• Title/Summary/Keyword: Enzyme Linked Immunosorbent Assay(ELISA)

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The Effect of Mineral Trioxide Aggregate on the Production of Growth Factors and Cytokine by Human Periodontal Ligament Fibroblasts (Mineral trioxide aggregate (MTA)가 치주인대 섬유아세포에서 분비되는 cytokine과 성장인자 TGF-β1, FGF-2 발현에 미치는 영향)

  • Kwon, Ji-Yoon;Lim, Sung-Sam;Baek, Seung-Ho;Bae, Kwang-Shik;Kang, Myung-Hoe;Lee, Woo-Cheol
    • Restorative Dentistry and Endodontics
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    • v.32 no.3
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    • pp.191-197
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    • 2007
  • Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 $(TGF-\beta1)$, fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of $1\times10^5$ cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of $TGF-\beta1$, FGF-2 and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA) The data were analyzed using one-way ANOVA. The level of $TGF-\beta1$ was down-reg ulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated co)Is were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.

ELISA of rat sera infected with Paragonimus iloktsuenensis (이락촌폐흡충 감염 쥐의 혈청에 대한 효소면역 반응(ELISA))

  • Im, Byeong-Gil;Lee, Ok-Ran;Nam, Hae-Seon
    • The Korean Journal of Parasitology
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    • v.28 no.4
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    • pp.207-212
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    • 1990
  • Enzyme-linked immunosorbent assay(ELISA) of paragonimiasis iloktsuenensis rat sera was performed using crude antigens of Paragonimus iloktsuenensis(PIA), P. westermani (PWA) and Clonorchis sinensis(CSA). Three crude antigens(PIA, PWA, CSA) were prepared to saline homogenated supernatants of whole adult worms. Infected rat sera were obtained biweekly from the albino rats fed 50∼.80 metacercariae of P. iloktsuenensis through gastric catheter. Experimental groups were divided into 4 groups: GI(controls), GII, GIII and GIV according to 1∼7 worms as GII, 10∼19 worms as GIII and 22∼40 worms as GIV, respectively, In ELISA, the mean OD values of each group for the homologous antigen(PIA) were increased significantly compared to the control sera at the 4th week of infection. With the progress of duration of infection, the mean OD values of infected sera of GII & GIV continuously increased up to the 12th week(last week), but in GIII the mean OD value increased until the loth week. No significance was noted among the infection dose groups (GII, GIII and GIV), after the 6th week of infection. Also, the OD values of all infected rats did not show any Proportional relytionships to the number of worms recovered. In brief, the antibody productivity of individual rats were strongly different. The rat sera infected with p. iloktsuenensis cross-reacted with those infected with P. westermani or C. sinensis, as identified by OD values.

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Effect of Wood Vinegar Produced from Morus alba on Hypersecretion of Airway Mucus (상지(桑枝) 목초액이 호흡기 객담 과다분비에 미치는 영향)

  • Kim, Ho;Jung, Hye-Mi;Kim, Sol-Li;Seo, Un-Kyo
    • The Journal of Internal Korean Medicine
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    • v.31 no.3
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    • pp.650-666
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    • 2010
  • Objectives : In this study, the author tried to investigate whether wood vinegar produced from Morus alba (MA) significantly affects the increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats, and in vitro airway mucin secretion and PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production / gene expression from human airway epithelial cells. Materials and Methods : For the in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to SO2 over 3 weeks. Effect of orally-administered MA over 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using histopathological analysis after staining the epithelial tissue with alcian blue. For the in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of MA to assess the effect of MA on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of MA and treated with PMA (10 ng/ml), EGF (25 ng/ml) or TNF-alpha (0.2 nm) for 24 hrs, to assess both effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicities of MA in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of MA were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering MA orally. Results : 1. MA decreased the amount of intraepithelial mucosubstances of rats exposed to sulfur dioxide inhalationally. 2. MA decreased in vitro mucin secretion from cultured RTSE cells. 3. MA significantly inhibited PMA-, EGF-, and TNF-alpha-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. 4. MA did not show either in vitro or in vivo hepatic or renal toxicities. Conclusion : The results from this study suggests that MA can regulate the secretion, production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and does not show in vivo toxicity to liver and kidney functions after oral administration. Effects of MA should be further studied using animal experimental models that simulate the diverse pathophysiology of respiratory diseases via future research.

