• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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• pp.120-122
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• 2007

All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

• 생명과학회지
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• 제17권2호통권82호
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• pp.235-240
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• 2007

본 연구는 Affymetrix Arabidopsis DNA chip을 이용하여 비 병원성 인자인 AvrRpt2 단백질에 의해서 특이적으로 전사 과정이 조절되는 애기장대 유전자들을 분리하고 병 저항성 방어체계와 관련한 이들 유전자들의 기능 분석을 시도하였다. 그 중에서 먼저 식물 호르몬인 ethylene의 신호 조절에 관여하는 ERFs (ethylene-responsive element binding factors) 전사조절 유전자 family 중에서 Bla subfamily 그룹으로 알려져 있는 AtERF11 유전자의 병 저항성 관련 기능을 규명하였다. 저항성 유전자 RPS2가 없는 경우에는 비 병원성 인자인 AvrRpt2 단백질은 기주 식물체내의 기초 병저항성을 감소시키고 병원성 세균의 증식을 향상시켜서 병증을 증대시키는 effector로 작용한다는 기존의 연구결과와 유사하게, 저항성 유전자 RPS2가 없는 조건에서 AtERF11 유전자의 발현이 AvrRpt2 단백질의 작용에 의해서 특이적으로 감소되는 것을 확인하였다. 이러한 결과를 바탕으로 AtERF11 유전자는 식물체의 병 저항성 방어기작에 있어서 positive regulator로서 작용하기 때문에 effector로 작용하는 AvrRpt2 단백질에 의해서 조절되는 것으로 추측하였다. 본 가설을 증명하기 위해 AtERF11의 발현을 증폭시킨 애기장대 형질전화체를 제작하고 P. syringae pv. tomato DC 3000에 대한 병저항성을 실험하였다. AtERF11 유전자가 대량 발현하는 형질전화 된 애기장대에서는 야생종에 비해 대략 100배 이상 세균의 증식이 억제되는 강력한 병저항성을 가진다는 것을 검증하였다.

• 한국응용곤충학회지
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• 제35권4호
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• pp.315-320
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• 1996

Drosophila의 actin 5C 유전자 promoter에 쥐의 DNA polymerase $\beta$cDNA를 도입시킨 형질전환 초파리가 고감수성 환경성 변이원 검출계로 사용할 수 있는지를 조사하였다. 체세포 염색체 재조환과 체세포 염색체 돌연변이의 검출을 위해서는 geterozygous(mwh/+) 계통을 사용하였다. 염색체상의 결실이나 비분리 등에 의한 small mwh spot의 자연 발생적 빈도는 non-transgenic w 계통과 transgenic p[pol $\beta$]-130 계통에서 각각 0.351 및 0.606 정도였다. 체세포 염색체 재조환에 의한 large mwh spot의 자연 발생적 빈도의 경우는 transgenic p[pol $\beta$]-130 계통(0.063)이 non-transgenic w 계통(0.021)에 비해 약 3배 정도 높게 나타났다. IQ, Glu-P-1 및 {TEX}$AFB_{1}${/TEX} 등의 돌연변이원의 처리에 의한 경우, 두 종류의 mutant clone의 발생 빈도는 쥐의 DNA polymerase $\beta$가 도입된 transgenic p[pol $\beta$]-130 계통이 non-transgenic w 계통에 비하여 모두 약 2-3배 정도 높게 나타났다. 본 연구의 결과는 쥐의 DNA polymerase $\beta$가 최소한 체세포 염색체 돌연변이 유발이나 체세포 염색체 재조환의 생성 과정에 관여함을 의미하며, 형질전환 초파리 계통이 환경성 변이원 검출계로서 충분한 응용가능성이 있음을 보여 주었다.

