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Stem Rot on Ligularia fischeri Caused by Sclerotium rolfsii in Korea (Sclerotium rolfsii에 의한 곰취 흰비단병)

  • Moon, Youn-Gi;Kim, Se-Won;Choi, Jun-Keun;Kwon, Soon-Bae;Shim, Hong-Sik;Ju, Ho-Jong;Choi, In-Young
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.36-39
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    • 2015
  • In June 2012 and 2013, a destructive stem rot symptoms of Ligularia fischeri occurred sporadically in Hoengseong-gun and Pyeongchang-gun Gangwon-do, Korea. The typical symptom included water-soaking on the main stem, rotting, wilting and blighting, which eventually leads to death of the plant. White mycelial mats were spread over lesions and brown sclerotia were formed on stems and near soil surface. The sclerotia were white to brown, spherical or irregular, 1-3 mm in size on potato dextrose agar (PDA), The optimum temperature range of hyphal growth was $25-30^{\circ}C$ and the hyphal diameter was $4-10{\mu}m$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. The resulting sequence of 695 bp was deposited in GenBank. A BLAST search revealed that sequences of the this isolates showed >99% identity with those of Sclerotium rolfsii. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers ITS rDNA, the fungi were identified as S. rolfsii. A pathogenicity test was carried out to fulfill Koch's postulates. To our knowledge, this is the first report of S. rolfsii on Ligularia fischeri in Korea.

Study on Formation of FePd Nano-dot Using Agglomeration of Fe/Au Bilayer (Fe/Au 이중층의 응집현상을 이용한 FePd 나노 점 형성에 관한 연구)

  • Koo, J.K.;Kim, J.M.;Ryua, D.H.;Choi, B.J.;Kim, D.W.;Lee, D.H.;Kim, U.I.;Mitani, S.;J.G., M. Kamiko;Ha, J.G.
    • Journal of the Korean Vacuum Society
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    • v.20 no.1
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    • pp.7-13
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    • 2011
  • [ $L1_0$ ]phase FePd nano-dot structures were successfully fabricated on self-organized Fe/Au bilayers. With atomic force microscopy, it is determined that surface morphologies of initially flat Fe/Au bilayer films were agglomerated and transformed their shape into nano-dots structures with increasing annealing temperature. With this bilayer as a template, FePd multilayers were deposited at various temperatures, i.e. $300^{\circ}C$, $350^{\circ}C$, $400^{\circ}C$, and $450^{\circ}C$. Surface morphologies of FePd superlattice had a near resemblance to self-organized bilayer. According to X-ray diffraction results, it is confirmed that $L1_0$ superlattice structures of FePd were obtained from samples which were annealed above $350^{\circ}C$. Results of X-ray photoelectron spectroscopy depth-profile analysis showed that chemical composition is identical to deposition sequence. As a result, without additional etching processes, fabrication of chemically ordered FePd superlattice nano-dots was achieved.

Bioequivalence of Flucona Capsule to Diflucan Capsule (Fluconazole 50 mg) (디푸루칸 캅셀(플루코나졸 50 mg)에 대한 플루코나 캅셀의 생물학적 동등성)

  • Cho, Hea-Young;Kang, Hyun-Ah;Lee, Suk;Oh, In-Joon;Lim, Dong-Koo;Moon, Jai-Dong;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.33 no.2
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    • pp.135-140
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    • 2003
  • Fluconazole is an orally active bis-triazole antifungal agent, which is used in the treatment of superficial and systemic candidiasis and in the treatment of cryptococcal infections in patients with the acquired immuno deficiency syndrome (AIDS). The purpose of the present study was to evaluate the bioequivalence of two fluconazole capsules, Diflucan (Pfizer Pharmaceuticals Korea Inc.) and Flucona (Korean Drug Pharmaceuticals Co., Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). The fluconazole release from the two fluconazole capsules in vitro was tested using KP VII Apparatus II method at 0.1 M hydrochloride dissolution media. Twenty normal male volunteers, $23.60{\pm}1.88$ years in age and $63.57{\pm}6.17\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After three capsules containing 50 mg as fluconazole was orally administered, blood was taken at predetermined time intervals and the concentrations of fluconazole in serum were determined using HPLC method with UV detector. The dissolution profiles of two fluconazole capsules were very similar at 0.1 M hydrochloride dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$ and $C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AUC_t,\;C_{max}\;and\;T_{max}$ between two capsules based on the Diflucan were 4.96%, 5.65% and -13.76%, respectively. There were no sequence effects between two capsules in these parameter. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log(0.8) to log(1.25) $(e.g.,\;log(1.01){\sim}log(1.08)\;and\;log(1.00){\sim}log(1.12)\;for\;AUC_t\;and\;C_{max},\;respectively)$, indicating that Flucona capsule is bioequivalent to Diflucan capsule.

