• Title/Summary/Keyword: double antibody

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Flatfish Vitellogenin Detection Using Optical Waveguide Lightmode Spectroscopy-based Immunosensor

  • Kim, Nam-Soo;Ryu, Hyung-Seok;Kim, Woo-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1445-1451
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    • 2007
  • A sensitive optical waveguide lightmode spectroscopy-based immunosensor was developed to detect vitellogenin in seawater flatfish (Paralichthys olivaceus). For this purpose, anion-exchange column chromatography with DE-52 resin was used to purify flatfish vitellogenin from flatfish serum containing vitellogenin that had been induced using an intraperitoneal $17{\beta}$-estradiol injection. The anti-flatfish vitellogenin antibody used as the biological component of the above immunosensor was prepared using the purified flatfish vitellogenin. The change in the incoupling angle according to the complexation between the flatfish vitellogenin and its antibody, immobilized over an optical grating coupler sensor chip, was measured to calculate the sensor response. The immunosensor was quite specific to flatfish vitellogenin binding, based on no sensor response in the case of bovine serum albumin immobilization. When plotted using double-logarithmic scales, the sensor responses increased linearly in flatfish vitellogenin concentrations of 0.00675-67.5 nM, with a detection limit of 0.0675 nM. The reusability during seven repetitive measurements was reasonably fair for the preliminary screening of flatfish vitellogenin.

Risk assessment of the potential for a classical swine fever outbreak in Korea based on a herd immunity

  • Lee, Changhee;Yoon, Hachung;Park, Choi-Kyu
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.429-435
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    • 2007
  • The risk of an outbreak of classical swine fever (CSF) was evaluated in relation to the vaccination and seroprevalence of antibodies. Blood samples were collected from 60 piggeries throughout Korea and information regarding CSF vaccination habits was also obtained via in-person interviews with pig farmers. The results of the survey indicated that 51 out of 60 farms were regularly performing CSF vaccinations in reproductive herds. Farmers preferred to vaccinate their reproductive pigs before weaning (41 farms) than on (9 farms) or after weaning (1 farm). In growing herds, however, double vaccinations as recommended were implemented for only 40 farms (66.7%) and vaccine schedules were identified as being frequently ignored. Moreover, many farms (18/40) were found to vaccinate earlier or later than the recommended time. According to antibody seroprevalence levels, only 36 farms (60%) were considered to be safe regarding potential risk for a CSF outbreak. Among the remaining pig farms, 6 were at low-risk (10%), 12 were at medium-risk (20%), and another 6 were at high-risk (10%). Antibody levels were found to be consistent with vaccination status obtained from personal interview surveys. Our findings demonstrate the importance of vaccinations regarding the prevention of a CSF outbreak, suggesting that vaccinations must be promoted toward pig raisers and practitioners.

Detection of Carp Vitellogenin with Piezoelectric Immunosensor (압전류적 면역센서에 의한 잉어 Vitellogenin 검출)

  • Kim, Nam-Soo;Park, In-Seon;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.254-258
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    • 2006
  • Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.

Effects of coffee intake on airway hypersensitivity and immunomodulation: an in vivo murine study

  • Ying-Chi Wong;Wen-Cheng Hsu ;Tzee-Chung Wu ;Ching-Feng Huang
    • Nutrition Research and Practice
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    • v.17 no.4
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    • pp.631-640
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    • 2023
  • BACKGROUND/OBJECTIVES: Coffee is a complex chemical mixture, with caffeine being the most well-known bioactive substance. The immunomodulatory and anti-inflammatory properties of coffee and caffeine impact health in various aspects, including the respiratory system. The objective is to investigate the effects of coffee and caffeine on airway hyperresponsiveness and allergic reactions, as well as to analyze and compare associated cytokine profiles. MATERIALS/METHODS: BALB/c mice were intraperitoneally sensitized with ovalbumin (OVA) and given OVA inhalation to induce airway hypersensitivity. Two weeks after sensitization, they were intragastrically gavaged with coffee or caffeine, both containing 0.3125 mg caffeine, daily for 4 weeks. Control mice were fed with double-distilled water. Serum OVA-specific antibody levels were measured beforehand and 5 weeks after the first gavage. Airway hyperresponsiveness was detected by whole body plethysmography after gavage. Cytokine levels of bronchoalveolar lavage and cultured splenocytes were analyzed. RESULTS: Coffee effectively suppressed T helper 2-mediated specific antibody response. Airway responsiveness was reduced in mice treated with either coffee or caffeine. Compared to the control, coffee significantly reduced OVA-specific immunoglobulin (Ig) G, IgG1 and IgE antibody responses (P < 0.05). Caffeine also attenuated specific IgG and IgG1 levels, though IgE level was unaffected. Coffee significantly reduced interleukin (IL)-4 and increased IL-10 concentration in spleen cells and bronchoalveolar lavage fluid (P < 0.05). CONCLUSIONS: Coffee effectively attenuated airway hyperresponsiveness and systemic allergic responses induced by OVA food allergen in mice. As a complex composition of bioactive substances, coffee displayed enhanced immunomodulatory and anti-inflammatory effects than caffeine.

