• Title/Summary/Keyword: Detection,

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Incorporation of RAPD linkage Map Into RFLP Map in Glycine max (L, ) Merr (콩의 RAPD 연관지도를 RFLP 연관지도와 합병)

  • Choi, In-Soo;Kim, Yong-Chul
    • Journal of Life Science
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    • v.13 no.3
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    • pp.280-290
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    • 2003
  • The incorporation of RAPD markers into the previous classical and RFLP genetic linkage maps will facilitate the generation of a detailed genetic map by compensating for the lack of one type of marker in the region of interest. The objective of this paper was to present features we observed when we associated RAPD map from an intraspecific cross of a Glycine max$\times$G. max, 'Essex'$\times$PI 437654 with the public RFLP map developed from an interspecific cross of G. max$\times$G. soja. Among 27 linkage groups of RAPD map, eight linkage groups contained probe/enzyme combination RFLP markers, which allowed us the incorporation of RAPD markers into the public RFLP map. Map position rearrangement was observed. In incorporating L.G.C-3 into the public RFLP linkage group a1 and a2, both pSAC3 and pA136 region, and pA170/EcoRV and pB170/HindIII region were in opposite order, respectively. And, pk400 was localized 1.8 cM from pA96-1 and 8.4 cM from pB172 in the public RFLP map, but was localized 9.9 cM from i locus and 18.9 cM from pA85 in our study. A noticeable expansion of the map distances in the intraspecific cross of Essex and PI 437654 was also observed. Map distance between probes pA890 and pK493 in L.G.C-1 was 48.6 cM, but it was only 13.3 cM in the public RFLP map. The distances from the probe pB32-2 to pA670 and from pA670 to pA668 in L.G. C-2 were 50.9 cM and 31.7 cM, but they were 35.9 cM and 13.5 cM in the public RFLP map. The detection of duplicate loci from the same probe that were mapped on the same or/and different linkage group was another feature we observed.

Detection of Chemical Characteristics in Hamcho (Salicornia herbacea L.) according to Harvest Periods (함초(Salicornia herbacea L.)의 채취 시기별 이화학적 특성 탐색)

  • Cha, Jae-Young;Jeong, Jae-Jun;Kim, Yong-Taek;Seo, Won-Seok;Yang, Hyun-Ju;Kim, Jin-Sook;Lee, Yong-Soo
    • Journal of Life Science
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    • v.16 no.4
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    • pp.683-690
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    • 2006
  • The chemical compositions of amino acids, minerals, betaine, and polyphenolic compounds from Salicornia herbacea (Hamcho) according to harvest periods were analyzed. Changes of chemical characteristics in water and ethanol extracts from Hamcho were evaluated for titratable acidity, pH, soluble solid, and Hunter's color values. The antioxidative activity of water extract from Hamcho was also determined by DPPH $({\alpha},{\alpha}'-diphenyl-{\beta}-picrylhydrazyl)$ scavenging radical activity. Total polyphenolic compounds of Hamcho were shown the highest in August harvested by 201.6 ppm. The betaine of Hamcho water-extract was identified by high performance liquid chromatography (HPLC), which content was 0.248%, 0.269% and 0.204% in June, August, and October, respectively. Major compositional amino acids (mg%) were glutamic acid (582: 519: 664), proline (552: 471: 322), phenylalanine (480: 431: 424), aspartic acid (322: 297: 330), and arginine (282: 321: 483) in June, August, and October, respectively, and major free amino acids (mg%) were proline (9.7: 3.4), asparagine (6.7: 1.4), hydroxyproline (6.4: 2.8), valine (3.9: 2.5), arginine (1.7: 3.0) in June and August, respectively. Mineral contents (mg%) were Na (5,695: 7,536: 5,529), K (1,640: 963: 931), Mg (359: 428: 348), Ca (221: 234: 251), and P (207: 189: 259) in June, August, and October, respectively. Especially, K was high in June, Na and Mg were high in August, and Ca and P were high in October, respectively. DPPH scavenging radical activity was shown in the following order; 0.05% butylated hydroxytoluene (BHT)> August> June> October harvested Hamcho. The chemical components of polyphenolic compounds, betaine, amino acids and minerals were changed by harvest periods according to the growing season, and the highest concentrations of polyphenolic compounds and betaine of Hamcho were shown in August harvested.

Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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The NCAM Land-Atmosphere Modeling Package (LAMP) Version 1: Implementation and Evaluation (국가농림기상센터 지면대기모델링패키지(NCAM-LAMP) 버전 1: 구축 및 평가)

  • Lee, Seung-Jae;Song, Jiae;Kim, Yu-Jung
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.18 no.4
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    • pp.307-319
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    • 2016
  • A Land-Atmosphere Modeling Package (LAMP) for supporting agricultural and forest management was developed at the National Center for AgroMeteorology (NCAM). The package is comprised of two components; one is the Weather Research and Forecasting modeling system (WRF) coupled with Noah-Multiparameterization options (Noah-MP) Land Surface Model (LSM) and the other is an offline one-dimensional LSM. The objective of this paper is to briefly describe the two components of the NCAM-LAMP and to evaluate their initial performance. The coupled WRF/Noah-MP system is configured with a parent domain over East Asia and three nested domains with a finest horizontal grid size of 810 m. The innermost domain covers two Gwangneung deciduous and coniferous KoFlux sites (GDK and GCK). The model is integrated for about 8 days with the initial and boundary conditions taken from the National Centers for Environmental Prediction (NCEP) Final Analysis (FNL) data. The verification variables are 2-m air temperature, 10-m wind, 2-m humidity, and surface precipitation for the WRF/Noah-MP coupled system. Skill scores are calculated for each domain and two dynamic vegetation options using the difference between the observed data from the Korea Meteorological Administration (KMA) and the simulated data from the WRF/Noah-MP coupled system. The accuracy of precipitation simulation is examined using a contingency table that is made up of the Probability of Detection (POD) and the Equitable Threat Score (ETS). The standalone LSM simulation is conducted for one year with the original settings and is compared with the KoFlux site observation for net radiation, sensible heat flux, latent heat flux, and soil moisture variables. According to results, the innermost domain (810 m resolution) among all domains showed the minimum root mean square error for 2-m air temperature, 10-m wind, and 2-m humidity. Turning on the dynamic vegetation had a tendency of reducing 10-m wind simulation errors in all domains. The first nested domain (7,290 m resolution) showed the highest precipitation score, but showed little advantage compared with using the dynamic vegetation. On the other hand, the offline one-dimensional Noah-MP LSM simulation captured the site observed pattern and magnitude of radiative fluxes and soil moisture, and it left room for further improvement through supplementing the model input of leaf area index and finding a proper combination of model physics.

Inhibitory Effects of Ethanolic Extracts from Aster glehni on Xanthine Oxidase and Content Determination of Bioactive Components Using HPLC-UV (섬쑥부쟁이 에탄올 추출물의 잔틴산화효소 저해 효능 및 HPLC-UV를 이용한 유효성분의 함량 분석)

  • Kang, Dong Hyeon;Han, Eun Hye;Jin, Changbae;Kim, Hyoung Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1610-1616
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    • 2016
  • This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.

The Clinical Significance and Detection of Intraperitoneal Micrometastases by $ThinPrep^{(R)}$ Cytology with Peritoneal Lavage Fluid in Patients with Advanced Gastric Cancer (진행성 위암 환자에서 복강 내 미세전이 진단을 위한 복강 세척액 $ThinPrep^{(R)}$ 세포진 굄사의 임상적 의의)

  • Ryu, Chun-Kun;Park, Jong-Ik;Min, Jae-Seok;Jin, Sung-Ho;Park, Sun-Hoo;Bang, Ho-Yoon;Chae, Gi-Bong;Lee, Jong-Inn
    • Journal of Gastric Cancer
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    • v.8 no.4
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    • pp.189-197
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    • 2008
  • Purpose: Peritoneal lavage cytology is regarded as a useful diagnostic test for detecting intraperitoneal micrometastsis. However, there are currently no reports about cytological examination with $ThinPrep^{(R)}$ (CY), a newly introduced fluid-based diagnostic system, in patients with advanced gastric cancer (AGC). This study was performed to analyze the clinical significance of intraoperative peritoneal lavage for CY in AGC patients. Materials and Methods: 424 AGC patients were suspected to have serosal exposure macroscopically during surgery and they underwent intraoperative peritoneal lavage for CY between 2001 and 2006 at Korea Cancer Center Hospital. The clinical data, pathological data and CY results were collected and analyzed retrospectively. Results: The percentage of cytology positive results was 31.1%, and this was well correlated with the T-stage, N-stage and P-stage. The 3-year survival rates of CY0 and CY1 were 68.1% and 25.9%, respectively. According to the P-stage and CY, the 3-year survival rates were 71.1% in P0CY0, 38.9% in P0CY1, 38.5% in P1/2/3CY0 and 11.0% in P1/2/3CY1. Interestingly, both the P0CY1 and P1/2/3CY0 survival curves were similar figures, but they were significantly different from those of the other groups. Multivariate analysis indicated that CY was an independent, strong prognostic factor for survival, as well as sex, the T-stage, N-stage, P-stage, other metastasis and the serum CEA. CY1 was revealed as a risk factor for peritoneal recurrence in the curative resection group. Conclusion: The results certify indirectly that cytological examination using $ThinPrep^{(R)}$ is a very reliable diagnostic method for detecting intraperitoneal micrometastasis from the fact that it is not only a strong prognostic factor, but it is also a risk factor for peritoneal recurrence in AGC patients. Therefore intraoperative peritoneal lavage should be included in the routine intraoperative staging workup for AGC, and its result will provide a good target for the treatment of peritoneal micrometastasis.