Evaluation of the Estrogenic Activity by Yeast Two-hybrid Assay and Enzyme-linked Immunosorbent Assay in Sewage Treatment Plant (하수처리장의 내분비계장애물질에 대한 Yeast Two-hybrid Assay와 Enzyme-linked Immunosorbent Assay에 의한 에스트로겐활성도 평가)

  • Lee, Byoung-Cheun;Ra, Jin-Sung;Kim, Sang-Don;Kawai, Hukiko;Lee, Chul-Hee
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.771-777
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    • 2007
  • Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17$\beta$-estradiol, 17$\alpha$-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17$\beta$-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17$\beta$-estradiol 17$\alpha$-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17$\beta$-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.

Sedum sarmentosum Enhances Hyaluronan Synthesis in Transformed Human Keratinocytes and Increases Water Content in Human Skin (돌나물추출물에 의한 사람 각질형성세포에서의 Hyaluronan Synthesis 촉진과 인체 피부의 보습력 증진)

  • Sim, Gwan-Sub;Kim, Jin-Hwa;Lee, Dong-Hwan;Na, Young;Lee, Geun-Soo;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.1
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    • pp.17-22
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    • 2007
  • In this study, we investigated the effects of Sedum sarmentosum extract on the expression of hyaluronan synthase (HAS) genes and hyaluronan (HA) production in HaCaT keratinocytes. We also assessed water content (electrical capacitance) and transepidermal water loss (TEWL) in human skin after topical treatment with Sedum sarmentosum extract. Sedum sarmentosum extract increased expression of HAS-2 and HAS-3 genes in HaCaT cells, when assayed by real-time reverse-transcriptase polymerase chain reactions (RT-PCR). Sedum sarmentosum extract induced HA production in HaCaT cells, when determined by enzyme-linked immunosorbent assay (ELISA). Finally, treatment of Sedum sarmentosum extract on human skin increased the skin hydration and decreased TEWL when measured using Corneometer and Tewameter. Our study suggests that Sedum sarmentosum extract should be a very useful cosmetic ingredient, as a skin moisturizer.

A study on the early pregnancy diagnosis by changing of plasma progesterone concentration and morphology of ovary in pregnancy and non -pregnancy cows (소에서 비임신 및 임신 상태의 난소 형태와 혈중 progesterone 농도 변화에 의한 조기 임신진단)

  • Kim, Cheol-Ho;Bhak, Jong-Sik;Shin, Jung-Sub;Kang, Chung-Bo
    • Korean Journal of Veterinary Service
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    • v.31 no.3
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    • pp.397-414
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    • 2008
  • In order to evaluate conception rate of Hanwoo in northwestern region of Gyeongsang-nam-do, we investigated conception rate and reduction of reproductive disorder rate after artificial insemination (AI) in 1,000 heads of breeding cows, This study showed that 80.9% of cows were classified as fertility after 1st and 2nd AI. For a accurate pregnancy diagnosis with practicing ovariectomy and histeotomy, we comparatively investigated each of 80 slaughtered cows, including 30 of non-pregnancy, and used enzyme-linked immunosorbent assay (ELISA) for estimation of plasma progesterone concentration and serum luteal hormone. The mean diameter of non-pregnant corpus luteum is $18.9{\pm}4.2{\times}15.6{\pm}3.6 mm$ and that of pregnant corpus luteum is $22.5{\pm}2.7{\times}18.7{\pm}2.9 mm$. This indicates that corpus luteum is more developed in the ovary of pregnant than non-pregnant cows (P<0.05). The diameter of pregnant corpus luteum according to the stage of pregnancy showed $21.3{\pm}2.4{\pm}18.4{\pm}2.6 mm$ in early stage (1-3 month), $23.4{\pm}2.8{\times}19.1{\pm}2.7 mm$ in middle stage (4-6 month) and $22.8{\pm}3.0{\times}18.8{\pm}2.4mm$, in last stage (7-9 month). This indicates that corpus luteum in middle and last stage is more significantly developed than that of early stage(P<0.05). The mean plasma progesterone concentration of cows showing size of non-pregnant corpus luteum was $4.58{\pm}0.92ng/ml$ and that of pregnant corpus luteum $8.26{\pm}0.98ng/ml$. Thus, it was more significantly increased in pregnant corpus luteum(P<0.02).. However, it was low to $0.58{\pm}0.39ng/ml$. in estrus (corpus albicans). The plasma progesterone concentration according to gestation period was high in proportion to the degree of development in corpus luteum and more significantly increased (P<0.05) and maintained in middle and last state than early state. The concentration was sharply decreased to $0.56{\pm}0.32ng/ml$ at parturition. As a consequence, we can practice the early pregnancy diagnosis by confirming non-pregnancy when the mean plasma progesterone concentration is below 1ng/ml 19 to 22 days after AI and this can be available to diagnose reproductive disorder.

