• Molecular & Cellular Toxicology
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• 제4권4호
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• pp.348-354
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• 2008

Bisphenol A (BPA), an environmental endocrine disrupter, enters the human body continuously in food and drink. Young children are likely to be more vulnerable than adults to chemical exposure due to the immaturities of their organ systems, rapid physical development, and higher ventilation, metabolic rates, and activity levels. The direct effect of BPA on peripheral tissue might also be of importance to the development of insulin resistance. However, the influence that BPA has on insulin signaling molecules in skeletal muscle has not been previously investigated. In this study, we examined the effect of BPA on fasting blood glucose (FBG) in post-weaned Wistar rats and on insulin signaling proteins in C2C12 skeletal muscle cells. Subsequently, we investigated the effects of BPA on insulin-mediated Akt phosphorylation in C2C12 myotubes. In rats, BPA treatment (0.1-1,000 ng/mL for 24 hours) resulted in the increase of FBG and plasma insulin levels, and reduced insulin-mediated Akt phosphorylation. Furthermore, the mRNA expression of insulin receptor (IR) was decreased after 24 hours of BPA treatment in C2C12 cells in a dose-dependent manner, whereas the mRNA levels of other insulin signaling proteins, including insulin receptor substrate-1 (IRS-1) and 5'-AMP-dependent protein kinase (AMPK), were unaffected. Treatment with BPA increased GLUT4 expression and protein tyrosine phosphatase 1B (PTP1B) activity in C2C12 myotubes, but not in protein levels. We conclude that exposure to BPA can induce insulin resistance by decreasing IR gene expression, which is followed by a decrease in insulin- mediated Akt activation and increased PTP1B activity.

• Reproductive and Developmental Biology
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• 제34권2호
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• pp.89-94
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• 2010

Melatonin is induced by light information through the retina and leads to growth factor activation. Thus, we investigated the effects of melatonin by controlling the photoperiod of growing young rats. Male Sprague-Dawley rats (n=6; 4 weeks old) were divided into two experimental groups: the L/D group (normal photoperiod; light/dark: 12/12 h; lights on at 9:00 a.m.) and the L/L group (light/light: 24 h). Rat body weight and food consumption were measured daily for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of ketamine (50 mg/kg) and xylazine (10 mg/kg) and sacrificed. Tissue was then collected for RNA isolation (from brain, heart, liver, kidney, adrenal gland, testis, tibia, hind limb muscles). Also, serum was isolated from blood using a centrifugal separation. The L/L group had significantly lower body weight than the L/D group from 4 to 6 weeks (p<0.05). The L/D group had increased tissue mass, compared with the L/L group, but the difference was not statistically significant. The L/D group had a significantly higher melatonin concentration than the L/L group between the hours of midnight and 2:00 a.m (p<0.01). These results indicate that photoperiod length may affect the secretion of melatonin from the pineal gland. Also, the reduction of nocturnal melatonin secretion may retard the development of growing young rats. In future studies, we plan to compare exogenous melatonin administration with endogenous melatonin concentration induced by photoperiod control. Moreover, we will confirm whether the effects seen in pathological animal models can be reversed by controlling the photoperiod.

• 한국식품위생안전성학회지
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• 제33권3호
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• pp.207-213
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• 2018

본 연구는 까마귀쪽나무열매추출물(LJF-HE)의 유전독성을 평가하고자 하였다. 유전독성연구는 OECD와 MFDS(Korea Ministry of Food and Drug Safety) 지침에 따라 복귀돌연변이시험, 염색체이상시험, 마우스 골수세포를 이용한 소핵시험을 실시하였다. 세균을 이용한 복귀돌연변이시험은 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 S9 mix 존재유무에 상관없이 복귀돌연변이 콜로니 수는 음성 대조군과 비교하였을 때 증가 양상을 나타내지 않은 반면에 양성 대조물질에서 유발된 복귀돌연변이 콜로니 수는 대사활성계 미적용(S9-) 및 적용(S9+)의 모든 시험 균주에 대하여 음성(용매)대조 값의 2배를 넘어 증가한 것으로 나타났다. 염색체 이상 시험에서 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 모든 세포주의 처리시간 및 S9 mix 존재유무에 상관없이 5%미만의 비정상적인 염색체이상을 나타내었으나, 음성대조군에 비해 유의적인 변화는 없었다. 소핵시험은 까마귀쪽나무열매추출물(LJF-HE) 처리군에서 음성 대조군과 비교하여 소핵을 가진 다염성 적혈구의 증가는 볼 수 없었으며 통계학적인 유의성도 나타나지 않았다. 상기의 결과를 종합하면 까마귀쪽나무열매추출물(LJF-HE)은 유전독성을 유발하지 않는 것으로 판단되어진다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권8호
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• pp.1366-1372
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• 2018

