• Title/Summary/Keyword: Enzyme immunoassay

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Seroprevalence of abortion and stillbirth inducing disease in Hanwoo, in Jeonbuk eastern area (전북 동부지역 한우에서의 유사산 유발 질병 항체가 조사)

  • Song, Ji-Min;Shon, Ku-Rye;Koh, Won-Seok;Lee, Jeong-Won
    • Korean Journal of Veterinary Service
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    • v.37 no.3
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    • pp.179-183
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    • 2014
  • Reproductive losses in a herd can be huge. Fetal reabsorption or undefined infertility often remain undetected. Routine herds monitoring for exposure, controlling the introduction of potential agent carriers, appropriate biosafety procedures, and vaccination where possible are together the best security against abortion and stillbirth inducing disease. For biosecurity of local farms, we performed antibody titers of abortion and stillbirth related diseases such as bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBRV), Neospora caninum, Toxoplasma gondii and Campylobacter fetus subsp venerealis. The blood samples were collected from 500 female Hanwoo over 1 year old of 100 farms in Jeonbuk eastern area. Champhylobater serological test was evaluated by the standard tube agglutination test (STAT) and other pathogen's antibodies were detected by indirect-enzyme linked immunoassay (I-ELISA). The seroprevalence of abortion and stillbirth inducing disease were BVDV 72.4%, IBRV 13.0%, N. caninum 1.2%, T. gondii 10.4% and C. venerealis 0.6%, irrespectively.

Studies on the Improvement of Precision in Early Pregnancy Diagnosis by Milk Progesterone Test(EIA) in Dairy Cows (Milk Progesterone Test(EIA)에 의한 소의 임신조기판단 정확도 향상에 관한 연구)

  • 김정우
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.149-156
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    • 1989
  • These experiment was carried out to improve the precision of early pregnancy diagnosis in dairy cattle. Changes in progesterone concentration of milk were measured by Enzyme Immunoassay(EIA) in 73 cows up to 21 days after insemination. The average concentraton of progesterone in milk was 1.9ng/ml at eatrus ; it increased to 17.8ng/ml on day 14, and thereafter it declined to 4.3ng/ml on day 21 in nonpregnant cows. Whereas in pregnant animals, it was maintained and elevated further to 22.2ng/ml on day 21. The accuracy of the pregnancy diagnosis for animals classified as positive (pregnant ; over 10ng/ml and decreasing rate<1.5) and negative (non-pregnant ; under 7ng/ml and decreasing rate>1.5) were 95% and 100% respectively. The samples ranging between 7ng/ml and 10ng/ml were classified as positive (decreasing rate<1.5) and negative(>1.5), which accuracy was 54.5% and 100% respectively. However this range appears to be of the most interest for the veterinary practitioner since cows in proestrus or early interestrus tend to have milk progesterone levels within this value. Causes for the insufficient precision of false pregnancy diagnosis are discussed.

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In vitro and in vivo application of anti-cotinine antibody and cotinine-conjugated compounds

  • Kim, Hyori;Yoon, Soomin;Chung, Junho
    • BMB Reports
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    • v.47 no.3
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    • pp.130-134
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    • 2014
  • The combination of a high-affinity antibody to a hapten, and hapten-conjugated compounds, can provide an alternative to the direct chemical cross-linking of the antibody and compounds. An optimal hapten for in vitro use is one that is absent in biological systems. For in vivo applications, additional characteristics such as pharmacological safety and physiological inertness would be beneficial. Additionally, methods for cross-linking the hapten to various chemical compounds should be available. Cotinine, a major metabolite of nicotine, is considered advantageous in these aspects. A high-affinity anti-cotinine recombinant antibody has recently become available, and can be converted into various formats, including a bispecific antibody. The bispecific anti-cotinine antibody was successfully applied to immunoblot, enzyme immunoassay, immunoaffinity purification, and pre-targeted in vivo radioimmunoimaging. The anti-cotinine IgG molecule could be complexed with aptamers to form a novel affinity unit, and extended the in vivo half-life of aptamers, opening up the possibility of applying the same strategy to therapeutic peptides and chemical compounds.

Proinflammatory cytokine levels in oral lichen planus, oral leukoplakia, and oral submucous fibrosis

