• Title/Summary/Keyword: J48

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Synergistic action of pesticide mixtures using glutathione-s-transferase- and esterase-inhibiting properties in diamondback moth (Plutella xylostella L.) (Glutathione-S-transferase와 esterase 효소 저해특성을 이용한 농약의 혼합 상승효과)

  • Yu, Yong-Man;Hong, S.S.;Kim, S.;Hur, J.H.
    • The Korean Journal of Pesticide Science
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    • v.7 no.1
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    • pp.38-44
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    • 2003
  • In vitro inhibitory activity of 34 insecticides and 31 fungicides to glutathione-S-transferase and esterases extracted from rats was determined. Of tested pesticides, the pesticides with high activity on both detoxifying enzymes were mixed with pesticides that are known to be detoxified by detoxifying enzymes. Glutathione-S-transferase was inhibited by thiodicarb $(I_{50}:1.87\times10^{-4}M)$, thiocyclam $(7.40\times10^{-4}M)$, dithianon $(7.55\times10^{-5}M)$, and tolylfluanide $(8.66\times10^{-5}M)$, while esterases by dichlorvos $(8.95\times10^{-8}M)$, pirimicarb $(2.74\times10^{-6}M)$, pyrazophos $(3.31\times10^{-5}M)$, and benomyl $(4.96\times10^{-5}M)$. After acephate known to be detoxified by glutathione-S-transferase was mixed with glutathione-S-transferase-inhibiting pesticides and phenthoate known to be detoxified by esterases was mixed with esterases-inhibiting pesticides, insecticidal activities of such mixtures were determined against diamondback moth (PlutelLa xylostella L.). Synergistic action was observed in all pesticide combinations. The highest synergistic action was obtained when phenthoate was combined with dichlorvos, showing that co-toxicity coefficients were 1512 and 1877 after 24 and 48 hours of treatment, respectively. Several other combinations of pesticides, such as phenthoate with benomyl, and acephate with dithianon, also showed synergism, showing that their co-toxicity coefficients were about 1,000 and 500, after 24 hours of treatment, respectively. Our results showed that combinations of pesticides inhibited by detoxifying enzymes and ones detoxified by detoxifying enzymes resulted in increased toxicities of pesticides, suggesting that such combinations could be used to develop pesticide mixtures with more broad spectrum and high effectiveness.

Effect of Media, Synchronization of Fibroblast Cells, Culture Time, $\textrm{O}_2$ Concentration and Activation on Developmental Rate of Nuclear Transferred Porcine Oocytes (배양액, 섬유아세포, 배양시간, 산소 농도 및 활성화 처리가 돼지 핵이식 배의 체외발달에 미치는 영향)

  • Quan J. H.;Rhee M. H.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.191-199
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    • 2004
  • This study was conducted to examine in vitro development of porcine embryos constructed by the microinjection of cultured fetal fibroblast cells into porcine oocytes matured in vitro. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5% FBS, at 38.5$^{\circ}C$ for 6 to 8 days in 5% $CO_2$ and air. In experiment 1, fusion rates of nuclear transfer embryos did not differ for fetal fibroblast cells incubated in 5% FBS + NCSU-23 or 5% FBS + TL Heaps medium, nor did fusion rates of donor cells differ between 1-8 hr incubation durations. Fusion rates for the four treatment subclasses ranged from 72.1% to 78.0%. In experiment 2, Pre-synchronization in medium containing 0.1 $\mu\textrm{g}$/m Hoechst 33342 an increase from 0 and 8 versus 15 h culture an increased percentage of porcine fibroblast cells in G2/M at the end of the synchronization period (12.4%, 17.5% and 47.6%). Neither an increase in the concentration of H 33342 (0.2-1.6 $\mu\textrm{g}$/$m\ell$) nor a longer exposure time (12h, 18h and 24h) increased the proportion of porcine G2/M fibroblasts. In experiment 3, fusion rates did not differ significantly far nuclear transfer embryos constructed using donor cells cultured in 5% FBS + NCSU-23 medium for 1-2, 6-8 or 12-14 days (60.0%, 73.3% and 62.5%), respectively. The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1-2 days was 44.0%, significantly less than 56.7% and 50.0%. for 6-8 or 12-14 days duration of culture, respectively. In experiment 4, the proportions of nuclear transfer embryos that developed to the $\geq$2 cell and to the blastocyst stage were not affected significantly by culture medium (5% FBS + NCSU-23 or 5% FBS + TL-Heaps) or by $O_2$ concentration of the culture (5% vs 10%). Rates of development to the $\geq$2 cell stage ranged from 65.9% to 70.1%, and development rates to the blastocyst stage ranged from 9.8% to 12.5% for the four treatment subclasses. Developmental rate was highest for embryos cultured in 5% FBS + NCSU-23 under a gas atmosphere of 5% $O_2$ in air.

