Role of Leu188 in the Fatty Acid Hydroxylase Activity of CYP102A1 from Bacillus megaterium

Jang, H.H.,Shin, S.M.,Ma, S.H.,Lee, G.Y.,Joung, Y.H.,Yun, C.H. Elsevier 2016 Journal of molecular catalysis Enzymatic Vol.133 No.-

<P>P450 BM3 (CYP102A1) from Bacillus megaterium catalyzes the subterminal hydroxylation of fatty acids with 12-22 carbons at the omega-1, omega-2 and omega-3 positions. Several amino acids located at the substrate channel and active sites are known to be important for the catalytic activity of CYP102A1. The L188 residue at the C-terminus of alpha-helix F undergoes a large shift upon substrate binding and has frequently been found in different combinations of multiple mutations showing enhanced and altered activities. In this study, we examined the role of the L188 residue by comparing the catalytic activities of wild-type CYP102A1 and 19 mutants of L188. The mutants were made with site-directed mutagenesis and functionally expressed in Escherichia coll. The enzymatic properties of the mutants for a set of fatty acids (C-10-C-16) were compared to the properties of the wild-type. L188Q and L188 P mutants showed especially strong increases in hydroxylase activity toward C-10-C-13 fatty acids, although they did not have activity changes for C-14-C-16 fatty acids. Although most mutants showed very similar overall hydroxylation rates for myristic acid, 14 mutants showed apparent changes in the regioselectivity of hydroxylation with a preference for the omega-3 position over the omega-1 position. A possible role for the L188 residue has been discussed in the context of the structure and function of CYP102A1. (C) 2016 Elsevier B.V. All rights reserved.</P>

신제초제 Azimsulfuron 의 제초활성과 토양중 행동

마상용,전재철 한국환경농학회 1996 한국환경농학회지 Vol.15 No.4

Azimsulfuron [1H-pyrazole-5-sulfonamide,N-(((4,6-dimethoxy-pyridine-2-yl-aminocarbonyl-4-(2-methyl-2H-tetrazole-5-yl)] is a new sulfonamide herbicide that selectively controls a wide range of weeds in lowland rice (Oryza sativa). It effectively controlled Cyperus serotinus, Eleocharis kuroguwai, Sagittaria pygmaea, S. trifolia, and Scirpus juncoides at 7.5 - 30 g ai/㏊. In the tolerance test on grasses carried out in a nutrient solution containing 0.3 - 30 ppm of azimsulfuron, greater inhibition occurred in roots of both rice and barnyardgrass (Echinochloa crus galli) than in shoots. However, rice root was approximately 5-fold more tolerant than that of barnyardgrass. The downward movements as determined by 50% growth inhibition of S. juncoides were 4-㎝ in clay loam and 6.5-㎝ in sandy loam soil with 3-㎝/day leaching for 3 days. When incubated at 20 and 30℃, the residual effect in clay loam soil lasted for 30 and 21 days, respectively. In a soil column applied at 15 g ai/㏊ of azimsulfuron followed by 3-㎝/day leaching for 3 days, dry weights of S. trifolia emerging at 5, 10, and 15-㎝ depth were reduced to 87, 85, and 79% of the corresponding untreated control, respectively. Susceptibility of S. trifolia to azimsulfuron did not greatly vary with the emergence depth.

In Vitro Evaluation of the Genotoxic Activity and Apoptosis Induction of the Extracts of Roots and Leaves from the Medicinal Plant Coccoloba mollis (Polygonaceae)

Marcela S. Tsuboy,Juliana C. Marcarini,Rodrigo C. Luiz,Iuri B. Barros,Dalva T. Ferreira,Lu´cia R. Ribeiro,Ma´rio S. Mantovani 한국식품영양과학회 2010 Journal of medicinal food Vol.13 No.3

Coccoloba mollis (Family Polygonaceae) is a medicinal plant popularly used in cases of memory loss, stress, insomnia, anemia, impaired vision, and sexual impotence, but the scientific literature, to date, lacks studies on the biological effects of this species, particularly with regard to cytotoxicity and induction of DNA damage. The aim of the present study was to assess in vitro (in hepatic HTC cells) ethanolic extracts of the roots and leaves of C. mollis for cytotoxicity, genotoxicity, and induction of apoptosis. For these evaluations the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay, comet assay, micronucleus test with cytokinesis block, and an in situ test for detection of apoptotic cells with acridine orange staining were used. The results showed that the extract obtained from the roots of C. mollis is more cytotoxic than that obtained from the leaves and that the reduction in cell viability observed in the MTT assay was a result, at least in part, from the induction of apoptosis. Both extracts induced DNA damage at a concentration of 20μg/mL in the comet assay, but no genotoxicity was detected with any of the treatments carried out in the micronucleus test.

