• 한국수정란이식학회지
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• 제26권4호
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• pp.265-270
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• 2011

Procymidone is a fungicide with anti-androgenic properties widely used to protect fruits from fungal infection, which induces an excessive reactive oxygen species production in male reproductive organs. In this study, to clarify whether procymidone affect the cellular antioxidant system of prostate at onset of puberty, gene expression patterns of the representative antioxidant enzymes such as cytoplasmic glutathione peroxidase (GPx1), phospholipid hydroperoxide GPx (PHGPx), selenoprotein P (SePP), cytoplasmic copper/zinc superoxide dismutase (SOD1), and manganese SOD (SOD2) were investigated in the rat ventral prostates exposed to procymidone using real-time RT-PCR analyses. Seven-week-old Sprague-Dawley rats castrated at 6 weeks old were treated with procymidone (25, 50, or 100 mg/kg per day) orally for 7 consecutive days after testosterone propionate (0.4 mg/kg per day) administration by subcutaneous injection. As compared to normal control animals, GPx1 mRNA expression in prostates significantly increased by the administration with TP and/or procymidone. However, PHGPx and SOD1 mRNA levels significanatly decreased by over 25 mg/kg of procymidone treatment and SePP and SOD2 mRNA levels was significanatly reduced by over 50 mg/kg of procymidone treatment. These findings indicate that procymidone may affect the antioxidant system of prostatic cells in up-regulation mode of GPx1, but in down-regulation modes of PHGPx, SePP, SOD1, and SOD2, suggesting that procymidone may affect differently the cellular antioxidant system of prostate according to the exposure doses.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2000년도 춘계임시총회 및 학술발표대회
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• pp.52-56
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• 2000

느타리 버섯의 다양한 품종과 재배형태가 급변히 요구되는 시기에 충남농업기술원 버섯연구팀의 5년 (1996～2,000)에 걸쳐 국,내외 수집 20 균주를 공시하여 개발 육성한 청풍과 명월 신품종 느타리버섯을 현재 재배되고 있는 기존 느타리품종과 동일여부를 위해 PCR-RAPD분석과 배양적, 재배적 특성을 인공재배 시험한 결과를 요약하면 다음과 같다. 1. 청풍, 명월느타리버섯 균사배 양 온도는 청 풍은 20～$25^{\circ}C$,명 월은 25～3$0^{\circ}C$, 이고 배지는 PDA배지가 양호하고 균사생육은 청풍, 명월 모두 비슷하였으며 배지 pH는 6.5～7.5 였다. 2. 버섯재배가능은 청풍은, 5～26$^{\circ}C$ 명 월은, 7～28$^{\circ}C$로 온도범위가 넓어 재배가 용이하나 적정 생육온도는 청풍, 명월 공히 15～19$^{\circ}C$가 양호 하였다 3. 청풍과 명월의 배양온도 23$^{\circ}C$에서 균사배양기간 23～24일 (850cc/pp),폐면 균상 재배에서 균배양 기간은 청풍, 명월 공히 11～12일, 초발이 소요일수는 2～3일 소요 되었다. 4.초발이시 청풍과 명월 갓 색택은 진청색이고, 생육후기 청풍은 회갈색, 명월은 진회색으로 조직이 치밀하고 저장성이 양호하여 경제성이 기대된다. 5. 다발수는 춘추2호 (13개/다발) 대 비 청 풍11개, 명월12개로 약간 적었으나 개체중은 춘추2호(14g/개) 대비 청풍 16g/개, 명월 14g/개로 비슷한 경향 이었다. 6. 수량성은 균상 폐면재배시 춘추2호 (40kg/3.3$m^2$)대비 청풍 느타리버섯은 (46kg/3.3$m^2$) 15%증수되고, 명월느타리버섯은(41kg/3.3$m^2$)로 3% 증수되었으며 병재배 겸용도 가능하였다. 7. PDA대치배양과 PCR-RAPD시험에서 청풍, 명월 신품종느타리 버섯은 기존 재배 되고 있는 느타리품종과 다른 품종으로 확인 되었다.

