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Study on In Vitro Aggregation and Culture of Mouse Embryos by Phytohemagglutinin-P (Phytohemagglutinin-P 첨가(添加)에 따른 생쥐배(胚)의 시험관내(試驗管內) 응집(凝集)과 배양(培養)에 관하여)

  • Park, Hang Kyun;Ryou, Zae Yoong
    • Current Research on Agriculture and Life Sciences
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    • v.7
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    • pp.83-97
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    • 1989
  • This study was carried out to obtain basic information necessary for aggregation and in-vitro culture of mouse embryos by treating phytohemagglutinin-p (PHA-P). The 4-, 8-cell and morula embryos were obtained from female mice of albino BALE/C, CBA and C57BL strains, those were injected 5 i.u pregenant mare serum gonadotrophin and 5 i.u human chorionic gonadotrophin to superovulation. The zona pellucidia was removed by placing the embryos in Acidic Tyrode solution containing 1.0% protease or/and 5 ug/ml PHA-P. The pairs of zona free embryos were subjected to aggregation by glassneedle in BMOC-3 containing 5 ug/ml PHA-P. The aggregation embryos were cultured in Brinster's mouse ova culture-3(BMOC-3) medium under the gas phase of 5% $CO_2$ in air $37^{\circ}C$ for 13 to 50 hours. The results obtained in this study are summarised as follows : 1. When 4-, 8-cell and morula embryos were zona-freed in acidic Tyrode solution containing 1.0% protease or/and 5 ug/ml PHA-P, and cultured in vitro to blastocysts, the 4- and 8-cell embryos showed slightly less development rates than the morula one did, and solution of 5 ug/ml PHA-P brought some higher development rate than negative control. 2. As 2, 5 or 10 ug/ml PHA-P was added to the solution to aggregate 4-, 8-cell or morula embryos, 2 ug/ml solution represented slightly lower aggregation rate than the higher levels solutions, and 4- and 8-cell embryos showed higher rates than morula one did (P<.05). 3. In respect to the development rates of aggregated embryos to morula no significant difference was found among PHA-P levels and between 4-and 8-cell embryos. With respect to those of aggregated embryos to blastocysts the different levels of PHA-P showed similar results, however, the 4- and 8-cell embryos represented higher rates than the morula one did (P<.05). 4. The mean time necessary for development of aggregated 4-, 8-cell and morula embryos to blastocysts were 38.5-40, 26-27 and 19-20hrs. Respectively in solution for aggregation. 5. The aggregation rates of embryos were 34-94%, when treated protease or/and PHA-P. Supplementation of 5 ug/ml PHA-P to the solution for aggregation showed a trend demonstrating higher aggregation rate compared to negative control, although no significance was found. However, 4- and 8-cell embryos represented significantly higher aggregation rates than the morula one did (P<.05). 6. The development rates of 4- and 8-cell embryos to morula were 52.7-84.7 and 73.8-87.2%, respectively, showing no significant difference between two cell stages. However, the aggregation rates of embryos treated with solution containing PHA-P were higher than negative control (P<.05). 7. The development rates of 4- and 8-cell and morula embryos to blastocysts were 41.7-77.7 78.7-83.0 and 0-19.2%, respectively. The rates of 4-cell embryos treated with PHA-P were significant higher than the negative control (P<.05). The 8-cell and morula embryos also showed more rates when treated PHA-P.

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Seedling - Emergence of Rice, Weedy Rice, and Echinochloa species Sown before Wintering and in the Early Spring (월동전(越冬前) 초춘(初春)에 파종(播種)한 재배(栽培)벼, 잡초성(雜草性)벼 및 피의 출현특성(出現特性))

