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Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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Finding Weighted Sequential Patterns over Data Streams via a Gap-based Weighting Approach (발생 간격 기반 가중치 부여 기법을 활용한 데이터 스트림에서 가중치 순차패턴 탐색)

  • Chang, Joong-Hyuk
    • Journal of Intelligence and Information Systems
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    • v.16 no.3
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    • pp.55-75
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    • 2010
  • Sequential pattern mining aims to discover interesting sequential patterns in a sequence database, and it is one of the essential data mining tasks widely used in various application fields such as Web access pattern analysis, customer purchase pattern analysis, and DNA sequence analysis. In general sequential pattern mining, only the generation order of data element in a sequence is considered, so that it can easily find simple sequential patterns, but has a limit to find more interesting sequential patterns being widely used in real world applications. One of the essential research topics to compensate the limit is a topic of weighted sequential pattern mining. In weighted sequential pattern mining, not only the generation order of data element but also its weight is considered to get more interesting sequential patterns. In recent, data has been increasingly taking the form of continuous data streams rather than finite stored data sets in various application fields, the database research community has begun focusing its attention on processing over data streams. The data stream is a massive unbounded sequence of data elements continuously generated at a rapid rate. In data stream processing, each data element should be examined at most once to analyze the data stream, and the memory usage for data stream analysis should be restricted finitely although new data elements are continuously generated in a data stream. Moreover, newly generated data elements should be processed as fast as possible to produce the up-to-date analysis result of a data stream, so that it can be instantly utilized upon request. To satisfy these requirements, data stream processing sacrifices the correctness of its analysis result by allowing some error. Considering the changes in the form of data generated in real world application fields, many researches have been actively performed to find various kinds of knowledge embedded in data streams. They mainly focus on efficient mining of frequent itemsets and sequential patterns over data streams, which have been proven to be useful in conventional data mining for a finite data set. In addition, mining algorithms have also been proposed to efficiently reflect the changes of data streams over time into their mining results. However, they have been targeting on finding naively interesting patterns such as frequent patterns and simple sequential patterns, which are found intuitively, taking no interest in mining novel interesting patterns that express the characteristics of target data streams better. Therefore, it can be a valuable research topic in the field of mining data streams to define novel interesting patterns and develop a mining method finding the novel patterns, which will be effectively used to analyze recent data streams. This paper proposes a gap-based weighting approach for a sequential pattern and amining method of weighted sequential patterns over sequence data streams via the weighting approach. A gap-based weight of a sequential pattern can be computed from the gaps of data elements in the sequential pattern without any pre-defined weight information. That is, in the approach, the gaps of data elements in each sequential pattern as well as their generation orders are used to get the weight of the sequential pattern, therefore it can help to get more interesting and useful sequential patterns. Recently most of computer application fields generate data as a form of data streams rather than a finite data set. Considering the change of data, the proposed method is mainly focus on sequence data streams.

Studies on Mycoplasma-Like Organism Associated with Witches' Broom of Rhus javanica (I) (Mycoplasma 성(性) 붉나무빗자루병(病)에 관(關)한 연구(硏究) (I))

  • Kim, Young Ho
    • Journal of Korean Society of Forest Science
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    • v.47 no.1
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    • pp.1-15
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    • 1980
  • The occurrence of witches' broom of Rhus javanica was first noticed in Korea by the author in 1979. Subsequently, studies were made on the symptomatology, etiology, and transmission of the disease, as well as the effect of some antibiotics on the disease development. The results of these studies are summarized as follows: 1. Symptoms of the infected plant were characterized by dwarfing of the tree accompanied by yellowing and brooming of the foliage. 2. Electron microscopy of witches' broom diseased Rhus javanica plant revealed the occurrence of numerous mycoplasma-like organisms (MLO's) in the phloem tissue cells (sieve tube elements and phloem parenchyma cells) of the rachis and midribs of infected leaves. 3. The MLO's were bounded by a single unit membrane and contained ribosome-like granules and strands presumed to be DNA. It also appears that the MLO multiply possibly by budding as well as binary and plurinary fission. 4. In the midrib of healthy leaves, vascular bundles were collaterally discontinuous. In the diseased leaves, however, xylems were connected to each other and phloem cells showed an atrophy. Granules, which were prominent in the normal abaxial epidermis, were not observed in the peidermis of diseased leaves. 5. Electron microscopy revealed crystals or osmopholic granules in the phloem parenchyma cells, and that normal stacks of grana were not developed in the chloroplasts of infected levels. 6. The disease was experimentally transmitted by grafting. Budding was more effective than crown grafting for transmitting the disease. The disease has been transmitted by grafting even when complete union of stocks and scions has not taken place. The disease agent was not transmitted by sap inoculation. Insect transmission has not been confirmed. 7. Dipping the roots of infected plants into the 500 ppm and 1,000 solutions of either tetracycline HCI or oxytetracycline, HCI was more effective on temporary remision of the symptoms than spraying the 100 ppm and 200 ppm solutions of the same antibiotics. A greater effect was achieved through dipping into 1,000 ppm than into 500 ppm.

