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Detection of Salmonella in Milk by Polymerase Chain Reaction

  • Park, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.15 no.3
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    • pp.262-266
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    • 2000
  • The polymerase chain reaction was used to selectively detect sequences within the fimbrial antigen of Salmonella enteritidis. Sterile milk was artificially inoculated with known amount of S. enteritidis and then DNA was extracted with guanidine thiocyanate/phenol/chloroform, followed by PCR. A detection limit of as few as 100 colony forming unit (cfu) per 0.5 ml milk was obtained with this method. For the whole procedure, it took only 5 h. A semi-quantitative polymerase chain reaction assay which allows an estimation of colony forming unit of S. enteritidis was developed. Known amount of standard plasmid pGem-4Z-Sef B(-) containing cloned S. enteritidis fimbrial antigen gene was co-amplified with Salmonella genomic DNA isolated from artificially inoculated milk. The same set of primers were used for the amplification and the products were cleaved with Bam HI. The concentration of the target DNA could be estimated by comparing the intensity of the two bands after electrophoresis. The PCR-based protocol described in this paper provides a rapid, simple, and sensitive method for detecting S. enteritidis in milk.

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Sequence Analysis of the Gene Encoding gp55 Protein of Suri Strain, an Attenuated Classical Swine Fever (Hog Cholera) Virus (Classical Swine Fever (Hog Cholera) Virus 약독순화주 (Suri 주)의 gp55 Gnen 염기서열 분석)

  • Kim, Kui-Hyun;Chang, Kyung-Soo;Kang, Kyong-Im;Lee, Byung-Hyung;Park, Jong-Hyeon;An, Soo-Hwan;Jun, Moo-Hyung
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.303-316
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    • 1998
  • An attenuated classical swine fever virus (CSFV), Suri strain, is a variant derived from a vaccine virus, LOM strain. This study was performed to elucidate the molecular biologcal properties of CSFV Suri strain, and to obtain the basic data for molecular epidemiological approaches for the disease. The truncated form of gp55 gene without the C-terminal transmembrane domain, in size of 1,023bp, was amplified by RT-PCR and sequenced by dye terminator cyclic sequencing method, and inserted into BamHI site of pAcGP67B baculovirus vector, establishing a cloned pAcHEG plasmid. By the nucleotide sequences determined, 341 amino acid sequences were predicted. As compared the nucleotide and amino acid sequences of gp55 of Suri with the various CSFV, Suri strain showed the high homology over 99.1% with ALD and LOM strains, but comparably the lower homology with Alfort and Brescia. In comparison of amino acid sequence in variable domain of gp55 protein, the similar tendency of homology was observed. In hydrophobicity analysis, all of four CSFV strains revealed the analogous patterns of hydrophobicity. The numbers and locations of N-glycosylation site and cysteine residues in gp55 were analyzed, those of Suri strain being coincident with ALD and LOM strains. The results suggest that gp55 in Suri strain has the high similarity to those in ALD and LOM strains in terms of the nucleotide and amino acid sequences and the functional properties of gp55 protein.

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Expression and Biochemical Characterization of Cold-Adapted Lipases from Antarctic Bacillus pumilus Strains

  • Litantra, Ribka;Lobionda, Stefani;Yim, Joung Han;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1221-1228
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    • 2013
  • Two lipase genes (bpl1 and bpl3) from Antarctic Bacillus pumilus strains were expressed in Bacillus subtilis. Both recombinant lipases BPL1 and BPL2 were secreted to the culture medium and their activities reached 3.5 U/ml and 5.0 U/ml, respectively. Their molecular masses apparent using SDS-PAGE were 23 kDa for BPL1 and 19 kDa for BPL3. Both lipases were purified to homogeneity using ammonium sulfate precipitation and HiTrap SP FF column and Superose 12 column chromatographies. The final specific activities were estimated to be 328 U/mg for BPL1 and 310 U/mg for BPL3. Both lipases displayed an optimum temperature of $35^{\circ}C$, similar to other mesophilic enzymes. However, they maintained as much as 70% and 80% of the maximum activities at $10^{\circ}C$. Accordingly, their calculated activation energy at a temperature range of $10-35^{\circ}C$ was 5.32 kcal/mol for BPL1 and 4.26 kcal/mol for BPL3, typical of cold-adapted enzymes. The optimum pH of BPL1 and BPL3 was 8.5 and 8.0, respectively, and they were quite stable at pH 7.0-11.0, showing their strong alkaline tolerance. Both lipases had a preference toward medium chain length ($C_6-C_{10}$) fatty acid substrates. These results indicate the potential for the two Antarctic B. pumilus lipases as catalysts in bioorganic synthesis, food, and detergent industries.