Changes in Urinary Nitric Oxide in Pediatric Renal Diseases (소아 신장질환에서 소변 내 산화질소의 변화)

  • Kim, Jong-Hwa;Jung, Ji-In;Yim, Hyung-Eun;Hong, Young-Sook;Lee, Joo-Won;Yoo, Kee-Hwan
    • Childhood Kidney Diseases
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    • v.11 no.1
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    • pp.24-31
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    • 2007
  • Purpose : Nitric oxide(NO) is a very potent vasodilator synthesized from L-arginine by endothelial cells. We investigated whether urinary NO excretion was altered in various renal diseases in children and whether urinary NO excretion could be used in predicting pathologic causes and fibrosis in renal diseases in children. Methods : We recruited 48 patients(32 minimal change nephrotic syndrome[MCNS] and 16 vesicoureteral reflux[VUR] patients from the pediatric renal clinic in Korea University Guro Hospital. We measured the concentration of nitrite$(NO_2)$ and nitrate$(NO_3)$ by Griess reaction and that of creatinine(Cr) by Jaffe method in randomized spot urines. We then analyzed the urinary$(NO_2+NO_3)/Cr$ ratios and compared the values between each patient group. Urinary $(NO_2+NO_3)/Cr$ ratios were also evaluated according to the recurrence and the degree of proteinuria at sampling in the MCNS group and compared according to the presence of renal scarring and the grade of reflux in the VUR group. Results : The ratios of urinary$(NO_2+NO_3)/Cr$ were significantly increased in the VUR and MCNS groups, as compared to the control group. In the MCNS group, a higher level of urine $(NO_2+NO_3)/Cr$ was observed In frequent relapse patients(relapse over four times within one year after first diagnosis) and the patients with severe proteinuria at sampling, respectively. The VUR group with renal scars also showed a higher level of urinary$(NO_2+NO_3)/Cr$ compared to that without scars. Conclusions : In summary, VUR may play a role in the pathogenesis of VUR and MCNS. NO also seems to affect proteinuria and renal scar formation. (J Korean Soc Pediatr Nephrol 2007;11:24-31)

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Studies on the Seroepidemiology of Helminthic Diseases in Korea (우리나라의 주요 기생충질환(寄生蟲疾患)에 대한 혈청역학적(血淸疫學的) 조사(調査))