Objective: A disintegrin and metallopeptidase with thrombospondin motifs type 8 (ADAMTS8) is crucial for diverse physiological processes, such as inflammation, tissue morphogenesis, and tumorigenesis. The chicken ADAMTS8 (chADAMTS8) gene was differentially expressed in the kidney following exposure to different calcium concentrations, suggesting a pathological role of this protein in metabolic diseases. We aimed to examine the molecular characteristics of chADAMTS8 and analyze the gene-expression differences in response to toll-like receptor 3 (TLR3) stimulation. Methods: The ADAMTS8 mRNA and amino acid sequences of various species (chicken, duck, cow, mouse, rat, human, chimpanzee, pig, and horse) were retrieved from the Ensembl database and subjected to bioinformatics analyses. Reverse-transcription polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR) experiments were performed with various chicken tissues and the chicken fibroblast DF-1 cell line, which was stimulated with polyinosinic-polycytidylic acid (poly[I:C]; a TLR3 ligand). Results: The chADAMTS8 gene was predicted to contain three thrombospondin type 1 (TSP1) domains, whose amino acid sequences shared homology among the different species, whereas sequences outside the TSP1 domains (especially the amino-terminal region) were very dif­ferent. Phylogenetic analysis revealed that chADAMTS8 is evolutionarily clustered in the same clade with that of the duck. chADAMTS8 mRNA was broadly expressed in chicken tissues, and the expression was significantly up-regulated in the DF-1 cells in response to poly(I:C) stimulation (p<0.05). These results showed that chADAMTS8 may be a target gene for TLR3 signaling. Conclusion: In this report, the genetic information of chADAMTS8 gene, its expression in chicken tissues, and chicken DF-1 cells under the stimulation of TLR3 were shown. The result suggests that chADAMTS8 expression may be induced by viral infection and correlated with TLR3-mediated signaling pathway. Further study of the function of chADAMTS8 during TLR3-dependent inflammation (which represents RNA viral infection) is needed and it will also be important to examine the molecular mechanisms during different regulation, depending on innate immune receptor activation.

• 농약과학회지
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• 제15권4호
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• pp.485-489
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• 2011

Bacillus amyloliquefaciens strain EXTN-1 처리에 의해 생육촉진 효과와 함께 광범위한 식물 병 방제효과가 보고되었다. EXTN-1의 PGPR 효과는 생육초기에 PR-1a, PDF1.2 등의 저항성 관련 유전자 발현에 의한 oxidative burst의 증가나 SA, JA나 ethylene 대사에 의한 유도저항성의 발현에 기인한다. 이 연구의 목표는 B. amyloliquefaciens EXTN-1가 기존에 보고된 다른 작물의 경우에서와 마찬가지로 벼의 생육촉진이나 벼줄무늬잎마름병에 대한 저항성에 관여하는지를 확인하기 위해 수행되었다. 벼 종자를 B. amyloliquefaciens EXTN-1에 침지한 후 파종하였을 때 생육촉진 효과와 병에 대한 저항성 발현이 확인되었다. B. amyloliquefaciens EXTN-1을 처리한 30일묘에서 벼의 초장, 생물중, 뿌리길이는 무처리구에 비해 각각 12.6%, 9.8%, 16.0% 증가하여 PGPR 효과가 나타남을 확인할 수 있었다. RSV 접종구에서도 B. amyloliquefaciens EXTN-1 20일묘는 초장, 생물중, 뿌리길이는 무처리구에 비해 각각 12.6%, 9.8%, 16.0% 증가하였다. 유도저항성 발현효과는 감수성 품종에서 저항성 품종에서 상대적으로 뚜렷하게 나타났다.

