• Title/Summary/Keyword: insertion

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Studies on the Function of the Rv2435c Gene of the Mycobacterium bovis BCG (Mycobacterium bovis BCG Rv2435c 유전자의 기능에 대한 연구)

  • Lee Seung-Sil;Bae Young-Min
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.415-422
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    • 2005
  • library of the mutants was prepared by transposon mutagenesis of the Mycobacterium bovis BCG. We screened this library for the resistance to an anti-tuberculosis antibiotic, PA-824. Most of the mutants resistant to the PA-824 were not able to synthesize the coenzyme $F_{420}$ which is normally produced by the wild type M. bovis BCG strains. HPLC analysis of the cellular extract showed that one of those mutants which lost the ability to synthesize $F_{420}$ still produced F0. The insertion site of the transposon in this mutant was determined by an inverse PCR and the transposon was found to be inserted in the Rv2435c open reading frame (ORF). Rv2435c ORF is predicted to encode an 80.3 kDa protein. Rv2435c protein appears to be bound to the cytoplasmic membrane, its N-terminal present in the periplasm and C-terminal in the cytoplasm. The C-terminal portion of this protein is highly homologous with the adenylyl cyclases of both prokaryotes and eukaryotes. There are 15 ORFs which have homology with the class III AC proteins in the genome of the M. tuberculosis and M. bovis. Two of those, Rv1625c and Rv2435c, are highly homologous with the mammalian ACs. We cloned the cytoplasmic domain of the Rv2435c ORF and expressed it with six histidine residues attached on its C-terminal in Escherichia coli to find out if this protein is a genuine AC. Production of that protein in E. coli was proved by purifying the histidine-tagged protein by using the Ni-NTA resin. This protein, however, failed to complement the cya mutation in E. coli, indicating that this protein lacks the AC activity. All of the further attempts to convert this protein to a functional AC by a mutagenesis with UV or hydroxylamine, or construction of several different fusion proteins with Rv1625c failed. It is, therefore, possible that Rv2435c protein might affect the conversion of F0 to $F_{420}$ not by synthesizing cAMP but by some other way.

Study on the Relationships between Rice Straw Degradation and Changes of Fibrolytic Bacteria Population by in Vitro Rumen Fermentation (In Vitro 반추위 발효를 통한 볏짚 분해와 섬유소 박테리아 군집 변화의 관계 연구)

  • Sung, Ha Guyn
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.1
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    • pp.35-43
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    • 2017
  • This study was to research the relationships between rice straw degradation and changes of fibrolytic bacteria population during the in vitro rumen fermentation. Dry matter(DM) digestion of rice straw and population of fibrolytic bacteria were measured at the 0. 4, 8, 12 and 48 hours during the incubation. The populations of F. succinogenes. R. albus and R. flavefaciens were defined as log copy number of 16S rDNA by technical method of Quantitative real-time PCR. Total population of F. succinogenes, R. flavefaciens and R. albus was sum of bactera attached on rice straw and suspended in medium. It's population was increased with incubation, reached top level of 29.0 Log copy No at the 24 hour and then decreased. In the meantime, DM digestion of rice straw showed the higher increasement from the 8 hour to the 24 hour than from the 0 hour to the 8 hour, and then a slowdown in increasing trend of digestibility. Attachments of F. succinogenes, R. flavefaciens and R. albus were detected immediately after start of in vitro rumen incubation. At the same time, the colonized bacterial share were respectively 34.5%, 84.4% and 67.9% in total population. All of them was reached the highest colonized bacterial share above 94.7% at the 4 hour incubation. However population of attached bacteria was shown the highest level at the 12 hour or the 24 hour incubation. Kinetics of colonization were formed area of top speed from the 12 hour to the 24 hour and respectively reached 10.33, 9.28 및 8.30 Log copy No/h/g DM at the 24 hour by F. succinogenes, R. flavefaciens and R. albus. The kinetics of rice straw degradation was formed top level of 0.95% DM/h at the 24 hour. The present results gave clear evidence that degradation of rice straw was increased with the development of total fibrolytic bacteria in process of rumen fermentation. Also, their attachment was largely occurred immediately after insertion of rice straw, the colonized bacteria was actively proliferated, and then degradation of rice straw was maximized.

