• Title/Summary/Keyword: Assay system

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Convenient Assay of O2- Generated on Potato Tuber Tissue Slices Treated with Fungal Elicitor by Electron Spin Resonance - No Secondary Oxidative Burst Induction by H2O2 Treatment

  • Park, Hae-Jun;Doke, Noriyuki
    • The Plant Pathology Journal
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    • v.21 no.3
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    • pp.283-287
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    • 2005
  • Since the discovery of generation of $O_2^-$ in plant, many evidence for the oxidative burst (OXB) has been accumulated in various combinations of plant and pathogen or elicitor systems. $O_2^-$ generating system responsible for the OXB was coupled with oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in microsomal fraction isolated from sliced aged potato tuber slices which were treated by hyphal wall components elicitor from Phytophthora infestans (HWC). We developed new assay method for quantitative measurement of oxygen radical $O_2^-$ by using electron spin resonance (ESR) analysis during elicitor­induced OXB on the surface of plant tissues. The ESR analysis using an $O_2^-$ trapper, Tiron (1,2-dihydroxy-3,5­benzenedisulfonic acid), provided a convenient assay for detecting only $O_2^-$ during elicitor-induced OXB producing various active oxygen species (AOS) on plant tissue surface. Tiron was oxidized to Tiron semiquinon radical by $O_2^-$. Quantity of the radical signal was measured by specific spectra on ESR spectroscopy. The level of $O_2^-$ was high in from surface of potato tuber tissue treated with hyphal cell wall elicitor (HWC) from Phytophthora infestans. There was no secondary OXB induction by $H_2O_2$ treatment in plant.

Characteristics and application of monoclonal antibody to progesterone II. Development of progesterone enzyme-linked immunosorbent assay(ELISA) (Progesterone의 단크론성 항체에 관한 특성 및 활용에 관한 연구 II. ELISA 기법의 개발)

  • Kang, Chung-boo;Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.403-409
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    • 1991
  • This experiment was carried out to develop a sensitive, rapid, solid-phase microtitre plate assay of progesterone using the monoclonal antibody to this hormone. Monoclonal antibody to progesterone was much higher titre and binding affinity about 10 times than conventional polyclonal antibody to progesterone. Dot-blot analysis of monoclonal antibody revealed a single precipitation band when reacted with anti-mouse IgM and anti-mouse K. A competitive reaction was used with a reaction time of 2 hours. The standard dose-response curve was linear through 1,000pg/well. This ELISA system approach is applicable to evaluation for the rapid assessment of luteal function and reproductive status in both clinical and research in a wide variety of species.

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Genotoxicity Test of Bojungikkeehapdaechilki-tang water extract (보중익기합대칠기탕(補中益氣合大七氣湯) 추출물의 유전독성 평가)

  • Hwang, Hui-Jeung;Byun, Joon-Seok;Heo, Jin-Il
    • Herbal Formula Science
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    • v.14 no.1
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    • pp.141-167
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    • 2006
  • The genotoxicity of water extract of Bojungikkeehapdaechilki-tang was tested by In Vitro Chromosome Aberration Test. Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines. The obtained results were as follows : 1. Chromosome Aberration Test: In Vitro Chromosome Aberration Test of Bojungikkeehapdaechilki-tang extracts was carried out using cultured Chinese hamster lung cells in the presence and absence of metabolic activation system(S-9 mix). No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in Bojungikkeehapdaechilki-tang extracts treated groups. 2. Bacterial Reveres Mutation Assay: Bojungikkeehapdaechilki-tang extracts was evaluated for its potential to induce reverse mutation in the histidine auxotroph strains of Salmonella typhimurium such as TA100, TA1535, TA98 and TAl537 and the tryptophan auxotroph strain of Escherichia coli WP2 uvrA. No significant changes in the number of revertant colonies compared to its negative control were detected in Bojungikkeehapdaechilki-tang extracts treated groups against all 5 strains. 3. Micronucleus test: Micronucleus test of Bojungikkeehapdaechilki-tang extracts were performed using specific pathogen free 7-week old male ICR mouse. No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all Bojungikkeehapdaechilki-tang extracts treated groups. In summarized above-mentioned results, it is concluded that Bojungikkeehapdaechilki-tang extracts have not genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

