• Title/Summary/Keyword: S1, S2 Detection

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Detection of Pectobacterium chrysanthemi Using Specific PCR Primers Designed from the 16S-23S rRNA Intergenic Spacer Region

  • Kwon, Soon-Wo;Myung, In-Sik;Go, Seung-Joo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.252-256
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    • 2000
  • The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

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An Enzyme-Linked Immunosorbent Assay for Detection of Milk proteins in Food (우유단백질의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Bae, Gun-Won;Kim, Soon-Mi
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.564-569
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    • 2000
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of milk proteins in processed foods. The ${\alpha}_{s1}-casein({\alpha}_{s1}-CN)$, a heat stable major milk protein, was immunized into rabbits to produce specific antibodies. When competitive indirect ELISA(ciELISA) using $anti-{\alpha}_{s1}-CN$ antibodies was established, its detection limit was $0.1\;{\mu}g/mL$. The reactivities of the specific antibodies toward ${\alpha}_{s1}-CN$, skim milk, ${\beta}-CN$ and whey protein isolate(WPI) were 100, 37, 0.14 and 0.04%, respectively, as determined by ciELISA. However $anti-{\alpha}_{s1}-CN$ antibodies did not have any reactivity to other milk proteins such as ${\beta}-lactoglobulin,\;{\alpha}-lactalbumin$, bovine serum albumin, and isolated soy protein. When sandwich ELISA was established, its detection limit was $0.01\;{\mu}g/mL$ which was 10 times more sensitive than that of ciELISA. In the spike test which was performed by adding 1-10% of whole CN to market milk, mean assay recovery as determined by sandwich ELISA was 94.8%(CV, 8.2%). Food stuffs and dairy products were assayed by sandwich ELISA to show 29, 0.13, 0.25, and 6.9% of whole CN in skim milk powder, WPI, semi-solid yoghurt, and processed cheese, respectively.

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Concurrent Hypermethylation of SFRP2 and DKK2 Activates the Wnt/β-Catenin Pathway and Is Associated with Poor Prognosis in Patients with Gastric Cancer

  • Wang, Hao;Duan, Xiang-Long;Qi, Xiao-Li;Meng, Lei;Xu, Yi-Song;Wu, Tong;Dai, Peng-Gao
    • Molecules and Cells
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    • v.40 no.1
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    • pp.45-53
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    • 2017
  • Aberrant hypermethylation of Wnt antagonists has been observed in gastric cancer. A number of studies have focused on the hypermethylation of a single Wnt antagonist and its role in regulating the activation of signaling. However, how the Wnt antagonists interacted to regulate the signaling pathway has not been reported. In the present study, we systematically investigated the methylation of some Wnt antagonist genes (SFRP2, SFRP4, SFRP5, DKK1, DKK2, and APC) and their regulatory role in carcinogenesis. We found that aberrant promoter methylation of SFRP2, SFRP4, DKK1, and DKK2 was significantly increased in gastric cancer. Moreover, concurrent hypermethylation of SFRP2 and DKK2 was observed in gastric cancer and this was significantly associated with increased expression of ${\beta}-catenin$, indicating that the joint inactivation of these two genes promoted the activation of the Wnt signaling pathway. Further analysis using a multivariate Cox proportional hazards model showed that DKK2 methylation was an independent prognostic factor for poor overall survival, and the predictive value was markedly enhanced when the combined methylation status of SFRP2 and DKK2 was considered. In addition, the methylation level of SFRP4 and DKK2 was correlated with the patient's age and tumor differentiation, respectively. In conclusion, epigenetic silencing of Wnt antagonists was associated with gastric carcinogenesis, and concurrent hypermethylation of SFRP2 and DKK2 could be a potential marker for a prognosis of poor overall survival.

Establishment of Quantitative Analysis Method for Genetically Modified Maize Using a Reference Plasmid and Novel Primers

  • Moon, Gi-Seong;Shin, Weon-Sun
    • Preventive Nutrition and Food Science
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    • v.17 no.4
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    • pp.274-279
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    • 2012
  • For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.