• Molecules and Cells
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• 제40권3호
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• pp.230-242
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• 2017

In the Arabidopsis genome, approximately 80 MAP3Ks (mitogen-activated protein kinase kinase kinases) have been identified. However, only a few of them have been characterized, and the functions of most MAP3Ks are largely unknown. In this paper, we report the function of MAP3K16 and several other MAP3Ks, MAP3K14/15/17/18, whose expression is salt-inducible. We prepared MAP3K16 overexpression (OX) lines and analyzed their phenotypes. The result showed that the transgenic plants were ABA-insensitive during seed germination and cotyledon greening stage but their root growth was ABA-hypersensitive. The OX lines were more susceptible to water-deficit condition at later growth stage in soil. A MAP3K16 knockout (KO) line, on the other hand, exhibited opposite phenotypes. In similar transgenic analyses, we found that MAP3K14/15/17/18 OX and KO lines displayed similar phenotypes to those of MA3K16, suggesting the functional redundancy among them. MAP3K16 possesses in vitro kinase activity, and we carried out two-hybrid analyses to identify MAP3K16 substrates. Our results indicate that MAP3K16 interacts with MKK3 and the negative regulator of ABA response, ABR1, in yeast. Furthermore, MAP3K16 recombinant protein could phosphorylate MKK3 and ABR1, suggesting that they might be MAP3K16 substrates. Collectively, our results demonstrate that MAP3K16 and MAP3K14/15/17/18 are involved in ABA response, playing negative or positive roles depending on developmental stage and that MAP3K16 may function via MKK3 and ABR1.

• Restorative Dentistry and Endodontics
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• 제20권2호
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• pp.508-526
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• 1995

This experiment was performed to study mechanisms of desensitization by chemical desensitizing agents in hypersensitive dentin and compare effects of these agents by measuring the activity of intradental nerves and observing their occluding aspects on dentinal tubules with SEM over time after application of chemical desensitizing agents to the exposed dentinal surfaces. Canines of adult cats weighing 2-3 kg were cross-sectioned at 1.5 mm from incisal apex, and the smear layer of the exposed dentinal susface was removed by 32 % $H_3PO_4$ for 15 sec. Chemical desensitizing agents such as 10% $SrCl_2$, 5% $KNO_3$ and 30% $K_2C_2O_4$, were applied to the exposed dentin surfaces for 2 minutes. Intradental nerve activity was measured immediately after application of the agents, at 15 minutes and at 30 minutes by stimulating with 4M NaCl. To compare occluding ability of desensitizing agents on dentinal tubules in vivo and in vitro, the structures of the exposed dentinal surfaces of nonvital and vital teeth were morphologically observed by SEM. The results obtained were as follows : 1. Intradental nerve activity was decreased immediately after the application of 10 % $SrCl_2$, 5% $KNO_3$ and 30% $K_2C_2O_4$. (p<0.01), among which 30% $K_2C_2O_4$. showed the highest desensitizing effect(p<0.01). 2. The immediately decreased intradental nerve activity after application of 10 % $SrCl_2$ and 5% $KNO_3$ was increased over time. 10% $SrCl_2$ and 5% $KNO_3$ showed no desensitizing effect respectively at 30 minutes and at 15 minutes after application. 3. The immediately decreased intradental nerve activity after application of 30 % $K_2C_2O_4$ was persistently continued during the period of observation (p<0.01). 4. Precipitates of $SrCl_2$ and $KNO_3$ were not noted on the exposed dentinal surfaces and within dentinal tubules by SEM examination. On the other hand, 30 % $K_2C_2O_4$ produced precipitates on the exposed dentinal surfaces and openings of dentinal tubules without any formed preciptates within dentinal tubules. 5. Ten percent $SrCl_2$, 5 % $KNO_3$ and 30 % $K_2C_2O_4$ showed no differences in their occluding aspects on dentinal tubules either in vivo or in vitro studies and either immediately following application or at 30 minutes. These results suggest that the desensitizing effect of $SrCl_2$ and $KNO_3$ is resulted from their reducing effect on the intradental nerve activity rather than from their precipitates' occluding the dentinal tubules. However, desensitizing effect of 30 % $K_2C_2O_4$, is probably resulted from its precipitates' occluding the openings of the dentinal tubules as well as from it's reducing effect on the intradental nerve actibity.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.456-464
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• 2020

