bovine ifnγ  (Kingfisher Biotech)


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    Name:
    Bovine IFN gamma Do It Yourself ELISA
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    DIY0665B-003
    Price:
    700.0
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    1 Set
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    Structured Review

    Kingfisher Biotech bovine ifnγ
    Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) <t>IFNγ</t> (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for

    https://www.bioz.com/result/bovine ifnγ/product/Kingfisher Biotech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bovine ifnγ - by Bioz Stars, 2021-09
    93/100 stars

    Images

    1) Product Images from "Pathogenesis of percutaneous infection of goats with Burkholderia pseudomallei: clinical, pathologic, and immunological responses in chronic melioidosis"

    Article Title: Pathogenesis of percutaneous infection of goats with Burkholderia pseudomallei: clinical, pathologic, and immunological responses in chronic melioidosis

    Journal: International Journal of Experimental Pathology

    doi: 10.1111/iep.12068

    Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) IFNγ (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for
    Figure Legend Snippet: Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) IFNγ (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for

    Techniques Used: Infection

    2) Product Images from "Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells"

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    Journal: Molecules and Cells

    doi: 10.14348/molcells.2016.2358

    IFN-γ downregulates CSN2 synthesis and decreases the triglyceride content via an autophagy-dependent mechanism. (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P
    Figure Legend Snippet: IFN-γ downregulates CSN2 synthesis and decreases the triglyceride content via an autophagy-dependent mechanism. (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Techniques Used: Expressing

    IFN-γ enhances autophagic flux in BMECs. (A) The cells were treated with IFN-γ in the presence or absence of E64d (10 μg/ml). After 24 h, cell lysates were prepared and analyzed via immunoblotting using anti-MAP1LC3, anti-SQSTM1 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. Oneway ANOVA; * P
    Figure Legend Snippet: IFN-γ enhances autophagic flux in BMECs. (A) The cells were treated with IFN-γ in the presence or absence of E64d (10 μg/ml). After 24 h, cell lysates were prepared and analyzed via immunoblotting using anti-MAP1LC3, anti-SQSTM1 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. Oneway ANOVA; * P

    Techniques Used:

    IFN-γ activates autophagy in primary BMECs in vitro . (A) The BMECs were treated with IFN-γ, anti-IFNGR1, and anti-IFNGR2 alone or in combination for 6, 24 and 48 h. The MAP1LC3, ATG12-ATG5 and GAPDH expression levels were analyzed by Western blot analysis with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P
    Figure Legend Snippet: IFN-γ activates autophagy in primary BMECs in vitro . (A) The BMECs were treated with IFN-γ, anti-IFNGR1, and anti-IFNGR2 alone or in combination for 6, 24 and 48 h. The MAP1LC3, ATG12-ATG5 and GAPDH expression levels were analyzed by Western blot analysis with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Techniques Used: In Vitro, Expressing, Western Blot

    IFN-γ downregulates milk synthesis in primary BMECs in vitro . (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P
    Figure Legend Snippet: IFN-γ downregulates milk synthesis in primary BMECs in vitro . (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Techniques Used: In Vitro, Expressing

    Arginine supplementation assists in recovering milk synthesis that had been reduced by IFN-γ-induced autophagy. (A) Arginine supplementation partially inhibits the effects of IFN-γ on BMECs. The BMECs were incubated with 10 ng/ml IFN-γ alone or with the indicated concentration of arginine or glycine (as control) or were left untreated. The expression levels of the t-GCN2, p-GCN2, t-EIF2S1, p-EIF2S1, ATF4, MAP1LC3, STAT5, ELF5, CSN2 and GAPDH proteins were evaluated by immunoblotting. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (B) Effects of 15 different amino acid supplements on LC3II accumulation. The BMECs were incubated with 10 ng/ml IFN-γ alone or with a 4-fold increase in the endogenous level of Asp, Thr, Ser, Gln, Gly, Ala, Val, Met, Ile, Leu, Tyr, Phe, Lys, His and Pro for 24 h. At the end of the treatment, the expression levels of MAP1LC3-II and β-actin (loading control) were analyzed by immunoblotting with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (C) The CSN2 content was detected by ELISA. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P
    Figure Legend Snippet: Arginine supplementation assists in recovering milk synthesis that had been reduced by IFN-γ-induced autophagy. (A) Arginine supplementation partially inhibits the effects of IFN-γ on BMECs. The BMECs were incubated with 10 ng/ml IFN-γ alone or with the indicated concentration of arginine or glycine (as control) or were left untreated. The expression levels of the t-GCN2, p-GCN2, t-EIF2S1, p-EIF2S1, ATF4, MAP1LC3, STAT5, ELF5, CSN2 and GAPDH proteins were evaluated by immunoblotting. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (B) Effects of 15 different amino acid supplements on LC3II accumulation. The BMECs were incubated with 10 ng/ml IFN-γ alone or with a 4-fold increase in the endogenous level of Asp, Thr, Ser, Gln, Gly, Ala, Val, Met, Ile, Leu, Tyr, Phe, Lys, His and Pro for 24 h. At the end of the treatment, the expression levels of MAP1LC3-II and β-actin (loading control) were analyzed by immunoblotting with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (C) The CSN2 content was detected by ELISA. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Techniques Used: Incubation, Concentration Assay, Expressing, Enzyme-linked Immunosorbent Assay

