• Title/Summary/Keyword: IgG concentration

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Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder

  • Kim, Yesol;Shukla, Shruti;Ahmed, Maruf;Son, Seokmin;Kim, Myunghee;Oh, Sejong
    • Food Science of Animal Resources
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    • v.32 no.6
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    • pp.706-712
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    • 2012
  • The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

The Effect of He-Ne IR Laser Therapy on The Healing of Acute Septic Knee Arthritis in Rat (흰쥐의 화농성 슬관절염 치유에 He-Ne IR레이저 처치의 효과)

  • 김재영;노민희;고진복
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.223-228
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    • 2000
  • The purpose of this study was to find out the effect of He-Ne, IR laser therapy for 4 and 10 minutes on healing of septic arthritis. Sprague-Bawley rats (212.1$\pm$10 g) were randomly assigned to one control and three arthritis groups. Septic arthritis groups were divided into arthritis control, 4 or 10 minute laser therapy groups. Arthritis was experimentally induced by intraperitoneally administration of staphylococcus aureus in citrate buffer(pH 4.3). These result were obtained as follows: AST, LDH, ALP, CK activities and IgG concentration in 10 minute laser group were signifantly increased compare with those in the arthritis group. AST, LDH and CK activities in 4 minute laser group were significantly decreased compare with those in the arthritis group. We think that laser therapy in 4 minute may effective on the healing of septic arthritis. And we will have to study of the effect of laser therapy on healing mechanism, appropriate time of septic arthritis.

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Inhibitory Effects on the Type I hypersensitivity and Inflammatory Reaction of Hwanggigunjung-tang (황기건중탕이 알레르기에 미치는 영향)

  • Lee Jeong Hoon;Hong Hyun Woo;Gam Chul-woo;Park Dong il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1293-1298
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    • 2003
  • HGJT has been used for the treatment of general weakness, digestive organ disease and so on. This study was carried out for the purpose of knowing the inhibitory effect on the Type I hypersensitivity and Inflammatory reactions of Hwanggigunjungtang(HGJT). The reasults were obtained as follows: HGJT(0.1. 0.5. 1, 2mg/g) concentration of dependently inhibited compound48/80 induced anaphylaxis reaction in mice. HGJT(2mg/g) also inhibited permeability of evans blue into peritoneal cavity in mice. HGJT reduced IgE, CRP. WBC and Platelets on egg albumin induced hypersensitivity. But serum NO was grown. According to above results, HGJT may be beneficial in the type I hypersensitivity and Inflammatory reactions by inhibition of histamine release from mast cells.

Development of an enzyme immunoassay for determination of steroid hormones to improve the reproductive efficiency of domestic animals (가축(家畜)의 번식효율증진(繁殖效率增進)을 위한 steroid hormones 의 효소면역분석법(酵素免疫分析法) 개발(開發))

  • Choi, Han-sun;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.611-615
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    • 1993
  • A rapid, solid-phase microtitre plate enzyme immunoassay(EIA) to determine the concentration of progesterone and testosterone in dairy cow is described. Both steroid hormones were analysed employing antibodies against $11{\alpha}$-hemisuccinate-progesterone bovine serum albumin and 4-androsten-$17{\beta}$-ol-3-one-carboxymethyloxime bovine serum albumin, respectively. as primary antibodies and sheep Ig G as secondary antibody. The conjugated used as labels for progesterone and testosterone was progesterone-$11{\alpha}$-hydroxy-hemisuccinate- horseradish peroxidase and 4 ${\alpha}$-androsten-$17{\beta}$-ol-3-hemisuccinate- horseradish peroxidase, respectively. Detection limit of microtitre plate EIA was 6.7 pg/well for progesteone and 1.0 pg/well for testosterone.