Mast cells (MCs) are systemically distributed and secrete several allergic mediators such as histamine and leukotrienes to cause type I hypersensitivity. Dasatinib is a type of anti-cancer agent and it has also been reported to inhibit human basophils. However, dasatinib has not been reported for its inhibitory effects on MCs or type I hypersensitivity in mice. In this study, we examined the inhibitory effect of dasatinib on MCs and MC-mediated allergic response in vitro and in vivo. In vitro, dasatinib inhibited the degranulation of MCs by antigen stimulation in a dose-dependent manner (IC₅₀, ~34 nM for RBL-2H3 cells; ~52 nM for BMMCs) without any cytotoxicity. It also suppressed the secretion of inflammatory cytokines IL-4 and TNF-α by antigen stimulation. Furthermore, dasatinib inhibited MC-mediated passive cutaneous anaphylaxis (PCA) in mice (ED₅₀, ~29 mg/kg). Notably, dasatinib significantly suppressed the degranulation of MCs in the ear tissue. As the mechanism of its effect, dasatinib inhibited the activation of Syk and Syk-mediated downstream signaling proteins, LAT, PLCγ1, and three typical MAP kinases (Erk1/2, JNK, and p38), which are essential for the activation of MCs. Interestingly, in vitro tyrosine kinase assay, dasatinib directly inhibited the activities of Lyn and Fyn, the upstream tyrosine kinases of Syk in MCs. Taken together, dasatinib suppresses MCs and PCA in vitro and in vivo through the inhibition of Lyn and Fyn Src-family kinases. Therefore, we suggest the possibility of repositioning the anti-cancer drug dasatinib as a treatment for various MC-mediated type I hypersensitive diseases.

• Molecules and Cells
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• 제28권5호
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• pp.431-439
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• 2009

Rice Oryza sativa accelerated cell death and resistance 1 (OsACDR1) encodes a putative Raf-like mitogen-activated protein kinase kinase kinase (MAPKKK). We had previously reported upregulation of the OsACDR1 transcript by a range of environmental stimuli involved in eliciting defense-related pathways. Here we apply biochemical, gain and loss-of-function approaches to characterize OsACDR1 function in rice. The OsACDR1 protein showed autophosphorylation and possessed kinase activity. Rice plants overexpressing OsACDR1 exhibited spontaneous hypersensitive response (HR)-like lesions on leaves, upregulation of defense-related marker genes and accumulation of phenolic compounds and secondary metabolites (phytoalexins). These transgenic plants also acquired enhanced resistance to a fungal pathogen (Magnaporthe grisea) and showed inhibition of appressorial penetration on the leaf surface. In contrast, loss-of-function and RNA silenced OsACDR1 rice mutant plants showed downregulation of defense-related marker genes expressions and susceptibility to M. grisea. Furthermore, transient expression of an OsACDR1:GFP fusion protein in rice protoplast and onion epidermal cells revealed its localization to the nucleus. These results indicate that OsACDR1 plays an important role in the positive regulation of disease resistance in rice.

• 대한한방부인과학회지
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• 제27권4호
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• pp.57-68
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• 2014