• 한국식품저장유통학회지
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• 제13권4호
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• pp.524-529
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• 2006

곡류분말의 위생화 및 물리적 특성 개선을 위해 감마선 조사기술의 이용 가능성이 높아짐에 따라 이들의 안전성을 확보할 목적으로 30 kGy의 고선량 감마선조사 곡류분말의 유전독성학적 안전성 평가를 실시하였다. 감마선 조사 및 비조사 곡류분말의 S. typhimurium TA 98, TA100, TA1535 및 TA1537에 대한복귀변이 집락수를 조사한 결과, 대사활성계 도입 및 부재시 모두 시험적용 농도인 0.625-10 mg/plate의 범위에서 복귀변이 집락수의 농도 의존적인 증가 혹은 감소를 보이지 않아 감마선 조사 곡류분말(30 kGy)은 돌연변이원성이 없는 것으로 나타났다. 또한, 설치류 망상적혈구를 이용하여 감마선조사 곡류분말의 소핵 형성 시험을 수행한 결과, 30 kGy 감마선 조사 곡류분말은 시험적용 용량인 625-5,000 mg/kg의 범위에서 소핵을 가진 망상 적혈구의 출현율이 음성대조군과 유의한 차이를 나타내지 않아 소핵을 유발하지 않음을 확인하였다. 포유류 배양세포를 이용한 염색체 이상시험에서도 30 kGy 감마선 조사곡류분말은 시험적용 용량에서 염색체이상 유발능이 5% 미만이었다.

• 한국식품저장유통학회지
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• 제9권4호
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• pp.429-433
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• 2002

Candida sp. L-16 균주가 생산하는 D-xylulokinase는 배양균체를 초음파 파쇄한 조효소액으로 하여 황산암모늄 염석, DEAE-cellulose, chromatography, Sephadex G-100과 Sephadex G-200 gel filtration 과정으로 정제하여 최종 수율 11.2%로 약 23.2배 정제하였다. 정제 효소의 분자량은 SDS-PAGE로 분석한 결과 분자량은 75,000 dalton으로, Sephadex G-200겔 여과에 의해 150,000 dalton으로 나타나 dimer로 확인되었다. 효소 활성에 미치는 최적 반응 온도는 4$0^{\circ}C$로 나타났고, 온도안정성은 비교적 불안정하여 3$0^{\circ}C$이상에서는 빠르게 실활되었다. 정제 효소의 최적 반응 pH는 pH 8.0이었고, pH 7.0에서 pH 9.0 사이에서 비교적 효소활성이 높았다. 본 효소는 D-xylulose, D-arabinose, D-ribose등에서는 높은 기질특이성을 가지고 있었으나 D-xylose, D-glucose, L-arabinose 등은 기질로서 작용하지 못하였다. 정제 효소의 활성화 에너지값(Ea)은 $25^{\circ}C$ 내지 4$0^{\circ}C$$^{\circ}C$의 온도 범위에서 4.75kcal/mol이었다. 효소의 활성화제로는 EDTA, cysteine-HCl, DTT, glutathione 등이 존재하며 억제제로는 6-phosphogluconic acid, 2-koeto-gluconic acid 등으로 나타났다.

