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Development of an Efficient Screening System for Resistance of Watermelon Plants to Didymella bryoniae (수박 덩굴마름병에 대한 효율적인 저항성 검정 방법 개발)

  • Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.22 no.2
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    • pp.72-80
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    • 2016
  • Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.

Isolation and Identification of Oceanisphaera sp. JJM57 from Marine Red Algae Laurencia sp. (Ceramiales: Rhodomelaceae) (해양 홍조류 Laurencia sp. (Ceramiales: Rhodomelaceae)에서 분리한 Oceanisphaera sp. JJM57의 분리 및 동정)

  • Kim, Man-Chul;Dharaneedharan, S.;Moon, Young-Gun;Kim, Dong-Hwi;Son, Hong-Joo;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.58-63
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    • 2013
  • A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

Hematologic Changes and Factors Related to Postoperative Hemorrhage Following Cardiopulmonary Bypass (체외순환에 따른 혈액학적 변화와 술후 출혈에 관계하는 인자에 관한 연구)

  • 김하늘루;황윤호;최석철;최국렬;김승우;조광현
    • Journal of Chest Surgery
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    • v.31 no.10
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    • pp.952-963
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    • 1998
  • Background: Cardiopulmonary bypass(CPB)-induced hemostatic defects may result increased possibility of excessive hemorrhage and additional multiple transfusion reactions or reoperation. Particularly, fibrinolytic activation and decreased platelet count and function by CPB were proposed as a predictor of hemorrhage during postoperative periods in several reports. Materials and methods: Present study, which was conducted in 20 adult patients undergoing CPB, was prospectively designed to examine the hematologic changes, including fibrinolytic activation during and after CPB and to clarify the relationships between these changes and the magnitude of the postoperative nonsurgical blood loss. The serial blood samples for measurment of hematologic parameters were taken during operation and postoperative periods. Blood loss was respectively counted via thoracic catheter drainage at postoperative 3, 6, 12, 24, 48 hours and total period. Results: The results were obtained as follows:Platelet count rapidly declined following CPB(p<0.01), which its decreasing rate was an inverse proportion to total bypass time(TBT, r=0.55, p=0.01), And platelet count in postoperative 7th day was barely near to its control value. Fibrinogen degradating product(FDP) and D-dimer level significantly increased during CPB(p<0.0001, p<0.0001, respectively), and both of fibrinogen and plasminogen concentration correlatively decreased during CPB(r=0.57, p<0.01), implying activation of fibrinolytic system. Postoperative bleeding time (BT), postoperative activated partial thromboplastin time(aPTT) and postoperative prothrombin time (PT) were significantly prolonged as compare with each control value (p=0.05, p<0.0001, p<0.0001, respectively). Total blood loss was positively correlated with patient's age, aortic clamping time (ACT) and TBT, while there was negative correlation between platelet count and blood loss at pre-CPB, CPB-off and the 1st postoperative day, and in some periods. Postoperative aPTT and postoperative PTwere positively related to postoperative 6 hr and 48 hr blood loss(r=0.53, p=0.02; r=0.43, p=0.05) but not to total blood loss, whereas there was no relationship between postoperative BT and blood loss at any period. Conclusions: These observations suggest that CPB results various hematologic changes, including fibrinolytic activation and severe reduction in platelet count. Diverse factors such as age, platelet count, ACT, TBT and postoperative aPTT and PT may magnify the postoperative bleeding. This study will be a basic reference in understanding CPB-induced hemostatic injuries and in decreasing the postoperative hemorrhage

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Study on Antioxidant Potency of Green Tea by DPPH Method (DPPH 방법을 통한 녹차의 항산화 활성에 대한 연구)