    Activation of the GCN2/eIF2α pathway mediated IFN-γ-induced autophagy. (A) IFN-γ does not interfere with mTOR signaling. The BMECs were incubated with 10 ng/ml of IFN-γ or 10 nM rapamycin or were left untreated for 24 or 48 h. Whole-cell lysates were prepared and analyzed via Western blot analysis using anti-mTOR, anti-phospho-mTOR, anti-phospho-4E-BP1, anti-MAP1LC3 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P
    Figure Legend Snippet: Activation of the GCN2/eIF2α pathway mediated IFN-γ-induced autophagy. (A) IFN-γ does not interfere with mTOR signaling. The BMECs were incubated with 10 ng/ml of IFN-γ or 10 nM rapamycin or were left untreated for 24 or 48 h. Whole-cell lysates were prepared and analyzed via Western blot analysis using anti-mTOR, anti-phospho-mTOR, anti-phospho-4E-BP1, anti-MAP1LC3 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Techniques Used: Activation Assay, Incubation, Western Blot

    3) Product Images from "Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves"

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves

    Journal: Journal of Animal Science

    doi: 10.1093/jas/skaa252

    SCFP treatment does not alter BRSV-specific T cell responses in the blood. Peripheral blood was collected on day 10 after BRSV infection. PBMCs were isolated and cryopreserved. (A) PBMCs were thawed and labeled with CellTrace Violet proliferation dye, and 5 × 10 5 cells/well were cultured for 6 d in the presence or absence of 0.01 MOI BRSV strain 375. CD4 (left) and CD8 (right) T cell proliferation was analyzed by flow cytometry for CellTrace dilution. Control wells remained unstimulated (Mock). T cell proliferation was analyzed using the gating strategy presented in Supplementary Figure S1 . Background proliferation was subtracted, and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data are presented as means ± SEM. P -values were determined by student’s t- test. (B) Virus-specific IFNγ production was analyzed in PBMCs on day 10 postinfection; 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S2 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (C and D) Cell culture supernatants were collected from the PBMCs cultures in (A) and analyzed by commercial ELISA kit for (C) IFNγ and (D) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.
    Figure Legend Snippet: SCFP treatment does not alter BRSV-specific T cell responses in the blood. Peripheral blood was collected on day 10 after BRSV infection. PBMCs were isolated and cryopreserved. (A) PBMCs were thawed and labeled with CellTrace Violet proliferation dye, and 5 × 10 5 cells/well were cultured for 6 d in the presence or absence of 0.01 MOI BRSV strain 375. CD4 (left) and CD8 (right) T cell proliferation was analyzed by flow cytometry for CellTrace dilution. Control wells remained unstimulated (Mock). T cell proliferation was analyzed using the gating strategy presented in Supplementary Figure S1 . Background proliferation was subtracted, and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data are presented as means ± SEM. P -values were determined by student’s t- test. (B) Virus-specific IFNγ production was analyzed in PBMCs on day 10 postinfection; 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S2 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (C and D) Cell culture supernatants were collected from the PBMCs cultures in (A) and analyzed by commercial ELISA kit for (C) IFNγ and (D) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Techniques Used: Infection, Isolation, Labeling, Cell Culture, Flow Cytometry, In Vitro, Staining, Expressing, Enzyme-linked Immunosorbent Assay

