• Title/Summary/Keyword: T/R probe

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Development of a real-time PCR method for detection and quantification of the parasitic protozoan Perkinsus olseni

  • Gajamange, Dinesh;Yoon, Jong-Man;Park, Kyung-Il
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.387-393
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    • 2011
  • The objective of this study was to develop a real-time PCR method for the rapid detection and quantification of the protozoan pathogen Perkinsus olseni using a TaqMan probe. For the standard, genomic DNA was extracted from $10^5$ in vitro-cultured P. olseni trophozoites, and then 10-fold serial dilutions to the level of a single cell were prepared. To test the reliability of the technique, triplicates of genomic DNA were extracted from $5{\times}10^4$ cells and 10-fold serial dilutions to the level of 5 cells were prepared. The standards and samples were analyzed in duplicate using an $Exicycler^{TM}$ 96 real-time quantitative thermal block. For quantification, the threshold cycle ($C_T$) values of samples were compared with those obtained from standard dilutions. There was a strong linear relationship between the $C_T$ value and the log concentration of cells in the standard ($r^2$ = 0.996). Detection of DNA at a concentration as low as the equivalent of a single cell showed that the assay was sensitive enough to detect a single cell of P. olseni. The estimated number of P. olseni cells was similar to the original cell concentrations, indicating the reliability of P. olseni quantification by real-time PCR. Accordingly, the designed primers and probe may be used for the rapid detection and quantification of P. olseni from clam tissue, environmental water, and sediment samples.

Cytogenetic and Molecular Genetic Studies on Duchenne Muscular Dystrophy (Duchenne Muscular Dystrophy에 관한 세포유전학 및 분자유전학적 연구)

  • Hong, Hea-Sook
    • Journal of Korean Biological Nursing Science
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    • v.7 no.1
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    • pp.29-46
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    • 2005
  • Purpose ; 본 연구는 X-염색체와 관련된 장애 중에서 가장 흔하고 심한 Duchenne Muscular Dystrophy(DMD)의 세포유전학 및 분자유전학적 특성을 설명하기 위해서 DMD에 영향을 받고 있는 두 가계의 13명을 대상으로 가계도 분석과 염색체 분석 및 DNA 분석을 하였다. Method ; DNA분석은 DNA probe을 이용한 Southern blotting method로써 RFLPs와 DMD유전자 부위의 exon소실 유무를 조사하여 아래와 같은 결과를 얻었다. Conclusion ; A 염색체 분석 : 말초혈액과 양수를 표본으로 High-Resolution GTG염색에서 A가계와 B가계의 염색체 분석에서 12명의 염색체는 정상 X-염색체였으나 B가계의 I-2(DMD여성)에서 46, x,-x,+t(2:x)(q 21.1 : p21.2)로 나타난다. B. DNA분석3 : 1) RFLPs의 분석 J66,XJ-1.1,754-11로써 B가계의 RELPs(Restriction Fragment Length Polymorphisms)에서 J66/Pst I은 1.7hb(E), 1.6kb(e)을 보여 주었고 XJ-1.1/Taq I은 3.6kb(F), 3.0kb(f), 754-11/EoR I은 4.2kb(G), 2.0kb(g)의 대립인자를 나타내었다. 이상의 결과를 바탕으로 영향을 받고 있는 남자 (II-2)의 haplotype는 보인자인 어머니의 한쪽 인자를 받았으며 어머니와 딸은 보인자이고 임산부의 태아는 남아였고 태아의 인자들은 그의 할아버지로부터 물려받아 DMD에 영향을 받지 않은 것으로 진단되었다. 2) DMD 유전자의 exon 소실에 대한 분석 cDNA probe 8과 cDNA probe 2b-3으로써 소실에 대한 진단은 영향을 받은 남자(II-2)는 cDNA probe 8에서 12, 7.3, 6.6, 4.2kb에 소실이 있고 cDNA 2b-3은 1.7kb에 소실에 나타났다.

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Myth of 61.8% rule and the practical application notices (접지저항 측정의 61.8%법칙 적용의 맹점과 현실적 접근 방법)

  • Lee, Sang-Mu;Cho, Pyung-Dong
    • 한국정보통신설비학회:학술대회논문집
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    • 2005.08a
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    • pp.259-262
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    • 2005
  • Ground resistance measurement is an elementary technique for the evaluation of grounding system. There are main environmental factors to consider for correct measurement but the problem is that it is practically most cases to measure ground resistance unable to know the factors. This paper presents a methodology toward true value of resistance in the unknown circumstances, utilizing the defined term 'variation rate' of potential difference curve appearing in the distance to a current probe as in the three point fall-of-potential method which comprises the characteristics of environmental factors. This methodology is a induced result from the previous demostrated studies.