  • Kwon, Y.W.;Lee, B.W.;Kim, D.S.
    • Korean Journal of Weed Science
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    • v.16 no.2
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    • pp.88-99
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    • 1996
  • In recent years dry direct-seeding of rice has been encouraged by the government and increasingly practiced by farmers in Korea. This has been bringing up an increased occurrence of weedy rites. Some farmers in the southern region dare to sow the rice before winter after harvest, while most farmers wish to sow as early as possible in the spring to secure the growing period, and to disperse the intensive labor in early May. The purpose of this study was to determine the feasibility of moving the sowing of rice to an earlier date under dry direct-seeding, and to elucidate the nature of emergence of an old strain of rice, weedy rites, and barnyardgrasses tinder this farming practice and their adaptive competence over present cultivate. The presently recommended rice cultivar, Dongjinbyo and an old rice strain, Dadajo which prevailed in early 1900s, almost could not emerge from soil deeper than 6cm and could emerge to only 5.3% at best from 1cm deep loamy soil field when the seeds were sown on Nov. 28. However, two strains of weedy rites being weedy for over 200 years emerged by 17.0 to 63.0% from the loamy and sandy clay loam field 1 to 6cm deep. Emergence of the weedy rites was greater in the loamy soil and at a shallow depth, and negligible from the soil depth of 9cm. Barnyardgrasses sown on Nov. 28 emerged by 13.4 to 51 % from the 1 to 3cm deep loamy soil, and 8.6 to 46.7% from the 1cm deep sandy clay loam. Echinochloa crus-galli var. crus-galli emerged more than var. praticola, and var. oryzicola least. Most of the non-emerged barnyardgrasses seem to have entered secondary dormancy. Seeding rice a month earlier than the season lowered the emergence of Dongjinbyo by ca. 10, 18, and 26%, respectively at 1, 3, and 6cm soil depths, indicating that moving the seeding date a month earlier is impractical. The old strain, Dadajo sown in the soil at a depth of 6cm responded similarly. However, the strain has shown a significantly higher ability in emergence from 9cm deep soil. Weedy rices sown a month earlier A month earlier sown weedy rices have shown very similar emergence rates at various soil depths to those sown on May 1. Barnyardgrasses have also shown similar emergence rates when sown between April 3 and May 1. Like barnyardgrasses, the old strain and weedy rices apparantly posessed a greater adaptability to emerge under lower temperatures, and from deeper soil ; Dongjinbyo${\leq}$ Echinochloa species in that order. However, emergence- speed under lower temperature(sown on April 3) was faster in the order of weedy rice

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Studies on the N-compounds during Chung-Kook-Jang Meju Fermentation (1) -Changes of Soybean Protein during Chung-Kook-Jang Meju Fermentation- (청국장(淸國醬) 메주 발효과정중(醱酵過程中)의 질소화합물(窒素化合物)의 소장(消長)에 관(關)한 연구(硏究)(I)-대두단백질(大豆蛋白質)의 소장(消長)에 관(關)하여-)