Physicians' Attitude towards Accreditation of Ambulatory Health Care Organization and Related Characteristics (의원 자율인증제에 대한 개원의사 태도 및 관련 특성 분석)

  • Kim, Min-Ji;Shin, Eui-Chul;Park, Seong-Hi;Kim, Jee-Yoon
    • Quality Improvement in Health Care
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    • v.19 no.2
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    • pp.36-46
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    • 2013
  • Objectives: The purpose of this study is to find a way to effectively introduce accreditation for ambulatory health care organization by identifying the physicians' attitude for accreditation and characteristics that affect thei r attitude. Methods: A web survey was conducted from February 15 to March 4, 2011 for 183 physicians who work in ambulatory health care organizations throughout the nation. Self-reported questionnaire was used for this study. SPSS WIN(version 12.0) was utilized for statistical analysis. Results: Physician's attitude towards accreditation for ambulatory health care organization was positive(25.7%), moderate(39.3%), and negative(35.0%). Clinics that practice as a group or which treat more patients per day showed more positive attitude(p<0.05). The result of ordinal regression analysis indicates the groups with daily patients over 100 showed 36.3 times more positive attitude than the one under 75(p<0.05). Conclusion: The accreditation for ambulatory health care organization has not been throughly discussed yet. Many physicians did not have knowledge about accreditation. Clinics that see more patients per day had more positive attitude for accreditation. It indicates the cost may play important role in voluntary participation in accreditation.

MARGINAL DISCREPANCY AND RETENTION FORCE OF CONICAL TELESCOPE OUTER CROWN WITH CO-CR-TI ALLOY (Co-Cr-Ti 합금으로 제작한 conical telescope 외관의 변연적합도와 유지력에 관한 연구)

  • Jung, Hi-Chan;Jeong, Chang-Mo;Jeon, Young-Chan;Lim, Jang-Seop
    • The Journal of Korean Academy of Prosthodontics
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    • v.38 no.2
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    • pp.214-225
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    • 2000
  • The purpose of this study was to investigate the effects of investing conditions on the marginal discrepancy of conical telescope outer crown with Co-Cr-Ti alloy(Dentitan) and to compare the marginal discrepancy and the retention force of outer crowns using different pattern materials(plastic foil, casting wax, pattern resin). To evaluate the effects of investing conditions on the marginal discrepancy, patterns with plastic foil were invested under three different liquid/powder ratio conditions using phosphate bonded investment(Univest-nonprecious): standard, 10% decreased and 10% increased. At each liquid/powder ratio condition, metal ring was lined with single or double layers of ceramic ring liner. The marginal discrepancy of outer crown at different investing conditions was measured by ${\times}100$ compact measuring microscope(STM5, Olympus, Japan). For measurement of the marginal discrepancy and the retention force of outer crown using different pattern materials, the investing condition of 10% decreased liquid/powder ratio and double layers of ring liner was selected because this investing condition resulted in the best fit of outer crown. Marginal discrepancy was measured in the same way above and retention force on universal testing machine. Under the conditions of this study, the following conclusions were drawn: 1. The thickness of ring liner had more influence on the marginal discrepancy of outer crown than the liquid/powder ratio, and the acceptable marginal fitness could not be expected at the investing condition directed by investment manufacturer 2. There were no differences in the marginal discrepancy of outer crown among three different pattern materials(P>0.05). 3. Casting wax showed the greatest retention force(1640g) of outer crown, followed by pattern resin(1110g), plastic foil(820g). However, there was no significant difference between plastic foil and pattern resin(P>0.05). 4. Plastic foil showed the least variation in marginal discrepancy and retention force.

