• Title/Summary/Keyword: Enzyme immunoassay

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Comparative study on the antimicrobial and antiinflammatory activity of commercially available toothpastes (수종의 상용 세치제들의 향균 및 향염효과 비교연구)

  • Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.557-566
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    • 1996
  • It is known that some natural extracts from plants have a various range of antimicrobial and antiinflammatory activity. There are lots of clinical trials to develop toothpastes containing natural extracts for prevention of dental caries and gingival inflammation. The purpose of this study was to evaluate antimicrobial and antiinflammatory activity of magnolol containing toothpastes and other commercial toothpastes. Eleven kinds oftoothpastes were used. They include magnolol, sanguinarine, Myrrha, Mori radicis cortex,Cimicifugae rhizoma, sodium fluoride, aminocaprolactic acid etc. Six strains of bacteria were used for this test, ego Porphylomonas gingivalis, Prevotellain-termedia, Actinobacillus actinomy cetemcomitans, Streptococcus mutans, Stretococcus sanguis, and Actinomyces species. Antimicrobial activity was determined by an agar dillution method and a broth microdillution method. Antiinflammatory activity was assessed by the inhibition of $PGE_2$ production from gingival fibroblast with the addition of rHIL-1 and centrifuged solution of toothpastes. Control group was only rHIL-1 additive sample. $PGE_2$ enzyme immunoassay systemfAmersham, In. Buckinghamshire, U.K). $PGE_2$ level was measured by ELISA reader with 450 nm, The results from the study revealed that toothpastes containing natural extracts generally had high antimicrobial and antiinflammatory activity. Especially magnolol containing toothpaste showed higher antimicrobial activity than other toothpastes, and sanguinarine containing toothpaste showed particularly high antimicrobial activity in A. actinomicetemcomitans and A. viscosus. In some degree all toothpastes inhibited $PGE_2$ production, but magnolol containing toothpaste was potent inhibitor of $PGE_2$. Sodium chloride containing toothpaste had also effective result. The results suggested that toothpastes containing natural extracts were promising in plaque control and prevention of dental caries and gingivitis.

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Increased Interlenkin-2 Serum Level in Male Schizophrenic Patients (남자 정신분열증 환자에서 혈청 Interlenkin-2 농도의 증가)

  • Kim, Yong-Ku;Kim, Sa-Jun;Lee, Min-Soo
    • Korean Journal of Biological Psychiatry
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    • v.3 no.1
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    • pp.109-114
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    • 1996
  • We have previously reported that Korean schizophrenic patients hove low production of IL-2 in vitro suggestive of autoimmunity to the pathogenesis of the disorder. In an attempt to further explore this issue, we measured in vivo serum levels of interleukins(IL-$1{\beta}$, IL-2, and IL-6) using a quantitative "sandwich" enzyme immunoassay(ELISA) in 26 male schizophrenic patients and in 26 age-matched normal controls. Patients met DSM-IV criteria for schizophrenia and were drug free for at least six months. The severity of symptoms was assessed by SANS and SAPS. We found a significant increase of IL-2 level(p<0.05) in schizophrenic patients as compared with normal controls. There were significant positive correlations between IL-2, IL-6 levels and negative symptom scores. There were no correlations between age, age at onset, duration of illness and interleukin levels. Our results may support the hypothesis of viral-autoimmune dysfunction in schizophrenia. IL-2 or IL-6 may be associated with specific clinical feature in schizophrenic syndrome, especially negative symptom.