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Analysis of Human Serum Amyloid A-1 Concentrations Using a Lateral Flow Immunoassay with CdSe/ZnS Quantum Dots (Human Serum Amyloid A-1 단백질 농도 분석을 위한 CdSe/ZnS 양자점 기반의 Lateral Flow Immunoassay 방법 개발)

  • Fajri, Aidil;Goh, Eunseo;Lee, Sanghyuk;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.30 no.4
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    • pp.429-434
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    • 2019
  • A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

Effect of Mercury Chloride on Humoral and Cell-mediated Immune Responses in Mice (수은이 마우스의 체액성 및 세포성 면역반응에 미치는 영향)

  • Youm, Jung-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.28 no.1 s.49
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    • pp.27-42
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    • 1995
  • The in vivo and in vitro humoral and cell-mediated immune responses of lymphocytes of BALB/c mouse exposed to mercury chloride$(HgCl_2)$ were investigated. In vitro exposure of the splenocytes to mercury chloride produced overt cytotoxicity in 3 hours period. The $IC_{50}$(the concentration required to inhibit a splenocyte viability by 50%) for mercury chloride was >0.1mM for cytotoxicity. In vivo mercury chloride exposed mice were significantly depressed delayed type hypersensitivity(DTH) response to sheep red blood cells(SRBC) in a dose-dependent manner compared with control group. Mercury chloride inhibited the proliferative responses of splenocytes to lipopolysaccharide, pokeweed mitogen, concanavalin A and phytohemagglutinin in a dose-dependent manner. Hemagglutinin response to SRBC in mercury chloride exposed mice was significantly depressed in a dose-dependent manner compared with control group. After 7 weeks of mercury chloride exposure in vivo, mercury chloride induced an increase of nonspecific serum $IgG_1$ and IgE levels in BALB/c mice.

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Reevaluation of the Necessity of Iron Injection to Newborn Piglets

  • Yu, I.T.;Lin, J.;Wu, J.F.;Yen, H.T.;Lee, S.L.;Yang, T.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.1
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    • pp.79-83
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    • 2002
  • The growth and immune responses to endotoxin lipopolysaccharide (LPS) challenge ( $20{\mu}g/kg$) of piglets with and without a iron dextran injection (Fe, 200 mg/head) two days after birth are compared. Sixty-four newborn piglets from eight litters were allocated randomly to one of four treatments. The control received no iron dextran and only saline (Sal) injection on the second and fifteenth day after birth (Sal-Sal). The remaining three groups received Fe-Sal, Sal-LPS, Fe-LPS treatments respectively. On fifteen days of age, blood samples of piglets were taken at 0 h, 1 h, 2 h and 4 d after saline or LPS injection to determine immune functions and blood characteristics. The trial terminated when the pig reached 56 days and the average daily gain of piglets was then measured. Daily gain, serum immunoglobulin G (IgG) concentration and red blood cell counts did not vary significantly among the four groups at any measuring times. Serum tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) concentration increased sharply 1 h after LPS injection. However, iron injection did not change TNF-${\alpha}$ concentration responds to LPS injection. White blood cell counts of two LPS injection groups were significantly lowered 1 h following the injection. In contrast, serum lactoferrin concentration had increased significantly 1 and 2 h postinjection. Furthermore, iron injection produced no further effects on these two criteria. Iron injection increased the hemoglobin (Hb) concentration of piglets at any measuring time, and LPS injection lowered Hb concentration. In conclusion, a 200 mg/head of iron dextran injection on the second day after birth increased Hb concentration, had no detrimental effect on the immune responses and growth of piglets. Moreover, if creep feed (175 mg Fe/kg feed) is provided from d 7 after birth, the Fe-injection does not contribute to overall performance of piglets and may not be a necessity in practice.

Anti-inflammatory Effects of Gamiyunjo-tang on Lipopolysaccharide Induced Inflammatory Responses in RAW 264.7 Cells (가미윤조탕(加味潤燥湯)이 LPS로 유도된 RAW 264.7 대식세포에서의 항염 효과 연구)

  • Choi, Jong-Min;Kim, Yong-Min;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.28 no.1
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    • pp.23-31
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    • 2015
  • Objectives : Allergic disease has been well known as an IgE-dependent immunologic response. Recently, interest about the late inflammatory reaction has grown up as well as early allergic reaction characterized by IgE and mast cell. The purpose of this study was to find the anti-inflammatory effect of Gamiyunjo-tang(GMYJT) in allergic reaction. Methods : The experiment was performed using Raw 264.7 cells pretreated with GMYJT extracts. In this study, we observed the toxicity of cells by MTT analysis and measured the production of LPS-induced NO, $PGE_2$, IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ at a concentration of 50, 100, 200 and $400{\mu}g/ml$. Results : No toxicity of GMYJT (50, 100, 200, $400{\mu}g/ml$) on RAW 264.7 cells was found after 24 hours incubation. LPS-induced NO production was reduced after treatment with GMYJT (100, 200, $400{\mu}g/ml$)(P<0.05). $PGE_2$ was reduced after treatment with GMYJT (100, 200, $400{\mu}g/ml$)(P<0.05). IL-$1{\beta}$ did not decrease at any dose. IL-6 decreased at 200, $400{\mu}g/ml$(P<0.05). TNF-${\alpha}$ production decreased only at $400{\mu}g/ml$(P<0.05). Conclusions : These data suggest that GMYJT has anti-inflammatory effects in late allergic reaction.

