• 농약과학회지
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• 제13권2호
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• pp.105-110
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• 2009

감 과수원으로부터 분리한 탄저병균(Colletotrichum gloeosporioides)에 대한 carbendazim, thiophanate-methyl, mancozedb 및 propineb 등 총 4종의 살균제의 약제반응을 조사하였다. Carbendazim($415{\mu}g/m{\ell}$)과 thiophanate-methyl($750{\mu}g/m{\ell}$)에 대하여 90%이상의 균사생육 억제율을 보이는 균주의 비율은 각각 68.2와 35.5%로 나타났으며 두 약제 사이의 교차 저항성을 조사한 결과, 상관관계가 높은 것으로 나타났다($r^2=0.7862$). Mancozeb와 propineb에 대하여 91%이상의 균사생육 억제율을 보인 균주의 비율은 각각 90%과 53.6%이상으로 조사되었으며, 두 약제가 교차 저항성의 위험성은 낮은 것으로 나타났다($r^2=0.0174$). 시험에 사용한 두 종의 benzimidazole계 살균제와 mancozeb와 propineb 사이의 교차 저항성은 조사한 결과, 상관관계가 없는 것으로 나타났다.

• Biomolecules & Therapeutics
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• 제13권3호
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• pp.190-197
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• 2005

Advanced glycation end products (AGE) have been implicated in the pathogenesis of diabetic complications including nephropathy. However, the role of AGE in the activation of mesangial cells cultured under high glucose has not been elucidated. The effects of aminoguanidine, which prevents formation of AGE and protein cross-linking, on the synthesis of $TGF-{\beta}1$ and fibronectin by rat mesangial cells cultured under high glucose for 2 weeks were examined and compared with the effects of $N^G$-nitro-L-arginine methyl ester (NAME), a selective nitric oxide synthase inhibitor, because aminoguanidine also inhibits the inducible nitric oxide synthase. Culture of mesangial cells in 30 mM (high) glucose for 2 weeks induced 1.5-fold (ELISA) and 1.9-fold (Western blot analysis) increase in AGE in the culture media compared to 5.6 mM (control) glucose. Northern blot analysis revealed 1.5-fold increase in $TGF-{\beta}1$ and 1.7-fold increase in fibronectin mRNA expression in cells cultured under high glucose compared to control glucose. Increases in mRNA expression were followed by increased protein synthesis. Mink lung epithelial cell growth inhibition assay revealed 1.4-fold increase in $TGF-{\beta}1$ protein in high glucose media compared to control. Fibronectin protein also increased 2.1-fold that of control glucose by Western blot analysis. Administration of aminoguanidine suppressed AGE formation in a dose dependent manner and at the same time suppressed $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells cultured in both control and high glucose. In contrast, NAME did not affect high glucose-induced changes. These findings support a role for AGE in high glucose-induced upregulation of $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells.

• 대한치과의료관리학회지
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• 제7권1호
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• pp.50-55
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• 2019

The purpose of this study was to compare the number of bacteria before and after the use of surface disinfectants, demonstrate the bactericidal effect of surface disinfectants, and emphasize on the importance of surface disinfectants by recognizing the importance of infection control in dentistry. Chlorhexidine, hydrogen peroxide, ethanol, and chemical disinfectants are commonly used in dentistry. NaOCl was selected as the experimental group, and the bacterium test results obtained by comparing the table without surface sterilization as a control group showed that all disinfectants had an effective bactericidal effect (p<0.05). In the growth inhibition test comparing the experimental and control groups, all results were 100%, proving the effectiveness of surface disinfectants. The results showed that all surface disinfectants preferred and used by medical institutions were effective. Therefore, all surface disinfectants used in the experimental group were effective for surface disinfection for infection control. Dental clinicians should be aware of the necessity of disinfection of surfaces, such as table, chairs, and unit chairs, and make an active effort to ensure that both clinicians and patients are safe from infection.

• 미생물학회지
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• 제17권3호
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• pp.97-130
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• 1979

Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

• 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.

• 생명과학회지
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• 제25권1호
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• pp.93-100
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• 2015