  • Kaur, Jasdeep;Jacobs, Reinhilde
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.41 no.4
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    • pp.171-175
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    • 2015
  • Objectives: The objective of this study was to identify salivary and serum concentrations of interleukin (IL)-8, IL-6, and tumor necrosis factor alpha ($TNF-{\alpha}$) in patients with oral lichen planus, oral leukoplakia, oral submucous fibrosis, and healthy controls. Materials and Methods: Patients selected included 54 oral lichen planus (41 to 65 years), 50 oral leukoplakia (42 to 65 years), 51 oral submucous fibrosis (41 to 65 years), and 50 healthy controls (42 to 65 years). Oral lichen planus, oral leukoplakia, and oral submucous fibrosis cases were diagnosed using histopathological analysis. Salivary and serum cytokine concentrations were measured using enzyme-linked immunoassay kits in all subjects. Results: The levels of serum and salivary $TNF-{\alpha}$, IL-6, and IL-8 were statistically significantly increased in oral leukoplakia, submucous fibrosis, and lichen planus in contrast to normal healthy subjects (P<0.05). Serum and salivary correlation analysis revealed strong and highly significant correlations for $TNF-{\alpha}$, IL-6, and IL-8 in all groups (r=0.72-0.82, P<0.05). Conclusion: Salivary and serum cytokines were also elevated when analyzed in oral precancerous lesions. Thus, salivary and serum IL-8, IL-6, and TNF-${\alpha}$ levels might act as diagnostic markers for detection of oral precancer.

Detecting Activated Thrombin Activatable Fibrinolysis Inhibitor (TAFIa) and Inactivated TAFIa (TAFIai) in Normal and Hemophilia A Plasmas

  • Hulme, John P.;An, Seong Soo A.
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.77-82
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    • 2009
  • Thrombin activatable fibrinolysis inhibitor (TAFI) also known as plasma procarboxypeptidase B or U is a 60 kD glycoprotein, which is the major modulator of fibrinolysis in plasma. TAFI is a proenzyme, which is activated by proteolytic cleavage to an active carboxypeptidase B-like enzyme (TAFIa, 35.8 kD) by thrombin/thrombomodulin and plasmin. Modulation of fibrinolysis occurs when TAFIa enzymatically removes C-terminal lysine residues of partially degraded fibrin, thereby inhibiting the stimulation of tissue plasminogen activator (t-PA) modulated plasminogen activation. TAFIa undergoes a rapid conformational change at $37{^{\circ}C}$ to an inactive isoform called TAFIai. Potato tuber carboxypetidase inhibitor (PTCI) was shown to specifically bind to TAFIa as well as TAFIai. In this study, a novel immunoassay TAFIa/ai ELISA was used for quantitation of the two TAFI activation isoforms TAFIa and TAFIai. The ELISA utilizes PTCI as the capture agent and a double antibody sandwich technique for the detection. Low levels of TAFIa/ai antigen levels were detected in normal plasma and elevated levels were found in hemophilia A plasmas. TAFIa/ai antigen represents a novel marker to monitor fibrinolysis and TAFIa/ai ELISA may be a valuable assay for studying the role of TAFI in normal hemostasis and in pathological conditions.

Anti-inflammatory Effects of Gagamtongsoon-San Extract on Lipopolysaccharide(LPS)-Induced Inflammation in RAW264.7 Cells (LPS로 유발한 RAW264.7 세포 염증반응에 대한 가감통순산(加減通順散)의 억제 효과)

  • Lee, Soo-Hwan;Kim, Soon-Joong
    • Journal of Korean Medicine Rehabilitation
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    • v.28 no.2
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    • pp.37-45
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    • 2018
  • Objectives This study was designed to investigate whether the Gagamtongsoon-San (GT) has an inhibitory effect and its mechanisms are associated with the iNOS and COX-2. Methods Cytotoxic activity of GT extract on RAW264.7 cells was evaluated by using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) solution. Inflammatory condition was induced by LPS. NO production was measured using Griess reagent system. The expressions of iNOS and COX-2 mRNA and protein were determined by realtime PCR. The concentrations of PGE2 were measured by an enzyme immunoassay (EIA). Results The GT does not impair the cell viability in tested concentration $500{\mu}g/ml$ or below. GT significantly reduced the NO production in a dose-dependent manner. GT $500{\mu}g/ml$ also suppressed LPS-induced mRNA expressions of iNOS and COX-2. GT $500{\mu}g/ml$ reduced the PGE2 secretion in LPS induced RAW264.7 cells. Conclusions These outcomes show that GT extract has an anti-inflammatory activities. And also this conclusion can be the data that supports the GT's anti-inflammatory effect objectively.

Serological Diagnosis for Swine Cysticercus Cellulosae Infection in Korea (돼지의 유구낭충(有鉤囊蟲)에 관(關)한 혈청학적시험(血淸學的試驗))

  • Jean, Y.;Lee, T.J.
    • Korean Journal of Veterinary Research
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    • v.25 no.1
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    • pp.77-89
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    • 1985
  • The agar gel precipitation(AGP), indirect hemagglutination(IHA) and indirect enzyme immunoassay(IEIA) tests were used to detect antibodies in pigs naturally infected with Cysticercus cellulosae in Jeju. The results obtained were summarized as follows: 1. Sera collected from pigs naturally infected with Cysticcrcus cellulosae did not react in AGP test. 2. In the IHA test for swine cysticercosis, the peak titers observed were between 1:20 and 1:160 and non-specific reaction was recognized with a few samples among control sera. 3. In the IEIA test, optical density(OD) values were obtained the best results under the condition of OPD-substrats are reacted at room temperature for 5minutes. The OD values of greater than 0.2 were determined as positive and the high titers in positive sera ranged from 1:40 to 1:1,080. 4. Antibodies to swine cysticercosis were detected by IHA and IEIA tests but the latter was more sensitive and specific than the former. 5. In the preparation of Cystisercus antigens, saline extract which was prepared the precipitate of internal membrane treated ultrasonicator were better results than other antigens for serological tests. 6. Amounts of protein in antigens was not related in direct proportion to results of serological reaction.

