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PCR in diagnosis of pneumocystosis of rats (중합반응을 이용한 흰쥐 페포자충증의 진단)

  • 홍성태
    • Parasites, Hosts and Diseases
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    • v.34 no.3
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    • pp.191-196
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    • 1996
  • Polymerase chain reaction (PCR) is a powerful technique to detect scanty amount of DNA from living organisms. The present study intended to develope specific primers for PCR diagnosis of pneumocystosis and to evaluate diagnostic efficacy by preparation of template DNAs from invasive BAk fluid and also to screen serum or blood as a non-invasive specimen. Albino rats of Wistar or Fischer strains were experimentally infected by Pneumocwstis ccrinii. Extracted DNAs or cell Iysates of their blood, bronchoalveolar lavage fluid, and lung homogenate were used as the tenlplate DNA. Primers were synthetic oligonucleotides among 16s rDNA sequences. All of the primer combinations gave PCR products, but the primer pair of #24 and #27 gave best quality product of 666 bp. The sensitivity of PCR with Iysates of BAk fluid was 57.7% but it increased to 84.6% with extracted DNAs. None of BAL Iysate or DNA was positive among 13 microscopically negatives. The serum DNAs were positive only in 2 cases out of 20 morphologically positive rats. DNAs of human, rat, other parasites, yeast, and microorganisms were negative. The findings suggest that the present primers are specific but simple Iysate of BAL fluid is not sensitive. PCR may be used as a routine diagnostic method of pneumocystosis if simple and rapid preparation of non-invasive clinical specimens are available.

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Evaluation of the Reliability of Salivary Testing Instrument (타액검사기의 신뢰도 평가)

  • Han, Keumah;Lee, Joonhaeng
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.3
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    • pp.363-369
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    • 2018
  • A salivary testing instrument has an advantage that the method is simple and can be performed in a short time. However, it is necessary to verify the factors that affect the reliability of the result, because the device is easy to use and even saliva collection is simple. The aim of this study was to compare the difference of the test results according to the measurement time in order to analyze the time factor of the external variable among the factors that may affect the measurement results of the salivary testing instrument. The relationship between the measured values of the salivary testing instrument to identify the internal variables was analyzed. Saliva was collected from 20 randomly selected patients regardless of age, sex, or diseases. The mean age was 46.6 years, 10 males and 10 females. The saliva collected was directly measured with the salivary testing instrument as group I. The saliva samples were placed in air in a paper cup for 10 minutes, and then measured as group III. Then group I was remeasured after 30 minutes and assigned as group II. Group III was remeasured after 30 minutes and called as group IV. As a result, all of the cariogenic bacteria, acidity, buffer capacity, blood, leukocyte, protein and ammonia, except buffer capacity, showed statistically significant changes in group II and IV. This means that the reliability of the test results is poor if the measurement time is not observed. Cariogenic bacteria were correlated with leukocyte and protein, buffer capacity was related to acidity, protein, and protein was related to buffer capacity and leukocyte. In conclusion, the result according to the measurement time as the external variable was different, which means that time must be strictly monitored when testing saliva. It is also necessary to take into account the relevance of the correlations between the internal variables and the clinical data.

Numerical Analysis for the Conjugate Heat Transfer of Skin Under Various Temperature Conditions of Contrast Therapy (냉온 자극의 다양한 온도경계조건들에 대한 피부 내 온도 분포의 수치해석)

  • Park, Da Ae;Oh, Han Nah;Jeon, Byoung Jin;Kim, Eun Jeong;Lee, Seung Deok;Choi, Hyoung Gwon
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.39 no.11
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    • pp.897-903
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    • 2015
  • In this paper, the contrast therapy of skin was numerically investigated by solving the conjugate heat transfer problem. A finite volume method based on the SIMPLE algorithm was adopted to solve the axisymmetric incompressible Navier-Stokes equations, coupled with an energy equation. These equations are strongly coupled with the Pennes bio-heat equation in order to consider the effect of blood perfusion rate. We investigated the thermal response of skin at some selected depths for various input temperature profiles of a stimulator for contrast therapy. From the numerical simulations, the regions with cold/hot threshold temperatures were found for five input temperature profiles. It was shown that the temperature varies mildly for different input profiles as the depth increases, owing to the Pennes effect. The input temperatures for effective hot/cold stimulation of dermis layer were found to be $47^{\circ}C$ and $7^{\circ}C$, respectively. The present numerical results will be used for finding an optimal temperature profile of a stimulator for contrast therapy.

Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR

  • Thanchomnang, Tongjit;Intapan, Pewpan M.;Tantrawatpan, Chairat;Lulitanond, Viraphong;Chungpivat, Sudchit;Taweethavonsawat, Piyanan;Kaewkong, Worasak;Sanpool, Oranuch;Janwan, Penchom;Choochote, Wej;Maleewong, Wanchai
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.645-650
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    • 2013
  • A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at $81.5{\pm}0.2^{\circ}C$, $79.0{\pm}0.3^{\circ}C$, $76.8{\pm}0.1^{\circ}C$, and $79.9{\pm}0.1^{\circ}C$, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.

The Character Factor of Myopia by the Living Styles (생활 환경에서 오는 후천적 근시의 성격요인)

  • Kim, Jeong-Sik;Kang, Hywang-Yoo
    • Journal of Korean Ophthalmic Optics Society
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    • v.4 no.2
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    • pp.129-133
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    • 1999
  • For the myopia eyes of the people which are ametropia, the classified distribution has showed % for the simple myopic, 50% for the compound myopic astigmatism, 15% for the simple myopic astigmatism and 19% for the mixed astigmatism. The myopic ametropia for the both eyes has the distribution of 35% for -0.50D~-2.00Dptr, 54% for -2.00~6.00Dptr, and 11% for over -6.00Dptr. The classifying distribution for the age for the myopic ametropia was 54% for 15~20, 22% for 21~40, 14% for 41~60 and 10% for 61. The occupational distribution for the myopic ametropia has showed 61.5% for the student(Middle, High, College), 13.5% for the office worker, and 15% for the house wives as well as the small business. The hour affecting the refraction most for a day was after P.M. 7 which recorded 45% as the highest value. The reason is that the myopia degree decreases in the morning as the cornea flats and the situation is reversed in the afternoon so that there is a difference of Sph -0.50D and as getting darker the refraction degree of the light coming through the enlarged pupil refraction around the cornea is high. For the seasons the highest myopic degree has been recorded for 68% in the summer due to the shortage of nutrition and the climination inside the body by the exhaustion of sweat. In the blood types A and B are distributed closely as 34% and more active man with O has recorded higher myopic degree than woman. However woman showed higher accommodation power than man regardless the blood types. In the characteristic factors of myopic eye, the character feels fatigue easily has showed the distribution for 42% which is the highest and it could be classified largely by two the.

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Association between Low-level Mercury Exposure and Neurobehavioral Functions in Korean Adults Living in a Coastal City

  • Kim, Rock Bum;Kim, Byoung-Gwon;Kim, Yu-Mi;Hong, Young-Seoub;You, Chang-Hun;Kim, Dae-Seon
    • Environmental Analysis Health and Toxicology
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    • v.28
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    • pp.15.1-15.8
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    • 2013
  • Objectives The aim of this study was to determine the association between low-level mercury exposure and neurobehavioral functions in adults living in coastal regions of Korea. Methods We selected 172 adults aged 20-65 years living in a city in the coastal region of Korea. A sociodemographic survey was conducted, mercury levels in the blood, urine, and hair were measured, and the associations according to computerized neurobehavioral tests were determined using univariate analysis. After adjustment for associated variables, a multivariate linear regression analysis was performed. Results The geometric mean mercury levels in the blood, urine, and hair were $5.41{\mu}g/L$ (range, $0.00-15.84{\mu}g/L$), $1.17{\mu}g/g$-creatinine (range, $0.00-32.86{\mu}g/g$-creatinine), and 1.37 mg/kg (range, 0.42-6.56 mg/kg), respectively. Variables that were associated with simple reaction time according to the neurobehavioral test results were age and urine mercury level. Variables associated with choice reaction time were the recent use of Korean traditional medicine and urine mercury level. Variables associated with the right-hand finger tapping speed test were age, gender, smoking behavior, education level, monthly household income, and urine mercury level. Variables associated with the left-hand finger tapping speed test were age, gender, education level, and urine mercury level. After adjustment for associated variables, there was no significant association between urine mercury level and simple reaction time (${\beta}=25.96$; p =0.47), choice reaction time (${\beta}=50.37$; p =0.32), or the number of left-hand finger taps (${\beta}=-1.54$; p =0.21). However, urine mercury level was significantly associated with the number of right-hand finger taps (${\beta}=-3.86$; p =0.01). Conclusions We found no evidence that low-level mercury exposure in adults is associated with deficits in neurobehavioral functions. A longer follow-up study is required to confirm this conclusion.

