• Title/Summary/Keyword: ELISA method

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Comparison of Analysis Methods for Detection of Replication Competent Virus and Functional Titers of HIV-l Based Lentivirus Vector (HIV-l 유래 렌티바이러스 벡터의 복제가능 바이러스 검출과 역가측정 분석방법 비교)

  • Chang Seok Kee;Oh Il Ung;Jeong Jayoung;Ahn Kwang Soo;Sohn Yeowon
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.217-224
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    • 2005
  • Human Immunodeficiency Virus type 1 (HIV-l) based lentivirus vector has demonstrated great potential as gene therapy vectors mediating efficient gene delivery and long-term transgene expression in both dividing and nondividing cells. However, for clinical studies it must be confirmed that vector preparations are safe and not contaminated by replication competent lentivirus (RCL) related to the parental pathogenic virus, HIV-l. In this study, we would like to establish the method for titration and RCL detection of lentivirus vector. The titration was determined by vector expression containing the green fluorescent protein, GFP in transduced cells. The titer was $1{\times}10^7$ Transducing Unit/ml in the GFP expression assay and $8.9{\times}10^7$ molecules/ml in the real-time PCR. Also, for the detection of RCL, we have used a combination method of PCR and p24 antigen detection. First, PBS/psi and VSV-G region in the genomic DNA of transduced cells was detected by PCR assay. Second, transfer and expression of the HIV-1 gag gene was detected by p24 ELISA. In an attempt to amplify any RCL, the transduced cells were cultured for 3 weeks (amplification phase) and the supernatant of amplified transduced cell was used for the second transduction to determine whether a true RCL was present (indicator phase). Analysis of cells and supernatant at day 6 in indicator phase were negative for PBS/psi, VSV-G, and p24 antigen. These results suggest that they are not mobilized and therefore there are no RCL in amplification phase. Thus, real-time PCR is a reliable and sensitive method for titration and RCL detection of lentivirus vector.

Detection of E.coli biofilms with hyperspectral imaging and machine learning techniques

  • Lee, Ahyeong;Seo, Youngwook;Lim, Jongguk;Park, Saetbyeol;Yoo, Jinyoung;Kim, Balgeum;Kim, Giyoung
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.645-655
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    • 2020
  • Bacteria are a very common cause of food poisoning. Moreover, bacteria form biofilms to protect themselves from harsh environments. Conventional detection methods for foodborne bacterial pathogens including the plate count method, enzyme-linked immunosorbent assays (ELISA), and polymerase chain reaction (PCR) assays require a lot of time and effort. Hyperspectral imaging has been used for food safety because of its non-destructive and real-time detection capability. This study assessed the feasibility of using hyperspectral imaging and machine learning techniques to detect biofilms formed by Escherichia coli. E. coli was cultured on a high-density polyethylene (HDPE) coupon, which is a main material of food processing facilities. Hyperspectral fluorescence images were acquired from 420 to 730 nm and analyzed by a single wavelength method and machine learning techniques to determine whether an E. coli culture was present. The prediction accuracy of a biofilm by the single wavelength method was 84.69%. The prediction accuracy by the machine learning techniques were 87.49, 91.16, 86.61, and 86.80% for decision tree (DT), k-nearest neighbor (k-NN), linear discriminant analysis (LDA), and partial least squares-discriminant analysis (PLS-DA), respectively. This result shows the possibility of using machine learning techniques, especially the k-NN model, to effectively detect bacterial pathogens and confirm food poisoning through hyperspectral images.

The Effects of Polygonum Cuspidatum on the Skin Functions (호장근추출물이 피부질환 및 피부미용에 미치는 영향)

  • Jung, Hyun-A;Roh, Seok-Seon;Oh, Min-Suck
    • Journal of Korean Medicine Rehabilitation
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    • v.19 no.1
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    • pp.73-89
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    • 2009
  • Objectives : This study was carried out to investigate the effects of Polygonum cuspidatum extract on several skin functions including inflammation and wrinkle formation. Methods : To investigate in vitro anti-oxidant activity assay, ethanol extracts of medicinal plants tested by DPPH method. In the next experiment, to investigate anti-inflammatory test, the RAW 264.7 macrophage cells was cultured using DMEM including the 10% FBS. To study anti-allergic effect, we blended cultured Human Mast Cells(HMC-1), and then observe $TNF-{\alpha}$, IL-8 by ELISA Results : Polygonum Cuspidatum extract has the effects of anti-inflammation and anti-allergy, which may be due to its inhibitory potential on the macrophage activation. Furthermore, Polygonum Cuspidatum extract has the anti-wrinkle effects through the inhibitory potential on the collagnease, elastase and gelatinase activities. Conclusions : The above results suggest that Polygonum Cuspidatum extract could be applicable for improvement of several skin functions.