The Significance of Serum $Beta_2-Microglobulin$ Measurement in Various Renal Diseases (각종(各種) 신질환(腎疾患)에서의 혈청(血淸) $\beta_2-microglobulin$ 측정(測定)의 의의(意義))

  • Koong, Sung-Soo;Oh, Ha-Yong;Han, Jin-Suk;Lee, Jung-Sang
    • The Korean Journal of Nuclear Medicine
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    • v.19 no.1
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    • pp.127-136
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    • 1985
  • To evaluate change of serum $beta_2-microglobulin$ concentration$(s\beta_2-MG)$ and the usefulness of $s\beta_2-MG$ and $s\beta_2-MG/serum$ creatinine concentration(sCr) ratio in various renal diseases, $s\beta_2-MG$ and sCr were measured in 25 normal controls and 90 patients of various renal diseases(16 cases of glomerulonephritis, 12 cases of acute renal failure, 8 cases of chronic renal failure, 24 cases of nephrotic syndrome, 15 cases of tubulointerstitial diseases and 15 cases of lupus nephritis) using $Phadebas^\circledR$ $Beta_2-Micro$ Test kits. The results were as follows; 1) In normal control, the mean value of $s\beta_2-MG$ was $1.65{\pm}0.41mg/l$ and the mean value of $s\beta_2-MG/sCr$ ratio was $0.14{\pm}0.05$. 2) In various renal diseases, the mean value of $s\beta_2-MG$ was $6.74{\pm}5.47mg/l$. The mean value of $s\beta_2-MG/sCr$ ratio was $0.24{\pm}0.11$ and significantly elevated than that of normal control. (p<0.05) 3) The correlation between $s\beta_2-MG$ and sCr in glomerular and tubulointerstitial disease was log $s\beta_2-MG-0.90$ log sCr-0.48 and its correlation coefficient was 0.78(p<0.05). 4) In glomerular disease, the correlation between $s\beta_2-MG$ and sCr was log $s\beta_2-MG-0.89$ log sCr-0.46(r - 0.76) and in tubulointerstitial disease, it was log, $s\beta2-MG-0.95$ log sCr-0.59 (r-0.87). There was no significant difference between the two groups(p<0.05). 5) Among 32 cases of glomerular and tubulointerstitial disease patients, whose sCr was within normal range, 17 cases showed elevated $s\beta_2-MG$. The mean values of $s\beta_2-MG/sCr$ ratio in these patients was $0.30{\pm}0.14$ and significantly elevated than that of normal control(p<0.05). 6) In 15 cases of lupus nephritis, 12 cases showed elevated $s\beta_2-MG$ with normal sCr and 12 cases showed elevated $s\beta_2-MG/sCr$ ratio. With above results, it was found that the $s\beta_2MG$ can be used as an index of glomerular filtration rate as in the case of sCr and that $s\beta_2-MG/sCr$ ratio can be used as a tool in early detection of slightly decreased glomerular filtration rate and in detection of the renal disease of increased $\beta_2-MG$ production.

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A Study on the Characteristics of an Optical Sensor Linear Fire Detection System with Miniature Model Fire Experiment (축소 모형실험을 통한 광센서 선형 화재 감지 시스템의 특성에 관한 연구)

  • Kim, Dong-Eun;Kim, Si-Kuk;Lee, Young-Sin;Lee, Chun-Ha;Lim, Woo-Sup
    • Fire Science and Engineering
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    • v.30 no.2
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    • pp.19-26
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    • 2016
  • In this study, we conducted a low temperature operating test and miniature tunnel model test to study the fire detection capability and properties of an early fire detection system using an optical sensor linear detector that can be installed in harsh environments such as tunnel or utility-pipe conduits which are becoming the major and national infrastructure facilities. The test showed that the optical sensor linear detector was the only one functioned properly among five thermal detectors installed at a low temperature of $-20^{\circ}C$ for 5 days. To study were analyzed adaptability of optical sensor linear detector in the windy tunnel, the operating properties of the optical sensor linear detector when the wind velocity was varied between 0 m/s and 1 m/s in a miniature tunnel model. The temperature change was high when the wind velocity was 0 m/s.

Downscaling Forgery Detection using Pixel Value's Gradients of Digital Image (디지털 영상 픽셀값의 경사도를 이용한 Downscaling Forgery 검출)

  • RHEE, Kang Hyeon
    • Journal of the Institute of Electronics and Information Engineers
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    • v.53 no.2
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    • pp.47-52
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    • 2016
  • The used digital images in the smart device and small displayer has been a downscaled image. In this paper, the detection of the downscaling image forgery is proposed using the feature vector according to the pixel value's gradients. In the proposed algorithm, AR (Autoregressive) coefficients are computed from pixel value's gradients of the image. These coefficients as the feature vectors are used in the learning of a SVM (Support Vector Machine) classification for the downscaling image forgery detector. On the performance of the proposed algorithm, it is excellent at the downscaling 90% image forgery compare to MFR (Median Filter Residual) scheme that had the same 10-Dim. feature vectors and 686-Dim. SPAM (Subtractive Pixel Adjacency Matrix) scheme. In averaging filtering ($3{\times}3$) and median filtering ($3{\times}3$) images, it has a higher detection ratio. Especially, the measured performances of all items in averaging and median filtering ($3{\times}3$), AUC (Area Under Curve) by the sensitivity and 1-specificity is approached to 1. Thus, it is confirmed that the grade evaluation of the proposed algorithm is 'Excellent (A)'.

