• Title/Summary/Keyword: Enzyme Linked Immunosorbent Assay Kit

Search Result 81, Processing Time 0.022 seconds

Fucoidan Suppresses Prostaglandin E2 Production and Akt Activation in Lipopolysaccharide-Stimulated Porcine Peripheral Blood Mononuclear Cells

  • Park, Geon-Tae;Ahn, Changhwan;Kang, Byeong-Teck;Kang, Ji-Houn;Jeung, Eui-Bae;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
    • /
    • v.34 no.3
    • /
    • pp.172-177
    • /
    • 2017
  • Fucoidan, a cell wall polysaccharide found in the brown seaweed, is reported to have broad-spectrum biological activities. The objectives of this study were to examine the effect of fucoidan on prostaglandin $E_2$ ($PGE_2$) and cyclooxygenase-2 (COX-2) expression in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs) and to determine whether these effects are involved in Akt activation. The levels of $PGE_2$ production in the culture supernatants from PBMCs were determined by the enzyme-linked immunosorbent assay (ELISA) kit and the levels of COX-2 mRNA were measured by real time polymerase chain reaction (RT-PCR). Akt activity was determined by Western blot analysis. Fucoidan in LPS-$na{\ddot{i}ve}$ PBMCs has no effect on $PGE_2$ production and COX-2 mRNA expression. Furthermore, fucoidan does not affect Akt activation in LPS- $na{\ddot{i}ve}$ PBMCs. However, $PGE_2$ production and COX-2 mRNA expression on PBMCs were remarkably enhanced by LPS stimulation. Akt activity was also increased by LPS. Increasing effects of $PGE_2$ production and COX-2 mRNA expression in PBMCs induced by LPS were suppressed by addition of fucoidan. In addition, fucoidan reduced an increase in Akt activity in LPS-stimulated PBMCs. These results suggested that fucoidan exerts potent anti-inflammatory properties by suppression of $PGE_2$ production, COX-2 mRNA expression and Akt activation in LPS-stimulated PBMCs.

Comparison of ELISA and HI titers in broiler chicks vaccinated with infectious bronchitis virus and Newcastle disease virus (전염성기관지염 및 뉴캣슬병 백신을 접종한육계에서 ELISA 및 HI 항체가 비교)

  • 고원석;이정원;곽길한;권정택;송희종
    • Korean Journal of Veterinary Service
    • /
    • v.24 no.1
    • /
    • pp.21-29
    • /
    • 2001
  • To compare of serum antibody titers using ELISA and HI, serum samples were collected from 100 breeders and their progeny 550 broilers. The breeders and broilers were vaccinated with infectious bronchitis(IB)- and Newcastle disease(ND)-viruses according to general vaccination program. The antibodies in serum samples against IB and ND viruses were detected by enzyme-linked immunosorbent assay(ELISA) using commercial ELISA kit and hemagglutination inhibition(HI) test. Geometric mean titer(GMT) of ELISA and In titers were monitored from 1-day-old to 35-day-old broilers and compared to those of breeder chickens. The antibody titers of breeders vaccinated with ]B virus showed 47,800, ELISA and 7.2, HI, respectively. Progeny chicks, 1-day-old, vaccinated with IBV showed high antibody titers than those of breed chickens. Those chicks were maintained protective antibody levels until 11-day-old. From 14-day-old, the antibody level decreased below protective levels. In ND, breeders serum antibody titers ELISA and Eiu were 30,200 GMT and 8.7 HI titer, respectively. On 1-day-old chicks, antibody levels was decreased to half in ELISA(16,270) compared with those of breeders, but In titers was 7.4. Progeny broilers, protective antibody level was maintained until 14- day-old by ELISA, but at 11-day-old by HI titers. After then, ND antibody titer was continuously decreased underdefense level. These result indicated that the ELISA method be more sensitive than HI titration to detect serum antibody level for IBV and NDV.

