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Saprolegnia parasitica Isolated from Rainbow Trout in Korea: Characterization, Anti-Saprolegnia Activity and Host Pathogen Interaction in Zebrafish Disease Model

  • Shin, Sangyeop;Kulatunga, D.C.M.;Dananjaya, S.H.S.;Nikapitiya, Chamilani;Lee, Jehee;De Zoysa, Mahanama
    • Mycobiology
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    • v.45 no.4
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    • pp.297-311
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    • 2017
  • Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

The Effect of Coptidis Rhizoma, Feaxini Cortex, Jinpisan(秦皮散) on Experimental Pseudomonas aeruginosa Keratitis. (黃連, 秦皮, 秦皮散이 綠膿菌性 角膜炎에 미치는 效果에 관한 實驗的 硏究)

  • Choi, Gyu-dong;Chae, Byung-yoon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.12 no.1
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    • pp.18-35
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    • 1999
  • Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) have been as eye washes of inflammatory eye disease in the oriental medicine. Especially Jinpisan(秦皮散) has been used for the disease which is similar to Peudomonas aeruginisa keratitis. This research was attempted to investigate the effect of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散), on Peudoronas aeruginisa keratitis. Pseudomonas aeruginosa keratitis causes a deep rapid intense ulceration which often leads to perforation of the cornea within 48 hours. In this research, we induced keratits in the rabbits by inoculating Pesudomonas aeruginosa(9027) and observed the effect on the keratitis and the irritation against the external eye. Also we mesured the minimum inhibitory consentration(MIC) of Coptidis Rhizoma, Fraxini Cortex, Jinpisan(秦皮散) by agar diliution method and the anti-bacterial activites by disk method. The tested bacteria were as follows : a) Pseudomonas aeruginosa (9027), b) Streptococcus pneumoniae(6303), c) Staphylococcus epidermidis(12228), d) Staphylococcus aureus(6538P). The results were as follows ; 1. The groups which were applied eye washes of Fraxini Cortex, Jinpisan reavealed a significant effect, but the group applied eye wash of Coptidis Rhizoma reveaded no effect on Pseudomonas aeruginosa keratitis. 2. Applying eye washes of Coptidis Rhizoma, Fraxini Cortex, Jinisan revealed an irritation against external eyes. 3. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylucoccus aureus by agar diliution method 4. Coptidis Rhizoma showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. 5. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method 6. Fraxini Cortex showed an anti-bacterial activity on Pseudomonas aeruginosa, Sireptococcus pneumoniae, Staphylococcus epidermidis, Staphy1ococcus aureus by disk method. 7. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by agar diliution method. 8. Jinpisan showed an anti-bacterial activity on Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus epidermidis, Staphylococcus aureus by disk method. According to the above results, Fraxini Cortex, Jinpisan(秦皮散) are recognized to have an effective treatment on the Pesudomonas aeruginosa keratitis, so this experiment is thought to be a basic ingredient in proving the effect of Fraxini Cortex, Jinpisan which is applied many in documents and clinical medicine. In the comparison of anti-bacterial activity and results of treatment on the Pesudomonas aeruginosa keratitis, Jinpisan(秦皮散) was more effective than Coptidis Rhizoma, Fraxini Cortex.

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Physiological and Anatomical Studies of Quinclorac Action (Quinclorac의 작용성(作用性)에 대한 연구(硏究))

  • Hong, S.Y.;Lee, I.J.;Kim, K.U.;Shin, D.H.;Lee, C.N.
    • Korean Journal of Weed Science
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    • v.13 no.1
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    • pp.62-70
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    • 1993
  • There was intraspecific variation in Echinochloa crus-galli var, crus-galli in response to quinclorac, showing that plan height and dry weight of a locally collected barnyardgrass(Chinjupi) from Chinju were 90.5 and 37.8% of the untreated control, while those of a locally collected one(Iripi) from Iri showed 19.1 and 14.4%, respectively. The normal distribution curve was obtained from frequency distribution of 89 rice cultivars as affected by the application rates of quinclorac at 30, 300, and 3,000g ai/ha. Protein patterns(SDS-PAGE) of two barnyardgrasses belonging to E, crus-galli var. crus-galli such as Iripi and Chinjupi were not affected by the quinclorac application, indicating that inhibition of enzyme and/or protein biosynthesis seems to be not the primary action target of quinclorac. Electronmicroscopic observation on the injured leaf of Iripi which is considered as a susceptible one showed prominent membrane disruption. Chuchungbyeo(rice variety) resulted in a greater inhibition of tomato growth than those from Chinjupi or Iripi, indicating a great amount of quinclorac discharged from rice root, Chinjupi which is relatively tolerant to quinclorac than Iripi, discharged more quinclorac causing a greater inhibition of tomato growth.

