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Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

Biological Activities of Essential Oil from Chamaecyparis obtusa (편백(Chamaecyparis obtusa) 정유의 항균, 항염, 항산화 효과)

  • Ahn Jeung-Youb;Lee Sung-suk;Kang Ha-young
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.503-507
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    • 2004
  • The essential oil from Chamaecyparis obtusa was investigated for biological activities in anti-oxidative, anti-inflammation and antibacterial method, respectively. The Growth inhibitory effect of C. obtusa oil on the bacteria was evaluated with MIC (minimum inhibitory concentration), $IC_{50}\;(50\%$ inhibitory concentration), and paper disc method. Two kinds of gram positive strains and two kinds of gram negative strains were used in this study. Gram positive strains were B. subtilis and S. aureus. and Gram negative strains were E. coli and P. aeruginosa. Gram positive strains showed much more intensive effect than gram negative strains. Anti-oxidative effect was investigated with DPPH (1,1-diphenyl-2-picrylhidrazyl) in methanol based and $IC_{50}\;was\;0.78\%.$ Our results suggest that the essential oil from Chamaecyparis obtusa has effects on anti-bacterial, anti- oxidative and anti-inflammation in in vitro and in uiuo. Then this material could be expect synergic effect with other candidated extracts and oils.

Magnetic Hardening of Rapidly Solidified $SmFe_{7+x}M_{x}(M=Mo,\;V,\;Ti)$ Compounds (급속냉각된 $SmFe_{7+x}M_{x}(M=Mo,\;V,\;Ti)$ 화합물에서 생성된 신 강자성상)

  • Choong-Jin Yang;E. B. Park;S. D. Choi
    • Journal of the Korean Magnetics Society
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    • v.4 no.3
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    • pp.226-232
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    • 1994
  • Rapidly solidified $SmFe_{7+x}M_{x}(M=Mo,\;V,\;Ti)$ compound were found to crystallize in the ${Sm(Fe,\;M)}_{7}$ based stable magnetic phase by introducing a second transition element into the Sm-Fe binary system. The ${Sm(Fe,\;M)}_{7}$ phase exhibits the highest Curie temperatuer ($T_{c}=355^{\circ}C$) ever Known in the Sm-Fe magnetic systems with a quite high intrinsic coercivity($_{i}H_{c}=3~6\;kOe $). The ${Sm(Fe,\;M)}_{7}$ phase remains stable even after annealing if once form during the rapid solidification. The primary reason for the high coercive force is due to the fine grain size($2000~8000\;{\AA}$)of the magnetic ${Sm(Fe,\;M)}_{7}$ matrix phase, and the enhanced Curie temperature is attributed to the extended solid-solubility of the additive transition elements in Fe matrix, which leads to volume expansion of the ${Sm(Fe,\;M)}_{7}$ cell causing an enhanced coupling constant of Fe atoms.

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The analgesic effect of combined electroacupuncture at Hoku (LI4) and Zusanli (ST36) using TFL (합곡혈(合谷穴)과 족삼리혈(足三里穴) 병용자극(倂用刺戟)이 TEL에 미치는 영향)

  • Baek, Kyong-Won;Ko, Eun-Sang;Min, Byung-Il;Park, Dong-Suk
    • Journal of Acupuncture Research
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    • v.18 no.1
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    • pp.76-87
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    • 2001
  • Objective : Zusanli(ST36) and Hoku(Li4) are analgesic acupuncture points frequently used for acupuncture in Oriental medicine. The present study was conducted to see the antinociceptive effects produced by electroacupuncture combined two frequencies(Low, High) and two different acupuncture points(LI4, ST36) in the rat tail flick test. Method : In this study the Rats (Sprague-Dawley, 250-300g) were partially anesthetized with thiopental sodium(40mg/kg, i.p.). The basal reaction time for the tail-flick was 3${\pm}$0.5 sec. Low frequency(3Hz, 5V, biphasic) and high frequency(100Hz, 5V, biphasic) were applied to the inserted needle for the period of insertion(twenty minutes). Experimental groups are divied as follow; a) electroacupuncture stimulation groups at Hoku with or high frequency(L-EA, H-EA), b) electroacupuncture stimulation groups at Zusanli with low or high frequency(1-EA, h-EA), c) low frequency at Hoku and Zusanli(LIEA), d) low frequency at Hoku and high frequency at Zusanli(LhEA), e) high frequency at Hoku and low frequency at Zusanli(HIEA), f) high frequency at Hoku and Zusanli(HhEA) Results : The individual stimulation at either Hoku or Zusanli with low frequency has stronger and longer analgesic effect than high frequency stimulation. In addition, the combined stimulation at Hoku and Zusanli with low frequency has superior effect to individual stimulation with low frequency. LhEA and LIEA have superior effect to other stimulation groups among the combined groups. In order to determine the involvement of opioid system on the different antinociceptive effects, Naloxone, an opioid antagonist, was used in the combined groups. LIEA is the most sensitive when naloxone was administrated among study groups. HhEA is the least sensitive in the administration of naloxone. Conclusion : From results, this study confirmed that the opioid system is involved in analgesic effect of low frequency stimulation of acupuncture point, and we also can suggest the stronger analgesic effect of combining stimulation points is due to the theory of spatial summation in the nervous system.