Immunocytochemical Localization of Nitric Oxide Synthase-containing Neurons in Mouse and Rabbit Visual Cortex and Co-Localization with Calcium-binding Proteins

  • Lee, Jee-Eun;Jeon, Chang-Jin
    • Molecules and Cells
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    • v.19 no.3
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    • pp.408-417
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    • 2005
  • Nitric oxide (NO) occurs in various types of cells in the central nervous system. We studied the distribution and morphology of neuronal nitric oxide synthase (NOS)-containing neurons in the visual cortex of mouse and rabbit with antibody immunocytochemistry. We also compared this labeling to that of calbindin D28K, calretinin, and parvalbumin. Staining for NOS was seen both in the specific layers and in selective cell types. The densest concentration of intense anti-NOS immunoreactive (IR) neurons was found in layer VI, while the weak anti-NOS-IR neurons were found in layer II/III in both animals. The NOS-IR neurons varied in morphology. The large majority of NOS-IR neurons were round or oval cells with many dendrites coursing in all directions. Two-color immunofluorescence revealed that only 16.7% of the NOS-IR cells were double-labeled with calbindin D28K in the mouse visual cortex, while more than half (51.7%) of the NOS-IR cells were double-labeled with calretinin and 25.0% of the NOS-IR cells were double-labeled with parvalbumin in mouse. By contrast, 92.4% of the NOS-IR neurons expressed calbindin D28K while only 2.5% of the NOS-IR neurons expressed calretinin in the rabbit visual cortex. In contrast with the mouse, none of the NOS-IR cells in the rabbit visual cortex were double-labeled with parvalbumin. The results indicate that neurons in the visual cortex of both animals express NOS in specific layers and cell types, which do not correlate with the expression of calbindin D28K, calretinin or parvalbumin between the two animals.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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Study on Vibration Energy Harvesting with Small Coil for Embedded Avian Multimedia Application

  • Nakada, Kaoru;Nakajima, Isao;Hata, Jun-ichi;Ta, Masuhisa
    • Journal of Multimedia Information System
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    • v.5 no.1
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    • pp.47-52
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    • 2018
  • We have developed an electromagnetic generator to bury in subcutaneous area or abdominal cavity of the birds. As we can't use a solar battery, it is extremely difficult to supply a power for subcutaneous implantation such as biosensors under the skin due to the darkness environment. We are aiming to test the antigen-antibody reaction to confirm an avian influenza. One solution is a very small generator with the electromagnetic induction coil. We attached the developed coil to chickens and pheasants and recorded the electric potential generated as the chicken walked and the pheasant flew. The electric potential generated with physical simulator is equal to or exceeds the 7 V peak-to-peak at maximum by 560/min of flapping of wings. Even if we account for the junction voltage of the diode (200 mV), efficient charging of the double-layer capacitor is possible with the voltage doubler rectifier. If we increase the voltage, other problems arise, including the high-voltage insulation of the double-layer capacitor. For this reason, we believe the power generated to be sufficient for subcutaneous area of birds. The efficiency, magnetic 2 mm in length and coil 15mm in length, if axial direction is rectified, the magnetic flux density given to the coil could calculated to 7.1 % and generated power average 0.47mW. The improvements in size and wire insulation are expected in the future.

