Journal: Journal of Animal Science

Article Title: Effects of increasing Fe dosage in newborn pigs on suckling and subsequent nursery performance and hematological and immunological criteria

doi: 10.1093/jas/skaa221

Figure Lengend Snippet: Effects of Fe dosage on immune function criteria at weaning (21-d postfarrowing) 1

Article Snippet: TNF-α response was measured by subjecting blood samples to 800 μL of lipopolysaccharide (LPS) and placing in 96-well enzyme-linked immunosorbent assay (ELISA) plates coated with anti-swine TNF-α polyconal antibody (Kingfisher, Biotech, Inc., St. Paul, MN) and absorbance was measured using a microtech spectrometer (BioTek EON, Winooski, VT).

Techniques: Clinical Proteomics

Journal: Frontiers in Veterinary Science

Article Title: Nexus Between Immune Responses and Oxidative Stress: The Role of Dietary Hydrolyzed Lignin in ex vivo Bovine Peripheral Blood Mononuclear Cell Response

doi: 10.3389/fvets.2020.00009

Figure Lengend Snippet: Interleukin (IL)-10 (A) , IL-12 (B) , IL-8 (C) , interferon-(IFN)-γ, (D) , tumor necrosis factor (TNF)-α, and (E) secretion by PBMCs unstimulated (UNC) and stimulated with LPS and ConA (SC, 1 and 5 μg/ml, respectively) and with 4 mM of H 2 O 2 for 24 h at 37°C/5% CO 2 . * P < 0.05 were considered significant among feeding strategy.

Article Snippet: Biotinylated secondary anti-bovine IFN-γ antibody (2 μg/ml, final concentration, AbD Serotec) and anti-bovine TNF-α antibody (2.5 μg/ml, final concentration in PBS, Kingfisher Biotech) dissolved in PBS were added for 45 min. For the IL-8 antibody conjunction, the rabbit anti-sheep interleukin-8 (2 μg/ml, final concentration in PBS, AbD Serotec) was used and added to the wells for 1 h. After washing four times, 100 μl of streptavidin–horseradish peroxidase (HRP) (1/500, AbD Serotec) was added for IFN-γ and TNF-α assay, whereas, for IL-8, a goat anti-rabbit IgG HRP-conjugated antibody (1/20,000 in PBS, Sigma-Aldrich, Italy) was added into wells.

Techniques:

Journal: Immunity, Inflammation and Disease

Article Title: Bovine leukemia virus reduces anti‐viral cytokine activities and NK cytotoxicity by inducing TGF‐β secretion from regulatory T cells

doi: 10.1002/iid3.93

Figure Lengend Snippet: Reduced anti‐viral cytokine production in BLV‐infected cattle. IFN‐γ (A) and TNF‐α (B) in the supernatants of cultured PBMCs from BLV‐infected aleukemic (AL: n = 9) or persistent lymphocytotic (PL: n = 7) cattle and uninfected cattle ( n = 4) were determined using ELISA. Detection of IFN‐γ producing CD4 + T cells in BLV‐uninfected and BLV‐infected cattle at different disease stages (C: BLV(−): n = 7, AL: n = 14, PL: n = 13). Correlations between IFN‐γ producing CD4 + T cells and lymphocyte numbers (D) and proviral loads (E), TNF‐α producing CD4 + T cells, lymphocyte numbers (F), and proviral loads (G) in BLV‐infected cattle ( n = 27). Lymphocyte numbers in BLV‐infected cattle were counted using Celltac α MEK‐6450 and animals were diagnosed with PL when at least two consecutive lymphocyte counts were >7,500/µL; proviral loads were quantified using real‐time PCR. IFN‐γ‐ and TNF‐α‐producing CD4 + T cells in BLV‐infected cattle were detected using flow cytometry.

Article Snippet: After 24 h, supernatants were harvested and IFN‐γ (Mabtech, Nacka Strand, Sweden) and TNF‐α (Kingfisher Biotech) production was determined using ELISA kits with a microplate reader MTP‐650FA (Corona Electric, Ibaraki, Japan) at an absorbance of 450 nm.

Techniques: Infection, Cell Culture, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Flow Cytometry

Journal: Immunity, Inflammation and Disease

Article Title: Bovine leukemia virus reduces anti‐viral cytokine activities and NK cytotoxicity by inducing TGF‐β secretion from regulatory T cells

doi: 10.1002/iid3.93

Figure Lengend Snippet: Inhibition of IFN‐γ and TNF‐α production from CD4 + T cells by TGF‐β. PBMCs from normal cattle (IFN‐γ: n = 12, TNF‐α: n = 9) and BLV‐infected cattle ( n = 11) were pretreated with TGF‐β for 2 h and were cultivated with IL‐2 alone (A and B) or synthesized peptides from the BLV envelop region (C and D). IFN‐γ‐ or TNF‐α‐producing CD4 + T cells were detected using flow cytometry.

Article Snippet: After 24 h, supernatants were harvested and IFN‐γ (Mabtech, Nacka Strand, Sweden) and TNF‐α (Kingfisher Biotech) production was determined using ELISA kits with a microplate reader MTP‐650FA (Corona Electric, Ibaraki, Japan) at an absorbance of 450 nm.