  • 오중학;김은희;김정례;문영인;강영희;강정숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.7
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    • pp.1079-1084
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    • 2004
  • The present study was conducted to compare antioxidant activity of green teas, fermented teas and other related common teas by examining radical scavenging activity using DPPH (2,2 diphenyl l-picryl hydrazyl). Scavenging activity ($SC_{50}$/) of epigallocatechin gallate (EGCG) for 0.1 mM DPPH radical was 5.5 $\mu$M or 4.2 mg/L by weight, then catechin, 14 $\mu$M or 2.5 mg/L and vitamin C, 22 $\mu$M or 3.9 mg/L, respectively. Kyokuro tea (okro) powder of 24.2 mg/L or green tea powder of 25.2 mg/L was used to reach $SC_{50}$/ for 0.1 mM DPPH. One serving of 2 g green tea provides antioxidant activity equivalent to 109∼147 mg EGCG, 145∼185 mg catechin or 131∼168 mg vitamin C. Teas from the first harvest had the highest radical scavenging activity when compared with later harvest green teas grown in the same region, but there is virtually no difference by the harvest time. A Chinese green tea, Dragon well had the highest antioxidant activity among other green teas tested providing antioxidant capacity equivalent to 168 mg EGCG or 188 mg vitamin C per 2 g serving, but partially fermented Chinese teas had much lower antioxidant activity than any green tea tested. Black tea which is fully fermented showed as strong antioxidant activity as green teas (76.3 mg vs 86.7∼67.6 mg per tea bag). One tea bag of green teas from market provided antioxidant capacity equivalent to 52∼86 mg EGCG, 70∼105 mg catechin or 63-96 mg vitamin C. Teas made of persimmon leaf, pine needle, mulberry leaf had comparatively low anti-oxidant activity equivalent to 2.5∼4.8 mg EGCG or 15∼21 mg vitamin C per teabag. The third brewed green tea still had enough antioxidant activity, while tea from tea bag brewed for 3 min or 5 min did not have any difference in their antioxidant activity. More systemic studies are needed to clarify the relationship between tea catechins and antioxidant capacity focusing on how growing, harvest time, fermentation and other processes can influence on this.

Cell Death Study in Embryonic Stem Cell-derived Neurons and Its Applications (배아줄기세포 유래 신경계세포에서의 세포사멸 연구와 그 응용)

  • Lee, Chul-Sang
    • Development and Reproduction
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    • v.12 no.1
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    • pp.1-8
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    • 2008
  • Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

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Effect of Sucrose Concentration on Survival After Frozen-thawed of Bovine IVF Blastocysts in Ethylene Glycol Based Freezing Medium for Slow-Cooling (소 체외수정란의 Slow Freezing을 위해서 Ethylene Glycol 동결보호제에 Sucrose 첨가 농도에 의한 동결효율)

  • 조상래;김현종;최창용;진현주;손동수;최선호
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.797-804
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    • 2006
  • The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 󰠏7°C, after 2 min, the straw was seeded, maintained at 󰠏7°C for 8 min, and then cooled to 󰠏35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.

Effects of Feeding Patterns of Concentrate on Growth Performance, Blood Parameters and Carcass Characteristics in Fattening Hanwoo Cows (한우 암소 비육시 배합사료 급여형태가 발육, 혈액성상 및 도체특성에 미치는 영향)

  • Lee, Sang Min;Chang, Sun Sik;Jung, Ki Yong;Kim, Hyeong Cheol;Choi, Sun Ho;Kwon, Eung Gi;Park, Byung Ki;Yang, Boh Suk;Lee, Sung Sill;Cho, Young Moo
    • Journal of Animal Science and Technology
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    • v.55 no.1
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    • pp.33-39
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    • 2013
  • This study was conducted to investigate the effects of feeding patterns of concentrates on growth performance, blood parameters and carcass characteristics in Hanwoo cows. Randomly-allocated groups, restricted concentrate feeding (T1), restricted concentrate feeding for 6 months and ad libitum 2 months (T2), restricted concentrate feeding for 4 months and ad libitum 4 months (T3), were contained with 9 animals. According to feeding patterns of concentrate, growth performance was not significantly different among the treatment groups. However, ADG tended to be higher in T2 group (0.75 kg/d) compared to the other groups (T1: 0.62 kg/d, T3: 0.72 kg/d). DMI was not significantly different among the treatment groups, interestingly, rice straw intake was significantly higher in T1 group compared with others (p<0.05). There were significant difference among feed conversion ratio, which are 17.8, 12.8, and 14.1 kg for T1, T2, and T3 (p<0.05), respectively. The serum level of albumin, triglyceride, glucose and GPT were greater in T3 group compared to other groups at fattening 6 to 8 months (p<0.05). The results of yield traits, carcass weight, back fat thickness and rib eye area were not differ among treatment groups, but yield index was significantly greater in T2 group compared to T1 group (p<0.05). The 'A' appearance rate (%) of meat yield grade was highest in T2 group for 78%. The marbling score, meat color, fat color, texture and maturity in quality traits were not differ among the treatment groups. However, marbling score and appearance rate (%) of over 1st meat quality grade were tend to be increased at T3 rather (4.0, 56%) group compared with other groups (T1: 3.4, 56%; T2: 3.6, 33%). Thus, the present study suggested that restricted concentrate feeding (1.6% of BW) for 4 months during early fattening periods and ad libitum feeding for 4 months during late fattening periods are recommendable.