Comparison of the Formula of PSA, Age, Prostate Volume and Race Versus PSA Density and the Detection of Primary Malignant Circulating Prostate Cells in Predicting a Positive Initial Prostate Biopsy in Chilean Men with Suspicion of Prostate Cancer

  • Murray, Nigel P;Reyes, Eduardo;Fuentealba, Cynthia;Orellana, Nelson;Morales, Francisca;Jacob, Omar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.13
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    • pp.5365-5370
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    • 2015
  • Background: Combining risk factors for prostate cancer into a predictive tool may improve the detection of prostate cancer while decreasing the number of benign biopsies. We compare one such tool, age multiplied by prostate volume divided by total serum PSA (PSA-AV) with PSA density and detection of primary malignant circulating prostate cells (CPCs) in a Chilean prostate cancer screening program. The objectives were not only to determine the predictive values of each, but to determine the number of clinically significant cancers that would have been detected or missed. Materials and Methods: A prospective study was conducted of all men undergoing 12 core ultrasound guided prostate biopsy for suspicion of cancer attending the Hospital DIPRECA and Hospital de Carabineros de Chile. Total serum PSA was registered, prostate volumecalculated at the moment of biopsy, and an 8ml blood simple taken immediately before the biopsy procedure. Mononuclear cells were obtained from the blood simple using differential gel centrifugation and CPCs identified using immunocytchemistry with anti-PSA and anti-P504S. Biopsy results were classed as positive or negative for cancer and if positive the Gleason score, number of positive cores and percent infiltration recorded. Results: A total of 664 men participated, of whom 234 (35.2%) had cancer detected. They were older, had higher mean PSA, PSA density and lower PSA-AV. Detection of CPCs had high predictive score, sensitivity, sensibility and positive and negative predictive values, PSA-AV was not significantly different from PSA density in this population. The use of CPC detection avoided more biopsies and missed fewer significant cancers.Conclusions: In this screening population the use of CPC detection predicted the presence of clinically significant prostate cancer better than the other parameters. The high negative predictive value would allow men CPC negative to avoid biopsy but remain in follow up. The formula PSA-AV did not add to the predictive performance using PSA density.

Isolation of Monocytes with High Purity and Yield from Peripheral Blood Mononuclear Cells by Flotation Density Gradient Centrifugation (부유밀도구배 원심분리를 이용하여 말초혈액단핵구로부터 고순도 및 고수율의 단세포 분리방법)

  • Bae, Jae-Ho;Son, Cheol-Hun;Park, You-Soo;Kim, Dong-Wan;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.19 no.6
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    • pp.728-734
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    • 2009
  • In this work, a simple, inexpensive and reproducible technique of flotation density gradient centrifugation was developed to isolate monocytes with high purity and yield from peripheral blood mononuclear cells (PBMC) using Histopaque solution, density and osmolarity of which were modified to 1.072 g/ml and 335 mOsm with phosphate-buffered saline (PBS) and sodium chloride (NaCl) solution, respectively. The average purity of monocytes was 74.75${\pm}$3.84%, with the individual purity ranging from 71.44% to 82.38%. The average yield of monocytes was 32.62${\pm}$11.16%, with the individual yields ranging from 21.02 to 53.63%. The monocytes isolated by floatation density gradient centrifugation could be successfully cultured into morphologically, phenotypically and functionally dendritic cells in vitro. In conclusion, the entire procedure seemed to be faster and more convenient, simple and cost-effective than other monocyte isolation methods, including plastic adherence and density gradient methods, and has the potential to be developed as a closed system for clinical scale generation of dendritic cells.