Elevated Level of Prostate Specific Antigen Among Prostate Cancer Patients and High Prevalence in the Gangetic Zone of Bihar, India

  • Nath, A.;Singh, J.K.;Vendan, S. Ezhil;Priyanka, Priyanka;Sinha, Shreya
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.1
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    • pp.221-223
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    • 2012
  • Prostate cancer (CaP) is a common reproductive cancer among men. This study was conducted to correlate the cancer incidence with Gangetic zone and to correlate the tumor marker prostate specific antigen (PSA) level in serum with different age groups and stage of malignancy. Patients suffering from CaP in the pathology unit of Mahavir Cancer Sansthan (Hospital and Research Centre), Patna, Bihar, India were studied from June 2009 to May 2010. PSA level in the serum of CaP patients was estimated by ELISA method. CaP incidence was highly recorded in Gangetic zone than the non-Gangetic zone. Maximum patients were in the 56 - 75 years age group with a marked predominance. Results of PSA examination showed that serum PSA level was not correlating with the age of patient and stage of malignancy. Significantly, elevated level of more than 10 ng/ml of PSA was recorded among the studied cancer patients. In this study, it is concluded that Gangetic zone habitat have high risk of CaP and elevated level of PSA was marked in Bihar, India.

Leptin potentiates Prevotella intermedia lipopolysaccharide-induced production of TNF-$\alpha$ in monocyte-derived macrophages

  • Kim, Sung-Jo
    • Journal of Periodontal and Implant Science
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    • v.40 no.3
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    • pp.119-124
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    • 2010
  • Purpose: In addition to regulating body weight, leptin is also recognized for its role in the regulation of immune function and inflammation. The purpose of this study was to investigate the effect of leptin on Prevotella (P.) intermedia lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-$\alpha$ production in differentiated THP-1 cells, a human monocytic cell line. Methods: LPS from P. intermedia ATCC 25611 was prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. The amount of TNF-$\alpha$ and interleukin-8 secreted into the culture medium was determined by enzyme-linked immunosorbent assay (ELISA). TNF-$\alpha$ and Ob-R mRNA expression levels were determined by semi-quantitative reverse transcription-polymerase chain reaction analysis. Results: Leptin enhanced P. intermedia LPS-induced TNF-$\alpha$ production in a dose-dependent manner. Leptin modulated P. intermedia LPS-induced TNF-$\alpha$ expression predominantly at the transcriptional level. Effect of leptin on P. intermedia LPS-induced TNF-$\alpha$ production was not mediated by the leptin receptor. Conclusions: The ability of leptin to enhance P. intermedia LPS-induced TNF-$\alpha$ production may be important in the establishment of chronic lesion accompanied by osseous tissue destruction observed in inflammatory periodontal disease.

Critical Messages on the Fashion Industry System and Fashion Consumption Culture in Critical Fashion Design (크리티컬 패션에 표현된 패션산업 시스템과 패션소비문화에 대한 비평적 메시지)

  • Jung, Junghee;Yim, Eunhyuk
    • Fashion & Textile Research Journal
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    • v.21 no.6
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    • pp.717-729
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    • 2019
  • This study defines critical fashion designs and investigate its critical messages on fashion itself. The critical messages on fashion are categorized into two major issues of a fashion industry system and fashion consumer culture. This study contributes to the understanding of meaning and value for critical fashion messages that match critical art. As the research method, this study combines a literature review and case studies and the research scope focuses on cases that have appeared in fashion media since the 2000s when social critical messages in fashion began to emerge. The results of the study are as follows. Critical designers such as Viktor & Rolf, Elisa van Joolen, Issey Miyake, and Mary Ping have delivered messages challenging the nature of fashion industry system that criticize the cycle and limitation of a fashion system and pursues changes in perception of sustainability. The critical message on fashion consumer culture articulated by designers such as Alexander McQueen, Vivienne Westwood, Hussein Chalayan, and Ricarda Bigolin & Nella Themelios insist on the formation of community while delivering a critical message on social, political, and cultural problems that raise the mechanism of social awareness through fashion design.

A Survey of the Prevalence of Rubella Antibodies in Teachers of Child Bearing Age on Cheju Island (제주도 가임 여교직원의 풍진 항체 보유율 조사)

  • Yang, Hyun-Jong;Hong, Seong-Chul;Bae, Jong-Myon
    • Journal of Preventive Medicine and Public Health
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    • v.33 no.3
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    • pp.280-284
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    • 2000
  • Background : Congenital rubella syndrome (CRS) can be controlled by vaccination. Because rubella is typically a childhood disease, occurring predominantly in the 5 to 14 year age group, female school teachers nay be a high-risk population for CRS. Objectives : To determine the prevalence rate of rubella antibodies in school teachers of child bearing age. Methods : The study population consisted of primary, middle and high school teachers of child bearing age. The subjects were aged 35 years and younger, and consented to immunoglobulin (Ig) level testing using the ELISA method. Results : The positive rate of IgG was 77.9% in the study subjects (n=314). Sixty-three teachers (21.4%) were susceptible to rubella infection. Thirty-seven teachers (11.8%) had a history of rubella vaccination. Among the female teachers with no vaccination history, the proportion of negative IgM and IgG was 21.7%, and the proportion of positive IgM was 2.9%. Seventy-nine percent of the study subjects did not know that they should not become pregnant for three months after receiving the rubella vaccine. Conclusion : School teachers of child bearing age should be considered a high risk group for CRS, and should be vaccinated if they are found to be seronegative.