  • PDF

Comparative analysis of serological tests and fecal detection in the diagnosis of Mycobacterium avium subspecies paratuberculosis infection

  • Park, Hong-Tae;Ha, Seungmin;Park, Hyun-Eui;Shim, Soojin;Hur, Tai Young;Yoo, Han Sang
    • Korean Journal of Veterinary Research
    • /
    • v.60 no.3
    • /
    • pp.117-122
    • /
    • 2020
  • Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. The most effective way to eradicate JD is to detect infected individuals as early as possible and remove them from the herd. However, it is difficult to detect infected individuals early with the currently using diagnostic methods. Two serological diagnostic kits commercially used worldwide and a fecal detection test were compared using 298 serum samples and feces of cattle in this study to present an efficient diagnostic method. Although there was a high correlation between the 2 serological diagnostic kits (R2 = 0.7473), kit A showed a higher serological positive rate. However, the correlation between fecal tests and serological diagnosis was very low. MAP was also detected in fecal tests in many serologically negative individuals. In the periodical diagnosis of JD, MAP was detected in the feces of only cows with the higher antibody titer to MAP. These results suggest that for effective eradication of JD, early detection of infected individuals by fecal tests together with the serological tests currently in use and by removal of infected individuals are needed.

A Study on Antibacterial Activity and Seroprevalence of Ornithobacterium rhinotracheale Isolated from the Domestic Chickens (국내 사육 닭에서 분리된 Ornithobacterium rhinotracheale (OR)균의 약제 감수성 및 항체보유율에 대한 연구)

  • 전우진;권용국;윤여성;김재홍
    • Korean Journal of Microbiology
    • /
    • v.39 no.3
    • /
    • pp.161-165
    • /
    • 2003
  • Ornithobacterium rhinotracheale (OR) is a recently described gram-negative rod-shaped bacterium associated with respiratory tract infection in poultry. In order to investigate current occurrence of OR infection and to evaluate antibiotic susceptibility, the prevalence of OR antibody in domestic chickens were examined and the minimal inhibitory concentrations(MICs) of 8 antibiotics for 11 OR isolates was determined. All isolates tested were mostly susceptible to three antibiotics, ampicillin (MICs ranging from 0.38 ${\mu}g$/ml to 2 ${\mu}g$/ml), tetracycline (MICs 0.094~3 ${\mu}g$/ml) and doxycycline (MICs 0.047~4 ${\mu}g$/ml) but resistant to genatmicin. Ciprofloxacin, norfloxacin, enrofloxacin, and ofloxacin gave most isolates inhibition only in case of a higher concentration (MICs ranged in most cases from 3 ${\mu}g$/ml to 48 ${\mu}g$/ml). Out of 188 chicken flocks including broilers, broiler breeders, and layers, seropositive flock to OR were detected in 5 broilers (4%), 17 broiler breeders (50%), and 16 layers (55.2%), using commercial OR enzyme-linked immunosorbent assay(ELISA) kits. It suggested that OR infection was widespreaded in poultry farms in Korea.

Codium fragile Ethanol Extraction Inhibited Inflammatory Response through the Inhibition of JNK Phosphorylation

  • Han, Sin-Hee;Kim, Young-Guk;Lee, Su-Hwan;Park, Chung-Berm;Choi, Han-Gil;Jang, Hye-Jin;Lee, Young-Seob;Kwon, Dong-Yeul
    • Preventive Nutrition and Food Science
    • /
    • v.15 no.3
    • /
    • pp.206-212
    • /
    • 2010
  • Codium fragile (CF) is an edible green alga consumed as a traditional food source in Korea. In this study, the ethanol extract of CF was evaluated to determine if it has anti-inflammatory activity. Lipopolysaccharide (LPS), a toxin from bacteria, is a potent inducer of inflammatory cytokines, such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Therefore, we studied whether CF extracts have an anti-inflammatory effect in LPS-induced murine macrophage cell lines (RAW 264.7). In the present study, IL-6 production was measured using an enzyme-linked immunosorbent assay (ELISA), prostaglandin $E_2$($PGE_2$) production was measured using the EIA kit, and cyclooxygenase (COX)-2 and mitogen-activated protein kinase (MAPK) activation were determined by Western blot analysis. IL-6 mRNA, COX-2 mRNA and iNOS mRNA expression were measured using reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that CF extracts inhibit LPS-induced IL-6, NO and PGE2 production in a dose-dependent manner, as well as expression of iNOS and COX-2. CF extracts significantly inhibited LPS-induced c-Jun N-terminal kinase (JNK) 1/2 phosphorylation. Taken together, these findings may help elucidate the mechanism by which CF modulates RAW 264.7 cell activation under inflammatory conditions.

