• The Plant Pathology Journal
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• v.20 no.1
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• pp.52-57
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• 2004

Sclerotinia sp. (isolate BWC98-105) causes stem blight and root rot in Leghum sp., and is presently being evaluated as a potential mycoherbicide for the control of Trifoliorium repens. Bioassays have shown that Sclerotinia sp. produces phytotoxic substance which is biologically active against T. repens. Two biologically active compounds, designated as compoundsI and II, were produced in vitro from the culture filtrate of BWC98-105 isolate Sclerotium sp. Compounds I and II were purified by means of liquid-liquid extraction and $C_{18}$ open column chromatography (300 ${\times}$ 30 mm, i.d). To determine the purity, the purified compounds were analyzed by RP-HPLC. The analytical RP-HPLC column was a TOSOH ODS-120T (150 ${\times}$ 4.6 mm i.d, Japan), of which the flow rate was set at 0.7 mL/min using the linear gradient solvent system initiated with 15 % methanol to 85 % methanol for 50 min with monitoring at 254 nm. Under these RP-HPLC conditions, compounds I and II eluted at 3.49 and 4.13 min, respectively. Compound II was found to be most potent and host specific. However, compound I had a unique antibiotic activity against phytopathogenic bacteria like bacterial leaf blight (Xanthomonas oryzae) on rice, where it played a less important role in producing toxicity on T. repens. No toxin activity was detected in the water fraction after partitioning with several organic solvents. However, toxin activity was detected in the ethyl acetate and butanol fractions. In the leaf bioassay using compound II, the disease first appeared within 4-5 h as water soaked rot, which subsequently developed into well-defined blight affecting the whole plant.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.5
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• Radiation Oncology Journal
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• v.37 no.3
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• pp.215-223
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• 2019

Purpose: To determine prognostic significance of lymphovascular invasion (LVI) in prostate cancer patients who underwent adjuvant or salvage postoperative radiotherapy (PORT) after radical prostatectomy (RP) Materials and Methods: A total of 168 patients with prostate cancer received PORT after RP, with a follow-up of ≥12 months. Biochemical failure after PORT was defined as prostate-specific antigen (PSA) ≥0.2 ng/mL after PORT or initiation of androgen deprivation therapy (ADT) for increasing PSA levels regardless of the value. We analyzed the clinical outcomes including survivals, failure patterns, and prognostic factors affecting the outcomes. Results: In total, 120 patients (71.4%) received salvage PORT after PSA levels were >0.2 ng/mL or owing to clinical failure. The 5-year biochemical failure-free survival (BCFFS), clinical failure-free survival (CFFS), distant metastasis-free survival (DMFS), overall survival, and cause-specific survival rates were 78.3%, 94.3%, 95.0%, 95.8%, and 97.3%, respectively, during a follow-up range of 12-157 months (median: 64 months) after PORT. On multivariate analysis, PSA level of ≤1.0 ng/mL at the time of receiving PORT predicted favorable BCFFS, CFFS, and DMFS. LVI predicted worse CFFS (p = 0.004) and DMFS (p = 0.015). Concurrent and/or adjuvant ADT resulted in favorable prognosis for BCFFS (p < 0.001) and CFFS (p = 0.017). Conclusion: For patients with adverse pathologic findings, PORT should be initiated as early as possible after continence recovery after RP. Even after administering PORT, LVI was an unfavorable predictive factor, and further intensive adjuvant therapy should be considered for these patients.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.234-247
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• 2021

We used a heterozygous gene deletion library of fission yeasts comprising all essential and non-essential genes for a microarray screening of target genes of the antifungal terbinafine, which inhibits ergosterol synthesis via the Erg1 enzyme. We identified 14 heterozygous strains corresponding to 10 non-essential [7 ribosomal-protein (RP) coding genes, spt7, spt20, and elp2] and 4 essential genes (tif302, rpl2501, rpl31, and erg1). Expectedly, their erg1 mRNA and protein levels had decreased compared to the control strain SP286. When we studied the action mechanism of the non-essential target genes using cognate haploid deletion strains, knockout of SAGA-subunit genes caused a down-regulation in erg1 transcription compared to the control strain ED668. However, knockout of RP genes conferred no susceptibility to ergosterol-targeting antifungals. Surprisingly, the RP genes participated in the erg1 transcription as components of repressor complexes as observed in a comparison analysis of the experimental ratio of erg1 mRNA. To understand the action mechanism of the interaction between the drug and the novel essential target genes, we performed isobologram assays with terbinafine and econazole (or cycloheximide). Terbinafine susceptibility of the tif302 heterozygous strain was attributed to both decreased erg1 mRNA levels and inhibition of translation. Moreover, Tif302 was required for efficacy of both terbinafine and cycloheximide. Based on a molecular modeling analysis, terbinafine could directly bind to Tif302 in yeasts, suggesting Tif302 as a potential off-target of terbinafine. In conclusion, this genome-wide screening system can be harnessed for the identification and characterization of target genes under any condition of interest.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.207-215
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• 2004