Objectives: The purpose of this research is to investigate the relationship between Sasang constitution and cold hypersensitivity. Methods: We investigated 391 outpatients who visited Dong-Eui Oriental Hospital OB & GY from June 12, 2013 to April 18, 2014. Among 134 patients who complained feeling of cold, we analyzed 107 patients whose Sasang constitution is confirmed. Results: 1. There were 21 persons 19.8% of under age 27, 28 persons 26.4% of age 28-34, 23 persons 21.7% of age 35-41, 18 persons 17% of age 42-48, 16 persons 15.1% of age over 49. 2. Among 107 patients, there were 52 persons 48.6% of Soeumin, 29 persons 27.1% of Taeeumin, 25 persons 23.4% of Soyangin, 1 person 0.9% of Taeyangin. And Taeyangin interior disease was 1 case 100%, Soyangin exterior disease was 22 cases 88% and interior disease was 3 cases 12%, Taeeumin exterior disease was 11 cases 37.9% and interior disease was 18 cases 62.1%, Soeumin exterior disease was 19 cases 36.5% and interior disease was 33 cases 3.5%. 3. In the distribution of cold-hypersensitive part, hands and feet with chills were 58 cases 4.7% by largest number, and the following was hands and feet in 24 cases 22.6%. 4. In the distribution of chief complaint, there were 25 cases 23.58% of menstrual pain, 19 cases 17.92% of postpartum symptoms, 17 cases 16.04% of oligomenorrhea and hypomenorrhea, 14 cases 13.21% of cold hypersensitivity. 5. In the distribution of chief complaint depending on age, menstrual pain of under age 27, postpartum symptoms of age 35-41 and cold hypersensitivity of over age 49 were at a high rate. Conclusions: Results suggest that patients with symptom of cold hypersensitivity have some tendencies in age, Sasang constitution, chief complaint, cold-hypertensive part.

• 디지털융복합연구
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• 제19권6호
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• pp.429-438
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• 2021

본 연구는 국내 산후 복과 산후 우울증 및 스트레스 등의 실태조사를 통해 원인을 분석하고, 산모의 사회적 고립를 예방하고자 시도한 서술적 조사연구이다. 연구방법-전국에 거주하는 1살 이하의 자녀를 둔 주 대상인 20-40대 여성을 표본으로 선정하였고, 총 111부를 최종 분석 자료로 사용하였다. 연구결과-산모들의 일반적 특성 -일반적 특성으로 사회 경험을 묻는 말에서는 91.1%(102명)가 사회 경험이 있다고 응답하였고, 8%(9명)만이 사회 경험이 없다고 응답하였다.(사회경험유 91.1%(102명), 사회경험 무 8%(9명) 사회 복귀를 여부를 묻는 질문에서는 17.9%의 산모가 복귀하였다고 응답하였고, 54.5%의 산모는 복귀하지 않았으며, 18.8%의 산모는 출산 휴가 중인 것으로 나타났다. 출산 전과 후의 BMI 변화와 일반적 특성의 교차분석(카이제곱 검정) 산모들의 BMI 증가 수준을 살펴본 결과 Table 3과 같이 BMI가 평균 이하로 증가한 산모가 55%, 평균 이상으로 증가한 여성이 45%로 조사되었다. 연구결과- 임신과 출산으로 인해 여성들은 신체적 변화로 인해 과체중이 될 확률이 높다. 연구결과-임신과 출산으로 산후비만은 산후 정신적, 신체적 문제에 영향을 미치는 개인의 중요한 원인으로 확인되었다. 연구 결과를 바탕으로 산모비만관리와 우울증관리에 적극적인 정부 정책이 필요하다고 사료된다.

• 생명과학회지
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• 제33권2호
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• pp.176-182
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• 2023

석곡의 줄기는 전통 동양의학에서 위를 보하고, 진액을 보충하며, 열을 내리는 것에 사용되어 왔다. 본 연구에서는 RBL-2H3 세포에서 비만세포 탈과립과 TNF-α, IL-4, histidine decarboxylase (HDC) 발현에 미치는 석곡 열수추출물(DME)의 효과를 조사하였다. DME는 PMA와 Calcium ionophore 병행처리(PMACI)에 의해 유도되는 β-hexosaminidase 분비와 TNF-α, IL-4, HDC 발현을 현저히 억제하였다. 또한 PMACI에 의해 유도되는 NF-κB, AP-1 활성과 p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2)과 c-Jun N-terminal kinase (JNK)의 인산화가 DME 전처리에 의해 저해되었다. 이러한 결과들은DME가 비만세포 탈과립을 억제하고, MAPKs/NF-κB/AP-1 신호전달 경로를 통해 TNF-α, IL-4, HDC 발현을 억제한다는 것을 시사한다. 본 연구결과들로 보아 DME는 과민반응과 염증성 질환을 치료하는 약물로 개발될 가능성을 가지는 것으로 사료된다.