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Solar Cell Efficiency Improvement using a Pre-deposition Temperature Optimization in The Solar Cell Doping Process (도핑 공정에서의 Pre-deposition 온도 최적화를 이용한 Solar Cell 효율 개선)

  • Choi, Sung-Jin;Yoo, Jin-Su;Yoo, Kwon-Jong;Han, Kyu-Min;Kwon, Jun-Young;Lee, Hi-Deok
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2010.06a
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    • pp.244-244
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    • 2010
  • Doping process of crystalline silicon solar cell process is very important which is as influential on efficiency of solar. Doping process consists of pre -deposition and diffusion. Each of these processes is important in the process temperature and process time. Through these process conditions variable, p-n junction depth can be controled to low and high. In this paper, we studied a optimized doping pre-deposition temperature for high solar cell efficiency. Using a $200{\mu}m$ thickness multi-crystalline silicon wafer, fixed conditions are texture condition, sheet resistance($50\;{\Omega}/sq$), ARC thickness(80nm), metal formation condition and edge isolation condition. The three variable conditions of pre-deposition temperature are $790^{\circ}C$, $805^{\circ}C$ and $820^{\circ}C$. In the $790^{\circ}C$ pre-deposition temperature, we achieved a best solar cell efficiency of 16.2%. Through this experiment result, we find a high efficiency condition in a low pre-deposition temperature than the high pre-deposition temperature. We optimized a pre-deposition temperature for high solar cell efficiency.

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Analysis of An Anomalous Hump Phenomenon in Low-temperature Poly-Si Thin Film Transistors (저온 다결정 실리콘 박막 트랜지스터의 비정상적인 Hump 현상 분석)

  • Kim, Yu-Mi;Jeong, Kwang-Seok;Yun, Ho-Jin;Yang, Seung-Dong;Lee, Sang-Youl;Lee, Hi-Deok;Lee, Ga-Won
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.24 no.11
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    • pp.900-904
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    • 2011
  • In this paper, we investigated an anomalous hump phenomenon under the positive bias stress in p-type LTPS TFTs. The devices with inferior electrical performance also show larger hump phenomenon. which can be explained by the sub-channel induced from trapped electrons under thinner gate oxide region. We can confirm that the devices with larger hump have larger interface trap density ($D_{it}$) and grain boundary trap density ($N_{trap}$) extracted by low-high frequency capacitance method and Levinson-Proano method, respectively. From the C-V with I-V transfer characteristics, the trapped electrons causing hump seem to be generated particularly from the S/D and gate overlapped region. Based on these analysis, the major cause of an anomalous hump phenomenon under the positive bias stress in p-type poly-Si TFTs is explained by the GIDL occurring in the S/D and gate overlapped region and the traps existing in the channel edge region where the gate oxide becomes thinner, which can be inferred by the fact that the magnitude of the hump is dependent on the average trap densities.

A 60 GHz Bidirectional Active Phase Shifter with 130 nm CMOS Common Gate Amplifier (130 nm CMOS 공통 게이트 증폭기를 이용한 60 GHz 양방향 능동 위상변화기)

  • Hyun, Ju-Young;Lee, Kook-Joo
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.22 no.11
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    • pp.1111-1116
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    • 2011
  • In this paper, a 60 GHz bidirectional active phase shifter with 130 nm CMOS is presented by replacing CMOS passive switchs in switched-line type phase shifter with Common Gate Amplifier(bidirectional amplifier). Bidirectional active phase shifter is composed of bidirectional amplifier blocks and passive delay line network blocks. The suitable topology of bidirectional amplifier block is CGA(Common Gate Amplifier) topology and matching circuits of input and output are symmetrical due to design same characteristic of it's forward and reverse way. The direction(forward and reverse way) and amplitude of amplification can be controlled by only one bias voltage($V_{DS}$) using combination bias circuit. And passive delay line network blocks are composed of microstrip line. An 1-bit phase shifter is fabricated by Dongbu HiTek 1P8M 130-nm CMOS technology and simulation results present -3 dB average insertion loss and respectively 90 degree and 180 degree phase shift at 60 GHz.