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Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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A Design of a circularly polarized small UHF RFID antenna (소형 원형편파 UHF RFID 대역 융합형 안테나 설계)

  • Chae, Gyoo-Soo
    • Journal of the Korea Convergence Society
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    • v.6 no.1
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    • pp.109-114
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    • 2015
  • A circularly polarized small UHF RFID reader antenna is presented. The antenna is composed of four elements and printed on the plastic substrate(${\varepsilon}_r=2.2$, t=5mm). Each element is fed by a probe which is sequentially connected to the feed line. The feed line is manufactured on the FR-4 substrate(t=1.0mm, ${\varepsilon}_r=4.7$). The simulation results shows that the antenna can be achieved a return loss of 12dB, gain of 3.46dBic over the UHF band of 902-928MHz. According to our simulation results, two prototype antennas are manufactured and measured. The obtained antennas operate in UHF RFID bands and can be adapted for various portable applications. In addition, a parametric study is conducted to facilitate the design and optimization processes.

A STUDY ON THE EFFECT OF DENTIN ADHESIVE TO FLUORIDE RELEASE OF COMPOMER (상아질 결합제가 컴포머의 불소유리에 미치는 영향에 관한 연구)

  • Yoon, Yeo-Sang;Kim, Jong-Soo;Kwon, Soon-Won;Kim, Yong-Kee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.2
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    • pp.228-237
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    • 2001
  • It is well known that glass-ionomer cement and compomer can release fluoride which can inhibit the progression of dental caries. The purposes of this study were to evaluate whether the fluoride from fluoride-containing filling materials can penetrate the dentin bonding adhesive and the dentin bonding adhesive can increase the bond strength of resin-modified glass ionomer. The amount of fluoride release from resin-modified glass ionomer and compomer was measured during 28 days of period and statistically analyzed by t-test. The bond strength of each material with/without dentin bonding adhesive was measured and also statistically analyzed by t-test. The distribution of fluoride from each material into the tooth was explored by electron probe microanalysis system(EPMA). The experimental teeth used were second primary molars about to exfoliate. The amount of fluoride release from each material was diminished by dentin bonding adhesive during 28 days(p<0.05) and the bond strength was not increased by dentin adhesive in resin-modified glass ionomer. The bond strength of group VI was not detectable. The distribution of fluoride from each material into teeth was according to dentino-enamel junction and dispersed into pulp from pulpal floor. The widest distribution was shown in resin-modified glass ionomer cement filled without the application of dentin bonding adhesive.

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Milk Fat Analysis by Fiber-optic Spectroscopy

  • Ohtani, S.;Wang, T.;Nishimura, K.;Irie, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.580-583
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    • 2005
  • We have evaluated the application of spectroscopy using an insertion-type fiber-optic probe and a sensor at wavelengths from 400 to 1,100 nm to the measurement of milk fat content on dairy farms. The internal reflectance ratios of 183 milk samples were determined with a fiber-optic spectrophotometer at 5$^{\circ}C$, 20$^{\circ}C$ and 40$^{\circ}C$. Partial least squares (PLS) regression was used to develop calibration models for the milk fat. The best accuracy of determination was found for an equation that was obtained using smoothed internal reflectance data and three PLS factors at 20$^{\circ}C$. The correlation coefficients between predicted and reference milk fat at 5$^{\circ}C$, 20$^{\circ}C$ and 40$^{\circ}C$ were r=0.753, r=0.796 and r=0.783, respectively. The predictive explained variances ($Q^2$) of the final model, moreover, were more than 0.550 at all temperatures, and the regression coefficients of determination ($R^2$) were more than 0.6 (60%). Our results indicate that milk has different internal reflectance measured in the range of visible and near infrared wavelengths (400 to 1,100 nm), depending on its fat content.