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Angiotensin Converting Enzyme Gene Polymorphism in Alport Syndrome (알포트증후군 환자에서 안지오텐신전환효소 유전자 다형성의 의의)

  • Kim Ji-Hong;Lee Jae-Seung;Kim Pyung-Kil
    • Childhood Kidney Diseases
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    • v.8 no.1
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    • pp.18-25
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    • 2004
  • Purpose : Alport syndrome is clinically characterized by hereditary progressive nephritis causing ESRD with irregular thickening of the GBM and sensory neural hearing loss. The mutations of type IV collagen gene(COL4A5) located on the long arm of X chromosome is considered responsible for most of the structural abnormalities in the GBM of Alport patients. Since no definite clinical prognostic predictor has been reported in the disease yet, we designed this study to evaluate the significance of genetic polymorphism of the angiotensin converting enzyme in children with Alport syndrome as a prognostic factor for disease progression. Methods : ACE I/D genotype were examined by PCR amplification of the genomic DNA in 12 patients with Alport syndrome and 12 of their family members. Alport patients were divided into two groups; the conservative group, those who had preserved renal function for more than 10 years of age, the early CRF group, those who had progressed to CRF within 10 years of age. Results : The mean age of onset was $3.45{\pm}2.4$ years in the conservative group, $4.4{\pm}1.2$ years in the early CRF group. Sex ratios were 5:3 and 2:1 in each group. Among 12 cases of patients, 4 cases were in early CRF group and their mean duration of onset to CRF was 4.5 yews(8.9 years of age). Eight patients(67%) were in the conservative group and they had normal renal function for more than 10 years of age(mean duration of renal preservation was 10.6 years). The incidence of II type ACE gene were in 25.0%(3 cases), ID type in 41.7%(5 cases), DD type in 33.3%(4 cases). There was no significant difference between Alport patient and normal control(II type 44.3%, ID type 40.9%, DD type 14.8%). The incidence of DD type of early CRF group were higher than that of the conservative group(75% vs 12.5%)(p<0.05). There was no difference in ACE gene polymorphism between normal Alport family members and control group. Conclusion : Even though there was no significant difference of ACE polymorphism between Alport patients and the normal control group, the incidence of DD type is significantly increased in early CRF group which means DD type of ACE polymorphism has a possibility of being a predictor for early progression to CRF in Alport patients.

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A Critical Evaluation of the Correlation Between Biomarkers of Folate and Vitamin $B_{12}$ in Nutritional Homocysteinemia (엽산과 비타민 $B_{12}$ 결핍에 의한 호모시스테인혈증 흰쥐의 조직내 비타민 지표간의 상관관계 분석)