  • Park, Ke-In
    • Applied Biological Chemistry
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    • v.15 no.2
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    • pp.93-109
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    • 1972
  • Three lots of Chung-Kook-Jang were prepared by the use of 2 strains of Bacillus subtilis and Bacillus natto. For four samples taken from each lot in 12 hrs interval changes of nitrogenous compounds, insoluble protein, water soluble protein, peptides, free amino acids, amino and ammonia nitrogens during Chung-Kook-Jang fermentation, were studied together with the changes of moisture, pH, proteolytic enzyme activity. In addition the average peptide length of the peptides of a Bacillus subtilis lot was determined by the method of molecular sieving using ion exchange resin. The results were as follows: 1. The contents of moisture and total-nitrogen changed little in all samples throughout the fermentation as it would be expected. 2. In all three experimental lots the pH became higher gradually from the initial value of 6.65 to the final $7.5{\sim}7.85$ during the fermentation. Proteolytic enzyme activities, in accordance with this pH change, steadily increased up to $48{\sim}60$ hrs. of fermentation and then slightly decreased, probably affected by the high pH. The most strong proteolytic activity was observed in the experimental Chung-Kook-Jang fermentation lot using the Bacillus subtilis K-27 isolated by the author. 3. The contents of insoluble protein nitrogen in soybeans increased markedly (5%) by the cooking, after steeping 12 hrs in water. During the Chung-Kook-Jang fermentation, however, it decreased from 1/2 to 1/10 of that of the cooked soybeans. 4. The contents of water soluble protein nitrogen (5%) whereas, greatly decreased to the value of 1.0% by the cooking; but little changed further during the fermentation, 5. The total contents (0.25%) of peptides, amino, and ammonia-nitrogens, PAA-N., increased almost double by the cooking and steadily became higher as the fermentation proceeded, reaching finally up to$4{\sim}7%$ in 72 hrs fermentation. 6. The amounts of free amino acids of soybean generally decreased during the processing of cooking, even some of them like glutamic acid were destroyed completely, However in the subsequent Chung-Kook-Jang fermentation for 72 hrs., they showed from several to a few hundreds folds increases depending upon the kinds of amino acids. Valine which was contained in HCl-hydrolyzed steeped or cooked soybeans in amounts $220{\sim}267mg%$ was not detected at all as the free amino acid in all fermented samples. 7. Average peptide length (APL) of all fractions, eluted and fractionated by using the Dowex-50 ion exchange resin column, and fraction collector showed the highest value for the cooked soybean and then decreased as the fermentation proceeded. The APL value of effluent showed the highest in 12 hrs fermented sample, The value decreased thereafter by fermentation.

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Studies on the Induction of Available Mutants of Takju Yeast by UV light Irradiation (part 2) -On the Physiological Characteristics of the Mutants- (자외선조사(紫外線照射)에 의한 탁주효모(酵母)의 변이주육성(變異株育成)에 관한 연구 (제 2 보) -변이주(變異株)의 생리적성질(生理的性質)에 관하여)

  • Kim, Chan-Jo;Oh, Man-Jin;Kim, Seung-Yul
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.16-22
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    • 1975
  • This experiment was carried out to investigate the physiological characteristics of two original yeasts, 5-Y-5 and 6-Y-6, which selected from 24 Takju yeasts and three mutants, 30-24,30-81 and 40-27. induced from two original yeasts by the irradiation of UV light. The results were summarized as follows. 1) Alcohol tolerances of three mutants were decreased in some degree as compared with those of original yeasts. 2) Tolerances of lactic and citric acids of acid producing mutant 30-81, was increased than those of original yeasts. 3) In the case of using ammonium sulfate as a nitrogen source, two original yeasts and three mutants required Ca-pantothenate as a essential growth factor and four strains of yeasts except the mutant, 30-81, required biotin as a stimulated growth factor, When asparagine was used as a nitrogen source, two original yeasts and three mutants showed the same as above result but the stimulated effect of biotin was far less. 4) Propagation powers of the mutants were weaken than those of original yeasts, particular that of acid producing mutant, 30-81, was the weakest in the three mutants. 5) The optimum temperature for fermentation of original yeasts were $30^{\circ}C\;to\;35^{\circ}C$ but three mutants were $25^{\circ}C\;to\;30^{\circ}C$. 6) The optimum pH for fermentation of original yeasts were pH 5 to 6, and there is no appreciable difference between original yeasts and three mutants. The fermentation power of mutant,30-81, was decreased more rapidly than those of other mutants according to approach neutral. Three mutants were more sensible to heat than original yeasts. 7) Two original yeasts and three mutants were inhibited more over 20 percent of sugar for fermentation and three mutants were more sensible to sugar concentration than original yeasts.