Clinical Significance of Joint Detection of Serum VEGF, SIL-2R and HGF in Patients with Primary Hepatocellular Carcinoma before and after Percutaneous Microwave Coagulation Therapy

  • Chen, Ji-Dong;Xiong, Yan-Qun;Dong, Ke;Luo, Jun;Yue, Lin-Xian;Chen, Qin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.11
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    • pp.4545-4548
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    • 2014
  • Objective: To investigate the changes of serum vascular endothelial growth factor (VEGF), soluble interleukin-2 receptor (SIL-2R) and hepatocyte growth factor (HGF) contents in patients with primary hepatocellular carcinoma (HCC) before and after percutaneous microwave coagulation therapy (PMCT) and determine their clinical significance. Materials and Methods: Fasting venous blood (3 mL) from 81 patients with primary HCC diagnosed by pathology was collected in the mornings 1 day before PMCT, and 1 day, 7 days and 1 month after PMCT, and then the serum was separated and stored in $-70^{\circ}C$. The contents of VEGF, SIL-2R and HGF were detected by enzyme linked immunosorbent assay (ELISA). Results: The serum VEGF, SIL-2R and HGF contents in 81 patients with primary HCC had obviously dynamic changes before and after PMCT. By comparison to 1 day after PMCT with pre-operation, there was no statistical significance regarding VEGF and SIL-2R contents (P>0.05), but HGF content showed significant difference (P<0.01). Compared with pre-operation, VEGF, SIL-2R and HGF contents 7 days and 1 month after PMCT all manifested significant differences (P<0.01). By comparison to 7 days with 1 month after PMCT, there was no statistical significance regarding the VEGF content (P>0.05), whereas SIL-2R and HGF contents showed significant change (P<0.01). Conclusions: The contents of serum VEGF, SIL-2R and HGF have obviously dynamic changes in primary HCC before and after PMCT, and their joint detection is expected to be an effective hematologic evaluation index of PMCT for primary HCC.

A Ground Penetrating Radar Detection of Buried Cavities and Pipes and Development of an Image Processing Program (지반 공동 및 매립관의 지반 투과 레이더 탐사 및 이미지 처리 프로그램 개발)

  • Lee, Hyun-Ho
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.5 no.2
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    • pp.177-184
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    • 2017
  • Many ground subsidence accidents have happened in Korea. The accident was caused by the subsidence and leakage of the deteriorated sewage pipe. This study aims to establish the empirical data of the ground penetration radar(GPR) detection for ground subsidence. A test bed was also manufactured for the same purpose. The GPR detection variables are embedment depth and horizontal distance of embedded cast iron pipe and expanded polystyrene(EPS). From the detection results, the EPS embedded by a depth of 1.5m was difficult for detection. The EPS closely embedded to the cast iron pipe within a 0.5m distance had a very strong cast iron pipe signal. Therefore, the detection was impossible. This study developed an image processing program, called the GPR image processing program(GPRiPP), to process the GPR detection results. Its major function is the gain function, which amplifies the wiggle wave signal. Compared to the existing programs, the GPRiPP is capable of showing a similar image processing performance.

Detection of Colletotrichum acutatum and C. gloeosporioides by Real Time PCR (Real Time PCR을 이용한 Colletotrichum acutatum과 C. gloeosporioides의 검출)

  • Kim, Seung-Han;Kwon, Oh-Hun
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.219-222
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    • 2008
  • Real time PCR was used to discriminate Colletotrichum acutatum and C. gloeosporioides for analysis of population density. Two primers, caInt2 and cgint, used for conventional PCR to discriminate two species were modified with fluorescent dye to make probe for real time PCR. Fluorescence signals were successfully detected by fCaInt2 and vCgint probe coupled with primer pair Unicon and Unicor1 resulting in discrimination of C. acutatum and C. gloeosporioides by comparison of delta Rn value.