Studies on the Rapid Plasma Reagin(RPR) Card Test for the Diagnosis of Syphilis (매독진단(梅毒診斷)을 위(爲)한 신속(迅速)한 혈장항체검사법(血漿抗體檢査法)에 관(關)한 연구(硏究))

  • Kim, Joo-Deuk;Lew, Joon;Kim, Hyun-Joo
    • The Journal of the Korean Society for Microbiology
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    • v.3 no.1
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    • pp.15-23
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    • 1968
  • For the effective control of Syphilis, many investigators have developed a more rapid, simple and economical screening serological test which is adequately sensitive and specific. To fulfill the requirements of a more rapid serologic test for syphlis, a substitute for the conventional serum specimen was needed since considerable time and labor are involved in the processing of blood to serum. Burdon suggested the use of plasma in the serologic tests for syphilis as a substitute for serum. He noticed that plasma was more sensitive than serum in the Kline and Kahn tests, and attributed this to the presence of more antibody-like substance, "reagin" in plasma than in serum. However, to make plasma sufficiently sensitive, it was necessary to inactivate plasma by heating at a temperature of $56^{\circ}C$ for about 30 minutes. Heating of plasma resulted in the precipitation of fibrinogen which made centrifugation necessary to obtain dear plasma. Since the chief disadvantage to the use of unheated plasma(or serum) was a reduction in sensitivity of results-which probably was due to a labile factor such as complement-Portnoy et al began to consider rapid chemical methods of inactivation of plasma(or serum). They experienced that choline chloirde was shown to be anticomplementary which suggested its use as an inactivating agent for unheated plasma(or serum). In 1959 Portnoy et al reported the Rapid Plasma Reagin(RPR) Test for syphilis which is a more rapid, economical and simple. But still this test has many disadvantages as a rapid performing, field and office procedure, because it requires the usual laboratory equipments such as centrifuge, rotating machine, microscope etc. To substitute these disadvantages of the RPR test, in 1962, Portnoy et al developed the Rapid Plasma Reagin(RPR) card test for syphilis, which has the following advantages: a) Simplicity and rapidity of performance, b) Requires no laboratory equipments, c) Stable antigen suspension, d) Adequate sensitivity and specificity. This RPR card test can be used as a rapidly performing and screening test in field investigation, outpatient clinics, small laboratories and hospitals doing limited syphilis serology, and predonor in blood bank. Private clinic which has limited laboratory equipment and technic for syphilis serology can also use this RPR card test as a tool in the rapid diagnosis of syphilis. It was thought that this RPR card test is a useful tool in Korea for private physician and mass survey for syphilis diagnosis. But Portnoy patented the reagents needed for the performing the RPR card test. Therefore authors developed newly the reagents and according to Portnoy's method evaluated the newly developed. RPR card test compared with the VDRL, Kolmer CF, and RPCF tests. The RPR card and VDRL tests were performed plasma and serum from the total 1,132 cases. Among these 1,131 cases, 521 were syphilis suspected laboratory specimens, and 611 were syphilis unsuspected healthy young men. After screening with these two tests, the RPR card and VDRL tests, reactive specimens to the above one or both tests were retested by the Kolmer CF and RPCF tests.

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Genome Survey and Microsatellite Marker Selection of Tegillarca granosa (꼬막(Tegillarca granosa)의 유전적 다양성 분석을 위한 드래프트 게놈분석과 마이크로새틀라이트 마커 발굴)

  • Kim, Jinmu;Lee, Seung Jae;Jo, Euna;Choi, Eunkyung;Kim, Hyeon Jin;Lee, Jung Sick;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.1
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    • pp.38-46
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    • 2021
  • The blood clam, Tegillarca granosa, is economically important in marine bivalve and is used in fisheries industry among western Pacific Ocean Coasts especially in Korea, China, and Japan. The number of chromosomes in the blood clam is known as 2n=38, but the genome size and genetic information of the genome are not still clear. In order to predict the genomic size of the T. granosa, the in-silico analysis analysed the genomic size using short DNA sequence information obtained using the NGS Illumina HiSeq platform. As a result, the genomic size of T. granosa was estimated to be 770.61 Mb. Subsequently, a draft genome assembly was performed through the MaSuRCA assembler, and a simple sequence repeat (SSR) analysis was done by using the QDD pipeline. 43,944 SSRs were detected from the genome of T. granosa and 69.51% di-nucleotide, 16.68% trinucleotide, 12.96% tetra-nucleotide, 0.82% penta-nucleotide, and 0.03% hexa-nucleotide were consisted. 100 primer sets that could be used for genetic diversity studies were selected. In the future, this study will help identify the genetic diversity of T. granosa and population genetic studies, and further identify the classification of origin between homogenous groups.