    Impact of SCFP treatment on BRSV-specific T cell responses in the BAL. BAL samples were collected from all calves (SCFP treated and controls) on day 10 postinfection and cryopreserved. (A) BAL cells were thawed and 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S3 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (B and C) BAL cells were thawed and plated at 1 × 10 6 cells/well and stimulated in vitro with 0.01 MOI BRSV strain 375 for 72 h. Culture supernatants were collected and analyzed by commercial ELISA kit for (B) IFNγ and (C) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.
    Figure Legend Snippet: Impact of SCFP treatment on BRSV-specific T cell responses in the BAL. BAL samples were collected from all calves (SCFP treated and controls) on day 10 postinfection and cryopreserved. (A) BAL cells were thawed and 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S3 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (B and C) BAL cells were thawed and plated at 1 × 10 6 cells/well and stimulated in vitro with 0.01 MOI BRSV strain 375 for 72 h. Culture supernatants were collected and analyzed by commercial ELISA kit for (B) IFNγ and (C) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Techniques Used: In Vitro, Staining, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    4) Product Images from "Increased Susceptibility of Cattle to Intranasal RVFV Infection"

    Article Title: Increased Susceptibility of Cattle to Intranasal RVFV Infection

    Journal: Frontiers in Veterinary Science

    doi: 10.3389/fvets.2020.00137

    Interferons alpha, beta and gamma in nasal and oral swabs. IFN-α (A,D) , IFN-β (B,E) , and IFN-γ (C,F) were measured in nasal and oral swabs by ELISA; each value represents an individual animal. The horizontal dashed line indicates the diagnostic detection limit of the RT-PCR assay.
    Figure Legend Snippet: Interferons alpha, beta and gamma in nasal and oral swabs. IFN-α (A,D) , IFN-β (B,E) , and IFN-γ (C,F) were measured in nasal and oral swabs by ELISA; each value represents an individual animal. The horizontal dashed line indicates the diagnostic detection limit of the RT-PCR assay.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Reverse Transcription Polymerase Chain Reaction

    Related Articles

    other:

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells
    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Article Title: Increased Susceptibility of Cattle to Intranasal RVFV Infection
    Article Snippet: The inoculum was then removed and the cells were overlayed with 2 ml 1.75% carboxymethylcellulose (CMC).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Effects of increasing Fe dosage in newborn pigs on suckling and subsequent nursery performance and hematological and immunological criteria
    Article Snippet: .. Interferon-gamma response was measured by subjecting blood samples to 100 μL of phytohemagglutinin (PHA) and placing in 96-well ELISA plates coated with IFN-γ (Kingfisher, Biotech, Inc.) and absorbance measured using a microtech spectrometer (BioTek EON). ..

    Article Title: Immunization-Induced Anaplasma marginale-Specific T-Lymphocyte Responses Impaired by A. marginale Infection Are Restored after Eliminating Infection with Tetracycline
    Article Snippet: .. ELISAs were performed to measure bovine IL-2 and TNF-α with DuoSet ELISA kits from R & D Systems, and bovine IFN-γ was measured with an ELISA VetSet kit from Kingfisher Biotech, Inc., according to manufacturer's specifications. ..

    Article Title: Pathogenesis of percutaneous infection of goats with Burkholderia pseudomallei: clinical, pathologic, and immunological responses in chronic melioidosis
    Article Snippet: .. Analysis was performed using commercially available enzyme-linked immunosorbent assay (ELISA) kits for bovine CCL2 (VS0083B; Kingfisher Biotech, Inc., St. Paul, MN, USA), bovine IFNγ (VS0257B; Kingfisher Biotech, Inc.), bovine IL-10 (BV1495; TSZ ELISA, Framingham, MA, USA) and TGF-β1 (MB100B; R & D Systems, Minneapolis, MN, USA). ..

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves
    Article Snippet: .. The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN. ..

    Concentration Assay:

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves
    Article Snippet: .. The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN. ..

    Cell Culture:

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves
    Article Snippet: .. The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN. ..

    Intra Assay:

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves
    Article Snippet: .. The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN. ..

    Inter Assay:

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves
    Article Snippet: .. The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN. ..