Effect of Multiple Superovulation and Parity on Embryo Production in Hanwoo (한우의 반복 과배란 및 산차가 수정란 생산에 미치는 영향)

  • Choi S. H.;Ryu I. S.;Son D. S.;Cho S. R.;Han M. H.;Kim H. J.;Choe C. Y.;Kim Y. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.185-190
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    • 2005
  • This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

In Vivo Embryo Production and Embryo Transfer in Hanwoo and Jeju Black Cattle Using CIDR (CIDR를 이용한 제주 한우 및 흑우의 체내 수정란 생산과 이식)

  • Kim, Y.H.;Koo, J.C.;Oh, C.W.;Kang, S.Y.;Yang, B.S.;Oh, S.J.;Kim, C.N.;Song, J.Y.;Kim, I.H.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.191-198
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    • 2006
  • The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

Comparative efficacy of 5% EMLA cream and 20% Benzocaine gel during topical anesthesia (5% EMLA cream과 20% Benzocaine gel의 도포마취 효능 비교)

  • Lee, Dae-Woo;Baik, Byeong-Ju;Kim, Jae-Gon;Yang, Yeon-Mi;So, Yu-Ryeo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.38 no.1
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    • pp.1-8
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    • 2011
  • The objective of this study was to determine the effectiveness of topical 5% EMLA cream versus 20% Benzocaine gel in reducing pain from intra oral needle insertion alone as well as injection of anesthetic. The 2 topical anesthetics were tested against each other bilaterally using a randomized, controlled, single blinded, split mouth design. Phase I was conducted to find out the rapidity of onset action of the two agents on anterior/posterior vestibules and anterior/posterior palatal mucosa. Phase II was conducted to evaluate the efficacy of the two topical anaesthetic agents in reducing the pain of intraoral injections. The agents were left in anterior/posterior vestibules and anterior/posterior palatal mucosa for the amount of time recorded in phase I. Subjects recorded pain on a 100-mm modified visual analog scale(VAS). A pulse oximeter was used to recorded the preoperative and postoperative pulse rates. In phase I of the study, two topical agents showed the longer onset of action at anterior part and vestibules than posterior part and palatal mucosa. EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. In phase II of the study, the VAS grading of the pain for anesthetic administration showed EMLA cream was significantly(P<.05) better in elimination or reducing the pain on the anterior/posterior palatal mucosa. In conclusion, EMLA cream showed the rapidest onset of action compared to benzocaine gel except on anterior palatal mucosa. EMLA cream was found to be superior to Benzocaine gel with regards to pain reduction for anesthetic administration especially on anterior and posterior palatal mucosa.

How Reliable is Sputum PCR Test in the Diagnosis of Pulmonary Tuberculosis When Sputum Smear is Negative? (객담 결핵균 도말검사가 음성일때 중합효소연쇄반응검사와 진단적 신뢰도에 관한 연구)

  • Baek, Seung-Hoon;Lee, Jae-Myung;Kang, Min-Jong;Son, Jee-Woong;Lee, Seung-Joon;Kim, Dong-Gyu;Lee, Myung-Goo;Hyun, In-Gyu;Jung, Ki-Suck;Lee, Kyung-Wha;Joe, Hyun-Chan
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.222-228
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    • 2001
  • Backgrounds : Recent technological developments have introduced a new method to identifying M. tuberculosis complex DNA in clinical samples directly. The direct amplification test (DAT) is approved for identifying M. tuberculosis complex in respiratory specimens that are smear-positive for acid-fast bacilli (AFB). When there is a discrepancy between the AFB smear and DAT, no information on their clinical utility is currently available. In this study, the diagnostic reliability of DAT was investigated in suspected pulmonary tuberculosis patients whose sputum AFB smear was negative. Methods : From June 1, 1998 through May 30, 1999, 909 patients with presumed active pulmonary tuberculosis were enrolled. A sputum AFB stain, culture, DAT and/or biopsy were performed. Using the criteria of clinical tuberculosis or confirmed tuberculosis, the positive predictive value of DAT in diagnosing pulmonary tuberculosis was investigated. Results : The positive predictive value of DAT was 82.1% by the clinically active tuberculosis criteria. However, it decreased to 61.5% when diagnosis was restricted to only to culture positive or biopsy proven cases. The false positive rate of DAT was 18.0%. Conclusion : The DAT is a valuable diagnostic method in suspected patients whose sputum AFB is was negative.

