the plants with BP amendment and drought treatment. These results suggest that the soil amended with BP has a positive effect on the transport of nutrients and waters into Camelina by improving water retention in soil under drought condition.

Analysis of Stress Response of Domestic Chicken Breeds for the Development of a New Synthetic Parent Stock (국산 종계 개발을 위한 토종 계통들의 스트레스 반응 정도 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Ji Ae;Hong, Young Ho;Kim, Chong Dae
    • Korean Journal of Poultry Science
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    • v.42 no.2
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    • pp.157-167
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    • 2015
  • We compared the degrees of stress response of 12 domestic purebred chicken strains that have been bred at National Institute of Animal Science, RDA, Korea since 1980. As a physiological marker of stress response, the expression levels of heat shock protein (HSP)-70, HSP-$90{\alpha}$, HSP-$90{\beta}$, hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes and telomere length were measured by quantitative real-time polymerase chain reaction using the lymphocytes of 1,101 chickens. There was significant difference in HSP-70, HSP-$90{\alpha}$, HMGCR expression and telomere length among the strains. There was also significant difference in HSP-$90{\alpha}$, HSP-$90{\beta}$, and HMGCR expression between male and female chickens. Different age groups of chicken exhibited different expression levels of HSP-70, HSP-$90{\alpha}$ and telomere length. The results of the HSPs expression level suggested that, the strains of R, L and Y were highly resistant to stress, whereas the strains of S, O and W were susceptible to stress. Although the statistical differences in some of HSPs gene expression existed between genders, the HSP expression results varied in different strains that some opposed to the others, and there might be interaction between strains and genders, which conclude that there was no difference in stress response between male and female chickens. Moreover, despite of significant difference in some of HSPs expression level, it was considered that there was no difference in stress response between ages due to the inconsistent trends among HSP markers.

Comparison of Stress Response between Korean Native Chickens and Single Comb White Leghorns subjected to a High Stocking Density (닭의 품종 간 스트레스 반응 정도 비교 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Dhan Bee;Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.41 no.2
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    • pp.115-125
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    • 2014
  • With Single Comb White Leghorn (WL) and Korean Native Chicken (KNC) breeds, we compared the stress response with chicken breeds that were subjected to a high stocking density. Stress response was analyzed by the quantity of telomeric DNA, the rate of DNA damage and the expression levels of heat shock proteins (HSPs) and hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes on tissues and blood. The telomere length and telomere shortening rates were analyzed by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes and tissues. The DNA damage rate of lymphocytes was quantified by the comet assay. The expression levels of HSP70, HSP90-${\alpha}$, HSP90-${\beta}$ and HMGCR genes were measured by quantitative real-time polymerase chain reaction in lymphocytes. There was no significant difference between KNC and WL in body weight, weight gain, telomere shortening rate and DNA damage rate. However, the growth rate significantly decreased in chickens raised under high stocking density conditions, as compared to the control group. The telomere-shortening rate, DNA damage and HSPs expression of the lymphocytes were significantly higher in the high stocking density group than the control. The stress condition and breeds had a significant effect on the expressions of HSP70, HSP90-${\alpha}$ and HSP90-${\beta}$ in lymphocytes, except HMGCR. The stress response of WL was higher than that of KNC, as analyzed to the expression of HSP70 and HSP90-${\alpha}$. Therefore, we concluded that the chickens which were exposed to a high stocking density had increased the individual physiological stress response regardless of breeds, and White Leghorns are more susceptible to stress condition than Korean Native Chickens.