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Effects of Nelumbo Folium and Hayopokchun-san on Diabetes Mellitus in Streptozotocin Treated Rats (하엽(荷葉) 및 하엽옥천산(荷葉玉泉散)이 streptozotocin으로 유발된 당뇨(糖尿)에 미치는 영향)

  • Yang, Woong-Mo;Chang, Mun-Seog;Park, Wan-Su;Lee, Byong-Hee;Kim, Won-Nam;Kim, San-Woong;Choi, Dong-Gi;Lee, Hak-Chul;Park, Seong-Kyu
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1497-1501
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    • 2006
  • The therapeutic anti-diabetic effect of Nelumbo Folium and a poly herbal formula (NOC) was evaluated in the streptozotocin (STZ 60 mg/kg, I.p.) induced diabetic rats. For this study, test articles were orally administrated once a day from 7 d after STZ-injection for 3 weeks. The fasting blood glucose, body weight, pancreas weight changes, oral glucose tolerance test, hemoglobin Alc (Hb A1c) level changes and the histopathological changes were observed. WNL-treated rat had lower fasting blood glucose levels compared to control group and NOC-treated rat had significantly lower fasting blood glucose levels, compared to control group (p < 0.05). After 21 days of extract treatment, body weight in control was reduced (p < 0.001). WNL and NOC groups were also reduced compared to control group but insignificantly. Pancreas weight of control group was reduced (p < 0.001), but the weight of NOC group were increased (P< 0.05). During the 2 h oral glucose tolerance test (OGTT), WNL group were improved compared to control group (P < 0.05). NOC group were improved compared to control group but insignificantly. WNL and NOC groups had lower Hb Alc level compared to control group. In addition, atrophy of islet and decrease of insulin-producing cells were detected in STZ induced diabetes rats. However, these diabetic changes were decreased in WNL and NOC groups. These results suggest that N. Folium and NOC have favorable effects to inhibit the changes on the fasting blood glucose levels, pancreas weight, glucose tolerance, hemoglobin Alc and the histopathological changes of pancreas in STZ-induced diabetes.

Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine (소변 중 4-hydroxyproline 분석에 관한 연구)

  • Lee, Keou-Weon;Cho, Young-Bong;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.282-286
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    • 2002
  • Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

Pigment Distribution Analysis of High Speed Fan for Dusting Large Pasture at Livestock Farms (축산농가를 위한 대규모 목초지 방제용 고속 팬의 실험을 통한 색소 분포에 따른 분석)

  • Kim, C.S.;Min, B.R.;Seo, K.W.;Lee, D.W.
    • Journal of Animal Environmental Science
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    • v.15 no.2
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    • pp.99-106
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    • 2009
  • We conducted a comparison experiment of our wide-area spraying high speed fan with a fan which was developed by Japan MARUYAMA Inc. have been much recognized for it's history and skills in a field of spraying machine. In result, MARUYAMA Inc. fan sprayed up to 120 m and, on the other hand, wide-area spraying high speed fan of our own making was able to spray up to more than 160 m. Wide-area spraying machine have been supplied to domestic demands by importing from Japan, but it is determined to be possible that home products will occupy market for it's ability, price and A/S environment in future. The main research results are below. Some plastic cups which are in the open market are used for our experiment. After distributing 90 cups in a range of 10m vertically with localizing 5 cups horizontally and 18 cups vertically, spraying machine was moved and finally we make distribution charts by estimating the sprayed amounts on each cup. Vertical distance was set up to 5m and we calculated average values by using sprayed amount and data of 4 observers. In result our fan showed much sprayed amounts than MARUYAMA Inc. all over the field except situations of vehicle departing and stopping.