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Development of One-step Simultaneous Immunochromatographic Assay for Rapid Analysis of Aflatoxin B1 and Ochratoxin A

  • Shim, Won-Bo;Kim, Gyeong-Yeol;Ryu, Hee-Jung;Nam, Min-Ji;Chung, Duck-Hwa
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.641-648
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    • 2009
  • A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

Purification and Characterization of Authentic Human Growth Hormone Converted from Methionyl Human Growth Hormone by Immobilized Aminopeptidase M (고정화 Aminopeptidase M에 의해 메치오닐 인간성장호르몬으로부터 전환된 천연형 인간성장호르몬의 정제 및 특성 확인)

  • 이성희;조영우
    • KSBB Journal
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    • v.10 no.3
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    • pp.271-282
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    • 1995
  • The authentic hGH converted from met-hGH by immobilized ApM was purified by successive chromatographic processes based on the differences in isoelectric points, hydrophobicities and charges. The final recovery yield was about 14.1% and the specific activity of the purified hGH was 2.75IU per mg when assayed by enzyme immunoassay. The purified hGH was verified to be authentic hGH through the analysis of amino acid composition, amino-terminal amino acid sequence, carboxy-terminal amino acid and tryptic peptide map. The purity of purified hGH was higher than that of commercial hGH when assessed by SDS-PAGE, PAGE, IEF and HSGF. In weight-gain assay and tibia test with hypophysectomized rats, the hGH produced in this study showed the same growth effect as the commercial hGH.

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Detection of Serum Anti-Extracellular Protein Kinase a Autoantibodies as a Potential Tumor Marker

  • Lee, Seung-Ho;Kim, Ki-Nam;Seo, Sang-Hui;Sohn, Sung-Hwa;Kim, Yu-Ri;Kim, Hye-Won;Choi, Chul-Won;Kim, Jun-Suk;Kim, Meyoung-Kon
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.67-73
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    • 2006
  • In previous studies, it has been discovered that cancer cells not only overexpress regulatory subunit I (Rl)/protein kinase type I (PKA-I) but also secrete outside the cell an extracellular form of PKA (ECPKA) and that the ECPKA secretion detected in patients' serum is obviously greater than that found in non-cancer patients or healthy subjects. We now found that ECPKA elicits the formation of serum autoantibodies that can serve as a cancer diagnostic and prognostic marker. To measure the presence of anti-ECPKA autoantibody in the human sera, basic methodology for ECPKA assay was established an enzyme-linked immunosorbent assay (ELISA). We obtained serum samples from 199 patients with different types of cancer, and also obtained 31 serum samples to compare with ECPKA concentrations from non-cancer patients and 119 normal volunteers. Compared with normal or non-cancer patient sera, we found that the frequency of anti-ECPKA autoantibody was significantly higher in cancer patients (88%) than in those without cancer (17%). Furthermore the presence of anti-ECPKA autoantibodies in the serum of cancer patients was highly correlated with the site of metastasis. The immunoassay developed for anti-ECPKA antibodies is highly sensitive and specific. Therefore, this discovery of an autoantibody-based cancer diagnostic may have serious clinical application and may become an important advance over current technology.

NFATc Mediates Lipopolysaccharide and Nicotine-Induced Expression of iNOS and COX-2 in Human Periodontal Ligament Cells (사람 치주인대세포에서 Lipopolysaccharide와 니코틴으로 유도된 iNOS와 COX-2 발현에 NFATc의 관여)

  • Lee, Sang-Im;Yu, Ji-Su
    • Journal of dental hygiene science
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    • v.15 no.6
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    • pp.753-760
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    • 2015
  • Although nuclear factor of activated T cell (NFAT) plays a key role in inflammation, its anti-inflammatory effects and mechanism of action in periodontitis are still unknown. This study aimed to identify the effects of NFAT on the proinflammatory mediators activated by lipopolysaccharide (LPS) plus nicotine stimulation in human periodontal ligament cells (hPDLCs). The production of nitric oxide (NO) and prostaglandin $E_2(PGE_2)$ was evaluated using Griess reagent and an enzyme immunoassay, respectively. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and NFAT proteins was evaluated by Western blot analysis. LPS plus nicotine synergistically induced the production of NO and $PGE_2$ and increased the protein expression of iNOS, COX-2 and NFAT. Treatment with an NFAT inhibitor blocked the LPS plus nicotine-stimulated NO and $PGE_2$ release as well as the expression of iNOS and COX-2. Our data suggest that the LPS plus nicotine-induced inflammatory effects on hPDLCs may act through a novel mechanism involving the action of NFAT. Thus, NFAT may provide a potential therapeutic target for the treatment of periodontal disease associated with smoking and dental plaque.