Etiology and Clinical Features of Viral Lower-respiratory Tract Infections in Children in Winter, 2003 (2003년 겨울철 소아의 바이러스성 하기도 감염증에 대한 원인 및 바이러스별 임상양상)

  • Yun, Byung Ho;Lee, Hee Chul;Chun, Jung Mi;Yoon, So Young;Lee, Woo Gill;Shin, Son Moon
    • Clinical and Experimental Pediatrics
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    • v.48 no.7
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    • pp.723-730
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    • 2005
  • Purpose : This study was performed to characterize the etiology and clinical features of acute viral lower-respiratory tract infections(LRI). Methods : Etiologic agents and clinical features of acute viral LRI were studied from October. 2003 through March. 2004 in hospitalized children with LRI(253 cases) at Samsung Cheil Hospital. The viruses were identified by indirect immunofluorescent method. Medical records of patients with proven viral LRI were reviewed retrospectively. Results : Ninety two cases(36.4%) were confirmed as viral infections. The identified pathogens were respiratory syncytial virus(RSV, 76.0%), adenovirus(ADV, 12.0%), influenza virus type A(INFA, 9.8 %), influenza virus type B(INFB, 1.1%) and parainfluenza virus(PIV, 1.1%). Eight four point eight% of patients were younger than 2 years of age. Clinical diagnosis of LRI were pneumonia(56.5%), bronchiolitis(35.9%), tracheobronchitis(4.3%) and croup(3.3%). The clinical symptoms and signs were cough(98.8%), rhinorrhea(82.6%), fever(70.7%), rale(67.4%), wheezing(29.3%), chest retraction(28.3%) and cyanosis(4.3%). The severe respiratory symptoms and signs were more common in RSV-infected patients, even cyanosis could be observed. Seventeen point four percent of patient had fever of $38.5^{\circ}C$ or higher and their most common etiologic agent was INFA(66.7%). Twenty three point nine percent had fever more than 5 days and common etiologic agent was INFA(77.8%). The elevated WBC count($>14{\times}10^3/{\mu}L$) was in 14.1%, and common etiologic agents were INFA(22.2%) and ADV(18.2%). C-reactive protein(CRP >4.0 mg/dL) was increased in 13.0%, and common in ADV(63.6 %). Increased aspartate aminotransferase(AST)/alanine aminotransferase(ALT) was detected in 10.9%, and the most common etiologic agent was RSV(12.9%). Conclusion : The common agents of acute viral LRI were RSV, ADV and INF, respectively. Because the etiologic agents present variable clinical features, it may be helpful to treat and to evaluate acute viral LRI that we should understand their etiologic variability.

A Cyclin-Dependent Kinase Inhibitor, p16^{INK4A}, Induces Apoptosis in The Human Cancer Cells. (Cyclin-dependent Kinase저해 단백질 p16^{INK4A}의 인체 암세포에서의 세포사멸 유도 활성)

  • 김민경;이철훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.72-77
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    • 2004
  • Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

Determination of homogentisic acid in human plasma by GC-MS for diagnosis of alkaptonuria (GC-MS를 이용한 혈장 중 호모겐티식산의 분석;알캅톤뇨증의 진단)

  • Thapa, Maheshwor;Yu, Jundong;Lee, Wonjae;Islam, Fokhrul;Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.323-330
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    • 2015
  • Alkaptonuria, a rare inherited metabolic disease, is characterized by a lack of homogentisate dioxygenase and accumulation of homogentisic acid (HGA), leading to homogentisic aciduria, arthritis, and ochronosis. In this study, a rapid analytical method, without an expensive and tedious solid phase extraction step, was developed to quantify HGA in plasma using GC-MS. HGA-spiked pooled plasma samples were subjected to liquid-liquid extraction (LLE) with ethyl acetate, followed by trimethylsilyl derivatization (TMS) and GC-MS quantification using selected ion monitoring. The formation of TMS derivative of the 1 carboxylic and 2 hydroxyl functional groups was performed by reacting BSTFA (with 10% TMCS) for 5 min at 80 ℃. For selected ion monitoring, quantification and confirmation ions were determined based on specific ions (m/z 384, m/z 341 and m/z 252) of the TMS derivative of HGA. Calibration curves of pooled normal plasma specimens showed a linear relationship in the range of 1-100 ng/µL. The precision and accuracy were within a relative standard deviation (RSD) of 1 to 15% and a bias of -5 to 25%. Recoveries were obtained in the range of 99-125% and 95-115% for intra-day and inter-day assay, respectively, at 2, 20 and 80 ng/µL. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/µL and 4 ng/µL, respectively. No homogentisic acid was excreted from normal Korean plasma samples. Collectively, the results from the present study suggest that this method could be useful for routine diagnosis and therapeutic monitoring of alkaptonuria patients with excellent sensitivity and rapidity.