Pachymic acid는 복령에서 분리된 lanostane-type인 triterpenoid의 일종이다. 최근 pachymic acid가 항암 및 항염증 효능과 산화적 스트레스에 대한 항산화 효능 등과 같은 약리적인 효능이 있는 것으로 밝혀지고 있으나, 그에 대한 구체적인 분자생물학적 기전 연구는 매우 미비한 실정이다. 본 연구에서는 pachymic acid의 항암활성 및 관련 기전 조사의 일환으로 T24 인체 방광암세포 모델을 이용하여 pachymic acid에 의한 apoptosis 유발 여부를 검증하였다. 본 연구의 결과에 의하면 pachymic acid는 T24 세포의 증식을 유의적으로 억제시켰으며, 이는 apoptosis 유발과 연관성이 있음을 다양한 방법으로 확인하였다. Pachymic acid에 의한 apoptosis 유도에는 pro-apoptotic 인자들의 발현 증가와 anti-apoptotic 유전자 산물들의 발현 감소가 동반되었으며, MMP의 소실과 tBid의 발현 증가가 관찰되었다. 아울러 pachymic acid는 extrinsic 및 intrinsic apoptosis 경로의 개시에 관여하는 caspase-8 및 -9의 활성뿐 만 아니라, caspase-3의 활성도 증가시킴으로서 PARP와 같은 기질 단백질의 단편화를 초래하였다. 따라서 pachymic acid는 항암활성을 지니는 천연생리활성 물질로서의 잠재력이 매우 높음을 알 수 있었다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 춘계학술대회
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• pp.111-111
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• 2019

Glycyrrhizae radix is one of the most frequently prescribed ingredients in Oriental medicine, and G. radix extract has been shown to exert anti-cancer effects. However, the cellular and molecular mechanisms of apoptosis by G. radix are poorly defined. In the present study, it was examined the biochemical mechanisms of apoptosis by water extract of G. radix (WEGR) in human bladder T24 cancer cells. It was found that WEGR could inhibit the cell growth of T24 cells in a dose-dependent manner, which was associated with the induction of apoptotic cell death, as evidenced by the formation of apoptotic bodies, DNA fragmentation and increased populations of annexin-V positive cells. The induction of apoptotic cell death by WEGR was connected with an up-regulation of pro-apoptotic Bax protein expression and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL proteins, and inhibition of apoptosis family proteins (XIAP, cIAP-1 and cIAP-2). In addition, apoptosis-inducing concentrations of WEGR induced the activation of caspase-9, an initiator caspase of the mitochondrial-mediated intrinsic pathway, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose)-polymerase. WEGR also induced apoptosis via a death receptor-mediated extrinsic pathway by caspase-8 activation, resulting in the down-regulation of total Bid and suggesting the existence of cross-talk between the extrinsic and intrinsic pathways. Taken together, the present results suggest that WEGR may be a potential chemotherapeutic agent for the control of human bladder cancer cells.

• 한국육종학회지
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• 제50권4호
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• pp.387-395
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• 2018

다산 등 15개의 통일형 벼 품종을 이용하여 mesotrione을 처리한 후 5일부터 다산 등 13개 품종들은 신엽에서 백화현상이 발생하였으나, 밀양154호와 수원382호는 백화현상이 발생하지 않아 저항성 품종으로 선발되었다. Mesotrione 처리 후 다산 등 13개 품종들의 초장 및 건물중 억제율은 밀양154호와 수원382호보다 더 높았으며 약량에 따른 억제율이 증가하였다. 한아름2호/밀양154호의 $F_2$ 집단을 이용한 유전분석 결과 저항성이 149개체, 감수성이 41개체로 이론적 분리비 3:1($X^2=1.19$, P=0.31)에 적합하여, mesotrione 저항성 관련 유전자는 1개의 우성유전자에 지배되는 것으로 나타났다. BSA방법을 이용하여 mesotrione 저항성 관련 유전자를 탐색한 결과, 2번 염색체의 10.2 Mb에 위치한 SSR marker RM1358과 RM3501을 선발하였다. Fine mapping을 위해 선발된 5개의 저항성 개체 중에서 $F_2-137$은 RM12921부터 RM3501까지 재조환이 일어났고, $F_2-76$은 RM324부터 RM5101까지 재조환이 일어났다. 따라서, mesotrione 저항성 관련 유전자는 RM3501과 RM324 사이인 10.2 Mb~11.4 Mb에 존재하며, RM3501과 RM324의 조환률이 각각 0.53%과 2.65%이므로, RM3501을 mesotrione저항성 유전자 DNA 연관 marker로 선발하였다. 다산 등 20개 통일형 품종 및 mesotrione에 저항성을 나타내는 자포니카 품종인 운광과 감수성인 통일형 한아름의 $BC_2F_2$ 집단을 이용하여 RM3501의 MAS 이용 가능성을 검토한 결과, mesotrione 저항성 선발 marker로 활용은 가능하나, 정확한 선발 maker로 활용하기 위해서는 추가적인 실험이 필요할 것으로 판단된다.