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Anti-Allergic Inflammatory Effect of Bacteria Isolated from Fermented Soybean and Jeotgal on Human Mast Cell Line (HMC-1) (장류 및 젓갈 분리 균주 추출물의 비만세포 매개 항염증효과)

  • Ko, Yu-Jin;Kim, Hui-Hun;Kim, Eun-Jung;Kim, Jin-Yong;Kang, Sang-Dong;Son, Yong-Hwi;Choi, Sin-Yang;Cha, Seong-Kwan;Kim, Jong-Won;Lee, Jeong-Ok;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.21 no.3
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    • pp.393-399
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    • 2011
  • The mast cell is one of the major effector cells in inflammatory reactions and can be found in most tissues throughout the body. Activated mast cells can produce histamine, as well as a wide variety of other inflammatory mediators such as eicosanoids, proteoglycans, proteases, and several pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-${\alpha}$, and interleukins (IL-6), IL-8, IL-4, IL-13. In the present study, we isolated two bacterial strains (J80 and G147) from fermented soybean and Jeotgal, and investigated the inhibitory effects of their extracts which were prepared by several pretreatment methods (sonication for 20 min, heating at $100^{\circ}C$ for 30 min, autoclaving at $121^{\circ}C$ for 15 min) on the mast cell-mediated inflammatory response. The pretreated bacterial extracts had no cytotoxicity against Human Mast Cell (HMC-1). Among various pretreatments, the extracts treated at $100^{\circ}C$ showed highest inhibition of histamine release (J80, 28.46%; G147, 41.14%). The J80 and G147 extracts treated at $100^{\circ}C$ resulted in the inhibition of IL-6 secretion by 38.46% and 56.45%, respectively. The J80 extract treated at $100^{\circ}C$ resulted in the inhibition of TNF-${\alpha}$ secretion by 66.67%, but G147 extract showed the highest inhibition effect by 41.1% when treated with sonication. These results suggest that bacterial extracts treated at $100^{\circ}C$ have a higher level of anti-inflammatory effects than other treatments such as sonication or autoclaving.

Pulmonary Resection in the Treatment of Multidrug-Resistant Tuberculosis (다제 내성 폐결핵환자의 폐절제술에 관한 연구)

  • Kwon, Eun-Soo;Ha, Hyun-Cheol;Hwang, Su-Hee;Lee, Hung-Yol;Park, Seung-Kyu;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1143-1153
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    • 1998
  • Background : Recent outbreaks of pulmonary disease due to drug-resistant strains of Mycobacterium Tuberculosis have resulted in significant morbidity and mortality in patients worldwide. We reviewed our experience to evaluate the effects of pulmonary resection on the management of multidrug-resistant tuberculosis. Method : A retrospective review was performed of 41 patients undergoing pulmonary resection for multidrug-resistant tuberculosis between January 1993 and December 1997. We divided these into 3 groups according to the radiologic findings : (1) patients who have reasonably localized lesion (Localized Lesion Group ; LLG) (2) patients who have cavitary lesions after pulmonary resection on chest roentgenogram (Remained Cavity Group : RCG) (3) patients who have Remained infiltrative lesions postoperatively (Remained infiltrative group : RIG). We evaluated the negative conversion rate after resection and overall response rate of the groups. Then they were compared with the results of the chemotherapy on the multi drug-resistant tuberculosis which has been outcome by Goble et al. Goble et al reported that negative conversion rate was 65% and overall response rate, 56% over a mean period of 5.1 months. Results : Seventy five point six percent were men and 24.4% women with a median age of 31 years (range, 16 to 60 years). Although the patients were treated preoperatively with multidrug regimens in an effort to reduce the mycobacterial burden, 22 of 41 were still sputum culture positive at the time of surgery. 20 of 22 patients(90.9%, p<0.01) responded which is defined as negative sputum cultures within 2 months postoperative. Of 26 patients with the sufficient follow up data, 19 have Remained sputum culture negative for a mean duration of 25.7 months (73.1%, p<0.05). The bulk of the disease was manifest in one lung, but lesser amounts of contralateral disease were demonstrated in 15, consisted of 8 in RIG and 7 in RCG, of 41. 12 of 12 patients (100%, p<0.01) who were sputum positive at the time of surgery in LLG converted successfully. 14 of 15 patients (93.3%, p<0.05) with the follow up have completed treatment and not relapsed for a mean period of 25. 7 months. The mean length of postoperative drug therapy of LLG was 12.2 months. In RIG, postoperative negative conversion rate was 83.3% which was not significant statistically. There was a statistical significance in overall response rate (100%, p<0.05) of RIG for a mean period of 24.4 months with a mean length of postoperative chemotherapy, 11.8 months. In RCG a statistically lower overall response rate (14.3%, p<0.01) has been revealed for a mean duration of follow up, 24.2 months. A negative conversion rate of RCG was 75% which was not significant statistically. Conclusion : Surgery plays an important role in the management of patients with multidrug-resistant Mycobacterium tuberculosis infection. Aggressive pulmonary resection should be performed for resistant Mycobacterium tuberculosis infection to avoid treatment failure or relapse. Especially all cavitary lesions on preoperative chest roentgenogram should be resected completely. If all of them could not be resected perfectly, you should not open the thorax.