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  • 93
    Kingfisher Biotech bovine ifnγ
    Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) <t>IFNγ</t> (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for
    Bovine Ifnγ, supplied by Kingfisher Biotech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bovine ifnγ/product/Kingfisher Biotech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bovine ifnγ - by Bioz Stars, 2021-09
    93/100 stars
      Buy from Supplier

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    Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) IFNγ (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for

    Journal: International Journal of Experimental Pathology

    Article Title: Pathogenesis of percutaneous infection of goats with Burkholderia pseudomallei: clinical, pathologic, and immunological responses in chronic melioidosis

    doi: 10.1111/iep.12068

    Figure Lengend Snippet: Cytokine production in goats infected with B. pseudomallei . Responses of (a) TGF-β1 (log pg/ml), (b) IFNγ (log ng/ml) and (c) CCL2 (log ng/ml) in aerosol ( ) and percutaneous ( ) groups. Significant elevations were only present for

    Article Snippet: Analysis was performed using commercially available enzyme-linked immunosorbent assay (ELISA) kits for bovine CCL2 (VS0083B; Kingfisher Biotech, Inc., St. Paul, MN, USA), bovine IFNγ (VS0257B; Kingfisher Biotech, Inc.), bovine IL-10 (BV1495; TSZ ELISA, Framingham, MA, USA) and TGF-β1 (MB100B; R & D Systems, Minneapolis, MN, USA).

    Techniques: Infection

    IFN-γ downregulates CSN2 synthesis and decreases the triglyceride content via an autophagy-dependent mechanism. (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: IFN-γ downregulates CSN2 synthesis and decreases the triglyceride content via an autophagy-dependent mechanism. (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques: Expressing

    IFN-γ enhances autophagic flux in BMECs. (A) The cells were treated with IFN-γ in the presence or absence of E64d (10 μg/ml). After 24 h, cell lysates were prepared and analyzed via immunoblotting using anti-MAP1LC3, anti-SQSTM1 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. Oneway ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: IFN-γ enhances autophagic flux in BMECs. (A) The cells were treated with IFN-γ in the presence or absence of E64d (10 μg/ml). After 24 h, cell lysates were prepared and analyzed via immunoblotting using anti-MAP1LC3, anti-SQSTM1 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. Oneway ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques:

    IFN-γ activates autophagy in primary BMECs in vitro . (A) The BMECs were treated with IFN-γ, anti-IFNGR1, and anti-IFNGR2 alone or in combination for 6, 24 and 48 h. The MAP1LC3, ATG12-ATG5 and GAPDH expression levels were analyzed by Western blot analysis with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: IFN-γ activates autophagy in primary BMECs in vitro . (A) The BMECs were treated with IFN-γ, anti-IFNGR1, and anti-IFNGR2 alone or in combination for 6, 24 and 48 h. The MAP1LC3, ATG12-ATG5 and GAPDH expression levels were analyzed by Western blot analysis with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques: In Vitro, Expressing, Western Blot

    IFN-γ downregulates milk synthesis in primary BMECs in vitro . (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: IFN-γ downregulates milk synthesis in primary BMECs in vitro . (A) IFN-γ induces the expression of the CSN2 and ACACA mRNAs. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques: In Vitro, Expressing

    Arginine supplementation assists in recovering milk synthesis that had been reduced by IFN-γ-induced autophagy. (A) Arginine supplementation partially inhibits the effects of IFN-γ on BMECs. The BMECs were incubated with 10 ng/ml IFN-γ alone or with the indicated concentration of arginine or glycine (as control) or were left untreated. The expression levels of the t-GCN2, p-GCN2, t-EIF2S1, p-EIF2S1, ATF4, MAP1LC3, STAT5, ELF5, CSN2 and GAPDH proteins were evaluated by immunoblotting. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (B) Effects of 15 different amino acid supplements on LC3II accumulation. The BMECs were incubated with 10 ng/ml IFN-γ alone or with a 4-fold increase in the endogenous level of Asp, Thr, Ser, Gln, Gly, Ala, Val, Met, Ile, Leu, Tyr, Phe, Lys, His and Pro for 24 h. At the end of the treatment, the expression levels of MAP1LC3-II and β-actin (loading control) were analyzed by immunoblotting with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (C) The CSN2 content was detected by ELISA. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: Arginine supplementation assists in recovering milk synthesis that had been reduced by IFN-γ-induced autophagy. (A) Arginine supplementation partially inhibits the effects of IFN-γ on BMECs. The BMECs were incubated with 10 ng/ml IFN-γ alone or with the indicated concentration of arginine or glycine (as control) or were left untreated. The expression levels of the t-GCN2, p-GCN2, t-EIF2S1, p-EIF2S1, ATF4, MAP1LC3, STAT5, ELF5, CSN2 and GAPDH proteins were evaluated by immunoblotting. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (B) Effects of 15 different amino acid supplements on LC3II accumulation. The BMECs were incubated with 10 ng/ml IFN-γ alone or with a 4-fold increase in the endogenous level of Asp, Thr, Ser, Gln, Gly, Ala, Val, Met, Ile, Leu, Tyr, Phe, Lys, His and Pro for 24 h. At the end of the treatment, the expression levels of MAP1LC3-II and β-actin (loading control) were analyzed by immunoblotting with specific antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. (C) The CSN2 content was detected by ELISA. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques: Incubation, Concentration Assay, Expressing, Enzyme-linked Immunosorbent Assay