• Plant Resources
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• 제3권2호
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• pp.105-109
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• 2000

This study was carried out to find a useful mushroom at Chungnam Agricultural Research And Extention Service. Twenty materials used were collected from domestic and exotic area. These races were compared bontanical characteristics to leading varieties by PCR-RAPD methods. Mycelial growth temperature of Chongpung and Myongwol were at 20 to 25 $^{\circ}C$ and 25 to 30 $^{\circ}C$ at PDA medium, respectively mycelial growth of these varieties were similiar at pH 6.5 to 7.5. In case of mushroom cultivation temperature ranges, Chongpung was at 5 to 26$^{\circ}C$ and Myongwol was at 7 to 28$^{\circ}C$, but the optimum temperature range of these were appeared at 15 to 19$^{\circ}C$. Culture temperature of these was 23$^{\circ}C$ and period of mycelial culture was needed 23 to 24 days under 850cc/pp, while was needed 11 to 12 days at waste cotton medium. Cap color of these at first inducing mushroom was all dark blue, but at late growing stages Chongpung was shown as grey, and Myongwol was shown as dark grey. Yield of Chongpung was appeared as 46kg/3.3$m^2$ and that of Myongwol was 41kg /3.3$m^2$, while Chunchu No2 as check was 40kg/3.3$m^2$. Results from PDA medium and PCR-RAPD analysis two of these were different from others.

• 대한본초학회지
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• 제27권6호
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• pp.55-61
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• 2012

Objectives : Allium Hookeri (AH) is a traditional herb to treat inflammatory diseases in India and Myanmar. Recently, AH cultivation was succeeded in South Korea. This study was performed to evaluate the anti-inflammatory effects of Korean AH in RAW264.7 cells, mouse macrophage cell line. Methods : To evaluate the anti-inflammatory effects of root of AH, we prepared the 70% ethanol extract, then we examined the productions of nitrite, and pro-inflammatory cytokines. To examine the nitrite, and cytokines, the RAW264.7 cells were treated with AH, then stimulated with lipopolysaccharide (LPS, 500 ng/ml) for 24 h. Then the cells were harvested for griess assay, ELISA and real-time reverse transcription polymerase chain reaction (RT-PCR). Also to detect the ability of AH to induce heme oxygenase-1 (HO-1), we examined the HO-1 expression using real time RT-PCR and western blot. Furthermore, we examined the mitogen activated-protein kinases (MAPKs) and nuclear factor kappa B (NF-${\kappa}B$) activation to find out the underlying mechanisms. Results : AH ethanol extract significantly inhibited the productions of nitrite and interleukin (IL)-$1{\beta}$. AH treatment increased the HO-1 expression dramatically at 1 h, then peaked at 3 h. When the HO-1 was inhibited by tin (Sn) protoporphryin-IX (SnPP), the anti-inflammatory action of AH was reversed. AH treatment inhibited the activation of p38, but not extracelluar signal-regulated kinase (ERK 1/2) and c-Jun $NH_2$-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-$B{\alpha}$) in the LPS-stimulated RAW 264.7 cells. Conclusions : These data could suggest that AH exerts anti-inflammatory influences through up-regulation of HO-1 and deactivation of p38.

• 대한본초학회지
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• 제37권5호
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• pp.83-88
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• 2022

Objectives : Jakyakgamcho-tang (JGT) has been traditionally used to treat muscular convulsion and pain in South Korea. According to recent studies, JGT has been reported to have anti-depression, anti-inflammation, anti-oxidative, anti-diabetics, anti-spasm and analgesic effects, but studies on its anti-neuroinflammatory and neuroprotective effect have not been deeply conducted. Thus, we investigated the anti-neuroinflammatory activity of JGT on lipopolysaccharide (LPS)-stimulated mouse microglia cells. Methods : To investigate the anti-neuroinflammatory effects of JGT on BV2 microglial cells, we examined the production of nitric oxide (NO) using griess assay, and mRNA expressions of pro-inflammatory cytokines such as interleukin (IL)-1𝛽, IL-6, and tumor necrosis factor (TNF)-𝛼 using real time RT-PCR. Furthermore, to determine the regulating mechanisms of JGT, we investigated the heme oxygenase (HO)-1 by real time RT-PCR. Results : Pre-treatment of JGT effectively decreased NO production in LPS-stimulated BV2 cells at concentrations without cytotoxicity. Additionally, JGT significantly suppressed the production of IL-1𝛽, IL-6, and TNF-𝛼 in LPS-stimulated BV2 cells. Furthermore, JGT activated the HO-1 expression, which is one of the immunomodulatory signaling molecules. And the abolishment of HO-1 by tin protoporphyrin IX (SnPP, the HO-1 inhibitor) reversed the anti- inflammatory activity of JGT in LPS-stimulated BV2 cells. Conclusions : Our results suggest that the JGT has anti-neuroinflammatory effect through the activation of HO-1 in LPS-stimulated BV2 cells. Thereby, JGT could expected to be used for the prevention and treatment of neurodegenerative disease related to neuroinflammation.