Targeted Resequencing of 30 Genes Improves the Detection of Deleterious Mutations in South Indian Women with Breast and/or Ovarian Cancers

  • Rajkumar, Thangarajan;Meenakumari, Balaiah;Mani, Samson;Sridevi, Veluswami;Sundersingh, Shirley
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5211-5217
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    • 2015
  • Background: We earlier used PCR-dHPLC for mutation analysis of BRCA1 and BRCA2. In this article we report application of targeted resequencing of 30 genes involved in hereditary cancers. Materials and Methods: A total of 91 patient samples were analysed using a panel of 30 genes in the Illumina HiScan SQ system. CLCBio was used for mapping reads to the reference sequences as well as for quality-based variant detection. All the deleterious mutations were then reconfirmed using Sanger sequencing. Kaplan Meier analysis was conducted to assess the effect of deleterious mutations on disease free and overall survival. Results: Seventy four of the 91 samples had been run earlier using the PCR-dHPLC and no deleterious mutations had been detected while 17 samples were tested for the first time. A total of 24 deleterious mutations were detected, 11 in BRCA1, 4 in BRCA2, 5 in p53, one each in RAD50, RAD52, ATM and TP53BP1. Some 19 deleterious mutations were seen in patients who had been tested earlier with PCR-dHPLC [19/74] and 5/17 in the samples tested for the first time, Together with our earlier detected 21 deleterious mutations in BRCA1 and BRCA2, we now had 45 mutations in 44 patients. BRCA1c.68_69delAG;p.Glu23ValfsX16 mutation was the most common, seen in 10/44 patients. Kaplan Meier survival analysis did not show any difference in disease free and overall survival in the patients with and without deleterious mutations. Conclusions: The NGS platform is more sensitive and cost effective in detecting mutations in genes involved in hereditary breast and/or ovarian cancers.

Evaluation of enteral formulas for nutrition, health, and quality of life among stroke patients

  • Kang, Yun-Kyeong;Lee, Ho-Sun;Paik, Nam-Jong;Kim, Woo-Sub;Yang, Mi-Hi
    • Nutrition Research and Practice
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    • v.4 no.5
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    • pp.393-399
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    • 2010
  • Enteral nutritional support has been used via tube feeding for dysphagic stroke patients. We performed long and short term trials to evaluate the effects of commercial enteral nutritional supports on nutrition and health in stroke patients (mRS = 3~5) and quality of life in their caregivers. For a long term study, we recruited chronic (${\geq}$ 1 yrs) stroke patients (n = 6) and administered them 6 cans/day (1,200 kcal) of the commercial enteral formula N for 6 months according to IRB-approved protocol. We collected peripheral blood at 0, 2, 4 and 6 months. For a short term study, we recruited acute (${\leq}$ 3 months) stroke patients (n = 12) and randomly administered them two different commercial enteral formulas, N or J, for 2 weeks. We collected their blood at 0, 4, 7 and 14 day of the administration. Blood samples were analyzed to quantify 19 health and nutritional biomarkers and an oxidative stress biomarker, malondialdehyde (MDA). In order to evaluate quality of life, we also obtained the sense of competence questionnaire (SCQ) from all caregivers at 'before' and 'after trials'. As results, the enteral formula, N, improved hemoglobin and hematocrit levels in the long term trial and maintained most of biomarkers within normal ranges. The SCQ levels of caregivers were improved in the long term treatment (P < 0.05). In a case of the short term study, both of enteral formulas were helpful to maintain nutritional status of the patients. In addition, MDA levels were decreased in the acute patients following formula consumption (0.05 < P < 0.1). Most of health and nutrition outcomes were not different, even though there is a big difference in price of the two products. Thus, we evaluate the formula N has equal nutritional efficacy compared to the formula J. In addition, long term use of enteral formula N can be useful to health and nutrition of stroke patients, and the quality of life for their caregivers.