    Activation of the GCN2/eIF2α pathway mediated IFN-γ-induced autophagy. (A) IFN-γ does not interfere with mTOR signaling. The BMECs were incubated with 10 ng/ml of IFN-γ or 10 nM rapamycin or were left untreated for 24 or 48 h. Whole-cell lysates were prepared and analyzed via Western blot analysis using anti-mTOR, anti-phospho-mTOR, anti-phospho-4E-BP1, anti-MAP1LC3 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Journal: Molecules and Cells

    Article Title: Arginine Supplementation Recovered the IFN-γ-Mediated Decrease in Milk Protein and Fat Synthesis by Inhibiting the GCN2/eIF2α Pathway, Which Induces Autophagy in Primary Bovine Mammary Epithelial Cells

    doi: 10.14348/molcells.2016.2358

    Figure Lengend Snippet: Activation of the GCN2/eIF2α pathway mediated IFN-γ-induced autophagy. (A) IFN-γ does not interfere with mTOR signaling. The BMECs were incubated with 10 ng/ml of IFN-γ or 10 nM rapamycin or were left untreated for 24 or 48 h. Whole-cell lysates were prepared and analyzed via Western blot analysis using anti-mTOR, anti-phospho-mTOR, anti-phospho-4E-BP1, anti-MAP1LC3 and anti-GAPDH antibodies. The relative levels of the target proteins were estimated using densitometry, and the ratios were calculated relative to the GAPDH control. The data represent the mean ± SD of 3 independent experiments. Each bar represents the mean of three independent experiments. One-way ANOVA; * P

    Article Snippet: Reagents Bovine IFN-γ was purchased from the Kingfisher Group (King-fisher Biotech, Inc., USA).

    Techniques: Activation Assay, Incubation, Western Blot

    SCFP treatment does not alter BRSV-specific T cell responses in the blood. Peripheral blood was collected on day 10 after BRSV infection. PBMCs were isolated and cryopreserved. (A) PBMCs were thawed and labeled with CellTrace Violet proliferation dye, and 5 × 10 5 cells/well were cultured for 6 d in the presence or absence of 0.01 MOI BRSV strain 375. CD4 (left) and CD8 (right) T cell proliferation was analyzed by flow cytometry for CellTrace dilution. Control wells remained unstimulated (Mock). T cell proliferation was analyzed using the gating strategy presented in Supplementary Figure S1 . Background proliferation was subtracted, and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data are presented as means ± SEM. P -values were determined by student’s t- test. (B) Virus-specific IFNγ production was analyzed in PBMCs on day 10 postinfection; 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S2 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (C and D) Cell culture supernatants were collected from the PBMCs cultures in (A) and analyzed by commercial ELISA kit for (C) IFNγ and (D) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Journal: Journal of Animal Science

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves

    doi: 10.1093/jas/skaa252

    Figure Lengend Snippet: SCFP treatment does not alter BRSV-specific T cell responses in the blood. Peripheral blood was collected on day 10 after BRSV infection. PBMCs were isolated and cryopreserved. (A) PBMCs were thawed and labeled with CellTrace Violet proliferation dye, and 5 × 10 5 cells/well were cultured for 6 d in the presence or absence of 0.01 MOI BRSV strain 375. CD4 (left) and CD8 (right) T cell proliferation was analyzed by flow cytometry for CellTrace dilution. Control wells remained unstimulated (Mock). T cell proliferation was analyzed using the gating strategy presented in Supplementary Figure S1 . Background proliferation was subtracted, and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data are presented as means ± SEM. P -values were determined by student’s t- test. (B) Virus-specific IFNγ production was analyzed in PBMCs on day 10 postinfection; 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S2 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (C and D) Cell culture supernatants were collected from the PBMCs cultures in (A) and analyzed by commercial ELISA kit for (C) IFNγ and (D) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Article Snippet: The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN.