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Molecular Cloning and Function Analysis of an Anthocyanidin Synthase Gene from Ginkgo biloba, and Its Expression in Abiotic Stress Responses

  • Xu, Feng;Cheng, Hua;Cai, Rong;Li, Lin Ling;Chang, Jie;Zhu, Jun;Zhang, Feng Xia;Chen, Liu Ji;Wang, Yan;Cheng, Shu Han;Cheng, Shui Yuan
    • Molecules and Cells
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    • v.26 no.6
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    • pp.536-547
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    • 2008
  • Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase), a 2-oxoglutarate iron-dependent oxygenase, catalyzed the penultimate step in the biosynthesis of the anthocyanin class of flavonoids, from the colorless leucoanthocyanidins to the colored anthocyanidins. The full-length cDNA and genomic DNA sequences of ANS gene (designated as GbANS) were isolated from Ginkgo biloba for the first time. The full-length cDNA of GbANS contained a 1062-bp open reading frame (ORF) encoding a 354-amino-acid protein. The genomic DNA analysis showed that GbANS gene had three exons and two introns. The deduced GbANS protein showed high identities to other plant ANSs. The conserved amino acids (H-X-D) ligating ferrous iron and residues (R-X-S) participating in 2-oxoglutarate binding were found in GbANS at the similar positions like other ANSs. Southern blot analysis indicated that GbANS belonged to a multi-gene family. The expression analysis by real-time PCR showed that GbANS expressed in a tissue-specific manner in G. biloba. GbANS was also found to be up-regulated by all of the six tested abiotic stresses, UV-B, abscisic acid, sucrose, salicylic acid, cold and ethylene, consistent with the promoter region analysis of GbANS. The recombinant protein was successfully expressed in E. coli strain with pET-28a vector. The in vitro enzyme activity assay by HPLC indicated that recombinant GbANS protein could catalyze the formation the cyanidin from leucocyanidin and conversion of dihydroquercetin to quercetin, suggesting GbANS is a bifunctional enzyme within the anthocyanidin and flavonol biosynthetic pathway.

The Change of Compositions and Antioxidant Effect in Soybean Cultivars Pickled in Persimmon Vinegar (콩 품종별 감식초 절임 중 성분의 변화와 항산화 효과)

  • Bang, Hyo-Pil;Choi, One-Kyun;Cho, Gyu-Seong;Son, Jong-Youn;Ryu, Gi-Hyung
    • The Korean Journal of Food And Nutrition
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    • v.19 no.4
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    • pp.398-409
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    • 2006
  • Changes of component(amino acid, free sugar, fatty acid and mineral) and antioxidant effect by pickling of soybeans we.e investigated. Soybean cultivars, Daewon, Pungsannamul, Hwaeum(yellow bean), Ilpum Black and Zynuni(black bean) were soaked in persimmon vinegar for 10 days at $20^{\circ}C$. The major amino acids of raw soybeans were glutamic acid, aspartic acid, lysine and leucine. The content of total amino acid decreased by pickling. The major free sugar of raw soybeans were glucose, fructose and sucrose. Sucrose decreased and glucose and fructose increased by pickling. Maltose was found only in pickled Daewon and Ilpum Black. Linoleic and linolenic acid content of raw soybeans were $49.3%{\sim}57.1%\;and\;7.8{\sim}8.9%$, and the fatty acid contents did not change by pickling. The major mineral elements were K, P and Mg, and their compositions except for Na did not change by pickling. Total phenolic compound, vitamin $B_1$ and C content increased during pickling. The antioxidant effects, electron donating abilities, nitrite-scavenging abilities increased by pickling.