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Phototaxis of Fish 6. Responce of Sea-bass to the White Lights (어류의 주광성에 관한 연구 6. 백색광에 대한 능성어의 반응)

  • Yang, Yong Rhim
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.23 no.3
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    • pp.27-27
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    • 1987
  • The purpose of this study is to find the light intensity which induced maximum gathering rate and to observe the variation of the gathering rate both at daytime and night by using Sea-bass, Epinephelus septemfasciatus (Thunberg). An experimental tank (360 L×50 W×55H cm) was set up in a dark room. An illumination system was attached to the end of one side of the tank to control horizontal light intensity. Eight artificial light sources were prepared by combination of three with white light bulbs (10W, 60W, 100W) and eight filters. During the experiment water depth was maintained 50cm level in the tank. The tank was marked into six longitudinal sections each being 60cm long to observe the distribution of fish. The fish were acclimatized in dark condition for 50 minutes before the main experiment. Upon turning on the light, the number of fish in each section was counted 60 times every 30 seconds, and the gathering rate was obtained from the average number of fish in each section. The light intensity inducing maximum gathering rate was 24.13 lux (15.25~35.93 lux) at daytime and 0.41 lux (0.25~0.63 lux) at night. The variation of the gathering rate of fish in illumination time was great and did not show any definite pattern but fluctuated irregularly. Its different between daytime and night is remarkable.

Phototaxis of Fish 6. Responce of Sea-bass to the White Lights (어류의 주광성에 관한 연구 6. 백색광에 대한 능성어의 반응)

  • 양용림
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.23 no.3
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    • pp.137-143
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    • 1987
  • The purpose of this study is to find the light intensity which induced maximum gathering rate and to observe the variation of the gathering rate both at daytime and night by using Sea-bass, Epinephelus septemfasciatus (Thunberg). An experimental tank (360 L$\times$50 W$\times$55H cm) was set up in a dark room. An illumination system was attached to the end of one side of the tank to control horizontal light intensity. Eight artificial light sources were prepared by combination of three with white light bulbs (10W, 60W, 100W) and eight filters. During the experiment water depth was maintained 50cm level in the tank. The tank was marked into six longitudinal sections each being 60cm long to observe the distribution of fish. The fish were acclimatized in dark condition for 50 minutes before the main experiment. Upon turning on the light, the number of fish in each section was counted 60 times every 30 seconds, and the gathering rate was obtained from the average number of fish in each section. The light intensity inducing maximum gathering rate was 24.13 lux (15.25~35.93 lux) at daytime and 0.41 lux (0.25~0.63 lux) at night. The variation of the gathering rate of fish in illumination time was great and did not show any definite pattern but fluctuated irregularly. Its different between daytime and night is remarkable.

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Parish Nursing : A New Challenge for Primary Health Care (지역교회간호(Parish Nursing) - 일차건강간호를 위한 새로운 도약)