NMR Solution Structure of HP0827 (O25501_HELPY) from Helicobacter pylori: Model of the Possible RNA-binding Site

Jang, S.-B.,Ma, C.,Lee, J.-Y.,Kim, J.-H.,Park, S. J.,Kwon, A.-R.,Lee, B.-J. Oxford University Press 2009 The Journal of biochemistry Vol.146 No.5

<P>The HP0827 protein is an 82-residue protein identified as a putative ss-DNA-binding protein 12RNP2 Precursor from Helicobacter pylori. Here, we have determined 3D structure of HP0827 using Nuclear Magnetic Resonance. It has a ferredoxin-like fold, beta1-alpha1-beta2-beta3-alpha2-beta4 (alpha; alpha-helix and beta; beta-sheet) and ribonucleoprotein (RNP) motifs which are thought to be important in RNA binding. By using structural homologues search and analyzing electrostatic potential of surface, we could compared HP0827 with other RNA-binding proteins (sex-lethal, T-cell restricted intracellular antigen-1, U1A) to predict RNA-binding sites of HP0827. We could predict that beta sheets of HP0827, especially beta1 and beta3, are primary region for RNA binding. Consequently, similar to other RNA-binding proteins, RNP motifs (Y5, F45, F47), positively charged and hydrophobic regions (K32, R37, K40, K41, K43, R70, R73) are proposed as a putative RNA-binding sites. In addition, differences in amino acids composition of RNP motifs, N, C-terminal residues, loop-region fold and the orientation of alpha1-helix with other RNA recognition motif proteins could give specific biological functions to HP0827. Finally, the study on natural RNA target is also important to completely understand the biological function of HP0827.</P>

Clinical features of Senior-Loken syndrome with IQCB1/NPHP5 mutation in a Filipino man

Harold Henrison C,Chiu,Ma,Sergia Fatima P,Sucaldito,Ebner Bon G,Maceda,Jan Andre S,Montemayor,Diana R,Tamondong-Lachica 대한의학유전학회 2020 대한의학유전학회지 Vol.17 No.1

The Senior-Loken syndrome was first described in 1961 as an oculo-renal disease consisting of familial juvenile nephro-nophthisis and Leber congenital amaurosis. It is a rare autosomal recessive disorder with a prevalence of 1:1,000,000 caused by mutations in nine genes (NPHP 1-8 and NPHP 10). Ocular manifestations (e.g., photophobia, nystagmus, and extreme hyperopia) occur within the ἀrst few years of life while renal manifestations (e.g., formation of multiple cysts impairing kid-ney function and end-stage renal disease) appear in late childhood to adolescence. Here, we report a case of a Filipino male presenting with rotatory nystagmus and progressive deterioration of vision since childhood. He had congenital amaurosis and juvenile nephronophthisis that progressed to end stage renal disease by age 19. All laboratory and imaging ἀndings were consistent with chronic kidney disease. Molecular genetic testing of ciliopathy-related genes was performed revealing a ho-mozygous mutation in exon 11 of the IQCB1/NPHP5 gene, c.1090C>T (p.Arg364*). This sequence change created a prema-ture translational stop signal resulting in a truncated protein product, nephrocystin-5 and its consequent loss of function. His symptoms eventually improved with initiation dialysis. The prognosis of Senior-Loken syndrome remains dismal and a high index of suspicion, early diagnosis and timely intervention of renal complications are warranted.