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Effective Method for Extraction of Cell-Free DNA from Maternal Plasma for Non-Invasive First-Trimester Fetal Gender Determination: A Preliminary Study

  • Lim, Ji-Hyae;Park, So-Yeon;Kim, Shin-Young;Kim, Do-Jin;Kim, Mee-Jin;Yang, Jae-Hyug;Kim, Moon-Young;Kim, Min-Hyoung;Han, Ho-Won;Choi, Kyu-Hong;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.53-58
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    • 2010
  • Purpose: To find the most effective method for extraction of cell-free DNA (cf-DNA) from maternal plasma, we compared a blood DNA extraction system (blood kit) and a viral DNA extraction system (viral kit) for non-invasive first-trimester fetal gender determination. Materials and Methods: A prospective cohort study was conducted with maternal plasma collected from 44 women in the first-trimester of pregnancy. The cf-DNA was extracted from maternal plasma using a blood kit and a viral kit. Quantitative fluorescent-polymerase chain reaction (QF-PCR) was used to detect the SRY gene and AMEL gene. The diagnostic accuracy of the QF-PCR results was determined based on comparison with the final delivery records. Results: A total of 44 women were tested, but the final delivery record was only obtained in 36 cases which included 16 male-bearing and 20 female-bearing pregnancies. For the blood kit and viral kit, the diagnostic accuracies for fetal gender determination were 63.9% (23/36) and 97.2% (35/36), respectively. Conclusion: In non-invasive first-trimester fetal gender determination by QF-PCR, using a viral kit for extraction of cf-DNA may result in a higher diagnostic accuracy.

Primary Synchronous Lung Cancer Detected using Autofluorescence Bronchoscopy (자가 형광 기관지 내시경을 이용한 동시성 원발성 폐암의 진단)

  • Kwon, Sun Jung;Lee, Yun Seun;Joung, Mi Kyong;Lee, Yu Jin;Jang, Pil Soon;Lee, Jeung Eyun;Chung, Chae Uk;Park, Hee Sun;Jung, Sung Soo;Kim, Sun Young;Kim, Ju Ock
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.6
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    • pp.645-652
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    • 2006
  • Objective: Patients with lung cancer have a relative high risk of developing secondary primary lung cancers. This study examined the additional value of autofluorescence bronchoscopy (AFB) for diagnosing synchronous lung cancers and premalignant lesions. Methods: Patients diagnosed with lung cancer from January 2005 to December 2005 were enrolled in this study. The patients underwent a lung cancer evaluation, which included white light bronchoscopy (WLB), followed by AFB. In addition to the primary lesions, any abnormal or suspicious lesions detected during WLB and AFB were biopsied. Results: Seventy-six patients had non-small cell lung cancer (NSCLC) and 23 had small cell lung cancer (SCLC). In addition to the primary lesions, 84 endobronchial biopsies were performed in 46 patients. Five definite synchronous cancerous lesions were detected in three patients with initial unresectable NSCLC and in one with SCLC. The secondary malignant lesions found in two patients were considered metastatic because of the presence of mediastinal nodes or systemic involvement. One patient with an unresectable NSCLC, two with a resectable NSCLC, and one with SCLC had severe dysplasia. The detection rate for cancerous lesions by the clinician was 6.0% (6/99) including AFB compared with 3.0% (3/99) with WLB alone. The prevalence of definite synchronized cancer was 4.0% (4/99) after using AFB compared with 2.0% (2/99) before, and the staging-up effect was 1.0% (1/99) after AFB. Since the majority of patients were diagnosed with advanced disease, the subjects with newly detected cancerous lesions did not have their treatment plans altered, except for one patient with a stage-up IV NSCLC who did not undergo radiotherapy. Conclusions: Additional AFB is effective in detecting early secondary cancerous lesions and is a more precise tool in the staging workup of patients with primary lung cancer than with WLB alone.

Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

Influences of Rate of Artificial Insemination Following Estrus Induction in Dog (개에서 발정유도가 인공수정효율에 미치는 영향)

  • 이영락;강태영;최상용
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.61-68
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    • 2003
  • Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to improve reproductive efficiency of artificial insemination with fresh- and frozen-semen following estrus induction in dog. Fifty infertilie dogs (age 2~3 years) were selected fur the study and divided into three different estrus induction treatment groups. Group 1 : dogs (n=15) were given clomifene (0.1 mg/kg) orally f3r five days at 12 hr intervals. Croup 2: dogs (n=15) were given bromocriptine (50 $\mu$g/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Croup 3, n=20) when pro-estrus occurred. After being treated, the dogs were evaluated fur the rates of estrus induction and time interval lapses from treatment to beginning of the pro-estrus. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The ejaculated semen was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM; TES, 209 mM; citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 co above the surface of liquid nitrogen (L$N_2$) for 23 min. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, there was no significant differences in the pregnancy rate of dogs between group treated with a combination of GnRH and bromocriptine and group treated clomifene or bromocriptine only. However, frozen-thawed semen can be used successfully fur artificial insemination in dog.