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Seasonal Variations of Water Quality in the Lower Part of the Nagdong River (낙동강 하류수질의 계절적 변화)

  • KIM Yong-Gwan;SHIM Hye-Kung;CHO Hak-Rae;YOU Sun-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.6
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    • pp.511-522
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    • 1984
  • The Nagdong is one of the biggest rivers in Korea, which is very important water source not only for tap water of Pusan city but also for the industrial water. Therefore, authors tried to check the water quality year by year. In this experiment one hundred and twenty water samples collected from August 1983 to July 1984 were analyzed bacteriologically and physiologically. Fifteen sampling stations were established between near Samrangjin and estuary of the river. To evaluate the water quality, temperature, pH, chloride ion, salinity, chemical oxygen demand (COD), electrical conductivity, nutrients, total coliform, fecal coliform, fecal streptococcus, viable cell count and bacterial flora were observed. The variation of water temperature was ranged $-1.5{\sim}29.0^{\circ}C$ (Mean value $13.9{\sim}16.5^{\circ}C$), it in spring was higher as $10{\sim}15^{\circ}C$ about $10^{\circ}C$ than in winter and it in autumm was very stabilized as about $20^{\circ}C$ at each station. The pH variation of the samples was ranged $6.68{\sim}9.15$. The range of concentration of chloride ion and salinity varied $7.4{\sim}l,020.5$ mg/l and $1.05{\sim}33.0\%0$, respectively. Especially, salinity of the 3rd water war was the higher than others as $25.76{\sim}31.58\%0$. COD was ranged $1.45{\sim}14.94$ mg/l and the lower part of the Nagdong River was heavily contaminated by domesitc sewage and waste water from the adjacent factor area. The range of electrical conductivity was $1.360{\times}10^2{\sim}5.650{\times}10^4{\mu}{\mho}/cm$ and that was by far higher the estuary than the upper. Concentration of nutrients were $0.008{\sim}0.040$ mg/l (Mean value $0.019{\sim}0.068$ mg/l) for $NO_2-N,\;0.038{\sim}5.253$ mg/l ($0.351{\sim}2.347$ mg/l) for $NO_3-N,\;0.100{\sim}2.685$ mg/l($0.117{\sim}1.380$ mg/l) for $NH_4-N,\;0.003{\sim}0.084$ mg/l($0.014{\sim}0.065$ mg/l) for $PO_4-P$ and $0.154{\sim}6.123$ mg/l ($1.165{\sim}3.972$ mg/l) for $SiO_2-Si$, respectively. Usually nutrients contents of the water in the upper part(included station 1 to 5) were higher than those of the estuarine area. The bacterial density of the samples ranged 7.3 to 460,000/100 ml for total coliforms, 3.6 to 460,000/100 ml for fecal coliform, $0{\sim}46,000/100ml$ for fecal streptococcus and $<30{\sim}1.2{\times}10^5/ml$ for viable cell count. Composition of coliform was $28\%$ Escherichia coli group, $18\%$ Citrobacter freundii group, $31\%$ Enterobacter aerogenes group and $22\%$ others. Predominant species among the 659 strains isolated from the samples were Pseudomonas spp. ($42\%$), Flavobacterium spp. ($20\%$) and Moraxella spp. ($12\%$).