Paricalcitol prevents cisplatin-induced renal injury by suppressing apoptosis and proliferation

Park, J.W.,Cho, J.W.,Joo, S.Y.,Kim, C.S.,Choi, J.S.,Bae, E.H.,Ma, S.K.,Kim, S.H.,Lee, J.,Kim, S.W. North-Holland ; Elsevier Science Ltd 2012 european journal of pharmacology Vol.683 No.1

The present study was performed to examine whether paricalcitol may prevent the cisplatin-induced kidney injury. Furthermore, potential molecular mechanisms underlying the protective effect of paricalcitol were explored. Male Sprague-Dawley rats were treated with vehicle (n=12), cisplatin (n=12, 6mg/kg/day, i.p.), or cisplatin+paricalcitol (n=12, 0.2μg/kg/day, s.c.) for 4days. In another series of experiment, HK-2 cells were treated with cisplatin (50μM), with or without paricalcitol (0.2ng/ml). Paricalcitol counteracted the cisplatin-induced decline in renal function. Paricalcitol also suppressed the expression of TGF-β1, Smad signaling, and the subsequent epithelial-to-mesenchymal process in cisplatin-treated rats. The expression of P-p53 and p21 was increased in cisplatin-induced nephropathy. These changes were completely prevented or significantly attenuated with paricalcitol co-treatment. The expression of p27<SUP>kip1</SUP> was increased in cisplatin-treated rats, which was, however, further augmented by the paricalcitol co-treatment. In HK-2 cells, cisplatin increased the expression of p-ERK½ and P-p38. Cisplatin also increased the expression of fibronectin and CTGF. Cisplatin increased the expression of pro-apoptotic markers. The expression of CDK2 and Cyclin E as well as that of PCNA was increased. These changes were completely prevented or significantly attenuated by the paricalcitol pretreatment. In contrast, cisplatin increased the expression of p27<SUP>kip1</SUP>, which was further augmented by the paricalcitol-pretreatment. These results suggest that paricalcitol may ameliorate cisplatin-induced renal injury by suppressing the fibrotic, apoptotic and proliferative factors. Its underlying mechanisms may include inhibition of TGF-β1, mitogen-activated protein kinase signaling, p53-induced apoptosis, and augmentation of p27<SUP>kip1</SUP>.

Age-related differences in virtual histology-intravascular ultrasound findings in patients with coronary artery disease

Hong, Y.J.,Jeong, M.H.,Choi, Y.H.,Ma, E.H.,Ko, J.S.,Lee, M.G.,Park, K.H.,Sim, D.S.,Yoon, N.S.,Youn, H.J.,Kim, K.H.,Park, H.W.,Kim, J.H.,Ahn, Y.,Cho, J.G.,Park, J.C.,Kang, J.C. Japanese College of Cardiology 2010 Journal of cardiology Vol.55 No.2

Background: We assessed the age-related differences in pre-intervention virtual histology-intravascular ultrasound (VH-IVUS) findings at target lesion sites in patients with coronary artery disease. Methods: A total of 553 patients who underwent pre-intervention VH-IVUS imaging were grouped according to age: non-elderly (@?70 years, n=429) and elderly (>70 years, n=124); 191 had stable angina and 362 acute coronary syndrome. VH-IVUS classified the tissue into: fibrotic, fibro-fatty, dense calcium (DC), and necrotic core (NC). Results: Overall, the absolute and percent volumes of DC (11.0+/-11.0mm<SUP>3</SUP> vs. 9.7+/-11.9mm<SUP>3</SUP>, P=0.033; 11.7+/-8.1% vs. 9.8+/-7.2%, P=0.014, respectively) and NC (18.5+/-17.6mm<SUP>3</SUP> vs. 16.6+/-18.9mm<SUP>3</SUP>, P=0.020; 18.8+/-8.8% vs. 16.5+/-9.3%, P=0.026, respectively) were significantly greater in the elderly than in the non-elderly. In stable angina patients, the absolute and percent volumes of DC (10.4+/-9.9mm<SUP>3</SUP> vs. 7.2+/-7.6mm<SUP>3</SUP>, P=0.022; 13.4+/-10.0% vs. 9.2+/-6.5%, P=0.011, respectively) and NC (14.8+/-11.2mm<SUP>3</SUP> vs. 12.0+/-11.9mm<SUP>3</SUP>, P=0.035; 19.6+/-8.8% vs. 15.5+/-8.4%, P=0.006, respectively) were significantly greater in the elderly. However, in acute coronary syndrome patients, there were no significant differences in absolute and percent volumes of DC (11.4+/-11.6mm<SUP>3</SUP> vs. 10.9+/-13.4mm<SUP>3</SUP>, P=0.8; 10.7+/-6.5% vs. 10.1+/-7.5%, P=0.5, respectively) and NC (24.1+/-20.3mm<SUP>3</SUP> vs. 23.9+/-21.2mm<SUP>3</SUP>, P=0.9; 22.0+/-8.8% vs. 21.3+/-9.6%, P=0.6, respectively) between the elderly and non-elderly groups. Myocardial infarction (OR: 2.56, 95% CI: 1.45-4.12, P=0.003), diabetes mellitus (OR: 2.23, 95% CI: 1.30-3.53, P=0.009), and high-sensitivity C-reactive protein (OR: 1.44, 95% CI: 1.06-2.45, P=0.042), but not age, were independent predictors of percent NC volume >20% in lesion site. Conclusions: Myocardial infarction, diabetes mellitus, and high-sensitivity C-reactive protein, but not age, were associated with NC-rich lesions. Clinical presentation, risk factors, and inflammatory status, but not age, are important factors for plaque components.