  • Rim, Han-Jong;Lee, Joon-Sang;Joo, Kyoung-Hwan;Chung, Myung-Sook
    • Journal of agricultural medicine and community health
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    • v.16 no.1
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    • pp.48-60
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    • 1991
  • In a seroepidemiological study in several areas of Korea, the ELISA technique was performed to determine prevalence of some important helminthic diseases in our nation during March $15^{th}$ to June $30^{th}$. 1991. In this survey the serum antibody positive rates of anisakiasis, toxocariasis, clonorchiasis, paragonimiasis, cysticercosis, and sparganosis were measured. Among, 6,704 cases examined, 19.7% showed positive antibody titer at least one of the six items studied. Overall positive antibody rate was 8.1% in anisakiasis, 5.6% in toxocariasis, 3.6% in clonorchiasis, 1.7% in paragonimiasis, 4.5% in cysticercosis, and 2.6% in sparganosis respectively. In Pusan port southeastern part of Korea, antibody positive rate of anisakiasis was 2.9%, and clonorchiasis was 2.8% among 450 examine. In TaeJ$\check{o}$n city, central part of Korea. toxocariasis(6.7%) and anisakiasis(3.7%) showed high serologic positive rate. Of the 875 persons in Chunche$\check{o}$n gun(=province), northern central rural area of South Korea, anisakiasis was revealed as 3.4% seropositivity. In Tonghae port, eastern coast of South Korea. 9.9% of population examined showed positive antibody titer in anisakiasis. Of the 1,122 persons examined in Southern part of Cholla-Namdo(Southwestern coastal area of Korea), anisakiasis was 16.9%, cysticerocosis was 12.7% and the paragonimiasis was 3.3% respectively. In some localized area of Cholla-Pukdo, anisakiasis was 9.3% and cysticekosis was 4.3% among 702 cases examined. In some localized area of Kyungsang-Pukdo, anisakiasis was 10.6%. and toxocariasis was 16.1% among 900 cases examined. And finally, in Cheju-do, southern island of Korea, anisakiasis showed high positive rate(6.7%). Because cross reactions between related helminth group may disturb the analysis of these data, use of further developed techniques such as EITB(enzyme-linked immunoelectrotransfer blot) was considered as a essential tools for the study. We thought that probably most of the positive cases of cysticerosis were taeniasis cases. We can't rule out taeniasis even though EITB was employed as far as crude worm extract or cystic fluid of cysticercus was used as antigen. It was well Known that toxocariasis and anisakiasis also showed cross reactivity. However, the data presented here focus on seropositive rate of several helminthic diseases in Korea, not true prevalence rate of helminthiases, and to wait for more expensive purified antigen in sufficient amount for epidemiologic use is not necessary because increased immunologic sensitivity had little effect on epidemiologic sensitivity. We, here, suggest that ELISA should be applied as soon as possible to the evaluation of prevalence of tissue invading parasitic diseases, and a review of the antibody positive rate obtained in this study would be a basic data for controlling program of parasitic diseases in Korea.

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A QUALITATIVE AND QUANTITATIVE STUDY ON OCCLUSAL CONDITIONS IN HEALTH VOLUNTEERS AND ATHLETES WITH NORMAL OCCLUSION (정상인과 체육인의 교합상태에 대한 정상적.정량적 비교 연구)

  • Jang, Jung-Mi;Lee, Sung-Bok
    • The Journal of Korean Academy of Prosthodontics
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    • v.36 no.2
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    • pp.302-322
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    • 1998
  • This study was accompolished to analyze and compare the number and area of the occlusal contact points in healthy volunteers and athletes with normal occlusion. For this study, 15 athletes(13 amles and 2 females with average age 20) and 21 healthy volunteers(14 mles and 7 females with average age 20.09) were selected. The visual display acquired by scanning of occlusal record in maximal intercuspation was converted into 16 gray value image. Then, using computer program(J & Lee Occlusal Analyser), the pixel which was in definite range of the gray value was recognized, and the numbers of recognized pixel were calculated to area. The results were as follows ; 1. The average numbers of total occlusal contact feints were 31.05 in control group, and 34.67 in athlete group. The average area of total occlusal contacts was $100.25mm^2$ in control group, and $127.78mm^2$ in athlete group. 2. In control group, the average numbers of occlusal contact points were revealed in order as follows ; the first molar(8.48), the second molar(8.24), the second premolar(4.71), the lateral incisor(2.90), the first premor(2.43), the central incisor(2.19), and the canine(2.1). The least average in canine(2.1) was similar to the average(2.19) in central incisor and (2.09) in lateral incisor. In athlete group, the average numbers of occlusal contact points were revealed in order as follows ; the first molar(8.97), the second molar(8.47), the second premolar(5.60), the canine(3.80), the lateral incisor(3.33), the first premolar(2.67), and the central incisor(1.93). 3. In control group, the average areas of occlusal contact surface were revealed in order as follows ; the first molar($39.47mm^3$), the second molar($37.54mm^3$), the second premolar($9.54mm^3$) the first premolar($6.18mm^3$), canine($3.49mm^3$), the central incisor($2.76mm^3$), and the lateral incisor($1.28mm^3$). In athlete group, the average areas of occlusal contact surface were revealed in order as follows ; the first molar($44.11mm^3$), the second molar($40.69mm^3$), the second premolar($16.50mm^3$), the first premolar($9.39mm^3$), the canine($5.08mm^3$), the lateral incisor($3.7mm^3$), and the central incisor($2.25mm^3$). 4. With aging in both control and athlete group, there was a decreasing tendancy in average number of occlusal contact point, and was an increasing tendancy in average area of occlusal contact surface. In comparison at each age, both the numbers and area of occlusal contact were greater in athlete group than in control group. It was not significant in the numbers of occlusal contact points beween athlete and control group(p>0.1), but significant in the area of occlusal contact surface(p<0.03). 5. In comparision as to the kind of sports(Gymnastics : 2, Rugby : 3, Soccor : 5, Ice hocky : 5), the numbers of occlusal contact points were the most in ice hocky, and the area of occlusal contact surface was the greatest in gymnastics. With increasing a career in athlete group, there was a decreasing tendancy in average numbers of occlusal contact points and was an increasing tendancy in average area of occlusal contact surface. 6. By T-scan analyzing, the contact numbers on the anterior teeth were greater in control group than in athlete group, and on the posterior teeth were greater in athlete group than in control group. And the results acquired by T-scan were lesser than that caquired by the silicone bite records. It was not significant in the posterior teeth, but significant in the anterior teeth. In T-scan records, the numbers of occlusal contact points on second molar were the greatest, but in the silicone records, the numbers on first molar were the greatest.