Use of enzyme-linked immunosorbent assay (ELISA) for detection of toxoplasmosis in dogs (ELISA 법을 이용한 개 톡소플라즈마병의 조기진단에 관한 연구)

  • Suh, Myung-deuk;Joo, Hoo-don;Lee, Byung-hoon
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.491-500
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    • 1991
  • This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.

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Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
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    • v.6 no.2
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    • pp.141-149
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    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

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Serological survey of avian pneumovirus infection in laying hens of Gyeongbuk province (경북지역 산란계에서 avian pneumovirus 에 대한 항체가조사)

  • 김순태;김성국;조민희;김영환
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.51-56
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    • 2003
  • Avian pneumoviros(APV), also known as avian rhinotracheitis virus(ARTV), affects both turkeys and chickens and is known to be the primary causative agent of turkey rhinotracheitis (TRT). The aim of this study was to establish the presence or absence of antibodies to avian pneumovirus in the commercial poultry population of Korea. For this purpose, chicken serum samples were obtained and tested by an enzyme-linked immunosorbent assay (ELISA). The tested serum was collected in laying hens with reduction of egg production or normal in Gyeongbuk province. A total of 184 sera representing 42 different poultry farms of the Gyeongbuk region of Korea were included in this study. Laying hens of 16 different farms with reduction of egg production and laying hens of 26 different farms with clinically healthy at the time of serum sampling were considered positive to antibody against APV. In the farms with reduction of egg production, positive farm to antibody against avian pneumovirus were 11 of 16 different farms(68.8%) and positive sera were 47(58.8%) of 80 different serum. In the farms with clinically healthy flock, positive farm to antibody against avian pneumovirus were 12(46.2%) of 26 different farms and positive serum sample were 39(37.5%) of 104 different sera. According to the results tested to 42 different farms in 14 city, 8 of 14 city have flocks with antibody positive laying hens against APV, 1 of 14 city have antibody suspicious and 5 of 14 city shown antibody negative, respectively.

Characteristics of outbreak for the classical swine fever (CSF) at Incheon metropolitan area in 2002 (인천지역에서 발생한 돼지콜레라의 역학적 특성)

  • 권효정;변재원;이정구;김경호;박은정;이성모;황현순
    • Korean Journal of Veterinary Service
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    • v.26 no.1
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    • pp.1-9
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    • 2003
  • The aim of this survey was to investigate the characteristics of outbreak farm determined as the classical swine fever(CSF) at Gangwha-gun and Seo-gu, Incheon metropolitan area from October 7 to November 25 in 2002. Sixty pigs in six different farms were confirmed to the CSF and a total of 9,106 pigs containing 3,194 related epidemiologically was slaughtered to stop spreading of the disease. Clinical signs of pigs diagnosed with the CSF were high fever, anorexia, depression, paralysis of hindlimbs, cyanosis, etc and gross lesions were typically represented with hemorrhage of submandibular and superficial lymph node, infarction of spleen, and petechial (ecchymotic) hemorrhage of kidney and skin. But some outbreak farms had not shown remarkable symptoms, so they were confused with other bacterial diseases. White blood cell (WBC) counts, the classical swine fever virus(CSFV) antigen and antibody enzyme linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) results about six farms indicated that total 60 pigs were infected with the CSFV. Although the origin and infection route of the CSFV were not clear, but the transmissions between farms were mainly through indirect contact such as the movement of farm personal and vehicles from outbreak farm.