Gene Expression of Beta-Amyloid Precursor Protein (베타-아밀로이드 단백질 유전자 발현 연구)

  • Kim, Jong-Suk;Woo, Jong-Inn;Suh, Yoo-Hun;Kim, Ju-Han;Rhi, Bou-Yong
    • Korean Journal of Biological Psychiatry
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    • v.3 no.2
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    • pp.170-180
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    • 1996
  • Lowered immune function in the senile dementia patients may be related to the abnormal metabolism of amyloid precursor protein(APP). To investigate the passibility of an abnormal metabolism of APP in lymphocytes and the possible role of APP in the activation of lymphocytes in senile dementia patients, immunohistochemical study of rat spleen and fluorescence activated cell sorter analysis(FACS) of human lymphocytes with the specific antigen far each lymphocyte and double fluorescent marker with antibody to APP were performed. After stimulating lymphocyte with phytohemagglutinin(PHA), APP mRNA and protein were extracted and quantitfied and the influence of ${\beta}$-amyloid protein($A{\beta}$) specific antibody on lymphocyte division was investigated. In spleen, the majority of cells showing $A{\beta}$ immunoreactivity was found in the T-sell dependent zone. FACS indicated that around 90% $CD_4(+)$ T-cells and 60% of $CD_8(+)$ T-sell were immunoreactive to $A{\beta}$ specific antibody(mAb 4G8). Northern blot analysis shows that lymphocyte APP mRNA was gradually increased to reach a maximum at 3 days after activation with lectin mitogen PHA. However, the $A{\beta}$ immunoreactivity an cell surface remained constant during stimulation with PHA, indicating that the release of APP(secreted farm of APP) might be increased. A very large increase in soluble APP secretion was observed in T-lymphocyte upon activation, but only law levels in the resting stale. Immunoblot was carried out an the protein obtained from cell lysate after stimulating lymphocyte by applying PHA to the cultured lymphocyte, and the result was that $A{\beta}$ band of immature farm under 116 KDa marker decreased as the duration of culture was increased after PHA stimulation. The monoclonal $A{\beta}$ specific(4G8) and polyclonal APP antibodies did not inhibit the [$^3H$]-thymidine uptake of mitogen-treated lymphocytes significantly, suggesting that mitogenesis can not be inhibited by specific $A{\beta}$ and polyclonal APP antibody. These results suggest that APP is expressed in T-cell and might be closely associated with the function of T-cells.

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Biological, Physico-chemical and Serological Characteristics of TMV Strains Isolated from Tobacco, Tomato and Pepper Plants (담배, 토마토 및 고추에서 분리된 TMV 계통의 생물학적, 물리화학적 및 혈청학적 특성)

  • 박은경;이청호;이영기;김영호
    • Journal of the Korean Society of Tobacco Science
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    • v.19 no.1
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    • pp.5-10
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    • 1997
  • Three strains of W isolated from tobacco, tomato and Pepper plants in Korea were characterized based on biological response, serological relationship, and peptide mapping of the capsid Proteins. The strains designated as TMV-common, TMV-Pepper, and TMV-tomato could be distinguishable by different visual symptoms on 3 varieties of tobacco, one variety of tomato and Pepper for each among 27 plant specieces. Serological relationships were examined by agar gel double diffusion test. Only traceable or weak reaction was observed in the incompatible antigen-antibody combinations. The Pepper strain, however, showed trace in reaction with other two antisera. Peptide maps of the capsid proteins digested by V8 protease or by trypsin were also distinguishable, suggesting differences in composition and/or sequence of the amino acids among the strains.

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Multiple Forms of Serine-type Carboxypeptidase Produced by Absidia zychae (Absidia zychae가 생산하는 Serine-type Carboxypeptidase의 다양성)

  • 이병로;안병용
    • KSBB Journal
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    • v.8 no.4
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    • pp.405-408
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    • 1993
  • Absidia zychae NRIC 1199 produced two forms of carboxypeptidase(CPZ-1 and CPZ-2) which were distinguished in their isoelectric points but had almost identical properties(1). The amino acid sequences for the N-terminal of both enzymes were the same (Tyr-Thr-Ser-Pro-Lys-Leu-Xaa-Asp-Pro-Asp-Val) and any significant difference was not observed between amino acid compositions of the two enzymes. The ouchterlony double diffusion technique using antibody raised against the CPZ-2 protein demonstrated a good cross-reaction between CPZ-1 and CPZ-2 Genomic Southern analysis showed only one gene encoding CPZ in the genome of Absidia zychae. However, a significant difference between two enzymes was observed on peptide map using Staphylococcus aureus V8 protease, distinguishable only one band, indicating that multiple forms of CPZ are caused by post-translational modification, such as deamidation.

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