Gender differences in coronary plaque components in patients with acute coronary syndrome: Virtual histology-intravascular ultrasound analysis

Hong, Y.J.,Jeong, M.H.,Choi, Y.H.,Ma, E.H.,Cho, S.H.,Ko, J.S.,Lee, M.G.,Park, K.H.,Sim, D.S.,Yoon, N.S.,Youn, H.J.,Kim, K.H.,Park, H.W.,Kim, J.H.,Ahn, Y.,Cho, J.G.,Park, J.C.,Kang, J.C. Japanese College of Cardiology 2010 Journal of cardiology Vol.56 No.2

Background: The aim of this study was to evaluate the gender differences in plaque components in acute coronary syndrome (ACS) patients. Methods: We used virtual histology-intravascular ultrasound to evaluate the plaque components in culprit lesions in 362 ACS patients (254 men, 108 women). Results: Women were more likely to be diabetic (34% vs 23%, p=0.030), had greater percentage necrotic core (%NC) volume (19.0+/-12.7% vs 16.8+/-11.9%, p=0.040), and had trends toward higher high-sensitivity C-reactive protein (hs-CRP) (0.85+/-1.28mg/dl vs 0.53+/-0.48mg/dl, p=0.063), and higher incidence of thin-cap fibroatheroma (TCFA) (62% vs 52%, p=0.078) compared with men. %NC volume was significantly greater in diabetic patients compared with nondiabetic patients (20.4+/-10.2% vs 16.0+/-8.9%, p<0.001) and was significantly greater in patients with elevated hs-CRP (>=0.2mg/dl) compared with those with normal hs-CRP (<0.2mg/dl) (18.8+/-8.9% vs 16.6+/-9.7%, p=0.021). However, there were no differences in plaque components between diabetic women and men, and between women and men with elevated hs-CRP levels. Diabetes [odds ratio (OR): 2.44, 95% confidence interval (CI): 1.35-3.82, p=0.003] and hs-CRP (OR: 1.54, 95% CI: 1.08-2.65, p=0.032), but not female gender, were the independent predictors of TCFA. Conclusions: Although it seems likely that female ACS patients have more vulnerable plaque components compared with male ACS patients, these findings may result not from true gender differences in plaque components but higher prevalence of diabetes and hs-CRP elevation in women.

Zinc acetate를 precursor로 사용한 ZnO막의 제조 및 습도감지 특성

마대영,이수철,김상현,박기철,김기완 경북대학교 센서기술연구소 1995 센서技術學術大會論文集 Vol.6 No.1

Anhydrous zinc acetate [Zn(CH_(3)COOH)_(3)] has been used as a novel precursor for the growth of ZnO in a conventional thermal evaporator. Zinc acetate was directly heated in the laboratory made brass boat. Zinc acetate was sublimed at the boat temperature of about 220℃. Oxygen has been flowed into the deposition chamber to change the partial pressure of oxygen. X-ray diffraction patterns showed all the films to be amorphous. The films deposited at high oxygen pressure exhibit higher resistivity than films at low pressure. Energy dispersive spectroscopy(EDS) and rutherford backscattering spectrometry(RBS) were conducted on the films to reveal the composition of the ZnO films. The ZnO films deposited at the high oxygen pressure were extremly sensitive to the humidity of higher than 70%.

Effect of Temperature on the Decay Constant of the Axial Solid Holdup Profile in the Splash Region of a Gas Fluidized Bed

Choi, J.-H 한국화학공학회 1997 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.35 No.2