Anti-stress Effects of Natural Products from Jeju Island in Zebrafish (제브라피쉬에서의 제주도 천연추출물의 항스트레스 효과)

  • Lee, Jeongwon;Lee, Seungheon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.85-85
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    • 2019
  • Objective: In this study, the anti-stress effects of extract of Hydrangeae Dulcis Folium (EHDF) or ethalonic extract of Opuntiaficus-indica (EOF) of natural extracts from Jeju Island were investigated. Methods: We performed measurement of whole-body cortisol level and behavioral experiments including the novel tank test (NTT) or the open field test (OFT) to assess stress responses in zebrafish. To induce physical stress, we used the net handling stress (NHS). Fish were treated with EOF or EHDF for 6 min before they were exposed to stress. And then, we sacrificed fish for collecting body fluid from whole-body or conducted behavioural tests, including novel tank test and open field test, were evaluated to observe anxiety-like behaviours and locomotion. We used the cortisol enzyme-linked immunoassay kit to measure the amount of cortisol in each zebrafish sample. Results: The results indicate that increased anxiety-like behaviours in novel tank test and open field test under stress were prevented by treatment with both EOF and EHDF (P < 0.05). Moreover, compared with the unstressed group, which was not treated with NHS, the whole-body cortisol level was significantly increased by treatment with NHS. Compared with the NHS-treated stressed control group, pre-treatment with each EHDF and EOF for 6 min significantly prevented the NHS-increased whole-body cortisol level (P < 0.05). Conclusions: In conclusion these results suggest that both EOF and EHDF pretreatment may prevent stress responses and that its mechanism of action may be related to its positive effects on cortisol release.

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Effects of feather processing methods on quantity of extracted corticosterone in broiler chickens

  • Ataallahi, Mohammad;Nejad, Jalil Ghassemi;Song, Jun-Ik;Kim, Jin-Soo;Park, Kyu-Hyun
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.884-892
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    • 2020
  • Corticosterone is known as a biological stress index in many species including birds. Feather corticosterone concentration (FCC) has increasingly been used as a measure for chronic stress status in broiler chickens. As sample preparation is the first step of analytical process, different techniques of feather matrix disruption need to be validated for obtaining better result in analysing corticosterone extraction. The current study was a validation of pulverizing the feather by bead beater (BB) and surgical scissors (SS) processing prior to corticosterone extraction in feather of broiler chickens. The type of feather processing prior to the hormone extraction may alter the final output. Thereby, finding a standard method according to laboratory facilities is pivotal. This study carried out to determine the effects of feather pulverization methods on the extraction amount of corticosterone in broiler chickens. Feathers were sampled from four weeks old Ross 308 broiler chickens (n = 12 birds). All broiler chickens were kept under the same environmental condition and had access to feed and water. Feather samples were assigned to one of the following processing methods 1) using a BB for pulverizing and 2) using a SS for chopping into tiny pieces. Each sample was duplicated into two wells during enzyme immunoassay (EIA) analysis to improve the accuracy of the obtained data. The results showed lower standard errors and constant output of FCC by using the BB method compared with the SS method. Overall comparison of FCC showed a significantly higher (p < 0.001) amount of the FCC in the BB compared with the SS. Overall, using the BB method is recommended over the SS method for feather processing due to the ability to homogenize a large number of samples simultaneously, ease of use and greater extraction of feather corticosterone.