• 원예과학기술지
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• 제34권3호
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• pp.453-460
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• 2016

The parentage of the horticulturally important pear cultivar 'Niitaka' was confirmed by determining its S-genotypes based on the S-RNase and $PpSFBB^{-{\gamma}}$ genes, and genotyping using simple sequence repeat (SSR) markers. Previous reports suggested that the cultivars 'Amanogawa' and 'Imamuraaki' were the parents of 'Niitaka', although the cultivars 'Chojuro' and 'Shinchu' were also examined as candidate parents, along with two other cultivars. In the present study, the S-genotype of 'Niitaka' was determined to be $S^3S^9$. The $S^9$-RNase of 'Niitaka' was found to be likely inherited from the parent 'Amanogawa' ($S^1S^9$) and the $S^3$-RNase from 'Chojuro' ($S^3S^5$) or 'Shinchu' ($S^3S^5$). Based on the S-genotypes, the cultivar 'Imamuraaki' ($S^1S^6$) had no contribution to the parentage of 'Niitaka' ($S^3S^9$). A total of 67 polymorphic SSR markers were used to further confirm the parentage of 'Niitaka'. Discrepancies were found at several SSR loci between 'Niitaka' and the cultivars 'Imamuraaki' and 'Shinchu', whereas 'Niitaka' inherited alleles from 'Amanogawa' and 'Chojuro' at all SSR loci. Therefore, our findings established that 'Amanogawa' and 'Chojuro' are the parents of pear cultivar 'Niitaka', and not 'Imamuraaki' as previously reported.

• 농약과학회지
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• 제11권4호
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• pp.320-330
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• 2007

충북 진천군 이월면 장미재배지와 경남 밀양시 고추밭에서 채집한 담배가루이에 대해서 16S DNA 염기서열을 분석하였다. 각각의 PCR 산물을 EcoT14 I (Sty I)과 Stu I을 처리한 결과 경남 밀양시 고추밭에서 채집한 담배가루이는 Sty I 에서 555pp와 311bp 두 개의 단편이 만들어져 biotype Q임을 확인하였고, 충북진천군 장미재배지에서 채집한 담배가루이는 Stu I에서 560bp와 3060p 두 개의 단편이 만들어서 biotype B임을 확인하였다. 두 biotype에 대한 12종 살충제로 발육단계별 약제감수성, 침투이행성 및 잔효성을 비교하였고, esterase, acetylcholinesterase (AChE), glutathione S-transferase 등의 효소활성에 미치는 저해정도를 검토하였다. 두 biotype간에 약제감수성 차이가 있었고, biotype B가 Q보다 더 감수성이었다. 엽면침투이행성과 근부침투이행성 및 잔효성에서도 biotype B가 Q보다 더 감수성이었다. 유기인계인 fenitrothion과 카바메이트계인 fenothiocarb의 저해제에 대해서 esterase, acetylcholinesterase (AChE), glutathione S-transferase등의 효소활성에 미치는 영향을 검토한 결과, biotype Q가 B보다 활성이 높게 나타났다. 이상의 결과를 종합하여 볼 때 담배가루이 biotype Q가 B보다 약제에 대해서 저항성임을 알 수 있었다.

• KSBB Journal
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• 제22권1호
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• pp.16-21
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• 2007

Cellobiohyolase (CBH) I 유전자의 확보는 CBH I 유전자 cellulase 생산 균주인 Trichoderma reesei를 배양, 수거하고 액체 질소를 이용하여 세포를 파쇄 후 RT-PCR kit를 이용하여 CBH I gene을 합성하였다. 그 후 발현벡터인 pYGAL에 cloning하였다. CBH I 유전자 앞부분에는 CBH I 단백질이 cell 외부로 분비할 수 있도록 하는 ppL이 포함되었다. CBH I 단백질 발현은 Protein gel 결과를 통하여 발현을 확인하였다. Cellulase 활성을 증대시키기 위한 분자진화 방법 개발은 error prone PCR과 DNA shuffling을 수행하였다. 얻은 CBH I 유전자를 발현 벡터에 삽입하고 효모에 transformation하여 이것을 다시 screening하였다. 1차 screening 후 confirm test하기 위해 DNS (Dinitrosalicylic Acid) 환원당 측정법을 이용하였으며, 이 결과 121-D8, 228-G2, 389-E3, 412-B4, 456-D2의 cellulase 변이체를 획득할 수 있으며, 456-D2의 경우 original CBH I과 비교하여 약 510%의 활성이 증가된 것을 확인할 수 있었다. 분자 진화 된 cellulase sequence 분석결과 CBH I wild type과 비교하였을 때 121-D8의 경우 변경된 9개의 염기 중 아미노산의 변화에 영향을 준 염기는 6개, 228-G2의 경우 변경된 7개의 바뀐 염기 중 아미노산의 변화에 영향을 준 염기는 4개, 389-E3의 경우 변경된 13개의 바뀐 염기 중 아미노산의 변화에 영향을 준 염기는 9개, 412-B4의 경우 변경된 9개의 바뀐 염기 중 아미노산의 변화에 영향을 준 염기는 6개, 456-D2의 경우 변경된 10개의 바뀐 염기 중 아미노산의 변화에 영향을 준 염기는 7개이었다. Cellulase 생산 공정 최적화는 5 L 발효기를 이용하여 고농도 배양을 실험한 결과 최종 O.D. 120까지 진행할 수 있었으며, 약 1.2 g/L의 cellulase를 얻을 수 있었다. Cellulase 생산 공정 scale-up 5 L 규모에서 확립된 최적 fed-batch 발효 공정을 300 L로 scale-up하여 실험하였다. 최종 O.D.는 약 280 정도이며 cellulase의 발현양은 약 3.2 g/L 수준임을 확인하였다. Cellulase 정제 공정 최적화 결과 80%의 수율과 95%의 순도를 확보하였다.