Comparison of Growth, Milk Yield and Draughtability of Murrah-Philippine Crossbred and Philippine Native Buffaloes

  • Salas, R.C.D.;van der Lende, T.;Udo, H.M.J.;Mamuad, F.V.;Garillo, E.P.;Cruz, L.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.5
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    • pp.580-586
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    • 2000
  • Data collected between 1981 and 1991 at the Philippine Carabao Center at Central Luzon State University (PCC-CLSU) were used for the comparison of growth, milk yield and draughtability of Murrah-Philippine crossbred and Philippine native buffaloes. Body weights and body measurements were available at 3-month intervals from birth to 36 months of age for a total of 34 $Murrah{\times}Philippine$ native buffalo F1 crossbreds (CBB; 21 cows, 13 bulls) and 32 Philippine native buffaloes (PNB; 16 cows, 16 bulls). Lactation records were available for 14 CBB and 19 PNB cows. Data for draughtability under wet and dry ploughing conditions were available for 4 CBB and 4 PNB steers. The results indicate that crossbreds grow faster (0-9 months of age: cows $442{\pm}19$ vs. $301{\pm}21g/day$, bulls $305{\pm}23$ vs. $296{\pm}21g/day$; 9-36 months of age: cows $227{\pm}10$ vs. $147{\pm}12g/day$, bulls $282{\pm}13$ vs. $138{\pm}12g/day$), mature earlier and produce more milk (1st lactation: $1139{\pm}153$ vs. $450{\pm}112kg$; 2nd lactation: $1115{\pm}132$ vs. $488{\pm}136kg$) than native buffaloes, but have a poorer draughtability (wet ploughing; force as % of body weight $8.8{\pm}0.2$ vs. $12.2{\pm}0.6$; dry ploughing: cut depth $10.98{\pm}0.25$ vs. $11.92{\pm}0.13cm$, velocity $0.50{\pm}0.03$ vs. $0.60{\pm}0.02m/sec$, force as % of body weight $9.0{\pm}0.6$ vs. $11.3{\pm}0.7$). The correlation coefficients between body weight and body measurements at birth and at 3-month intervals indicate that heart girth has a relatively high correlation with body weight, especially in crossbreds. It is concluded that in Philippine smallholder farming systems in which meat and milk production are secondary to draught power, the native buffalo is preferable from the point of view of input needed to maintain the number of animals kept for a required draught force.

Quality of Mozzarella Cheese Analogues Prepared from Soy Milk with WP, WPC 34, WPC 80, DWP, or LP during the Storage Period (다양한 유청제품인 WP, WPC 34, WPC 80, DWP, LP를 Soymilk에 첨가하여 제조된 Mozzarella Cheese Analogue의 저장 중 품질 변화에 관한 연구)

  • Jin, Woo-Seung;Song, Kwang-Young;Seo, Kun-Ho;Yoon, Yoh-Chang
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.1
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    • pp.35-49
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    • 2013
  • The purpose of this study was to develop Mozzarella cheese analogues by using dairy products in the form of WPC 34, WPC 80, whey protein, demineralized whey powder, and lactose powder along with soy milk. Soy milk was separately blended with 5% WPC 34 (A), WPC 80 (B), DWP (C), WP (D), and LP (E) and also with 10% WPC 34 (F), WPC 80 (G), DWP (H), WP (I), and LP (J). Blending of soy milk and whey products showed that increase in the proportions of whey products (WPC 34, WPC 80, DWP, WP, and LP) led to increase in the protein, lactose, and SNF levels of the admixture. A decrease in fat content was observed for all cheeses prepared from mixtures, relative to those for the control cheese. The nitrogen content within analogue samples was higher than that in the control cheese and increased with increase in the proportions of whey products within soy milk. Higher water soluble nitrogen levels were observed in cheese prepared from whey-product-blended soy milk than in the control cheese. The non-protein nitrogen level within the control Mozzarella cheese was significantly lower than that in the Mozzarella analogues, and, in the case of cheese analogues, it increased with increase in the proportion of whey products in soy milk. With regard to the physicochemical and sensory qualities of the Mozzarella cheese analogues and control cheese, the pH of all analogue samples, with the exception of the cheese prepared from group G, was lower than that of the control Mozzarella cheese. Rheological studies showed that the hardness of Mozzarella cheese analogues was lower than that of the control Mozzarella, while the elasticity, cohesiveness, and brittleness of the analogues was higher. The control sample had a higher meltability level than any of the Mozzarella analogues. Mozzarella cheese prepared with the traditional method had higher browning and stretching levels than all the cheese analogues, but a lower oiling-off level.

