• Asian-Australasian Journal of Animal Sciences
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• 제26권11호
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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 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.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7043-7048
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• 2014

Inhibition of heat shock protein 90 (Hsp90) leads to inappropriate processing of proteins involved in DNA damage repair pathways after DNA damage and may enhance tumor cell radio- and chemotherapy sensitivity. To investigate the potentiation of antitumor efficacy of lidamycin (LDM), an enediyne agent by the Hsp90 inhibitorgeldanamycin (GDM), and possible mechanisms, we have determined effects on ovarian cancer SKOV-3, hepatoma Bel-7402 and HepG2 cells by MTT assay, apoptosis assay, and cell cycle analysis. DNA damage was investigated with H2AX C-terminal phosphorylation (${\gamma}H2AX$) assays. We found that GDM synergistically sensitized SKOV-3 and Bel-7402 cells to the enediyne LDM, and this was accompanied by increased apoptosis. GDM pretreatment resulted in a greater LDM-induced DNA damage and reduced DNA repair as compared with LDM alone. However, in HepG2 cells GDM did not show significant sensitizing effects both in MTT assay and in DNA damage repair. Abrogation of LDM-induced $G_2/M$ arrest by GDM was found in SKOV-3 but not in HepG2 cells. Furthermore, the expression of ATM, related to DNA damage repair responses, was also decreased by GDM in SKOV-3 and Bel-7402 cells but not in HepG2 cells. These results demonstrate that Hsp90 inhibitors may potentiate the antitumor efficacy of LDM, possibly by reducing the repair of LDM-induced DNA damage.

• Asian Pacific Journal of Cancer Prevention
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• 제13권7호
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• pp.3471-3476
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• 2012

Background and Objective: Histone deacetylase (HDAC) inhibitors represent a promising class of potential anticancer agents for treatment of human malignancies. In this study, we investigated the effect of trichostatin A (TSA), one such HDAC inhibitor, in combination with docetaxel (TXT), a cytotoxic chemotherapy agent or erlotinib, a novel molecular target therapy drug, on lung cancer A549 cells. Methods: A549 cells were treated with TXT, erlotinib alone or in combination with TSA, respectively. Cell viability, apoptosis, and cell cycle distribution were evaluated using MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide) assay, Hochst33258 staining and flow cytometry. Moreover, immunofluorescent staining and Western blot analysis were employed to examine alterations of ${\alpha}$-tubulin, heat shock protein 90 (hsp90), epidermal growth factor receptor (EGFR), and caspase-3 in response to the different exogenous stimuli. Results: Compared with single-agent treatment, co-treatment of A549 cells with TSA/TXT or TSA/erlotinib synergistically inhibited cell proliferation, induced apoptosis, and caused cell cycle delay at the $G_2/M$ transition. Treatment with TSA/TXT or TSA/erlotinib led to a significant increase of cleaved caspase-3 expression, also resulting in elevated acetylation of ${\alpha}$-tubulin or hsp90 and decreased expression of EGFR, which was negatively associated with the level of acetylated hsp90. Conclusions: Synergistic anti-tumor effects are observed between TXT or erlotinib and TSA on lung cancer cells. Such combinations may provide a more effective strategy for treating human lung cancer.

• Molecules and Cells
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• 제25권1호
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• pp.55-63
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• 2008

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

• 한국식품과학회지
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• 제35권6호
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• pp.1098-1103
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• 2003