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A Randomized, Double-Blind Study to Assess the Efficacy and Safety of Oral LB20304 (Gemifloxacin) at Doses of 160mg or 320mg (Equivalent to 200mg or 400mg of the Mesylate Salt) Once Daily for 7 Days for the Treatment of Acute Exacerbations of Chronic Bronchitis (AECB) in Korean Adult Population (한국인에서 만성기관지염의 급성악화를 치료하기 위한 LB20304(Gemifloxacin) 160mg 또는 320mg 1일 1회 7일간 투여의 유효성과 안전성에 대한 연구)

  • Kim, Young Whan;Shim, Young-Soo;Kim, Won Dong;Shim, Tae Sun;Kang, Hong Mo;Choi, ByoungWhui;Kim, Jae-Yeol;Kwon, O Jung;Kim, Hojoong;Kim, Ju Ock;Jung, Ki-Suck;Hyeon, In Gyou;Mo, Eun Kyung;Lee, Seung-Joon;Nam, Gui Hyun;Lee, Kye Young;Park, Jae Seuk
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.69-87
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    • 2003
  • Background : LB20304(gemifloxacin) is a new fluoroquinolone antibacterial agent with excellent activity against both Gram-negative and Gram-positive organisms. In vitro studies using clinical isolates have shown gemifloxacin to be highly active against penicillin-resistant strains of S. pneumoniae and in contrast to other reference quinolones, gemifloxacin retained good activity against clinical isolates of S. pneumoniae that were resistant to other members of the quinolone class. Therefore, gemifloxacin is thought to be effective in treating acute bacterial exacerbation of chronic bronchitis(AECB). The objective of this study was to evaluate the efficacy and safety of oral gemifloxacin at doses of 160mg or 320mg once daily for 7 days for the treatment of AECB in Korean adult population. Methods : This was a randomized, multicenter, double-blind, parallel group Phase II study to assess the clinical and antibacterial efficacy and safety of oral gemifloxacin for the treatment of AECB. Treatment Group A (67 patients) took oral gemifloxacin 160mg once daily for seven days and treatment Group B (70 patients) took oral gemifloxacin 320mg once daily for seven days. Results : The demographic profiles of the two treatment groups were similar. The clinical response at follow-up was 84.2% in the gemifloxacin 160-mg group, and 88.7% in the gemifloxacin-320 mg group, showing no statistically significant difference between two treatment groups(p=0.49). The clinical response at the end of therapy was 96.5% in the 160-mg group, and 96.4% in the 320-mg group. The bacteriological response at the end of therapy and follow-up were 81.8% and 78.9%, respectively, in the 160-mg group, and 86.4% and 84.2%, respectively, in the 320-mg group, showing no statistically significant difference between two treatment groups(p=0.68 and 0.68, respectively). S. pneumoniae(12 isolates) and H. influenzae(10 isolates) were the most prevalent pathogens. The MICs were lower for gemifloxacin than other quinolones against these key pathogens, and for S. pneumoniae, the MICs for gemifloxacin were considerably lower(${\leq}0.03$ ug/mL) than those for other quinolones, beta-lactams and macrolides. In the period on-therapy plus 30 days post-therapy, a total of 18 patients(26.9%) in the gemifloxacin 160mg group and 22 patients(31.4%) in the 320mg group reported at least one adverse event(AE). The most frequently reported AE was abdominal pain(3/67 patients, 4.5%) in the gemifloxacin 160mg group and increased level of hepatic enzyme(5/70 patients, 7.1%) in the 320mg group. The overall AE profiles for the two treatment groups were similar. Two out of 67 patients(3.0%) in the gemifloxacin 160mg group and 1/70 patients(1.4%) in the 320mg group reported at least one serious AE, however, none of which was considered by the investigator to be of suspected or probable relationship to study medication. Conclusion : The results of this study showed that gemifloxacin at doses of 160mg or 320mg once daily for 7 days in the treatment of acute exacerbations of chronic bronchitis(AECB) in adult Koreans was a very effective and safe treatment both clinically and bacteriologically.