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Antimutagenic and Cytotoxic Effects of Potato Vinegar (감자식초의 항돌연변이원성 및 세포독성 효과)

  • 함승시;김미남;정용진;이득식
    • Journal of the East Asian Society of Dietary Life
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    • v.11 no.1
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    • pp.19-25
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    • 2001
  • This study was performed to examine the antimutagenic and cytotoxic effects of potato vinegar and commercial vinegars(cider, brown rice, persimmon vinegars) on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and cytotoxicity assay, respectively. In Ames test, all vinegars did notexhibit any mutagenicity , but showed substantial inhibitory effects against N- methyl - N -nitro - N- nitrosog -uanidine(MNNG) , 4-nitroquinoline-1-oxide(4NQO), 3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indol(Trp-P-1)and benzo( $\alpha$ )pyrene(B( $\alpha$ )P). The number of revertants per plate decreased significantly when these vinegars(80 ug/plate) were added to the assay system using TA100 strain. Especially, potato vinegar(80 ug/plate) showed high inhibition rate of 69.9% against mutagenicity of B( $\alpha$ )P on TA100 strain. In the cytotoxicity assay, these vinegars also showed prominent cytotoxic activity against human cancer cell lines. Potato vinegar(10 ug/well) showed the strongest cytotoxic effect against HT1080 (fibrosacoma cell) andK562 ( myelogenous leukemia) at the same concentration when compared with other vinegars.

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Assays of Residual Antibiotics after Treatment of γ-ray and UV Irradiation (감마선과 UV 조사에 의한 항생제 분해)

  • Shin, Ji-Hye;Nam, Ji-Hyun;Yu, Seungho;Lee, Myunjoo;Lee, Dong-Hun
    • Journal of Radiation Industry
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    • v.4 no.1
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    • pp.39-45
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    • 2010
  • The pollution of antibiotics is a major cause of spreading antibiotics resistant bacteria in the environment. Applications of ozonation, UV, and ${\gamma}-ray$ irradiations have been introduced to remove antibiotics in the effluents from wastewater treatment system. In this study, we compared the chemical (HPLC) and biological (antimicrobial susceptibility test, AMS) assays in measuring of the concentrations of residual antibiotics after ${\gamma}-ray$ and UV irradiation. Most samples were degraded by ${\gamma}-ray$ irradiation (1~2 kGy). However, lincomycin and tetracycline were not degraded by UV irradiation. The concentration of residual antibiotics, that was treated with ${\gamma}-ray$ and UV irradiation, measuring by bioassay was similar to HPLC. The concentrations of ${\gamma}-ray$ irradiated cephradine measured by AMS test were 2 times higher than that of HPLC assay, indicating AMS test is more sensitive than HPLC assay. These results indicate that ${\gamma}-ray$ irradiation technique is more useful than UV irradiation, and biological assay is more useful to detect the antibiotics and toxic intermediates in antibiotics degradation.

Fissile Measurement in Various Types Using Nuclear Resonances

  • YongDeok Lee;Seong-Kyu Ahn
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.21 no.2
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    • pp.235-246
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    • 2023
  • Neutron resonance transmission technique was applied for assaying isotopic fissile materials produced in the pyro-process. In each process of the pyro-process, a different composition of the fissile material is produced. Simulation was basically performed on 235U and 239Pu assay for TRU-RE product, hull waste, and uranium addition. The resonance energies were evaluated for uranium and plutonium in the simulation, and the linearity in the detection response was examined on the fissile content variation. The linear resonance energies were determined for the analysis of 235U and 239Pu on the different fissile materials. For enriched TRU-RE assay, the sample condition was suggested; The sample density, content, and thickness are the key factors to obtain accurate fissile content. The detection signal is discriminated for uranium and plutonium in neutron resonance technique. The transmitted signal for fissile resonance has a direct relation with the content of fissile. The simulation results indicated that the neutron resonance technique is promising to analyze 235U and 239Pu for various types of the pyro-process material. An accurate fissile assay will contribute toward safeguarding the pyro-processing system.