열처리에 의한 대두 단백질의 용출 변화를 조사하기 위하여 만리, 태광, 은하, 풍산나물, 검정콩 1호와 일품검정의 6가지 장려품종 대두를 온도와 시간에 따라 열처리를 달리하여 침치액의 pH와 가용성 고형물함량과 단백질 특성을 살펴보았다. $60^{\circ}C$에서 90분 동안 열처리 후 침지액의 pH는 열처리 전보다 높았으며 $60^{\circ}C$에서 열처리 시간이 증가함에 따라 pH는 감소하였다. 침지액의 가용성고형물 함량은 열처리 온도에 따라서 $50^{\circ}C$ 이후 급격한 증가를 보였으며, 열처리 시간에 따라서 용출량의 증가를 보였다. 다른 대두에 비해 풍산나물과 은하의 경우 시간에 따라 비례적인 증가경향을 보였으며 검정콩 1호와 일품검정은 낮은 당도를 나타내었다. 단백질함량은 열처리 온도와 시간이 증가함에 따라 증가하였는데 검정콩1호와 일품검정은 다른 대두에 비해 각각 2배와 4배 정도 높은 단백질 농도를 나타내었다. 검정콩1호, 만리 및 은하 품종의 SDS-PAGE 특성은 모든 대두에서 비슷한 분리대를 보였다. 열처리 온도가 증가함에 따라 전체적으로 단백질 용출량이 증가하였고, $50^{\circ}C$ 및 $60^{\circ}C$에서 대략 31kDa과 16kDa의 새로운 단백질이 뚜렷하게 증가하였다. 열처리 시간이 증가함에 따라서도 단백질 용출량은 증가하였고 새로운 단백질 분획은 다른 단백질 보다 용출량이 더 증가하였다. 선행연구와의 비교로 열처리에 의해 나타난 31kDa와 16kDa의 단백질은 장려품종 대두에서 합성된 열충격 단백질(heat shock protein, HSP)이 용출된 것으로 판단하였다.養素) 이용율중(利用率中) 지방이용율은 대체로 일반식이 내의 륵지방(肋脂肪)보다는 지방질(脂肪質) 첨가식이내(添加食餌內)의 륵지방(肋脂肪)이 흡수율(吸收率)이나 이용률(利用率)이 더 우수함을 보여주었고, 근소축적률(筋素蓄積率)은 쥐에 있어서는 어유(魚油)군이 73.5%, 병아리에 있어서는 채종유군이 52.1%로 가장 높았으며 또한 쥐에서는 참기름군이 66.0%, 병아리에서는 대조구가 33.4로 가장 낮았으나 각 구별 통계적(統計的)인 차이는 없었다. 4. 쥐 실험(實驗)에서만 실시된 간지방(肝脂防) 함량측정(含量測定)은 채종유군의 그것이 다른 군보다 높았으며 옥수수기름, 콩기름, 참기름, 들기름, 동물유, 어유의 순서였으나 통계적(統計的)인 유의성(留意性)은 인정(認定)되지 않았고 일반적(一般的)으로 식물성 유지급여군의 간지방함량(肝脂肪含量)이 다른 군보다 높았다. 5. 혈청(血淸)콜레스테롤 함량(含量)은 쥐에 있어서는 채종유군이 가장 높았으며 참기름군이 비교적(比較的) 높은 수치(數値)를 나타냈고, 병아리실험에서는 동물유군이 어유, 콩기름, 참기름, 들기름, 옥수수기름, 채종유, 대조군보다 높았으나 통계적(統計的)인 유의차(留意差)는 없었다.6. 지방산조성(脂肪酸造成)은 동물성유지는 대체로 palmitic acid, myristic acid함량이 많았으며 식물성유는 Linoleic acid 와 oleic acid가 많았고 옥수수기름이 필수지방산인 Linoleic acid 함량이 54.7%로 가장많았으며 특히 들기름이 2중 결합 3개인 Linolenic acid가 58.4%로 다른 식물성기름보다 월등히 높았다. 한편 식물성 기름에는 Arachidonic acid 가 소량있으나 동물유(動物油)나 어유(魚油)에서는 분석(分析)되지 못했다.

• 한국환경농학회지
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• 제25권1호
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• pp.7-13
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• 2006

비닐하우스는 노지보다 UV-B 선량이 적고 외기와 차단되어 노지와 다른 생육환경 하에서 작물이 생육하게 된다. 본 연구는 UV-B 저선량 상태인 비닐하우스 재배 잎들깨에 UV-B를 처리하여 수량과 생리변화를 검토하고자 수행하였다. UV-B는 형광램프를 이용하여 잎들깨 수관으로부터 150, 120, 90 cm에서 처리하여 각각 노지자연량, 노지자연량+50% 증가량, 노지자연량+100% 증가량을 다른 광의 간섭이 없는 밤 9시부터 2시간씩 30일간 처리하였다. UV-B 처리에 의한 잎들깨 수량은 비닐하우스, 노지 자연량 150% 증가량, 노지자연량, 노지자연량+100% 증가량 처리 순이었다. 7월 20일의 자연 UV-B 선량은 노지 13.6 kJ/일, 비닐하우스 4.9 kJ/일로 비닐하우스의 UV-B 선량은 노지보다 64% 감소되는 것으로 측정되었다. UV-B가 균일하게 처리된 잎들깨의 단백질을 추출하여 2차원전기영동으로 분리하고, 이미지분석하여 발현량을 분석한 결과 UV-B에 의해 33개의 단백질 발현이 변화되었으며 이중 10개가 동정되었다. 동정된 단백질의 기능별로 분류하면 광합성과 관련된 것이 40%, 스트레스 및 스트레스 방어와 관련된 것이 60%였다. UV-B에 의해 발현이 감소한 단백질은 광합성과 관련된 ATP synthase CF1 alpha chain이었고 발현이 증가한 단백질은 DNA recombination and repair protein recF, Heat shock protein 21, Catalase, Galactinol synthase, S-adenosyl-L-methionine, Calcium-dependent protein kinase(CDPK)-like 로 주로 스트레스 및 스트레스 방어와 관련된 단백질들로서 UV-B는 잎들깨 세포내 DNA와 광합성기구를 손상시켜 광합성에 저해적으로 작용하여 수량을 감소시킨 것으로 판단되었다.