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Anti-inflammatory effect of SD-01 in RAW 264.7 cells (마우스대식세포주인 RAW 264.7에서 SD-01의 항염증 활성 연구)

  • Park, Sun-Dong
    • The Korea Journal of Herbology
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    • v.25 no.3
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    • pp.19-25
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    • 2010
  • Objective : The aim of this study was to investigate anti-inflammatory activity of SD-01 methanol extract in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods : Cytotoxic activity of SD-01 methanol extract on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. The nitric oxide (NO) production was measured by Griess reagent system. And proinflammatory cytokines and $PGE_2$ were measured by ELISA method. The levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), $I{\kappa}$-B-alpha and nuclear NF-${\kappa}$ B p65 expression were detected by western blot. Results : Our results indicated that methanol extract of SD-01 significantly inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of TNF-$\alpha$, IL-$1\beta$, IL-6 and MCP-1 production in RAW 264.7 cells. Moreover, methanol extract of SD-01 treatment also blocked LPS-induced NF-kB activation. Conclusion : These findings indicate that methanol extract of SD-01 inhibits the production of pro-inflammatory mediators and cytokines via suppression of NF-${\kappa}$ B activation. Take together, these results indicate that methanol extract of SD-01 has the potential for use as an agent of anti-chronic inflammatory diseases.

Study on Anti-thrombotic Activities of KIOM2003-080 (KIOM2003-080 처방에 대한 항혈전 효능 연구)

  • Jeon, Won-Kyung;Kim, Ho-Kyoung;Lee, Ju-Hyun;Ko, Byoung-Seob;Lee, A-Yeong;Lee, Kyung-Goo;Shin, Hyeun-Kyoo
    • Korean Journal of Oriental Medicine
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    • v.13 no.2 s.20
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    • pp.143-148
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    • 2007
  • The present study examined inhibitory effects of 20 efficient experience prescriptions on platelet aggregation induced by collagen in human whole blood using the impedance method of aggregometry. Among them, a hot water extract of KIOM 2003-080 was selected to be the most effective candidate. In an in vivo study using a mouse acute thrombosis model, the anti-thrombotic effects of the KIOM2003-080 crude extract were also observed. In addition, we accessed bio-marker of platelet activation using thromboxane B2 by ELISA assay. A significantly decrease in thromboxane B2 production was seen in the presence of KIOM2003-080. Consequently, the results from this experiment provide pharmacological evidence for the traditional use of KIOM2003-080 prescription, suggesting that its hot water extracts could be used to prevent platelet aggregation and thrombosis disease.

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The Effect of Injinchunggan-tang on Hepatitis C Virus Infection (인진청간탕이 C형 간염 바이러스의 임파구계 세포감염에 미치는 영향)

  • 김선민;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.22 no.1
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    • pp.53-62
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    • 2001
  • Objective : The aim of this study was to evaluate the efficacy of Injinchunggan-tang on Hepatitis C virus infection, and to clarify the mechanism of treatment by indentifying the effect of Injinchunggan-tang on cytokine secretion. Methods : In vitro model of HCV infection in MOLT 4 cell was used. The effect of Injinchunggan-tang on the attachment of HCV on MOLT 4 cell was studied by PCR method. The change of cytokine secretion according to Injinchunggan-tang treatment was investigated by ELISA. Results : Injinchunggan-tang inhibited the attachment of HCV on MOLT 4 in the concentration of $10-2{\mu\textrm{g}}/\mu\textrm{\ell}$ and $10-1{\mu\textrm{g}}/\mu\textrm{\ell}$. In cytokine assay, Injinchunggan-tang increased the secretion of IL-4 of mouse splenocytes and PBMC in 48 hour culture as well as the secretion of IL-12 of mouse splenocytes and PBMC in 48 hour culture, whereas it decreased the secretion of $IFN-{\gamma}$ of mouse splenocytes in 24 and 48 hour culture. Conclusion : The results of this study show that Injinchunggan-tang has an inhibitory effect on the attachment on HCV on Mo1t4 Cell, and that it increases the secretion of IL-4 and IL-12 of mouse splenocyte and PBMC.

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