  • No, Yu-Ja;Baek, Yeong-Mi
    • The Korean Nurse
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    • v.37 no.2
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    • pp.53-62
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    • 1998
  • ursing as a profession is characterized by its holistic, mind-body-spirit approach to the patient. Also, nurses have historically been the leaders in health education and promotion. Parish nursing has a great potential for providing primary preventive health care. services as well as assisting people to access the health care system. While working in the community, parish nurses see the church as the new arena for delivering health care services. The parish nurse program was introduced by Granger Westberg in 1984. The concept of parish nursing is based on several beliefs; health is multidimensional and affects all aspects of an individual-physical, psychological, social, and spiritaul being. Parish nursing is one model in which churches can cooperatively work with health care institutions to address the needs of their parishioners. The role of the parish nurse is emphasized in four basic area: a) health education, b) health counseling, c) referal services, and d) facilitation and organization of support groups within the congregation. The parish nurse programs work chiefly in congregation or commuity where a certain language of faith is ready at hand. This means that the parish nurse works in an ecology of meanings and care which encourages the drawing on the message of God's grace, the practices and habits it encourages. The parish nurse may be involved in the church's health ministries and may work on either paid or volunteer basis; however, one of the most important qualification of the parish nurse is to have the nursing knowledge and skills to practice within the standards of Nursing Practice Act. The completion of standards of practice for professional nurses practicing as parish nurses had been identified as a priority by the HMA Executive Board (1996, HMA). In conclusion, parish nursing promotes health and healing by empowering the faith community, family, or individual to incorporate health and healing practices. There are several preconditions that should proceed to establish the foundation for successful development of the parish nursing program in Korea. First, reciprocal relationship with home health nursing should be considered. Second, correct terms and concepts of parish nursing should be studied and understood. Third, systematic study and investigation should be followed for further development of parish nursing. Fourth, strengths and weaknesses of different models should be studied to develop proper model of parish nursing for Korean situation. Finally, consensus of standardized education program and corporation with various religious communities as well as health institutions should be established. When these preconditions are met, the role of parish nursing as a new program for the promotion of holistic health will be established.

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Practical Evaluation of Intellectual Capital (IC) Measurement Tool for Contract Foodservice Management Company (위탁급식전문업체 지적자본 측정도구의 운용시험 평가)

  • Park, Moon-Kyunkg;Yang, Il-Sun
    • Journal of Nutrition and Health
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    • v.38 no.10
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    • pp.880-894
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    • 2005
  • The purposes of this study were to a) measure the IC identified of CFMC (contract foodservice management company) ,b) examine IC circumstance of CFMC, c) evaluate practically IC measurement tool of CFMC, and d) present information for selecting an adequate CFMC to clients. The questionnaires of IC measurement were handed out to 108 CFMCs, there composing of main office employees, foodservice managers, customers, and clients of 207 school,38 hospital, and 86 husiness/industry foodservices. The statistical data analysis was completed using SPSS Win (ver 12.0) for descriptive analysis, t-test, Mann-Whitney U test. First, CFMCs had operational experience for an average of 8 years and 8 months, and served an average of 38,540 meals a day. Most of the respondent companies specialized in the school foodservice field and managed an average of 66 clients for the contract period of 2 years and 3 months. Second, the respondent companies had gotten a score of 77.78 points for the total average, 77.7 points in the large enterprise group and 78.1 points in the small and medium-sized enterprise group. Therefore, the minimum number of points for the accrediting license on Qualification is suggested to be over 70 out of a 100 point scale; this study would be serve as reference for the certification license on qualification. On the level of evaluation category, the scores were 14.15 to 20 points on $\ulcorner$finance$\urcorner$, 19.24 to 25 points on $\ulcorner$customer$\urcorner$, 19.33 to 25 points on $\ulcorner$process$\urcorner$, 14.31 to 20 points on $\ulcorner$human resource$\urcorner$, and 8.6 to 10 point on $\ulcorner$renewal and development$\urcorner$ . $\ulcorner$Renewal and development$\urcorner$ and $\ulcorner$customer focus$\urcorner$ received better grades than other evaluation categories. Third, $\ulcorner$Finance$\urcorner$ indicated similar distribution overall. Small and medium-sized companies had lower grades than large companies on 'market ability' of $\ulcorner$customer$\urcorner$ , but, clients of small and medium-sized companies had higher grade for 'client satisfaction' than large companies. Most of the companies supported 'infrastructure support for foodservice operation' of $\ulcorner$process$\urcorner$ by the main office of CFMCs, but, the branch chain offices of CFMCs were not applied efficiently. Large companies made more effort to improve the 'employee ability' of $\ulcorner$human focus$\urcorner$ than small and medium-sized CFMC. The 'research and development cost' of $\ulcorner$renewal and development$\urcorner$ was increased compared to the previous year. In conclusion, if CFMCs were to perform self-evaluation and a routine checkups by utilizing CFMC's IC measuring tool, improvements in CFMC operational capacities as well as foodservice quality can be noted. (Korean J Nutrition 38(10)'880$\sim$894,2005)