Separation and Purification of Angiotensin I-converting Enzyme Inhibitory peptide from Mackerel (고등어 유래 항고혈압 peptide의 분리 정제)

  • DO Jeong-Ryong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.2
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    • pp.153-157
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    • 2000
  • Hydrolysate which inhibit the Angiotensin I-converting enzyme (ACE) was prepared from mackerel muscle by pretense. The ACE inhibitory activity of mackerel muscle hvdrolysate (MMH) was $967 {\mu}g of IC_(50)$. ACE inhibitory peptides were isolated by ultrafiltration, gel permeation column chromatography (GPC), reversed phase column chromatography(RPC), reversed phasehigh performance liquid chromatography (RP-HPLC), and gel permeation high performance liquid chromatography (GP-HPLC) from the MMH. The amino acid sequence of the ACE inhibitory peptides was Tyr-Val-Ala. The $IC_(50)$ of this peptide for ACE from rabbit lung was $1.4 {\mu}M$.

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Predicting Parturition Time through Ultrasonic Measurement of Posture Changing Rate in Crated Landrace Sows

  • Wang, J.S.;Wu, M.C.;Chang, H.L.;Young, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.682-692
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    • 2007
  • This study presents an automatic system to predict parturition time in the crated sows. The system relies on ultrasonic transducers mounted from above along the length of the crate. Using a 40 kHz time of flight (TOF) single envelope wave, the momentary distances between the sensors are measured. Therefore, the local momentary height of the sow and the momentary posture, i.e. standing posture (SDP), kneeling posture (KP), sitting posture (STP) and lateral lying posture (LLP) are determined. Crated sows change their postures from standing to lying and vice versa which follows a characteristic pattern. As parturition approaches, sows exhibit uneasiness, restlessness and the stand up sequence (SUS, the posture transition from LLP to SDP) rate increases because of labor pains. In time series, the SUS rate demonstrates a peak and it happens approximately 0-12 h before parturition. In this paper, the basic parturition threshold value method (BPTVM) and the same hour method (SHM) are proposed for predicting parturition, both of which are based on the SUS rate. The BPTVM mainly detects the peak of the SUS rate. As the SUS rate exceeds the threshold value, the parturition becomes predictable. Moreover, the SHM calculates the difference in the SUS rates between a particular time of day and the corresponding time of the preceding day. Compared to the BPTVM, the SHM can eliminate the circadian rhythm of the SUS rate influenced by feeding behavior. Using the SHM the parturition can be approximately predicted within hours. In an attempt to define the threshold parameters of predicting parturition, a data set with 32 sows of the SUS rate are used to estimate assumable predicting probability. The results show the assumable probability of the parturition prediction within 9 h is 96.9% for the SHM and 84.4% for the BPTVM. Moreover, the SHM can even reach a 75% probability of prediction within three hours of parturition. We conclude that the SHM is more accurate and is more useful for parturition time prediction. When parturition is detected, the proposed algorithm generates a warning signal which can inform human personnel to protect the mother and newborn piglets.

Amplification of Porcine SRY Gene for Sex Determination

  • Choi, S.G.;Bae, M.S.;Lee, E.S.;Kim, S.O.;Kim, B.K.;Yang, J.H.;Jeon, C.E.;Kim, H.H.;Hwang, Y.J.;Lee, E.S.;Kim, D.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1107-1112
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    • 2009
  • The separation of X and Y chromosome-bearing sperm is of use in many aspects of livestock maintenance. In this study, we sought to determine the difference in DNA content between X- and Y-bearing sperm, separate sperm into X- and Y-enriched pools, and assess the efficacy of sorting. Sperm collected from Duroc and miniature pigs were stained with 20.8 $\mu{M}$ Hoechst 33342 and analyzed using a high-speed cell sorter. Measurement of the fluorescence intensity of stained sperm nuclei revealed that the X-bearing sperm of Duroc and miniature pigs respectively contain 2.75% and 2.88% more DNA than Y-bearing sperm. In total, 50.18% of the sperm were assigned to the X-sorted sample and 49.82% was assigned to the Y-sorted sample for Duroc pigs. For miniature pigs, the Xsorted sample represented 50.19% of the population and the Y-sorted represented 49.81% of the population. Duplex PCR was used to evaluate accuracy of sorting. A fast and reliable method for porcine sexing was developed through amplification of the sex-determining region of the Y chromosome gene (SRY). Oligonucleotide primers were designed to amplify the conserved porcine SRY high motility group (HMG) box sequence motif. We found that the primer pair designed in this study was 1.46 times more specific than previously reported primers. Thus, this study shows that the present method can be applied in porcine breeding programs to facilitate manipulation of the sex ratio of offspring and to achieve precise sexing of porcine offspring by amplification of the HMG box of the SRY gene.