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Development of Transgenic NT Embryos Using Bovine Fetal Fibroblasts Transfected with hFSH Gene (hFSH 유전자가 도입된 소 태아섬유아세포를 이용한 형질 전환 복제 수정란의 발달)

  • Yang B.C.;Im G.S.;Kim D.H.;Min K.S.;Yoon D.H.;Park H.S.;Kim S.W.;Hwang I.S.;Seo J.S.;Seong H.H.;Yang B.S.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.13-20
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    • 2006
  • The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.

Effects of Herbal Product (Miracle®) on the Performance, Nutrient Digestibility, Fecal Microflora and Immunoresponse in Weanling Pigs (생약제제(미라클®)첨가가 이유 자돈의 성장률, 영양소 이용율, 분내 미생물균총 및 면역기능에 미치는 영향)

  • Suk, J.C.;Lim , H.S.;Paik, I.K.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.767-776
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    • 2003
  • An experiment was conducted to investigate the effects of dietary herbal product(Miracle$^{\circledR}$) supplementation on the performance, nutrient digestibility, fecal microflora and blood parameters in pigs. Forty eight cross bred(Y${\times}$L${\times}$D) weanling pigs (average initial body weight 7.84kg${\pm}$0.17, 28 d old) were used in 35 d feeding trial. Pigs were allotted to 16 raised floor cages(W 35.5cm${\times}$L45cm${\times}$H 55cm). Each treatment had four replications of 3 pigs each. Treatments were: control (T1), CTC 100ppm (T2), Miracle$^{\circledR}$ 0.15% (T3) and CTC 100ppm + Miracle$^{\circledR}$ 0.15% (T4). The herbal product supplementation(T3) tended to improve the weight gain, feed intake, feed/gain but there were no significant differences among treatments. Nutrient digestibility was significantly(P〈0.05) affected by dietary treatments. The pigs fed T3 diet showed the lowest digestibility of DM, crude protein, NFE, P, and amino acids among treatments. Serum cholesterol level of pigs fed T3 diet was lowest and significantly(P〈0.05) lower than those fed antibiotic supplemented diet(T2). Triglyceride and HDL level of the pigs fed T3 diet were also lower than those of others but the differences were not significant. The level of serum IgG was significantly(P〈0.05) different among treatments. The level of serum IgG was highest in T4, followed by T1, T2 and T3. Serum AST(Aspartate aminotransferase) level of the control(T1) was lower than other treatments. Levels of serum total protein, albumin and ALT(Alanine aminotransferase) were not significantly different among treatments. The colony forming units (CFU) of Lactobacillus, Cl. perfringens and E.coli in feces were not significantly different among treatments but those of Cl. perfringens of T2 (CTC 100ppm) showed the lowest count at both 3rd and 5th wk. It is concluded that herbal product Miracle$^{\circledR}$ does not significantly affect growth performance of weanling pigs but it influenced the digestibility, serum IgG, cholesterol and AST level. The combination of the herbal product Miracle$^{\circledR}$ with antibiotic(CTC) showed no synergistic effects.

