• BMB Reports
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• 제40권6호
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• pp.911-920
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• 2007

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the leading infectious diseases to humans. It is urgent to discover novel drug targets for the development of antitubercular agents. The 2-C-methyl-Derythritol-4-phosphate (MEP) pathway for isoprenoid biosynthesis has been considered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammals. MEP cytidyltransferase (IspD), the third-step enzyme of the pathway, catalyzes MEP and CTP to form 4-diphosphocytidyl-2-C-methylerythritol (CDP-ME) and PPi. In the work, ispD gene from M. tuberculosis H37Rv (MtIspD) was cloned and expressed. With N-terminal fusion of a histidine-tagged sequence, MtIspD could be purified to homogeneity by one-step nickel affinity chromatography. MtIspD exists as a homodimer with an apparent molecular mass of 52 kDa. Enzyme property analysis revealed that MtIspD has high specificity for pyrimidine bases and narrow divalent cation requirements, with maximal activity found in the presence of CTP and $Mg^{2+}$. The turnover number of MtIspD is $3.4 s^{-1}$. The Km for MEP and CTP are 43 and $92{\mu}M$, respectively. Furthermore, MtIspD shows thermal instable above $50^{\circ}C$. Circular dichroism spectra revealed that the alteration of tertiary conformation is closely related with sharp loss of enzyme activity at higher temperature. This study is expected to help better understand the features of IspD and provide useful information for the development of novel antibiotics to treat M. tuberculosis.

• The Korean Journal of Physiology and Pharmacology
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• 제16권5호
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• pp.333-337
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• 2012

Gene expression of neuronal nitric oxide synthase (nNOS) changes in the hypothalamic paraventricular nucleus (PVN) depending on feeding conditions, which is decreased during food deprivation and restored by refeeding, and phosphorylated cAMP response element binding protein (pCREB) was suggested to play a role in its regulation. This study was conducted to examine if the fasting-induced down-regulation of the PVN-nNOS expression is restored by activation of cAMP-dependent protein kinase A (cAMP/PKA) pathway. Freely moving rats received intracerebroventricular (icv) injection of cAMP/PKA activator Sp-cAMP (40 nmol) or vehicle (sterilized saline) following 48 h of food deprivation. One hour after drug injections, rats were transcardially perfused with 4% paraformaldehyde, and the PVN tissues were processed for nNOS or pCREB immunohistochemistry. Sp-cAMP significantly increased not only nNOS but also pCREB immunoreactivities in the PVN of food deprived rats. Fastinginduced down-regulation of the PVN-nNOS was restored by 1 h after the icv Sp-cAMP. Results suggest that cAMP/PKA pathway may mediate the regulation of the PVN-nNOS expression depending on different feeding conditions.

• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.667-674
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• 2017

Angiotensin II (Ang II) is metabolized from N-terminal by aminopeptidases and from C-terminal by Ang converting enzyme (ACE) to generate several truncated angiotensin peptides (Angs). The truncated Angs have different biological effects but it remains unknown whether Ang-(4-8) is an active peptide. The present study was to investigate the effects of Ang-(4-8) on hemodynamics and atrial natriuretic peptide (ANP) secretion using isolated beating rat atria. Atrial stretch caused increases in atrial contractility by 60% and in ANP secretion by 70%. Ang-(4-8) (0.01, 0.1, and $1{\mu}M$) suppressed high stretch-induced ANP secretion in a dose-dependent manner. Ang-(4-8) ($0.1{\mu}M$)-induced suppression of ANP secretion was attenuated by the pretreatment with an antagonist of Ang type 1 receptor ($AT_1R$) but not by an antagonist of $AT_2R$ or $AT_4R$. Ang-(4-8)-induced suppression of ANP secretion was attenuated by the pretreatment with inhibitor of phospholipase (PLC), inositol triphosphate ($IP_3$) receptor, or nonspecific protein kinase C (PKC). The potency of Ang-(4-8) to inhibit ANP secretion was similar to Ang II. However, Ang-(4-8) $10{\mu}M$ caused an increased mean arterial pressure which was similar to that by 1 nM Ang II. Therefore, we suggest that Ang-(4-8) suppresses high stretch-induced ANP secretion through the $AT_1R$ and $PLC/IP_3/PKC$ pathway. Ang-(4-8) is a biologically active peptide which functions as an inhibition mechanism of ANP secretion and an increment of blood pressure.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.218.2-219
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• 2003

2-Hydroxymuconic semialdehyde dehydrogenase catalyzes the conversion of 2-hydroxymuconic semialdehyde (HMS) to an enol form of 4-oxalocrotonate which is a step in the catechol-meta cleavage pathway. A tomC gene encoding 2-HMS dehydrogenase of Burkholderia cepacia G4, a soil bacterium that can grow on toluene, cresol, phenol or tricholoro ethylene, is identified in between catechol 2,3-dioxygenase gene and HMS hydrolase gene, its sequence is analysed and the enzyme is characterised. (omitted)