VHL Gene Mutation Analysis of a Chinese Family with Non-Syndromic Pheochromocytomas and Patients with Apparently Sporadic Pheochromocytoma

  • Zhang, Bin;Qian, Jing;Chang, De-Hui;Wang, Yang-Min;Zhou, Da-Hai;Qiao, Gou-Mei
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.1977-1980
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    • 2015
  • Objective: The Von Hippel-Lindau syndrome (VHLD), an inherited neoplastic syndrome predisposing to central nervous system hemangioblastoma (CNS), pheochromocytoma (PCC), renal cell carcinoma(RCC), retinal hemangioma (RA) and renal cysts, is caused by mutations or deletions of the VHL tumor-suppressor gene. To assess VHL genotype-phenotype correlations with function of pVHL a gene mutation analysis of members in a Chinese family with non-syndromic PCCs and individuals with apparently sporadic pheochromocytoma (ASP) was performed. Materials and Methods: DNA samples of 20 members from the Chinese family with non-syndromic PCCs and 41 patients with ASP were analyzed by polymerase chain reaction and direct sequencing, confirmed by Taqman probe. Results: Three novel mutations (H125P, 623(^TTTGTtG) and R120T) were identified in the Chinese family and in 3 among 41 ASP patients. The mutations were all located in exon 2 of VHL gene encoding ${\beta}$-domain of pVHL. The tumor type in H125P carriers and R120T carriers was VHL type 2C. And 623(^TTTGTtG) carriers presented VHL type 2B or type 2C. Conclusions: VHL gene abnormalities were identified in the Chinese family with non-syndromic PCCs and patients with APS, resulting in dysfunction of pVHL. H125P and R120T could be associated with VHL type 2C, while 623(^TTTGTtG) might be linked with VHL type 2B or type 2C. Not only is the genetic analysis helpful for early diagnosis and treatment of patients with VHLD, it is also benefitial for research intoVHLD pathogenesis.

Relationship of Oral Bacterial Load Over One Year of Smoking Cessation

  • Kim, Sunghyun;Seo, Min-Seock;Hwang, Soo-Jeong
    • Journal of dental hygiene science
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    • v.19 no.4
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    • pp.213-219
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    • 2019
  • Background: Smoking exerts an adverse effect on the periodontal tissue by reorganizing the ecosystem of oral microorganisms and is considered to be an important factor in the development of periodontal disease. Although cross-sectional studies on smokers and non-smokers have been attempted to investigate the microbial differences in periodontal oral cavity, only few studies have been conducted to investigate the changes in oral microorganisms during smoking cessation. The purpose of this study was to investigate the changes of bacteria in saliva and gingival crevicular fluid (GCF) over a period of one year among 11 smokers trying to quit smoking. Methods: Eleven smokers trying to quit smoking visited the clinic at baseline, two weeks, two months, four months, six months, and 12 months to give saliva and GCF samples. The amounts of 16S rRNA, Porphyromonas gingivalis, Treponema denticola, Prevotella intermedia, Fusobacterium nucleatum subsp. nucleatum, Streptococcus mutans, and Streptococcus sobrinus in saliva and GCF were quantified using real-time polymerase chain reaction TaqMan probe assay. The results were analyzed by nonparametric statistical analysis using Friedman test and Spearman correlation coefficient. Results: After cessation of smoking, the amounts of 16S rRNA corresponding to P. gingivalis, F. nucleatum, P. intermedia, and T. denticola in saliva decreased and then again increased significantly. The amount of F. nucleatum 16S rRNA in GCF decreased significantly after smoking cessation. Positive correlations were observed between 16S rRNA and F. nucleatum and between F. nucleatum and T. denticola in saliva and GCF. Conclusion: Even if the number of subjects in this study was small, we suggest that smoking cessation may reduce the total bacterial amount and F. nucleatum in GCF. However, the results regarding changes in the microbial ecosystem due to smoking or smoking cessation were inconsistent. Therefore, further in-depth studies need to be carried out.

Compact 40 GHz Hairpin Band-Pass Filter (초소형 40 GHz Hairpin 대역통과 여파기)

  • Lee, Young Chul
    • Journal of Advanced Navigation Technology
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    • v.22 no.1
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    • pp.27-30
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    • 2018
  • In this study, a 40 GHz band pass filter(BPF) employing a hair-pin structure has been designed, fabricated, and characterized for millimeter-wave wireless communication applications. Using the 3 dimensional(3-D) electromagnetic(EM) tool and design equations of the hairpin BPF, the BPF was desgned on the 5 mil-thick Duroid substrate(RT5880) with a relative dielectric constant (${\varepsilon}_r$) of 2.2. The tapping point (t) of the U-shape resonator in the input and output port has been determined using extracted an external Q-factor ($Q_e$). The coupling coefficients between the other resonators are calculated by adjusting the physical dimensions for the desired response of the BPF. The fabricated BPF was characterized using probing method on a probe station. Its measured center frequency(fc) and fractional BW are 41.6 GHz and 7.43 %, respectively. The measured return loss is below -10 dB at the pass band and the insertion loss is 3.87 dB. The fabricated BPF is as small as $9.1{\times}2.8mm^2$.