    Techniques: Infection, Isolation, Labeling, Cell Culture, Flow Cytometry, In Vitro, Staining, Expressing, Enzyme-linked Immunosorbent Assay

    Impact of SCFP treatment on BRSV-specific T cell responses in the BAL. BAL samples were collected from all calves (SCFP treated and controls) on day 10 postinfection and cryopreserved. (A) BAL cells were thawed and 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S3 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (B and C) BAL cells were thawed and plated at 1 × 10 6 cells/well and stimulated in vitro with 0.01 MOI BRSV strain 375 for 72 h. Culture supernatants were collected and analyzed by commercial ELISA kit for (B) IFNγ and (C) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Journal: Journal of Animal Science

    Article Title: Supplementing a Saccharomyces cerevisiae fermentation product modulates innate immune function and ameliorates bovine respiratory syncytial virus infection in neonatal calves

    doi: 10.1093/jas/skaa252

    Figure Lengend Snippet: Impact of SCFP treatment on BRSV-specific T cell responses in the BAL. BAL samples were collected from all calves (SCFP treated and controls) on day 10 postinfection and cryopreserved. (A) BAL cells were thawed and 1 × 10 6 cells/well were stimulated in vitro with 0.01 MOI BRSV strain 375 for 16 h. Cells were then stained for intracellular IFNγ expression and analyzed by flow cytometry using the gating strategy outlined in Supplementary Figure S3 . The frequency of circulating CD4 + IFNγ + (left) and CD8 + IFNγ + (right) is shown. Background IFNγ + production was subtracted and results represent change over unstimulated samples; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM. P -values were determined by student’s t- test. (B and C) BAL cells were thawed and plated at 1 × 10 6 cells/well and stimulated in vitro with 0.01 MOI BRSV strain 375 for 72 h. Culture supernatants were collected and analyzed by commercial ELISA kit for (B) IFNγ and (C) IL-17; n = 10 control calves; n = 12 SCFP calves. Data represent means ± SEM.

    Article Snippet: The concentration of cytokines in cell culture supernatants was determined using bovine commercial ELISA kits for IFNγ (intra-assay coefficient of variation [CV] 3.6% and inter-assay CV 6.1%), interleukin ( IL)-17A (intra-assay CV 6.7% and inter-assay CV 7.6%), IL-6 (intra-assay CV 4.1% and inter-assay CV 5.0%), and TNFα (intra-assay CV 4.3% and inter-assay CV 17.0%) from Kingfisher Biotech, Inc. Minneapolis, MN.

    Techniques: In Vitro, Staining, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    Interferons alpha, beta and gamma in nasal and oral swabs. IFN-α (A,D) , IFN-β (B,E) , and IFN-γ (C,F) were measured in nasal and oral swabs by ELISA; each value represents an individual animal. The horizontal dashed line indicates the diagnostic detection limit of the RT-PCR assay.

    Journal: Frontiers in Veterinary Science

    Article Title: Increased Susceptibility of Cattle to Intranasal RVFV Infection

    doi: 10.3389/fvets.2020.00137

    Figure Lengend Snippet: Interferons alpha, beta and gamma in nasal and oral swabs. IFN-α (A,D) , IFN-β (B,E) , and IFN-γ (C,F) were measured in nasal and oral swabs by ELISA; each value represents an individual animal. The horizontal dashed line indicates the diagnostic detection limit of the RT-PCR assay.

    Article Snippet: ELISA for Interferons We used bovine interferon alpha (IFN-αA), beta and gamma ELISA kits (Kingfisher Biotech Inc., USA) to detect protein in nasal and oral swabs.

    Techniques: Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Reverse Transcription Polymerase Chain Reaction