• Bulletin of the Korean Chemical Society
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• 제28권12호
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• pp.2324-2328
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• 2007

Organic fluorine chemistry produces many useful products. This paper elucidates the reaction of ethyl-4,4- difluoro-2-iodo-5-oxo-5-phenylpentanoate (2) with primary amines in a one-pot scheme. The reaction produced a series of β-fluoropyrrole derivatives at ambient temperatures. In this reaction, the less bulky the primary amine the higher was the resultant yield. When (2) and aqueous methylamine (40%) were allowed to react below 0 oC, 5-(ethoxycarboxyl)-1-methyl-3,3-difluoro-2-hydroxy-2-phenylpyrrolidine, an intermediate molecule for 2-ethyl-4-flouro-1-methyl-5-phenylpyrrole-2-carboxylate (5), was isolated first. Then, (5) reacted with hydroperchloric acid and acetic anhydride to form 5-(ethoxycarboxyl)-1-methyl-3,3-difluoro-2- phenylpyrrolinium perchlorate (6), which was converted to 2-ethyl-4-flouro-1-methyl-5-phenylpyrrole-2- carboxylate gradually in the presence of a base. Our experiments demonstrate that the formation of 2-ethyl-4- flouro-1-methyl-5-phenylpyrrole-2-carboxylate occurs via both one-pot schemes and stepwise pathways, depending on the reaction conditions. The isolation and characterization of the isolated intermediate (6) suggest an anionic pathway for this reaction.

• Journal of Plant Biotechnology
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• 제44권4호
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• pp.462-471
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• 2017

Thiamine pyrophosphate (TPP), the active form of thiamine is a cofactor for enzymes involved in central metabolism pathways. However, it is also known to have a role as a stress signaling molecule in response to environmental changes. Anabaena sp. and N. oculata are microorganisms which are abundantly found in Malaysia's freshwater and marine ecosystem. However, not much studies have been done especially in regards to thiamine biosynthesis. This work aimed to amplify of gene transcripts coding for thiamine biosynthesis enzymes besides looking at the expression of thiamine biosynthesis genes upon stress application. Various stress inducers were applied to the cultures and RNA was extracted at different time points. The first two genes, ThiC and ThiG/Thi4 encoding enzymes of the pyrimidine and thiazole branch respectively in the thiamine biosynthesis pathway were identified and amplified. The expression of the genes were analysed via RT-PCR and the intensity of bands were analysed using ImageJ software. The results showed up to 4-fold increase in the expression of ThiC and ThiG gene transcript as compared to control sample in Anabaena sp. ThiC gene in N. oculata showed an expression of 6-fold higher as compared to control sample. In conclusion, stresses induced the expression of the gene coding for one of the most important enzymes in thiamine biosynthesis pathway. This is an agreement with the hypothesis that overexpression of thiamine is crucial in assisting plants to combat abiotic stresses.

• Genomics & Informatics
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• 제16권4호
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• pp.34.1-34.9
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• 2018

Cirsium japonicum belongs to the Asteraceae or Compositae family and is a medicinal plant in Asia that has a variety of effects, including tumour inhibition, improved immunity with flavones, and antidiabetic and hepatoprotective effects. Silymarin is synthesized by 4-coumaroyl-CoA via both the flavonoid and phenylpropanoid pathways to produce the immediate precursors taxifolin and coniferyl alcohol. Then, the oxidative radicalization of taxifolin and coniferyl alcohol produces silymarin. We identified the expression of genes related to the synthesis of silymarin in C. japonicum in three different tissues, namely, flowers, leaves, and roots, through RNA sequencing. We obtained 51,133 unigenes from transcriptome sequencing by de novo assembly using Trinity v2.1.1, TransDecoder v2.0.1, and CD-HIT v4.6 software. The differentially expressed gene analysis revealed that the expression of genes related to the flavonoid pathway was higher in the flowers, whereas the phenylpropanoid pathway was more highly expressed in the roots. In this study, we established a global transcriptome dataset for C. japonicum. The data shall not only be useful to focus more deeply on the genes related to product medicinal metabolite including flavolignan but also to study the functional genomics for genetic engineering of C. japonicum.