  • Min, Hye-Sun;Kim, Mi-Sook
    • Journal of Nutrition and Health
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    • v.42 no.5
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    • pp.423-433
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    • 2009
  • Folate and vitamin $B_{12}$ are essential cofactors for homocysteine (Hcy) metabolism. Homocysteinemia has been related with cardiovascular and neurodegenerative disease. We examined the effect of folate and/or vitamin $B_{12}$ deficiency on biomarkers of one carbon metabolism in blood, liver and brain, and analyzed the correlation between vitamin biomarkers in mild and moderate homocysteinemia. In this study, Sprague-Dawley male rats (5 groups, n = 10) were fed folatesufficient diet (FS), folate-deficient diet (FD) with 0 or 3 g homocystine (FSH and FDH), and folate-/vitamin $B_{12}$-deficient diet with 3 g homocystine (FDHCD) for 8 weeks. The FDH diet induced mild homocysteinemia (plasma Hcy 17.41 ${\pm}$ 1.94 nmol/mL) and the FDHCD diet induced moderate homocysteinemia (plasma Hcy 44.13 ${\pm}$ 2.65 nmol/mL), respectively. Although liver and brain folate levels were significantly lower compared with those values of rats fed FS or FSH (p < 0.001, p < 0.01 respectively), there were no significant differences in folate levels in liver and brain among the rats fed FD, FDH and FDHCD diet. However, rats fed FDHCD showed higher plasma folate levels (126.5 ${\pm}$ 9.6 nmol/L) compared with rats fed FD and FDH (21.1 ${\pm}$ 1.4 nmol/L, 22.0 ${\pm}$ 2.2 nmol/L)(p < 0.001), which is the feature of "ethyl-folate trap"by vitamin $B_{12}$ deficiency. Plasma Hcy was correlated with hepatic folate (r = -0.641, p < 0.01) but not with plasma folate or brain folate in this experimental condition. However, as we eliminated FDHCD group during correlation test, plasma Hcy was correlated with plasma folate (r = -0.581, p < 0.01), hepatic folate (r = -0.684, p < 0.01) and brain folate (r = -0.321, p < 0.05). Hepatic S-adenosylmethionine (SAM) level was lower in rats fed FD, FDH and FDHCD than in rats fed FS and FSH (p < 0.001, p < 0.001 respectively) and hepatic S-adenosylhomocysteine (SAH) level was significantly higher in those groups. The SAH level in brain was also significantly increased in rats fed FDHCD (p < 0.05). However, brain SAM level was not affected by folate and/or vitamin $B_{12}$ deficiency. This result suggests that dietary folate- and vitamin B12-deficiency may inhibit methylation in brain by increasing SAH rather than decreasing SAM level, which may be closely associated with impaired cognitive function in nutritional homocysteinemia.

Bacterial Blight Resistance Genes Pyramided in Mid-Late Maturing Rice Cultivar 'Sinjinbaek' with High Grain Quality (벼흰잎마름병 저항성 유전자 집적 고품질 중만생 벼 '신진백')

  • Park, Hyun-Su;Kim, Ki-Young;Baek, Man-Kee;Cho, Young-Chan;Kim, Bo-Kyeong;Nam, Jeong-Kwon;Shin, Woon-Chul;Kim, Woo-Jae;Ko, Jong-Cheol;Kim, Jeong-Ju;Jeong, Jong-Min;Jeung, Ji-Ung;Lee, Keon-Mi;Park, Seul-Gi;Lee, Chang-Min;Kim, Choon-Song;Suh, Jung-Pil;Lee, Jeom-Ho
    • Korean Journal of Breeding Science
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    • v.51 no.3
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    • pp.263-276
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    • 2019
  • 'Sinjinbaek' is a bacterial blight (BB)-resistant, mid-late maturing rice cultivar with high grain quality. To diversify the resistance genes and enhance the resistance of Korean rice cultivars against BB, 'Sinjinbaek' was developed from a cross between 'Iksan493' (cultivar name 'Jinbaek') and the F1 cross between 'Hopum' and 'HR24670-9-2-1' ('HR24670'). 'Jinbaek' is a BB-resistant cultivar with two BB resistance genes, Xa3 and xa5. 'Hopum' is a high grain quality cultivar with the Xa3 resistance gene. 'HR24670' is a near-isogenic line that carries the Xa21 gene, a resistance gene inherited from a wild rice species O. longistaminata, in the genetic background of japonica elite rice line 'Suweon345'. 'Sinjinbaek' was selected through the pedigree method, yield trials, and local adaptability tests. Using bioassay for BB races and DNA markers for resistance genes, three resistance genes, Xa3, xa5, and Xa21, were pyramided in the 'Sinjinbaek' cultivar. 'Sinjinbaek' exhibited high-level and broad-spectrum resistance against BB, including the K3a race, the most virulent race in Korea. 'Sinjinbaek' is a mid-late maturing rice cultivar tolerant to lodging. It has multiple disease resistance against BB, rice blast, and stripe virus. The yield of 'Sinjinbaek' was similar to that of 'Nampyeong'. 'Sinjinbaek' showed excellent grain appearance, good taste of cooked rice, and enhanced milling performance, and we concluded that it could contribute to improving the quality of BB-resistant cultivars. 'Sinjinbaek' was successfully introgressed with the Xa21 gene without the linkage drag negatively affecting its agronomic characteristics. 'Sinjinbaek' improved the resistance of Korean rice cultivars against BB by introgression of a new resistance gene, Xa21, as well as by pyramiding three resistance genes, Xa3, xa5, and Xa21. 'Sinjinbaek' would be suitable for the cultivation in BB-prone areas since it has been used in breeding programs for enhancing plants' resistance to BB (Registration No. 7273).