Knockdown of GCF2/LRRFIP1 by RNAi Causes Cell Growth Inhibition and Increased Apoptosis in Human Hepatoma HepG2 Cells

  • Li, Jing-Ping;Cao, Nai-Xia;Jiang, Ri-Ting;He, Shao-Jian;Huang, Tian-Ming;Wu, Bo;Chen, De-Feng;Ma, Ping;Chen, Li;Zhou, Su-Fang;Xie, Xiao-Xun;Luo, Guo-Rong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.6
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    • pp.2753-2758
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    • 2014
  • Background: GC-binding factor 2 (GCF2) is a transcriptional regulator that represses transcriptional activity of the epidermal growth factor receptor (EGFR) by binding to a specific GC-rich sequence in the EGFR gene promoter. In addition to this function, GCF2 has also been identified as a tumor-associated antigen and regarded as a potentially valuable serum biomarker for early human hepatocellular carcinoma (HCC) diagnosis. GCF2 is high expressed in most HCC tissues and cell lines including HepG2. This study focused on the influence of GCF2 on cell proliferation and apoptosis in HepG2 cells. Materials and Methods: GCF2 expression at both mRNA and protein levels in HepG2 cells was detected with reverse transcription (RT) PCR and Western blotting, respectively. RNA interference (RNAi) technology was used to knock down GCF2 mRNA and protein expression. Afterwards, cell viability was analyzed with a Cell Counting Kit-8 (CCK-8), and cell apoptosis and caspase 3 activity by flow cytometry and with a Caspase 3 Activity Kit, respectively. Results: Specific down-regulation of GCF2 expression caused cell growth inhibition, and increased apoptosis and caspase 3 activity in HepG2 cells. Conclusions: These primary results suggest that GCF2 may influence cell proliferation and apoptosis in HepG2 cells, and also provides a molecular basis for further investigation into the possible mechanism at proliferation and apoptosis in HCC.

Characterization of a Multimodular Endo-β-1,4-Glucanase (Cel9K) from Paenibacillus sp. X4 with a Potential Additive for Saccharification

  • Lee, Jae Pil;Kim, Yoon A;Kim, Sung Kyum;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.588-596
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

PKA-Mediated Regulation of B/K Gene Transcription in PC12 Cells

  • Choi, Mi-Hyun;Kim, Ho-Shik;Choi, Sung-Ho;Kim, Mi-Young;Jang, Yoon-Seong;Jang, Young-Min;Lee, Jeong-Hwa;Jeong, Seong-Whan;Kim, In-Kyung;Kwon, Oh-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.6
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    • pp.333-339
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    • 2005
  • B/K protein is a novel protein containing double C2-like domains. We examined the specific signaling pathway that regulates the transcription of B/K in PC12 cells. When the cells were treated with forskolin ($50{\mu}M$), B/K mRNA and protein levels were time-dependently decreased, reaching the lowest level at 3 or 4 hr, and thereafter returning to the control level. Chemicals such as dibutyryl-cAMP, cellpermeable cyclic AMP (cAMP) analogue and CGS21680, adenosine receptor $A_{2A}$ agonist, also repressed the B/K transcription. However, 1,9-dideoxyforskolin did not show inhibitory effect on B/K transcription, suggesting direct involvement of cAMP in the forskolin-induced inhibition of B/K transcription. Effect of forskolin, dibutyryl cAMP and CGS21680 was significantly reduced in PKA-deficient PC12 cell line (PC12-123.7). One cAMP-response element (CRE)-like sequence (B/K CLS) was found in the promoter region of B/K DNA, and electrophoretic mobility shift assay indicated its binding to CREM and CREB. Forskolin significantly suppressed the promoter activity in CHO-K1 cells transfected with the constructs containing B/K CLS, but not with the construct in which B/K CLS was mutated (AC:TG). Taken together, we suggest that the transcription of B/K gene in PC12 cells may be regulated by PKA-dependent mechanism.

Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.