• Journal of Ginseng Research
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• 제44권1호
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• pp.58-66
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• 2020

Background: The biological and pharmacological effects of BST204, a fermented ginseng extract, have been reported in various disease conditions. However, its molecular action in metabolic disease remains poorly understood. In this study, we identified the antiadipogenic activity of BST204 resulting from its inhibition of the S6 kinase 1 (S6K1) signaling pathway. Methods: The inhibitory effects of BST204 on S6K1 signaling were investigated by immunoblot, nuclear fractionation, immunoprecipitation analyses. The antiadipogenic effect of BST204 was evaluated by measuring mRNA levels of adipogenic genes and by chromatin immunoprecipitation and quantitative real-time polymerase chain reaction analysis. Results: Treatment with BST204 inhibited activation and nuclear translocation of S6K1, further decreasing the interaction between S6K1 and histone H2B in 10T1/2 mesenchymal stem cells. Subsequently, phosphorylation of H2B at serine 36 (H2BS36p) by S6K1 was reduced by BST204, inducing an increase in the mRNA expression of Wnt6, Wnt10a, and Wnt10b, which disturbed adipogenic differentiation and promoted myogenic and early osteogenic gene expression. Consistently, BST204 treatment during adipogenic commitment suppressed the expression of adipogenic marker genes and lipid drop formation. Conclusion: Our results indicate that BST204 blocks adipogenesis of mesenchymal stem cells through the inhibition of S6K1-mediated histone phosphorylation. This study suggests the potential therapeutic strategy using BST204 to combat obesity and musculoskeletal diseases.

• 한국식품저장유통학회지
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• 제24권1호
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• pp.100-106
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• 2017

본 연구는 환자용 균형영양식으로 제조하여 감마선 조사처리를 통해 살균한 모델식품의 유전독성 여부를 평가하기 위해 수행되었다. 4 kGy 이상의 흡수선량으로 감마선 조사 처리한 시료에서 생균수가 1 log CFU/g의 검출한계 이하로 나타났기 때문에 살균을 위한 조사처리 선량으로 4 kGy를 설정하였다. 복귀돌연변이 유발성 평가 결과, 모델식품의 열수 추출물과 메탄올 추출물은 처리한 용량과 대사활성계 존재 여부와는 상관없이 음성대조구(멸균증류수 또는 DMSO)와 유사한 수준의 복귀돌연변이 발생빈도를 나타내었다. 염색체 이상 시험에서 실험군들의 정상 염색체의 수는 음성대조군과 유사하였으며, 용량 및 대사활성계에 대한 비의존성을 나타내어 염색체 이상 유발능은 없는 것으로 판단하였다. 또한 최고 2,000 mg/kg 체중의 농도로 모델식품을 경구 투여한 마우스의 골수세포에서도 소핵 다염성적 혈구의 발생빈도는 음성대조군에 비해 유의적인 차이를 나타내지 않았다. 따라서 4 kGy의 흡수선량으로 감마선 조사처리한 환자용 균형영양식은 본 실험 조건하에서 유전독성을 나타내지 않는 것으로 사료된다.

• Journal of Applied Biological Chemistry
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• 제57권2호
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• pp.103-111
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• 2014

A1E is an extract from traditional Asian medicinal plants that has therapeutic activities against cancers, metabolic disease, and other intractable conditions. However, its mechanism of action on cervical cancer has not been studied. In order to ascertain if A1E would have pronounced anti-cervical cancer effect, cervical cancer cells were incubated with A1E and apoptosis was detected by nuclear morphological changes, annexin V-FITC/PI staining, cell cycle analysis, western blotting, Reverse-transcription polymerase chain reaction, and measurement of mitochondrial membrane potential. Expression of human papiloma virus E6 and E7 oncogenes was down-regulated in A1E-treated cervical cancer cells, while p53 and retinoblastoma protein levels were enhanced. A1E also perturbed cell cycle progression at sub-G1 and altered cell cycle regulatory factors in SiHa cervical cancer cells. A1E activated apoptotic intrinsic pathway markers such as caspase-9, caspase-3 and poly ADP-ribose polymerase, and down-regulated expression of Bcl-2 and Bcl-xl. A1E induced mitochondrial membrane potential collapse and cytochrome c release, and inhibited phosphatidylinositol 3-kinase (PI3K)/Akt, key factors involved in cell survival signaling. Taken all these results, A1E induced apoptosis via activation of the intrinsic pathway and inhibition of the PI3K/Akt survival-signaling pathway in SiHa cervical cancer cells. In conclusion, A1E exerts anti-proliferative action growth inhibition on cervical cancer cells through apoptosis which demonstrates its anti-cervical cancer properties.