• 한국자원식물학회지
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• 제32권4호
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• pp.255-263
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• 2019

본 연구에서는 다양한 약리학적 활성을 가지는 것으로 알려진 감초 열수추출물(GRW)의 항암효능을 알아보기 위하여 인체 방광암 T24 세포에서 생존율 및 증식억제에 미치는 영향과 이와 연관된 apoptosis 유발 여부 및 관련 인자들의 발현 변화를 조사하였다. 먼저 GRW의 처리에 따른 증식억제 정도를 조사한 결과, GRW 처리 농도 의존적으로 생존율 및 증식억제 현상이 나타났으며, 핵의 형태 변화, DNA 단편화 및 apoptosis 유발에 관하여 조사한 결과 역시 GRW 처리 농도 의존적으로 증가됨을 확인할 수 있었다. 이는 GRW의 처리에 의한 암세포의 증식억제 및 형태적 변형이 암세포의 apoptosis 유발과 밀접한 관련이 있음을 시사하여 주는 것으로 사료된다. GRW 처리에 의한 apoptosis 유발에 관여하는 유전자의 탐색을 위하여 apoptosis와 연관성을 가지는 Bcl-2 family에 속하는 유전자의 발현을 조사한 결과 GRW 처리 농도 의존적으로 Bax 단백질의 발현증가와 더불어 Bcl-2 및 Bcl-xL 단백질의 발현감소가 관찰되었다(Fig. 3A). 이는 GRW에 의한 T24 세포의 apoptosis 유발에 Bcl-2 family에 속하는 유전자의 발현 조절이 중요한 역할을 하는 것으로 사료된다. 또한 GRW의 처리에 따른 MMP의 소실은 미트콘드리아 막의 교란이 유발되었음을 의미하는 것으로, 이러한 MMP 값의 변동은 Bcl-2 family 단백질의 발현 변화에 의한 것이라 추정된다. 한편 Apoptosis에 중요한 역할을 하는 것으로 알려진 caspase(-3/-8/-9)의 발현과 이들의 활성을 억제하는 IAP family (XIAP, cIAP-1, cIAP-2)의 발현에 GRW이 어떠한 영향을 미치는지를 조사한 결과, caspase-3, -8 및 -9의 활성형 단백질 발현 및 정량적 활성증가를 확인하였으며, IAP family 속한 3가지 단백질 모두 발현이 감소하는 것이 관찰되었다. 이상의 결과에서 GRW은 외인적 및 내인적 경로의 개시에 핵심적인 역할을 하는 caspase-8 및 -9의 활성을 모두 증가시켰으며, 이에 따른 caspase-3의 활성증가에 의하여 apoptosis가 유발되었음을 알 수 있었다. 이러한 두 경로의 동시 활성화에는 미트콘드리아의 기능 소실과 Bcl-2 및 IAP family의 발현 변화가 관여하고 있었으며, 특히 Bid의 발현 감소는 GRW에 의한 내인적 경로를 증폭시키는 효과로 작용했을 것이라 추정된다. 방광암의 치료에 보다 효과적인 생리활성을 갖는 물질을 발굴하고 그와 관련된 분자 및 세포수준에서의 기전을 밝히는 것이 중요하기에 본 연구의 결과는 향후 GRW로 수행될 추가 실험을 위한 기초자료로서 그 가치가 매우 높을 것으로 사료된다.

• Journal of Nutrition and Health
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• 제54권6호
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• pp.618-630
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• 2021

본 연구에서는 생강을 효소처리하여 수용화율을 높인 후 고초균을 이용하여 발효시킨 생강발효물 (FGEs)을 제조하고, 이를 SH-SY5Y에 6-OHDA와 함께 처리하여 세포 보호 효과 및 AChE 저해 활성을 평가하였다. 그 결과, 6-OHDA로 자극된 신경세포에서 FGE를 처리한 모든 실험군에서 세포 생존율이 증가하고 LDH 농도가 감소하였다. 6-OHDA로 유발된 세포자멸사를 억제할 수 있는지 확인하기 위해 핵의 형태학적인 변화 및 caspase-3 활성을 확인하였다. FGE를 처리한 모든 실험군에서 핵의 손상 및 apoptotic body의 감소와 caspase-3 억제 활성을 확인할 수 있었다. 또한 FGE는 AChE의 활성을 유의적으로 감소시켰다. 시료 간의 활성 차이를 비교하였을 때, 생강에 효소 처리 후 고초균으로 발효한 추출물군 (E/BKG와 E/BCG)의 신경세포 보호 활성이 효소 처리하지 않은 발효생강군 (BKG와 BCG) 보다 유의적으로 크게 나타났다. 그러나 발효에 사용된 고초균 2종 간의 활성은 유사하였으며 처리군 간 유의적인 차이를 보이지 않았다. 본 연구 결과로부터 효소처리하여 수용화율을 높여 고초균으로 발효한 FGE는 신경세포 보호 및 AChE 저해 효과를 나타내어 향후 신경질환 연구를 위한 기초자료 제공 및 고부가가치 식품소재 개발에 이용 가능할 것으로 판단된다.