• 대한예방한의학회지
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• 제22권1호
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• pp.107-127
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• 2018

Objectives : The present study investigated the anti-obese and blood flow improvement activities of aqueous extracts of ginseng berry (GBe) on the mild diabetic obese mice as compared with metformin. Methods : After end of 56 days of continuous oral administrations of GBe 150, 100 and 50 mg/kg, or metformin 250 mg/kg, anti-obese and blood flow improvement effects - the changes of body weights, body and abdominal fat density by in live dual-energy x-ray absorptionmetry (DEXA), tail bleeding time, prothrombin time (PT), activated partial thromboplastin time (aPTT), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high density lipoprotein (HDL) levels, aorta and serum cyclic guanosine monophosphate (cGMP), nitric oxide (NO) and endothelin (ET)-1 levels, aorta phosphorylated PI3K (pPI3K), phosphorylated Akt (pAkt) and phosphorylated p38 MAPK (pp38 MAPK) levels were systemically analyzed. In addition, aorta vascular dilation and constriction related gene mRNA expressions - PI3K, Akt, eNOS, p38 MAPK and ET-1 were also analyzed by realtime RT-PCR. Results : The obesity and related blood flow impairment, induced by 84 days of continuous HFD supply, were significantly inhibited by 56 days of continuous oral treatment of GBe 150, 100 and 50mg/kg, dose-dependently, and they also dramatically normalized the changes of the aorta vascular dilation and constriction related gene mRNA expressions, also dose-dependently. Especially, GBe 150 mg/kg constantly showed favorable inhibitory activities against type II diabetes related obesity and vascular disorders through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities, as comparable to those of metformin 250 mg/kg in HFD mice. Conclusion : By assessing the key parameters for anti-obese and blood flow improvement activities on the HFD-induced mild diabetic obese mice, the present work demonstrated that GBe 150, 100 and 50 mg/kg showed favorable anti-obese and blood flow improvement effects in HFD-induced type II diabetic mice, through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities.

• Asian Pacific Journal of Cancer Prevention
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• 제16권14호
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• pp.5835-5842
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• 2015

Background: Melatonin, which is mainly produced by the pineal gland, has a good inhibitory effect on cell growth of multiple cancer types. However, the underlying molecular mechanisms of anti-tumor activity for colon cancer have not been fully elucidated. In this study, we investigated the effects of melatonin on migration in human colon cancer RKO cells and the potential molecular mechanisms. Materials and Methods: The viability of RKO cells was investigated by MTT assay after treatment with melatonin, SB203580 (p38 inhibitor) and phorbol 12-myristate 13-acetate (PMA, MAPK activator) alone or in combination for 48h. The effects of melatonin, and ML-7, a selective inhibitor of myosin light chain kinase (MLCK), and SB203580, and PMA on the migration of RKO cells were analyzed by in vitro scratch-wound assay. The relative mRNA levels of MLCK was assessed by real-time quantitative RT-PCR. Western blotting analysis was performed to examine the expression of MLCK, phosphorylation of myosin light chain (pMLC) and p38 (pp38). Results: The proliferation and migration of human colon cancer RKO cells were inhibited significantly after treatment with melatonin. The expression levels of MLCK and phosphorylation of MLC of RKO cells were reduced, and real-time quantitative RT-PCR showed that melatonin had significant effects on suppressing the expression of MLCK. Furthermore, the phosphorylation level of p38, which showed the same trend, was also reduced when cells were treated by melatonin. In addition, ML-7 (25umol/l) could down-regulate the phosphorylation of p38. Conclusions: Melatonin could inhibit the proliferation and migration of RKO cells, and further experiments confirmed that p38 MAPK plays an important role in regulating melatonin-induced migration inhibition through down-regulating the expression and activity of MLCK.