A Case of Canine Ectopic Parasitism of Heartworm in the Systemic Circulation (개에서 발생한 전신 순환계의 심장사상충 이소기생 증례)

  • Jang, Hyo-Mi;Lee, Hee-Chun;Jung, Dong-In
    • Journal of Veterinary Clinics
    • /
    • v.32 no.4
    • /
    • pp.347-351
    • /
    • 2015
  • A 3-year-old intact female Dachshund was referred due to paraplegia and cool extremities. History taking, physical examination, neurological examination, and blood profiling studies were evaluated to determine the diagnosis. Based on abdominal ultrasound and echocardiograph, features suggestive of adult heartworm were detected in aberrant places. The result of enzyme-linked immunosorbent assay (ELISA) testing with a commercial heartworm antigen kit was positive. The dog fell into a comatose state, and the client requested the dog be euthanized. On post-mortem examination, the patient was diagnosed with ectopic parasitism of heartworm in the left side of heart, aorta, aorta abdominalis, and iliac arteries, a circumstance that induced systemic thromboembolism. This case report describes the clinical, diagnostic imaging, and necropsy findings of canine ectopic parasitism of heartworm.

Prepartum body condition score affects milk yield, lipid metabolism, and oxidation status of Holstein cows

  • Zhao, Wei;Chen, Xue;Xiao, Jun;Chen, Xiao Hui;Zhang, Xue Feng;Wang, Tao;Zhen, Yu Guo;Qin, Gui Xin
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.32 no.12
    • /
    • pp.1889-1896
    • /
    • 2019
  • Objective: This study aimed to investigate the effects of prepartum body condition score (BCS) on the milk yield, lipid metabolism, and oxidative status of Holstein cows. Methods: A total of 112 multiparous Holstein cows were divided into 4 groups according to the BCS at 21 days before calving: medium BCS (3.0 to 3.25, MBCS), high BCS (3.5 to 3.75, HBCS), higher BCS (4.0 to 4.25, HerBCS), and highest BCS (4.5 to 5.0, HestBCS). Blood samples were collected on 21, 14, and 7 days before calving (precalving), on the calving day (calving), and on 7, 14, and 21 days after calving (postcalving). The indices of lipid metabolism and oxidative status were analyzed using bovine-specific enzyme-linked immunosorbent assay kit. Colostrum were taken after calving and analyzed by a refractometer and milk analyzer. The individual milk yield was recorded every 3 days. Results: The density and levels of immune globulin and lactoprotein of colostrum from Holstein cows in the HestBCS group were the highest (p<0.05). These animals not only had the highest (p<0.05) levels of serum non-esterified fatty acids and beta-hydroxybutyrate, but also had the highest (p<0.05) levels of malondialdehyde, superoxide dismutase, catalase, vitamin A, and vitamin E. In addition, greater (p<0.05) BCS loss was observed in the HestBCS cows. Conclusion: This study demonstrates that the milk yield, lipid metabolism, and oxidative status of Holstein cows are related to prepartum BCS and BCS loss during the transition period. HestBCS cows are more sensitive to oxidative stress and suffer greater loss of BCS after calving, whereas the MBCS animals had better milk yield performance.

The effect of light on follicular development in laying hens

  • Cheng, Shi Bin;Li, Xian Qiang;Wang, Jia Xiang;Wu, Yan;Li, Peng;Pi, Jin Song
    • Animal Bioscience
    • /
    • v.34 no.11
    • /
    • pp.1766-1775
    • /
    • 2021
  • Objective: The oxidative stress status and changes of chicken ovary tissue after shading were studied, to determine the mechanism of the effect of shading on follicular development. Methods: Twenty healthy laying hens (40 weeks old) with uniform body weight and the same laying rate were randomly divided into two groups (the shading group and normal light group). In the shading group, the cage was covered to reduce the light intensity inside the cage to 0 without affecting ventilation or food intake. The normal lighting group received no additional treatment. After 7 days of shading, oxidative stress related indicators and gene expression were detected. Results: Analysis of paraffin and ultrathin sections showed that apoptosis of ovarian granulosa cells (GCs) increased significantly after light shading. Enzyme linked immunosorbent assay results revealed that the levels of total antioxidant capacity, malondialdehyde, superoxide dismutase (SOD), glutathione, catalase (CAT), and other substances in the sera, livers, ovaries, and follicular GCs of laying hens increased significantly after shading for 7 days; and reactive oxygen species (ROS) levels in the livers of laying hens also increased significantly. ROS in the serum, ovarian and GCs also increased. After shading for 7 days, the levels of 8-hydroxy-2 deoxyguanosine in the sera and ovarian tissues of laying hens increased significantly. Cell counting kit-8 detection showed that the proliferation activity of GCs in layer follicles decreased after shading for 7 days; the expression level of the anti-apoptotic gene B-cell lymphoma-2 in ovarian tissue and follicular GCs was significantly reduced, and the expression levels of pro-apoptotic caspase 3 (casp3), and SOD, glutathione peroxidase 2 (GPX2), and CAT were all significantly increased. Conclusion: Oxidative stress induced by shading light has a serious inhibitory effect on follicular development during reproduction in laying hens.