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Methodological validation of measuring Hanwoo hair cortisol concentration using bead beater and surgical scissors

  • Nejad, Jalil Ghassemi;Ataallahi, Mohammad;Park, Kyu-Hyun
    • Journal of Animal Science and Technology
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    • v.61 no.1
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    • pp.41-46
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    • 2019
  • Different methodologies in hair cortisol extraction may alter the final output. Thus finding the standard methodology according to a laboratory facilities is pivotal. This study was carried out to validate the feasibility of two methods of grinding hair for cortisol extraction in Korean native (Hanwoo) cattle. Hair from nine cattle including mature cows, heifers, and calves were assigned to one of the following methods for grinding hair; 1) using bead beater (BB) and 2) using surgical scissors (SS). Hair samples (> 1 g) were harvested from forehead of each individual twice (first and second measurement) to validate the results. To improve the accuracy of the obtained data, each sample was duplicated into two wells during enzyme immunoassay (EIA) analysis. Overall comparison of hair cortisol concentration (HCC) showed that the data within the range (out of the range) of standards provided by the EIA kit were 88.9% (11.1%) and 66.7% (33.3%) for BB compared with SS, respectively. In the first measurement, application of BB was tended to show higher (p = 0.056) amount of HCC compared with SS. In the second measurement application of BB showed higher (p = 0.0028) amount of HCC compared with SS. Among the cattle, calves showed higher HCC using BB compared with SS (p < 0.05). Application of BB in hair grinding methodology for Hanwoo cattle may improve cortisol extraction in comparison to application of SS method, with more consistency. Thus, it would be the preferable method to use.

Mass Spectrometry-Based Analytical Methods of Amatoxins in Biological Fluids to Monitor Amatoxin-Induced Mushroom Poisoning

  • Choi, Jin-Sung;Lee, Hye Suk
    • Mass Spectrometry Letters
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    • v.13 no.4
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    • pp.95-105
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    • 2022
  • Amatoxin-induced mushroom poisoning starts with nonspecific symptoms of toxicity but hepatic damage may follow, resulting in the rapid development of liver insufficiency and, ultimately, coma and death. Accurate detection of amatoxins, such as α-, β-, and γ-amanitin, within the first few hours after presentation is necessary to improve the therapeutic outcomes of patients. Therefore, analytical methods for the identification and quantification of α-, β-, and γ-amanitin in biological samples are necessary for clinical and forensic toxicology. This study presents a literature review of the analytical techniques available for amatoxin detection in biological matrices, and established an inventory of liquid chromatography (LC) techniques with mass spectrometry (MS), ultraviolet (UV) detection, and electrochemical detection (ECD). LC-MS methods using quadrupole tandem mass spectrometry, time-of-flight mass spectrometry, and orbitrap MS are powerful analytical techniques for the identification and determination of amatoxins in plasma, urine, serum, and tissue samples, with high sensitivity, specificity, and reproducibility compared to LC with UV and ECD, enzyme-linked immunoassay, and capillary electrophoresis methods.

Effect of Dipsaci Radix Water Extract on LPS-induced Inflammatory Response in RAW264.7 Mouse Macrophages (속단(續斷)의 RAW264.7 세포에서 LPS에 의해 유도되는 염증반응에 대한 효과)

  • Min, Ji-Young;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.189-195
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    • 2009
  • Objectives : In this study, the effect of Dipsaci Radix(DR, Dipsacus asperoides C.Y. Cheng et T. M. Ai) water extract on LPS-induced inflammatory response in RAW264.7 cells were investigated. Methods : Dried roots of DR was extracted with water for 3 h(DR-W extract). RAW264.7 cells, a mouse macrophage line, were incubated with different concentrations of DR-W extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expression of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX)-2 mRNA and protein was determined by RT-PCR and Western blot, respectively. Results : DR-W extract was significantly inhibited LPS-induced productions of NO and PGE2 in RAW264.7 cells. DR-W extract was not suppressed the expressions of iNOS mRNA and protein in LPS-stimulated RAW264.7 cells. Conclusions : This study suggests that DR-W extract can attenuate inflammatory response via inhibition of the NO and PGE2 production in activated macrophages.