Effects of Amylase and Cellulase Supplementation in Sorghum-based Diets for Finishing Pigs

  • Park, J.S.;Kim, I.H.;Hancock, J.D.;Hines, R.H.;Cobb, C.;Cao, H.;Hong, J.W.;Kwon, O.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.70-76
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    • 2003
  • Three experiments were conducted to determine the effects of a sorghum-specific enzyme system, derived from an Aspergillus niger and Bacillus subtilis fermentation extract (carbohydrase activity of 1,650 $\alpha$-amylase units and cellulase activity of 30 fibrinolytic units/mL), on growth performance of finishing pigs. In Exp. 1,192 pigs (average initial BW of 46.1 kg) were fed sorghum-based diets without or with 360 mL of enzyme system per ton of sorghum in a 78 d growth assay. For d 0 to 39, gain/feed was improved (p<0.03) with enzyme supplementation, but ADG was not affected (p>0.15). For d 39 to 78 and overall (d 0 to 78), ADG, gain/feed, and digestibilities of DM and N were not affected (p>0.13) by enzyme supplementation. Backfat thickness, fat-free lean index, and scores for stomach keratinization and ulcers also were not affected (p>0.15) by the dietary treatments. In Exp. 2,168 pigs (average initial BW of 58.4 kg) were fed diets without or with 150, 300, or 450 mL/ton of the same enzyme system used in Exp. 1. Adding as much as 450 mL enzyme system / ton of sorghum did not affect (p>0.15) ADG or gain/feed for d 0 to 29 of the growth assay. However, during d 29 to 63, ADG increased by 11% (linear effect, p<0.02) and gain/feed increased by 10% (linear effect, p<0.06) as enzyme concentration was increased from none to 450 mL/ton of sorghum. For the overall period (d 0 to 63), ADG tended to increase (p<0.08) with enzyme supplementation, but gain/feed and digestibilities of DM and N were not affected (p>0.14). Carcass characteristics (dressing percentage, backfat thickness, and fat free lean index) also were not affected (p>0.20) by addition of the enzyme system. In Exp. 3,176 pigs (average initial BW of 46.7 kg) were fed diets without or with 450, 900, or 1,350 mL/ton of the same enzyme system used in Exp. 1 and 2 in a 71 d growth assay. Adding up to 1,350 mL/ton of enzyme had no effects (p>0.15) on ADG, gain/feed, digestibilities of DM and N, and carcass characteristics (dressing percentage, backfat thickness, and fat-free lean index). In conclusion, finishing pigs fed diets with a sorghum-specific enzyme system showed some positive trends for improved growth performance, but those effects were not large and (or) consistent.

Usefulness of the BACTEC MGIT 960 System for Mycobacterial Culture and TB Ag MPT64 Immunochromatographic Assay to Identify Mycobacterium tuberculosis (항산균 배양에서 BACTEC MGIT 960 System의 유용성 및 MGIT 양성배지에서 결핵균 진단을 위한 TB Ag MPT64 면역발색법의 유용성)

  • Lee, Seung Hun;Lee, Min Jeong;Lee, Jeong-Mi;Yim, Su Jin;Lee, Seung Jun;Kim, You Eun;Cho, Yu Ji;Jeong, Yi Yeong;Kim, Ho Cheol;Lee, Jong Deog;Kim, Sun Joo;Hwang, Young Sil
    • Journal of Yeungnam Medical Science
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    • v.29 no.2
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    • pp.83-88
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    • 2012
  • Background: This study was conducted to evaluate the usefulness of the BACTEC MGIT (Mycobacterium Growth Indicator Tube) 960 system for mycobacteria culture and immunochromatographic assay to identify Mycobacterium tuberculosis (MTB) in positive MGIT culture. Methods: Mycobacteria-culture-positive cases were retrospectively analyzed from December 2010 to July 2011. The detection rates and the recovery times of the mycobacteria between the Ogawa media and the MGIT were compared. An immunochromatographic assay (ICA) (SD BIO-LINE) was also performed in the positive MGIT culture for identification, and the results were compared with those of the Ogawa media in the Korea National Tuberculosis Association. Results: Among the 261 patients (M:F, 168:93; mean age, $61.6{\pm}17.16$ yrs), 450 specimens (sputa, 365; bronchial washing, 61; and pleural effusion, 24) were found positive with mycobacteria. Mycobacteria were grown both on the MGIT and Ogawa media in 310 cases (68.9%); only on the MGIT in 115 cases (22.6%); and only on the Ogawa media in 25 cases (5.5%) (p<0.05). The recovery time was $28.2{\pm}8.9$ days in the Ogawa media and $11.1{\pm}5.8$ days in the MGIT (p<0.05). Among the 127 cases from the positive MGIT culture, all 92 cases that were confirmed as MTB cases bythe Korea National Tuberculosis Association were identified as MTB by ICA, with 100% sensitivity. Conclusion: MGIT increases the detection rate and shortens the recovery time of mycobacteria in clinical respiratory specimens, and the TB Ag MPT64 kit using ICA is useful in identifying MTB in a positive MGIT culture.