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Polymorpshisms of XPC Gene and Risk of Primary Lung Cancer in Koreans (한국인에서 XPC 유전자의 다형성과 원발성 폐암의 위험도)

  • Kim, Kyung-Rock;Lee, Su-Yeon;Choi, Jin-Eun;Kim, Kyung-Mee;Jang, Sang-Soo;Jung, Chi-Young;Kang, Kyung-Hee;Jeon, Kyung-Neoyh;Cha, Sung-Ick;Kim, Chang-Ho;Kam, Sin;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.2
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    • pp.113-126
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    • 2002
  • Background : DNA repair plays a crucial role in protection from cancer-causing agents. Therefore, a reduced DNA repair capacity can increase the susceptibility to lung cancer. The XPC gene contains 15 exons and encodes a 940 amino acid protein that plays a central role in DNA damage recognition of the nucleotide excision repair pathway, which is a major DNA repair mechanisim removing the bulky-helix distorting DNA lesions caused by smoking. Recently several polymorphisms in the XPC gene were identified. In addition, it is possible that these polymorphisms may affect the DNA repair capacity, which modulate cancer susceptibility. The relationship between codon 499 and 939 polymorphisms, and a poly(AT) insertion/deletion polymorphism in the XPC gene, and the lung cancer risk were investigated. Materials and Methods : The genotypes were determined using either PCR or PCR-RFLP analysis in 219 male lung cancer patients and 150 healthy males controls. Results : The frequencies of the genotypes (Val499Ala, PAT and Lys939Gln) among the cases were not significantly different from those of the controls. There was no significant associantion between these polymorphi는 and the lung cancer risk when the analyses were stratified according to age, smoking status and the pack-years of smoking. Moreover, the genotypes had no apparent relationship with any of the histological types of lung cancer. There was a linkage disequilibrium among the Val499Ala, PAT and Lys939Gln polymorphisms. The PAT polymorphism had a strong linkage disequilibrium with the Lys939Gln polymorphism (kappa value=0.87). The XPC haplotypes showed no significant association with the lung cancer risk. Conclusion : These results suggest that XPC Val499Ala, PAT and Lys939Gln polymorphisms are not major contributors to the individual lung cancer susceptibility in Koreans.

Athermalization and Narcissus Analysis of Mid-IR Dual-FOV IR Optics (이중 시야 중적외선 광학계 비열화·나르시서스 분석)

  • Jeong, Do Hwan;Lee, Jun Ho;Jeong, Ho;Ok, Chang Min;Park, Hyun-Woo
    • Korean Journal of Optics and Photonics
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    • v.29 no.3
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    • pp.110-118
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    • 2018
  • We have designed a mid-infrared optical system for an airborne electro-optical targeting system. The mid-IR optical system is a dual-field-of-view (FOV) optics for an airborne electro-optical targeting system. The optics consists of a beam-reducer, a zoom lens group, a relay lens group, a cold stop conjugation optics, and an IR detector. The IR detector is an f/5.3 cooled detector with a resolution of $1280{\times}1024$ square pixels, with a pixel size of $15{\times}15{\mu}m$. The optics provides two stepwise FOVs ($1.50^{\circ}{\times}1.20^{\circ}$ and $5.40^{\circ}{\times}4.23^{\circ}$) by the insertion of two lenses into the zoom lens group. The IR optical system was designed in such a way that the working f-number (f/5.3) of the cold stop internally provided by the IR detector is maintained over the entire FOV when changing the zoom. We performed two analyses to investigate thermal effects on the image quality: athermalization analysis and Narcissus analysis. Athermalization analysis investigated the image focus shift and residual high-order wavefront aberrations as the working temperature changes from $-55^{\circ}C$ to $50^{\circ}C$. We first identified the best compensator for the thermal focus drift, using the Zernike polynomial decomposition method. With the selected compensator, the optics was shown to maintain the on-axis MTF at the Nyquist frequency of the detector over 10%, throughout the temperature range. Narcissus analysis investigated the existence of the thermal ghost images of the cold detector formed by the optics itself, which is quantified by the Narcissus Induced Temperature Difference (NITD). The reported design was shown to have an NITD of less than $1.5^{\circ}C$.