Production of a Specific Yolk Antibody against Enterotoxigenic E. coli F41 Fimbrial Antigen (장독성대장균 F41 섬모항원에 대한 특이난황항체 생산)

  • Shin, S.O.;Kim, J.W.
    • Journal of Animal Science and Technology
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    • v.44 no.5
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    • pp.633-642
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    • 2002
  • Enteric colibacillosis has economically become an important disease of young animals as a result of increasing intensification of farrowing management. The objective of this experiment is to isolate fimbrial antigen from enterotoxigenic E. coli F41, to develop specific polyclonal IgY which can effectively neutralize or reduce the proliferation of pathogens in feed or living animal system, and to apply IgY technologies to animal industry. The results obtained were as follows: The molecular weight of the purified F41 antigen was 29,500 dalton on sodium dodecyl sulfate-polyacrylamide gels. Fimbrial antigen was confirmed by the western blot method. It was observed that after immunization the level of serum antibody titer of laying hen was shown in two weeks and gradually increased. The antibody titer in egg yolk appeared two weeks after it was shown in serum antibody. The titers of egg yolk antibody were gradually increased to the maximum level of 320,000 (antigen 50${\mu}g$/$m\ell$), 450,000 (antigen 200${\mu}g$/$m\ell$), and 320,000 (antigen 600${\mu}g$/$m\ell$). According to the results of specificity test by ELISA, the anti-F41 antibodies from chicken serum and egg yolk reacted only with ETEC F41 antigen. There was no cross reaction with other ETEC strains (K88, K99, and 987P). In vitro condition, as a result of antigen binding ability of yolk antibodies, bacterial concentration was rapidly decreased to $10^5$ CFU/$m\ell$ from $10^9$ CFU/$m\ell$ when 2${\sim}$4 mg/$m\ell$ of freeze dried WSF (water soluble fraction) was added.

Effects of Bursectomy and Thyectomy on the Immunoglobulins in Korean Ogol Chickens (오골계의 Fabricius 낭과 흉선 적출이 Immunoglobulin 에 미치는 영향)

  • 김상근;김민수
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.119-125
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    • 1999
  • The concentration of serum proteins, lipoproteins, leukocytes and lymphocytes in bursectomy and thymectomy Ogol chickens were examined from 5 days to 150 days of age. Concentration of serum proteins and lipoproteins were measured by cellulose acetate electrophoresis. The results summarized as follows : 1. The gamma-globulin concentrations in growing Korean native chickens and Ogol chickens were increased 0.41$\pm$0.01~0.85$\pm$0.05 mg/㎗ and 0.50$\pm$0.01~0.98$\pm$0.08 mg/㎗ from 1 day after to 5 days of age and gradually decreased at 10 days of age, and thereafter increased 0.82$\pm$0.06 mg/㎗, 1.09$\pm$0.04 mg/㎗ at 100 days of age. 2. The serum apha-, beta-, gamma-lipoprotein concentrations of growing Ogol chickens were 74.1$\pm$6.8~240.2$\pm$9.7 mg/㎗, 89.7 5.7~225.8$\pm$11.3 mg/㎗ and 87.6$\pm$4.7-220.3$\pm$10 mg/㎗, respectively. The serum -lipoprotein concentrations from 5 days to 150 days of age Ogol chickens were significantly decreased. 3. The leukocyte and lymphocyte counts in bursectomy and thymectomy Ogol chickens slightly increased from 10 to 100 days of age. However the leukocyte and lymphocyte counts in thymectomy chickens were lower than in untreated chickens. 4. The Ig concentrations of 1 day to 50 days(0.30$\pm$0.03~0.58$\pm$0.04 g/㎗) in bursectomy chickens were similar to those found in untreated chickens and thereafer increased from 50 to 150 days of age(0.58$\pm$0.04~1.21$\pm$0.05 g/㎗). The Ig concentrations in thymectomy chickens increased significantly.

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