Effects of the cathepsin K inhibitor with mineral trioxide aggregate cements on osteoclastic activity

  • Kim, Hee-Sun;Kim, Soojung;Ko, Hyunjung;Song, Minju;Kim, Miri
    • Restorative Dentistry and Endodontics
    • /
    • v.44 no.2
    • /
    • pp.17.1-17.10
    • /
    • 2019
  • Objectives: Root resorption is an unexpected complication after replantation procedures. Combining anti-osteoclastic medicaments with retrograde root filling materials may avert this resorptive activity. The purpose of this study was to assess effects of a cathepsin K inhibitor with calcium silicate-based cements on osteoclastic activity. Methods: MC3T3-E1 cells were cultured for biocompatibility analyses. RAW 264.7 cells were cultured in the presence of the receptor activator of nuclear factor-kappa B and lipopolysaccharide, followed by treatment with Biodentine (BIOD) or ProRoot MTA with or without medicaments (Odanacatib [ODN], a cathepsin inhibitor and alendronate, a bisphosphonate). After drug treatment, the cell counting kit-8 assay and Alizarin red staining were performed to evaluate biocompatibility in MC3T3-E1 cells. Reverse-transcription polymerase chain reaction, tartrate-resistant acid phosphatase (TRAP) staining and enzyme-linked immunosorbent assays were performed in RAW 264.7 cells to determine the expression levels of inflammatory cytokines, interleukin $(IL)-1{\beta}$, IL-6, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and prostaglandin E2 (PGE2). Data were analyzed by one-way analysis of variance and Tukey's post hoc test (p < 0.05). Results: Biocompatibility results showed that there were no significant differences among any of the groups. RAW 264.7 cells treated with BIOD and ODN showed the lowest levels of $TNF-{\alpha}$ and PGE2. Treatments with BIOD + ODN were more potent suppressors of inflammatory cytokine expression (p < 0.05). Conclusion: The cathepsin K inhibitor with calcium silicate-based cement inhibits osteoclastic activity. This may have clinical application in preventing inflammatory root resorption in replanted teeth.

Sex hormones alter the response of Toll-like receptor 3 to its specific ligand in fallopian tube epithelial cells

  • Zandieh, Zahra;Amjadi, Fatemehsadat;Vakilian, Haghighat;Aflatoonian, Khashayar;Amirchaghmaghi, Elham;Fazeli, Alireza;Aflatoonian, Reza
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.45 no.4
    • /
    • pp.154-162
    • /
    • 2018
  • Objective: The fallopian tubes play a critical role in the early events of fertilization. The rapid innate immune defense is an important part of the fallopian tubes. Toll-like receptor 3 (TLR3), as a part of the innate immune system, plays an important role in detecting viral infections. In this basic and experimental study, the effect of sex hormones on the function of TLR3 in the OE-E6/E7 cell line was investigated. Methods: The functionality of TLR3 in this cell line was evaluated by cytokine measurements (interleukin [IL]-6 and IL-1b) and the effects of sex hormones on TLR3 were tested by an enzyme-linked immunosorbent assay kit. Additionally, TLR3 small interfering RNA (siRNA) and a TLR3 function-blocking antibody were used to confirm our findings. Results: The production of IL-6 significantly increased in the presence of polyinosinic-polycytidylic acid (poly(I:C)) as the TLR3 ligand. Using a TLR3-siRNA-ransfected OE-E6/E7 cell line and function-blocking antibody confirmed that cytokine production was due to TLR3. In addition, 17-${\beta}$ estradiol and progesterone suppressed the production of IL-6 in the presence and absence of poly(I:C). Conclusion: These results imply that sex hormones exerted a suppressive effect on the function of TLR3 in the fallopian tube cell line when different concentrations of sex hormones were present. The current results also suggest that estrogen receptor beta and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3, as these two receptors are the main estrogen and progesterone receptors in OEE6/E7 cell line.