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Detection of infectious canine hepatitis virus by TaqMan real-time PCR method (TaqMan 실시간 PCR법에 의한 개 전염성 간염 바이러스의 검출)

  • Wang, Hye-young;Choi, Jae-yong;Lee, Mi-jin;Park, Jin-ho;Cho, Mae-Rim;Han, Jae-cheol;Choi, Kyoung-seong;Chae, Joon-seok
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.655-662
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    • 2004
  • The aim of this work was the validation of a rapid real-time PCR assay based on TaqMan technology for the unequivocal identification of infectious canine hepatitis (ICH) virus, to be used directly on DNA purified from blood specimens. A real-time PCR system targeting at the E3 ORFA gene sequence of canine adenovirus type 1 was optimized and validated through comparative analysis of samples using conventional PCR system. The real-time PCR assay based on TaqMan technology could disclose 23 (37.7%) out of 61 samples as PCR positive. In contrast, 18 (29.5%) samples were found PCR positive when conventional PCR was applied on these samples. The use of the ABI Prism 7700 sequence detection system allowed the efficient determination of the amplified product accumulation through a fluorogenic probe. The entire real-time TaqMan PCR assay, including DNA extraction, amplification, and detection could be completed within 3 hours. The detection method of real-time TaqMan PCR assay was 1,000 times more sensitive than conventional PCR. Real-time TaqMan probe and primer set developed and optimized in this study is a sensitive, rapid and accurate method for detection of ICH virus and can be effective screening tool for the detection of ICH in a diagnostic laboratory routines.

DESIGN OPTIMIZATION OF RADIATION SHIELDING STRUCTURE FOR LEAD SLOWING-DOWN SPECTROMETER SYSTEM

  • KIM, JEONG DONG;AHN, SANGJOON;LEE, YONG DEOK;PARK, CHANG JE
    • Nuclear Engineering and Technology
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    • v.47 no.3
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    • pp.380-387
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    • 2015
  • A lead slowing-down spectrometer (LSDS) system is a promising nondestructive assay technique that enables a quantitative measurement of the isotopic contents of major fissile isotopes in spent nuclear fuel and its pyroprocessing counterparts, such as $^{235}U$, $^{239}Pu$, $^{241}Pu$, and, potentially, minor actinides. The LSDS system currently under development at the Korea Atomic Energy Research Institute (Daejeon, Korea) is planned to utilize a high-flux ($>10^{12}n/cm^2{\cdot}s$) neutron source comprised of a high-energy (30 MeV)/high-current (~2 A) electron beam and a heavy metal target, which results in a very intense and complex radiation field for the facility, thus demanding structural shielding to guarantee the safety. Optimization of the structural shielding design was conducted using MCNPX for neutron dose rate evaluation of several representative hypothetical designs. In order to satisfy the construction cost and neutron attenuation capability of the facility, while simultaneously achieving the aimed dose rate limit (< $0.06{\mu}Sv/h$), a few shielding materials [high-density polyethylene (HDPE)eBorax, $B_4C$, and $Li_2CO_3$] were considered for the main neutron absorber layer, which is encapsulated within the double-sided concrete wall. The MCNP simulation indicated that HDPE-Borax is the most efficient among the aforementioned candidate materials, and the combined thickness of the shielding layers should exceed 100 cm to satisfy the dose limit on the outside surface of the shielding wall of the facility when limiting the thickness of the HDPE-Borax intermediate layer to below 5 cm. However, the shielding wall must include the instrumentation and installation holes for the LSDS system. The radiation leakage through the holes was substantially mitigated by adopting a zigzag-shape with concrete covers on both sides. The suggested optimized design of the shielding structure satisfies the dose rate limit and can be used for the construction of a facility in the near future.