Techniques: Inhibition, Infection, Synthesized, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Transcriptomic Insights into Tumor Necrosis Factor α’s Role in the Fibrosis-Related Processes of Equine Endometrial Fibroblasts

doi: 10.3390/ijms262311344

Figure Lengend Snippet: ( A ) A dot plot illustrating Gene Ontology (GO) pathway enrichment analysis of the differentially expressed genes (DEGs; the established criteria: p adjusted < 0.05; log2 fold change (log2FC) ≥ 1.0/log2FC ≤ −1.0; determined by DESeq2 package [version 1.42.0] within R statistical software (version 4.4.2) identified in equine endometrial fibroblasts ( n = 5) after 96 h of TNFα treatment (10 ng/mL). The dot color depends on pathway enrichment significance, while the size of dots depends on the number of target genes assigned to particular processes. BP: biological process; CC: cellular components; MF: molecular function. ( B ) A dot plot illustrating Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the DEGs identified in equine endometrial fibroblasts ( n = 5) after 96 h of TNFα treatment (10 ng/mL). The dot color depends on pathway enrichment significance, while the size of dots depends on the number of target genes assigned to particular processes. Please note that the numbers inside the dots indicate the number of DEGs assigned to particular process.

Article Snippet: After the starvation time, the fibroblasts were treated with TNFα (10 ng/mL; equine recombinant, RP0137E; Kingfisher Biotech, St Paul, MN, USA) for 96 ( n = 5; treatments within particular experiments were run in technical triplicate).

Techniques: Software

Journal: International Journal of Molecular Sciences

Article Title: Transcriptomic Insights into Tumor Necrosis Factor α’s Role in the Fibrosis-Related Processes of Equine Endometrial Fibroblasts

doi: 10.3390/ijms262311344

Figure Lengend Snippet: Real-Time PCR validation of the selected differentially expressed genes (DEGs; the established criteria: p adjusted < 0.05; log2 fold change (log2FC) ≥ 1.0/log2FC ≤ −1.0; ( A ) IL-1α,( B ) IL-23, ( C ) IL-33, ( D ) FOS, ( E ) LIF, ( F ) FST, ( G ) CXCL3, ( H ) CXCL6, ( I ) CXCL8 identified by RNA-seq in equine endometrial fibroblasts ( n = 5) after 96 h of TNFα treatment (10 ng/mL). Data were expressed as mean ± SD and as a fold change to the control group within each endometrium category. Statistical analysis was performed using Student’s t -test or nonparametric Mann–Whitney U test. Asterisks denote statistical differences (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001) in comparison to the respective control group. IL-1α: interleukin 1α; IL-23: interleukin 23; IL-33: interleukin 33; FOS: Fos Proto-Oncogene, AP-1 Transcription Factor Subunit; LIF: leukemia inhibitory factor; FST: follistatin; CXCL3: C-X-C Motif Chemokine Ligand 3; CXCL6: C-X-C Motif Chemokine Ligand 6; CXCL8: C-X-C Motif Chemokine Ligand 8.

Article Snippet: After the starvation time, the fibroblasts were treated with TNFα (10 ng/mL; equine recombinant, RP0137E; Kingfisher Biotech, St Paul, MN, USA) for 96 ( n = 5; treatments within particular experiments were run in technical triplicate).

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, RNA Sequencing, Control, MANN-WHITNEY, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Transcriptomic Insights into Tumor Necrosis Factor α’s Role in the Fibrosis-Related Processes of Equine Endometrial Fibroblasts

doi: 10.3390/ijms262311344

Figure Lengend Snippet: The effect of TNFα (10 ng/mL) on the proliferation of equine endometrial fibroblasts ( n = 5) after 96 h of treatment. Data were expressed as mean ± SD and as a percentage of the control group, indicating normalized results from the BrdU assay measuring cell proliferation. Statistical analysis was performed using Student’s t -test. Asterisks denote statistical differences (** p < 0.01) in comparison to the control group.

Article Snippet: After the starvation time, the fibroblasts were treated with TNFα (10 ng/mL; equine recombinant, RP0137E; Kingfisher Biotech, St Paul, MN, USA) for 96 ( n = 5; treatments within particular experiments were run in technical triplicate).

Techniques: Control, BrdU Staining, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Transcriptomic Insights into Tumor Necrosis Factor α’s Role in the Fibrosis-Related Processes of Equine Endometrial Fibroblasts

doi: 10.3390/ijms262311344

Figure Lengend Snippet: The effect of TNFα (10 ng/mL) on the gene expression of ( A ) collagen type I ( COL1A1 ), ( B ) collagen type III ( COL3A1 ), ( C ) fibronectin 1 ( FN1 ), ( D ) actin alpha 2, smooth muscle ( ACTA2 ), ( E ) matrix metalloproteinase ( MMP ) 2, ( F ) MMP9 , ( G ) tissue inhibitor of matrix metalloproteinase ( TIMP ) 1 and ( H ) TIMP2 in equine endometrial fibroblasts ( n = 5) after 96 h of TNFα treatment (10 ng/mL). Data were expressed as mean ± SD and as a fold change to the control group. Statistical analysis was performed using Student’s t -test or nonparametric Mann–Whitney U test. Asterisks denote statistical differences (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001) within each category and time of treatment in comparison to the respective control group. Control: control cells, untreated; 24, 48, 96: time of treatment with TNFα.

Article Snippet: After the starvation time, the fibroblasts were treated with TNFα (10 ng/mL; equine recombinant, RP0137E; Kingfisher Biotech, St Paul, MN, USA) for 96 ( n = 5; treatments within particular experiments were run in technical triplicate).

Techniques: Gene Expression, Control, MANN-WHITNEY, Comparison