Coat Color Patterns and Genotypes of Extension and Agouti in Hanwoo and Jeju Black Cattle (제주흑우와 한우에서 Extension, Agouti 유전자형과 모색 출현 양상)

  • Han, Sang-Hyun;Cho, In-Cheol;Kim, Jae-Hwan;Ko, Moon-Suck;Kim, Young-Hoon;Kim, Eun-Young;Park, Se-Pill;Lee, Sung-Soo
    • Journal of Life Science
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    • v.21 no.4
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    • pp.494-501
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    • 2011
  • To understand the relationship between coat color inheritance patterns and genotypes of Extension (E) and Agouti (A) loci in cattle, the genotypes for melanocortin-1 receptor (MC1R) and agouti signaling protein (ASIP) were analyzed in Hanwoo, Jeju black cattle (JBC), and their crossbred progeny. Three MC1R alleles ($E^D$, $E^+$, and e) were found in the black-colored JBC population. JBC had no recessive homozygotes (e/e), but this genotype was predominant in the Hanwoo breed. However, MC1R $E^+$/e Hanwoo did not produce a black coat color as they appeared either as brown or solid red. For ASIP, three genotypes (A/A, A/$A^{Br}$, and $A^{Br}/A^{Br}$) were determined by insertion/deletion of an L1-BT element in Hanwoo. The ASIP $A^{Br}$ allele was rarely observed, and no ASIP $A^{Br}/A^{Br}$ homozygotes were detected in the JBC population. Cattle carrying ASIP $A^{Br}$ did not show any agouti-like brindle pigmentation patterns in either breed or their progeny. The coat colors of the crossbred progeny were discriminated by two colors, yellowish-brown versus dark-brown or black, and their coat colors were directly related to the genotypes of the Extension locus, yellowish-brown (e/e) and dark-brown or black ($E^+$/e), but not to the Agouti locus. ASIP genotypes probably did not affect coat color development in the Hanwoo or crossbred progeny. Our results suggest that the ASIP genotypes do not play key roles in coat color variation, but the MC1R genotypes do direct the phenotypes of Hanwoo, JBC, and their progeny.

Study on Iron-making and Manufacturing Technology of Iron Swords with Ring Pommel Excavated in Ipbuk-dong, Suwon (수원 입북동 출토 철제환두도의 제철과 제작기술 연구)

  • Kim, Soo-Ki
    • Journal of Conservation Science
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    • v.32 no.4
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    • pp.579-588
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    • 2016
  • This study analyzed nonmetallic inclusions in iron swords with a ring pommel excavated in the Ipbuk-dong, Suwon. Scanning electron microscopy with energy dispersive spectroscopy(SEM-EDS) was used to estimate the iron-making temperature, and we compared the oxide with $SiO_2$ to investigate the heat-treatment technology in the production of iron swords with a ring pommel by investigating the artificial insertion of a slag former and the metallurgical structure. From the wustite observed in most of the specimens, it is judged that these swords were produced by heating and forging iron smelted at a low temperature using the solid reduction method. In addition, judging from the partial presence of $P_2O_5$, it is assumed that they were smelted directly with natural ore, not calcined. From the ratios of $CaO/SiO_2$ and $TiO_2/SiO_2$, it is judged that the raw material for iron-making was iron ore and that a calcareous slag former was not artificially inserted. The structure of the blade part on the front end was pure iron. From the high carbon content of the blade part on the ring pommel and the formation of a martensitic structure and pearlite colony, it is judged that they were tempered after carburizing and that the back, handle part, and ring pommel were unintentionally carburized. Judging from the structure of these specimens, it was noted that they were produced by applying artificial partial heat-treatment technology. This study attempted to present a more scientific analysis by using the method of interpretation through component analysis of nonmetallic inclusions appearing in one relic by the ratio of the oxide divided by $SiO_2$. It is judged that reinterpreting the arguments by the results of the existing analysis and research in this way can obtain different interpretations.