The Standing Crops and Soil-borne Microfungal Flora of Phyllostachys reticulata in Korea (한국산(韓國産) 왕대나무의 현존량(現存量)과 토양(土壤) 미세균류상(微細菌類相))

  • Kim, Kwan-Soo
    • The Korean Journal of Mycology
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    • v.7 no.2
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    • pp.91-116
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    • 1979
  • This paper is to investigate the standing crops and microfungal flora in soil in Phyllostachys reticulata forests in both the Yesan area (A) and the Kwangsan area (B). The stand density of the bamboo revealed 17,250 shoots per ha in area A, and in area B 14,780 shoots which were 16.1% less in number than area A. In respect to the environmental factors between the two areas, the mean temperature during the growth period was $1.5{\sim}2^{\circ}C$ higher in area B than in area A, soil tempeature also was $1{\sim}2^{\circ}C$ higher in area B, and the total quantities of nitrogen, phosphoric acid and organic compounds contained in the soil of area B were also slightly higher than those of area A. In area B the quantities of dried leaf matter, humus, and vegetation in the bamboo forest were also larger than in area A. In addition, five more species of microfungi which playa role in the decomposition of the various organic materials in the bamboo forests were identified in area B: Mortierella elongata, Mucor circinelloides, Aspergillus japonicus, Penicillium waksmani and Trichoderma lignorum. The atmospheric temperature in the inner portions of the bamboo forests was lower than the outside temperature, but the humidity was higher. The rates of relative illuminance were measured in area A at 4.19%, and in area B at 2.7%. These values revealed that the photosynthetic acitivity in the lower part of the bamboo was lost but it was considered that lower illuminance increased the microfungal activities in the vicinity of the surface soil. Since the productive structure of the bamboo showed that the maximum amount of photosynthesis was located in the upper portion of the bamboo in area B, it was considered to be an effective structure in maintaining the high productivity of the bamboo. The allometric relation between $D^2H$ and dry weight of stems(Ws), branches(Wb) and leaves(Wl) of the bamboo in area A were appoximated by log Ws=0.5262 log $D^2H$+1.9546; log Wb=0.6288 log $D^2H$+1.5723; log Wl=0.5181 log $D^2H$+1.8732, and those of the bamboo in area B were approximated by log Ws=0.5433 log $D^2H$+1.8610; log Wb=0.1630 log $D^2H$+2.3475; log Wl=0.4509 log $D^2H$+2.0041. From the above, the standing crops in area A were measured thus: Ws was 1,128. 83kg; Wb, 689.05kg; Wl, 926.69kg and Wl, 2,744.57kg per 10a. In area B, Ws was 1,206. 66kg; Wb, 679.92kg; Wl, 1,112.51kg and Wt, 2.999kg per l0a. Significant differences from the result of t-test were for $D^2H$ Ws, Wl and Wt between areas A and B. But no significant difference was found for Wb. In order to record as completely as possible the microfungal flora of the areas, every possible means was tried, and 158 strains of fungi were isolated, and of these, the microfungi of 55 species were identified. The dominant species were Trichoderma viride, Penicillium janthinellum, P. commune, Aspergillus oryzae, A. niger, A. gigantus, A. fumigatus, Mortierella ramaniana, var. anguliFPora, Mucor hiemalis and Zygorhynchus moelleri. According to the above results, it was revealed that optimum soil, the increases of soil materials, more species of soil microfungi, and the atmospheric temperature during the growth period have made the bamboo flourish and bring more species and larger quantities of vegetation in the bamboo forests. The correlation between the standing crops and environmental factors in the bamboo forest is considered to be a complicated relationship of all the factors, but the stand density is thought to be the most important factor involved.

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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • Korean journal of applied entomology
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    • v.13 no.3 s.20
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    • pp.105-133
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    • 1974
  • The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

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