• Journal of Yeungnam Medical Science
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• 제14권2호
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• pp.337-349
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• 1997

세균 내독소에 의하여 발생하는 패혈성 쇼크와 혈관 반응성 감소의 원인을 관찰하였다. 혈관 절편이 고정된 실험조에 세균 내독소 0.2 mg 투여한 경우 $36{\pm}3.65$ nM NO가 발생되었고, NO 발생에 의한 혈관 이완 효과를 억제하기 위해 전처치한 L-MAME, methylene blue는 혈관 절편의 phenylephrine (PE) 유발 수축 반응을 증가시켰으며 methylene blue에 의해 더 강한 수축 반응의 증가가 관찰되었다. 이때 혈관 내피세포가 존재할 경우에 PE에 대한 혈관 반응성이 증가되는 경향을 나타내었다. 세균 내독소 투여에 의해 acetylcholine 유발 혈관 이완은 증가되는 경향을 나타내었고, 전처치한 L-NAME, methylene blue에 의해 혈관 절편의 acetylcholine (ACh) 유발 이완은 억제되었으며 methylene blue에 의해 현저히 억제되었다. 그러나 세균 내독소를 투여하지 않은 군의 ACh 유발 혈관 이완 반응은 methylene blue에 의해서만 억제되었다. 결론적으로 세균 내독소에 의한 혈관 반응성 감소와 혈관 이완 반응은 NO가 발생되어 guanylyl cyclase를 활성화하여 유발된다고 생각되며, 세균 내독소에 의한 효과는 L-arginine NO pathway 보다는 cyclic GMP 신호전달계를 경유한 경로에서 더 많은 영향을 받는것으로 사료된다.

• 생태와환경
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• 제40권1호
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• pp.93-102
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• 2007

남한강과 북한강 상류지역에 위치한 11개 하천을 대상으로 강우 시 발생하는 탁수의 생지화학적 특성을 조사하였다. 강우 시 하천에서 POM와 DOM의 유출특성은 유량변동에 따라 다르게 나타났으며, 특히 POM의 유출은 하천의 유역특성과 밀접한 관련이 있는 것으로 나타났다. 강우의 영향에 따라 DOM내 용존 부식물질(Humic substance)의 비율을 반영하는 SUVA값이 증가하였고, 이는 하천유역으로부터 난분해성유기물의 유입이 증가함을 의미한다. 강우 시 발생된 탁수의 생지화학적 특성을 분석한 결과 농경지역에서 유출되는 탁수가 삼림지역으로부터 유출되는 탁수보다 안정탄소동위원소비가 약 $1{\sim}2%_{\circ}$정도 높게 나타났다. 이는 상대적으로 안정탄소동위원소비가 높은 $C_4$계열의 작물이 농경지역에서 우세한 경우 또는 삼림지역으로부터 유출된 탁수에서 주로 $C_3$계열의 식물체의 리그닌(Lignin)의 함량이 높기 때문이다. 유기물의 기원에 따라 Isotopic mass balance를 적용한 결과 농경지나 경작지의 비율이 높아질수록 $C_4$계열 작물의 기여도가 높아지고 이에 따라 탄소동위원소비가 증가하는 경향을 보였다. 본 연구결과를 바탕으로 안정탄소동위원소를 이용한 탁수 연구는 유기물의 기원특성을 연구하는데 유용한 지표가 될 수 있다.

• 환경생물
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• 제40권2호
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• pp.214-223
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• 2022

멸강나방(Mythimna seperata)과 혹명나방(Cnaphalocrocis medinalis)은 중국 남부 양쯔강 유역에서 봄철 편서풍을 타고 국내로 유입되는 비래 해충(Migratory insect pests)으로 벼를 기주로 삼아 벼 잎을 가해하여 생육을 저해시킨다. 두 나방의 분포를 파악하기 위해서는 서식처의 온습도 뿐만 아니라 주변 환경 요소를 파악하는 것이 중요하다. 본 연구는 두 나방의 분포를 파악하기 위해서 SDM(Species Distribution Model) 중 Machine learning model인 MaxEnt (Maximum Entropy)에 출현 자료, SSPs (Shared Socio-economic Pathways) 시나리오, RCP (Representative Concentration Pathway) 시나리오를 적용하여 현재와 미래의 서식지 적합성 모형을 예측했다. 결과로 시기에 따른 서식처 면적이 큰 차이가 없었으며, SSPs 시나리오가 나빠짐에 따라 분포 면적이 넓어졌다. 두 나방은 중국으로부터 비래 후 생존하기 위한 최적의 장소가 기주가 있는 서해안과 남해안에 집중되어 있다. MaxEnt 결과 토지피복 자료, DEM (Degital Elevation Model) 순으로 기여도가 높게 나타났다. 이는 논에서의 출현 확률 높고 고도가 높아지면서 출현 확률이 낮아졌기 때문이다. 기후 변수에서 멸강나방은 BIO_4 (Temperature seasonality), 혹명나방은 BIO_2 (Mean Diurnal Range)가 높게 나타났다. 멸강나방은 계절에 의한 기온 차가 31.9℃ 이상일 때 서식처가 줄어들고, 혹명나방은 일교차 클수록 서식처가 넓어질 것으로 나타났다. 서식지 적합성 모형에서 두 나방은 대부분의 논에서 서식이 가능할 것이라 예측되었다. 하지만, 두 나방의 출현 위치를 정확하게 예측하는 데 한계가 있으므로, 서식지 적합성 지도를 기초로 조기에 대응하는 것이 중요하다고 판단된다.