The present study was carried out to evaluate the ISSR and RAPD markers for their efficiency as genetic marker systems to establish the relationships between 18 mulberry genotypes. A total of 36 from 56 (64%) RAPD primers and 12 from 48 (25%) ISSR primers produced reproducible amplification patterns. A high proportion of polymorphic bands ranging from 44 to 91% was observed respectively with RAPD and ISSR markers. The average Resolving Power (Rp) of ISSR primers was higher than RAPD primers. The ISSR primers, UBC 825, 868 and 873, and RAPD primers, UBC 712, 720 and 729, possessed the highest Rp values and could in each instance distinguish all the 18 genotypes. Similarity matrix values were estimated based on Jaccards coefficient, considering 109 polymorphic ISSR and 212 polymorphic RAPD bands and two dendrograms were constructed. The dendrograms obtained with ISSR and RAPD markers distinguished the eight exotic genotypes from the ten indigenous (Indian) genotypes. A significant correlation value (r=0.959; p=0.001) for the cophenetic matrix between the RAPD and ISSR matrices was observed. The results indicated that the ISSR and RAPD markers could assist in the differentiation of genotypes and permit the determination of genetic distances that might be exploited by mulberry breeders in improvement programs.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2005.10a
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• pp.334-337
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• 2005

The objective of this research works is to propose a rapid development methodology of small size deep drawing tools for electronic parts utilizing the technology combination of CAE and RP/RT. The technology is applied to the development of deep drawing tools with a drain shape. The final surface of tools is obtained from the evaluation of the formability using the CAE. In order to manufacture the physical prototype of tools fer try-out terming, several fabrication experiments are carried out for three types of rapid tool manufacturing technology. Through the fabrication experiments, the acceptable rapid manufacturing technologies of deep drawing tools with a small size have been proposed.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.6
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• pp.445-453
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• 2000

The present investigation tested the hypothesis that the activation of protein kinase G (PKG) leads to a phosphorylation of $Ca^{2+}-activated$ potassium channel $(K_{Ca}\;channel)$ and is involved in the activation of $K_{Ca}$ channel activity in cerebral arterial smooth muscle cells of the rabbit. Single-channel currents were recorded in cell-attached and inside-out patch configurations of patch-clamp techniques. Both molsidomine derivative 3-morpholinosydnonimine-N-ethylcarbamide $(SIN-1,\;50\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate $(8-pCPT-cGMP,\;100\;{\mu}M),$ a membrane-permeable analogue of cGMP, increased the $K_{Ca}$ channel activity in the cell-attached patch configuration, and the effect was removed upon washout of the drugs. In inside-out patches, single-channel current amplitude was not changed by SIN-1 and 8-pCPT-cGMP. Application of ATP $(100\;{\mu}M),$ cGMP $(100\;{\mu}M),$ ATP+cGMP $(100\;{\mu}M\;each),$ PKG $(5\;U/{\mu}l),$ ATP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l),$ or cGMP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ did not increase the channel activity. ATP $(100\;{\mu}M)+cGMP\;(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ added directly to the intracellular phase of inside-out patches increased the channel activity with no changes in the conductance. The heat-inactivated PKG had no effect on the channel activity, and the effect of PKG was inhibited by 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer $(Rp-pCPT-cGMP,\;100\;{\mu}M),$ a potent inhibitor of PKG or protein phosphatase 2A (PP2A, 1 U/ml). In the presence of okadaic acid (OA, 5 nM), PP2A had no effect on the channel activity. The $K_{Ca}$ channel activity spontaneously decayed to the control level upon washout of ATP, cGMP and PKG, and this was prevented by OA (5 nM) in the medium. These results suggest that the PKG-mediated phosphorylations of $K_{Ca}$ channels, or some associated proteins in the membrane patch increase the activity of the $K_{Ca}$ channel, and the activation may be associated with the vasodilating action.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.480-483
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• 2003

Fusion ferritin$(F_H+F_L)$, an iron-binding protein, was purified from recombinant E. coli by two-step sonications with urea. Unfolded ferritin was refolded by gel filtration chromatography with various concentration of urea. 50 mM Tris-HCl(pH 8.0) buffers with 1 M to 4 M urea were used in GFC. Objective was to characterize the structure change with urea concentration. Molecular weight was determined using GF-HPLC and RP-HPLC was used to quantify the unfolded and refolded proteins.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.677-683
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• 2008

The antioxidant activities of Wisteria floribunda flowers (WFF) were evaluated. The samples were prepared by extracting separately two different colored flowers (purple and white) with four different solvents (methanol, ethanol, acetone, and water). The antioxidant properties were evaluated by determining total phenolic contents (TPC), radical scavenging activity (RSA), and reducing power (RP). Water extract from purple WFF and ethanol extract of white WFF showed the highest total phenol contents (491 and 787 ${\mu}M$ gallic acid equivalents), respectively. Water extracts of purple and white WFF also showed higher RSA. In the case of RP, ethanol extract of purple WFF, methanol and water extracts of white WFF showed relatively higher values. The 200 ${\mu}M$ $H_2O_2$ induced oxidative DNA damage in human leukocytes was significantly inhibited with WFF extracts excluding ethanol and acetone extracts of purple flowers. These results suggest that W. floribunda flowers have significant antioxidative activity and protective effect against oxidative DNA damage.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.544-547
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• 2003

Fusion $ferritin(F_H+F_L),$ an iron-binding protein, was purified from recombinant E. coli by gel filtration chromatography after two-step sonications. Unfolded ferritin was refolded by GFC with various refolding enhancing additives. 50 mM Tris-HCI(pH 7.4) buffers containing 2 M urea and additive was used in GFC. Objective was to characterize the structure change at various conditions. Molecular weight was determined using GF-HPLC and RP-HPLC was used to quantify the unfolded and refolded proteins. Activity was confirmed by iron-uptake reaction.