Anti-inflammatory Effects of Ethanol Extract of Various Korean Compositae Herbs in LPS-induced RAW 264.7 Macrophages

  • Seo, Min-gyu;Kang, Yun-Mi;Chung, Kyung-Sook;Cheon, Se-Yun;Park, Jong Hyuk;Lee, Young-Cheol;An, Hyo-Jin
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.17-24
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    • 2017
  • Objective : This study was designed to evaluate candidate materials as anti-inflammation agent from extracts of various Korean Compositae herbs in Hwaak mountain. Among Korea medicinal herbs, Ainsliaea acerifolia (AA) belongs to the Compositae family, has been used for the treatment of rheumatic arthritis. However, AA has not been previously reported to have an anti-inflammatory effect. Therefore, we investigated the anti-inflammatory effects of AA and its underlying molecular mechanisms in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Methods : Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in RAW 264.7 macrophages. Nitric oxide (NO) was measured with Griess reagent and pro-inflammatory cytokines were detected by enzyme immunoassay (EIA) kits in LPS-stimulated RAW 264.7 macrophages. Protein expressions of inducible nitric oxide synthase, and cyclooxygenase-2 (COX-2) and p65 subunit of nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) were determined by Western blot analysis. Results : Among 8 extracts of Korean Compositae herbs tested, AA showed the inhibition of NO production without cytotoxicity. Consistent with the observation, AA reduced the expression levels of iNOS and COX-2 proteins in LPS-simulated RAW 264.7 macrophages in dose-dependent manner. In addition, AA inhibited the productions of $TNF-{\alpha}$ and IL-6 in LPS-simulated RAW 264.7 macrophages. However, AA did not inhibit activation of p65 $NF-{\kappa}B$ in LPS-simulated RAW 264.7 macrophages. Conclusion : These results suggest that down-regulation of iNOS, COX-2 protein expression and $TNF-{\alpha}$ and IL-6 production by AA are responsible for its anti-inflammatory effects.

Effect of Astragali Radix on SPS-induced PTSD in Rats (황기(黃芪)가 흰쥐에서 SPS로 유도된 PTSD에 미치는 효과)

  • Min, Ye-Eun;Lee, Tae-Hee
    • Herbal Formula Science
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    • v.30 no.3
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    • pp.109-121
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    • 2022
  • Objective : This study is conducted to investigate the effect of Astragali Radix on Post Traumatic Stress Disorder(PTSD) induced by the Single Proposed Stress(SPS). Methods : The experiment was conducted with five groups; SAL groups with only saline treatment, SPS group, SPS + ARX25 group, SPS + ARX50 group, and SPS + ARX100 group. After applying SPS, saline and ARX were administered for 14 days to identify the change of body weight, sucrose intake amount, and behavioral changes through Open Field Test(OFT) and Forced Swimming Test(FST). After the behavioral experiment, plasma corticosterone levels, serotonin, norepinephrine and dopamine concentrations were measured by enzyme-linked immunoassay in medical prefrontal cortex, hippocampus, and amygdala. Brain-derived neurotrophic factor(BDNF) in the hippocampus was measured using Reverse Transcription Polymerase Chain Reaction. Results : Weight change has significantly decreased in the SPS group compared to the SAL group(p<0.05). On day 14, the sucrose intake of rats has significantly increased in the SPS + ARX100 group compared to the SPS group(p<0.05). In OFT, the number of staying in the central space has significantly increased in the SPS + ARX100 group(p<0.01). In FST, immobility has significantly decreased in SPS + ARX50 group and SPS + ARX100 group(p<0.05). The concentration of serotonine, dopamine and BDNF expression has increased significantly in SPS + ARX100 group compared to SPS group(p<0.05) Conclusions : In the SPS-induced PTSD experiment, ARX increased sucrose intake and the numbers of crossing in the central zone space in OFT, decreased immobility time in FST, and increased concentration of serotonin, dopamine, and BDNF. It can be postulated that the ARX could be effective for the treatment of PTSD.