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Measuring Myun Health Worker's Performance by Time-Activity Approach (시간활동 접근법에 의한 면 보건요원의 업무량 분석)

  • Kim, Han-Joong;Kim, Moon-Shik
    • Journal of Preventive Medicine and Public Health
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    • v.10 no.1
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    • pp.34-43
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    • 1977
  • This study attempts to examine the performances of Myun Health Workers-the frontline workers in the Korean rural health care delivery system. The time-activity approach was mainly utilized as a measuring tool. This study was undertaken in September 1976 with 35 Myun Health Workers at the Kang Wha County. The pretested time-activity approach sheets were filled out daily for one month by those Myun Health Workers themselves. Statistical means and variances of analysis were utilized for statistical method in comparing some activities and functions converged into time distribution Findings: 1. The workers's average working hours derived in this study is 8 hours and 48 minutes per day, which takes half an hour longer than normal schedule. 2. They spend 56% working hour for direct services, in other words, the main function, 22% for supportive function, and 22% for other activities, the unrelated health services. 3. Considering the total working hours of main function, out-center activity is far more than in-center services with the ratio of 70% to 30% respectively, which proves, therefore, that the main activity of the workers is home visiting. 4. It takes 20 minutes purely for home visiting and takes 14 minutes for transportation. 5. This research also indicates that such factors as characteristics of the health workers and myun influence in shaping the structures of the worker's function and activity: a. The workers whose working site is located in myun office spend 15% among total working hours in carring out official myun activities, which is incidentally unrelated to health services, while the health subcenter have no rooms for administrative jobs for myun office. b. The workers whose office is in health subcenter contribute much time in doing main function and those working in special project distribute more time in performing supportive function. c. The types of workers are another dominant factor to influence the components of worker's functions and activities. MCH workers and MPW I spend much time for manipulating main function. d. MPW II, whose function is reorganized by special project in 2 myuns shows different pattern of time distribution compared to the TB worker orFP worker in the ordinary area. MPW II distributes their time evenly in performing MCH program, T.B. Program, F.P. program and education activity, while the unipurpose workers engage in carring out only their dominant role. e. Another variables which involve the variation of the worker's activity can be illustrated with the variables like target population, size of myun and convenience for transportation, among which the latter two are remarkable factors in determining the time for out-center service.

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A Study on Estimation of Failure Probability of Allowable Stress Design using Reliability Analysis to the Bearing Capacity the Deep Water Depth Large-diameter Drilled Shaft (대수심 대구경 현장타설말뚝의 지지력에 대한 신뢰성 해석을 이용한 허용응력 설계의 파괴확률 평가 연구)

  • Han, Yushik;Lee, Yunkyu;Choi, Yongkyu
    • Journal of the Korean GEO-environmental Society
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    • v.15 no.4
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    • pp.43-51
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    • 2014
  • A Large-diameter drilled shaft of deep water depth composite foundation supporting a high rise pylon of the test designed super long span bridge was designed by allowable stress design method and failure probability through reliability analysis to bearing capacity was estimated. The allowable stress design results for the bearing capacity of a drilled shaft were analyzed by reliability analysis and the probability of failure shows 0.12 % in case of CFEM, 0.0002 % in case of Korea Highway Corporation criterion, and 0.003 % in case of structure foundation design criterion. In the allowable stress design, the bearing capacity of a large-diameter drilled shaft was obtained by applying to safety factor 3 and reliability analysis for the results was done. If the failure probability suggested by AASHTO(2007) specification is set to 0.02 %, the socketed length of a drilled shaft shows an increase of 25 % in CFEM, decrease of 60 % in KHCC, and decrease of 89 % in SFDC.