• 동의생리병리학회지
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• 제17권1호
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• pp.241-246
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• 2003

Bambusae Caulis in Liquamen(BCL) has been used to relieve the cough and asthma, and remove the phlegm in traditional Oriental medicine. In recent years, it was studied for its antiinflammatory, antiallergenic, immune-modulating, and anticarcinogenic capabilities. This experiment was performed to evaluate the microarray profiles of CD genes in human mast cells before and after BCL treatment. The results are as follows: The expression of 51 of the genes studied was up-regulated in the Bel-treated group; they include the genes coding L apoferritin, beta-2-microglobulin, ferritin light polypeptide, CD63, monocyte chemotactic and activating fact, heme oxygenase 1, CD140a, integrin alpha M, colony stimulating factor 2 receptor, eukaryotic translation elongation factor, CD37, interleukin 18, NADH dehydrogenase 1 beta, CD48, 5-lipoxygenase activating protein, interleukin 4, ribosomal protein L5, GABA(A) receptor-associated protein, beta-tubulin, integrin beta 1, CD162, CD32, lymphotoxin beta, alpha-tublin, integrin alpha L, CD2, CD151, CD331, 90 kDa heat shock protein, CD59, CD3Z, microsomal glutathione S-transferase 2, CD33, CD162R, cyclophilinA, CD84, interleukin 9 receptor, interleukin 11, CD117, CD39-Like 2, and so forth. The expression of 7 of the genes studied was down-regulated in the BCL-treated group; they include the genes coding con, CD238, SCF, CD160, CD231, CD24, and CD130. Consequently, the treatment of BCL on the human mast cells increased the expression of 51 genes and decreased the expression of 7 genes. These data would provide a fundamental basis to the traditional applications of Bambusae Caulis in Liquamen.

• 생명과학회지
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• 제23권4호
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• pp.586-594
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• 2013

단백질 번역 후 O-GlcNAc 수식은 단백질 조절의 새로운 기전으로 대두되고 있다. 전통적인 당수식과 달리 O-GlcNAc 수식은 단 한번의 O-GlcNAc 전달로 이루어지며, 핵 및 세포질단백질 모두에 수식될 수 있다. O-GlcNAc은 이 분자를 끝으로 하는 최종수식으로 생각되어 왔으나, 최근의 논문(J Proteome Res. 2011 10:2725-2733)은 AP180 단백질에 O-GlcNAc-P가 존재함을 보고하였다. 이 논문은 O-GlcNAc-P가 일반적인 단백질수식인지에 대한 중요한 질문을 던진다. 이에 답하고자 저자들은 HEK293T 세포에 O-GlcNAc 인산화효소 NAGK를 DsRed2에 연결한 DsRed2-$NAGK_{WT}$ 혹은 효소활성이 없는 돌연변이 NAGK를 표현하는 DsRed2-$NAGK_{D107A}$를 표현시키고, 단백질 추출물을 얻어 2D-PAGE로 분리한 후 인산화 정도를 측정하여, $NAGK_{WT}$에 의하여 인산화가 증가되는 15개의 단백질 스폿을 선별하였다. 이 가운데 7개 스팟을 동정한 결과 2개의 스폿은 O-GlcNAc 수식 단백질인 $HSP90{\beta}$, 다른 2개의 스폿도 O-GlcNAc 수식 단백질인 ENO1로 동정되었으며, 나머지(dUTP nucleotidohydrolase mitochondrial isoform 2, glutathione S-transferase P, grp94)는 O-GlcNAc 수식 여부를 아직 모르는 단백질이였다. NAGK에 의하여 O-GlcNAc 단백질의 인산화가 증가된다는 사실은 O-GlcNAc이 인산화되어 O-GlcNAc-P로 수식됨을 시사하며, 따라서 본 연구의 결과는 O-GlcNAc이 최종 수식이 아님을 지지한다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권5호
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• pp.848-855
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• 2020

Objective: Shearing is one of the practices that is applied periodically to fiber producing animals, which can also alter resistance of animals to high temperatures in especially summer months. This study aimed to investigate effects of shearing on some physiological and hormonal parameters in Akkaraman sheep during summer season. Methods: This study was carried out on 39 non-pregnant Akkaraman ewes (aged 1.5 years at the beginning of experiment). The 39 ewes were chosen randomly from the flock belonging to the Erciyes University and they were assigned to two groups as follows: i) group A (n = 20) designed as the control group, they were shorn and group B (n = 19) designed as the experimental group, they were unshorn. Prior to the shearing (-1 day) and on days 1, 7, 15, 30, 45, 60, 75, and 90 following the shearing, blood samples were taken from the vena jugularis of each sheep. Cortisol, β-endorphin, growth hormone (GH), thyroxine (T₄), triiodothyronine (T₃), and heat shock protein 70 (HSP-70) concentrations were determined using the enzyme immunoassay method. Body weight (BW), rectal temperature (RT), pulse rate (PR), and respiratory rate (RR) of each sheep were recorded at the same time. The data obtained were analyzed using two-way repeated measures analysis of variance. Results: Statistical analysis showed a significant effect of shearing×period interaction (p<0.01) and a significant effect of period (p<0.01) on BW, HSP-70, cortisol, T₄ and RT, PR, GH, β-endorphin, T₃, respectively. Also these analysis showed no significant effect of shearing×period interaction or period on RR. Conclusion: The results showed that the thermoregulation abilities of sheep were affected by shearing treatment and the shorn ewes were less affected by heat stress. In conclusion, based on the data of this study, shearing can be considered as a necessary management practice that requires protection for sheep from the effect of heat stress.