Korean athlete's recognition of sports dentistry and condition of teeth wear (한국 운동선수의 스포츠 치의학에 대한 인지도 및 치아교모상태에 관한 연구)

  • Lee, Sung-Bok;Choi, Dae-Gyun;Han, Kwang-Heung
    • Journal of Dental Rehabilitation and Applied Science
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    • v.18 no.4
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    • pp.235-249
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    • 2002
  • This study was accompolished to analyze and compare the number and area of the occlusal contact points in healthy volunteers and athletes with normal occlusion. Before the study, 89 athletes(sports career:8.6 years, average age 20) at Kyung Hee University were selected, and survey was accomplished for athlete's recognition about sports dentistry. For this study, 15 athletes(13 amles and 2 females with average age 20) and 21 healthy volunteers(14 mles and 7 females with average age 20.09) at Kyung Hee University were selected. The visual display acquired by scanning of occlusal record in maximal intercuspation was converted into 16 gray value image. Then, using computer program(J & Lee Occlusal Analyser), the pixel which was in definite range of the gray value was recognized, and the numbers of recognized pixel were calculated to area. The results were as follows : (1) On the survey about sports dentistry, 28% of 89 athletes didn't agree that human occlusion may be important, and 30% didn't have any idea of the influence of human occlusion during their sports activities. (2) The average numbers of total occlusal contact points were 31.05 in control group, and 34.67 in athlete group. The average area of total occlusal contacts was $100.25mm^2$ in control group, and $127.78mm^2$ in athlete group. (3) In control group, the average numbers of occlusal contact points were revealed in order as follows; the first molar(8.48), the second molar(8.24), the second premolar(4.71), the lateral incisor(2.90), the first premor(2.43), the central incisor(2.19), and the canine(2.1). The least average in canine(2.1) was similar to the average(2.19) in central incisor and (2.09) in lateral incisor. In athlete group, the average numbers of occlusal contact points were revealed in order as follows; the first molar(8.87), the second molar(8.47), the second premolar(5.60), the canine(3.80), the lateral incisor(3.33), the first premolar(2.67), and the central incisor(1.93). (4) In control group, the average areas of occlusal contact surface were revealed in order as follows; the first molar($39.47mm^3$), the second molar($37.54mm^3$), the second premolar($9.54mm^3$) the first premolar($6.18mm^3$), canine($3.49mm^3$), the central incisor($2.76mm^3$), and the lateral incisor($1.28mm^3$). In athlete group, the average areas of occlusal contact surface were revealed in order as follows; the first molar($44.11mm^3$), the second molar($40.69mm^3$), the second premolar($16.50mm^3$), the first premolar($9.39mm^3$), the canine($5.08mm^3$), the lateral incisor($3.7mm^3$), and the central incisor($2.25mm^3$). (5). With aging in both control and athlete group, there was a decreasing tendancy in average number of occlusal contact point, and was an increasing tendancy in average area of occlusal contact surface. In comparison at each age, both the numbers and area of occlusal contact were greater in athlete group than in control group. It was not significant in the numbers of occlusal contact points beween athlete and control group(p>0.1), but significant in the area of occlusal contact surface(p<0.05). (6) In comparision as to the kind of sports(Gymnastics:2, Rugby:3, Soccor:5, Ice hocky:5), the numbers of occlusal contact points were the most in ice hocky, and the area of occlusal contact surface was the greatest in gymnastics. With increasing a career in athlete group, there was a decreasing tendancy in average numbers of occlusal contact points, and was an increasing tendancy in average area of occlusal contact surface.

A Study on Recognition and Practice of Teakyo by Pregnant Women (임부의 태교인식과 태교실천에 관한 조사연구)

  • Shin, Yong-Bun;Koh, Hyo-Jung
    • Women's Health Nursing
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    • v.6 no.1
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    • pp.142-152
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    • 2000
  • This study is a descriptive study to offer an actual basic data as Nursing intervention strategy of nurses before delivery in order that pregnant women in Korea may effectively practice Taekyo by examining the relation after apprehending level of recognition and practice of our pregnant women about Taekyo. This study collected questionnaires from 801 pregnant women who visited general hospitals in 10 areas(Seoul, Daejon, chunan, Daegu, Kummi, $Ky{\check{o}}ngju$, $P{\ddot{o}}hang$, Busan, $J{\ddot{o}}nju$, and $Y{\ddot{o}}nkwang$) for prenatal care through an outpatient obstetrics and gynecology from July 15 to August 30, 1999. This study used the tool of lee, Ki Young(1993) revised an complemented by investigator to measure recognition of Taekyo and the tool of Jang, Shun Buk and Park, Young Ju(1996) revised and complemented by investigator to measure practice of Taekyo. The Cronbach's alpha value of each tool was .88 in recognition of Taekyo, while the value was .90 in practice of Taekyo. For data analysis, this study used the descriptive and statistical analysis, Pearson correlation, t-test, ANOVA, Tukey's post hoc contrast, and Stepwise multiple regression in accordance with the purpose of this study with using SPSS Win 7.5 program. The results were as follows ; 1. The practice of Taekyo was low in comparison with recognition of Teakyo by showing average 4.28 points and standard deviation 0.48 at level of recognition of pregnant women about Taekyo on the basis of 5 points and showing to show average 2.81 points and standard deviation 0.36 at practice level on the basis of 4 points. 2. They showed the higher level of recognition on Taekyo at high educational background of pregnant woman(F=3.735, p=.005), Roman catholicism (F=4.570, p=.002), satisfied married life(F=5.448, p=.004), high monthly income(F=6.096, p=.000) and cases of hoping pregnancy(F=2.525, p=.012). 3. They showed the higher level of practice on Taekyo at high educational background of pregnant woman(F=2.883, P=.022), Roman catholicism(F=3.616, p=.032), satisfied married life(F=19.924, p=.000), good health condition(F=2.386, p=.017), cases of hoping pregnancy(F=0.677, p=.000), cases of planning pregnancy with husband(F=3.024, p=.001), cases of regular prenatal care before delivery(F=0.241, p=.005), cases of maternal breast feeding (F=9.132, p=.000), and the number of less children(F=2.763, p=.041). 4. In result of examining correlation between recognition and practice of Taekyo, they showed high level of practice on Taekyo under high level of recognition of pregnant women on Taekyo by showing the statistically significant correlation. 5. In result of examining the related factors that affect recognition and practice of Taekyo y the object, practice of Taekyo had 16.8 percents of explanatory range, purpose of practicing Taekyo 8.5 percents of explanatory range, and monthly income 1.9 percent of explanatory range as variables of affecting recognition of Taekyo. The total explanatory range was 27.2 percents, Recognition of Taekyo had 16.1 percents of explanatory ragne, time of starting Taekyo 3.2 percents, health condition 2.2 percents of explanatory range, condition of hoping pregnancy 1.1 percent of explanatory range, satisfaction of married life 0.8 percent of explanatory range, and religion 0.6 percent of explanatory range as variables of affecting practice of Taekyo. The total explanatory range was 24.0 percents.

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Microbiological and Enzymological Studies on Takju Brewing (탁주(濁酒) 양조(釀造)에 관(關)한 미생물학적(微生物學的) 및 효소학적(酵素學的) 연구(硏究))

  • Kim, Chan-Jo
    • Applied Biological Chemistry
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    • v.10
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    • pp.69-100
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    • 1968
  • 1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.

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