Journal: Theriogenology

Article Title: Estrogen-Responsive Genes Encoding Egg Yolk Proteins Vitellogenin and Apolipoprotein II in Chicken are differentially regulated by Selective Estrogen Receptor Modulators

doi: 10.1016/j.theriogenology.2015.08.015

Figure Lengend Snippet: Figure shows an ethidium bromide stained 2.5% agarose gel of the RT-PCR products of apo II cDNA. The in vitro transcribed apo II mRNA was used in mRNA stability assays to determine the effects of estrogen or the SERMs on the expression of E-RmRNASF in rooster livers. Lane 1 represents incubation of apo II mRNA in the absence of any added liver cytosolic extract, and lane 2 represent incubation of apo II with control liver cytosolic extract from roosters who received only the vehicle. Lanes 3-10, represent incubation of apo II mRNA with liver cytosolic extracts from roosters who were treated with estrogen (lane 3), resveratrol (lane 4), genistein (lane 5), catechin (lane 6), tamoxifen (lane 7), pterostilbene (lane 8), raloxifene (lane 9) or clomiphene (lane 10)

Article Snippet: Liver cytosolic extracts (S100) were prepared as previously described [ 9 ] from livers of 6-week-old white leghorn roosters (SPAFAS, Norwich, CT) who received two intramuscular injections of the vehicle (DMSO), 5μmol/kg of 17β-estradiol or 1000 μmol/kg of the respective SERMS (Sigma Chemical Co, Mo) or a combination of 5 μmol/kg of β-estradiol and 1000 μmol/kg of each of the respective SERMs, at 3-day intervals.

Techniques: Staining, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, In Vitro, Expressing, Incubation, Control

Journal: Theriogenology

Article Title: Estrogen-Responsive Genes Encoding Egg Yolk Proteins Vitellogenin and Apolipoprotein II in Chicken are differentially regulated by Selective Estrogen Receptor Modulators

doi: 10.1016/j.theriogenology.2015.08.015

Figure Lengend Snippet: The figure shows a chemiluminograph of a Western blot of liver cytosolic extracts from roosters who received treatment with the vehicle (lane 1, panels A, B, C & D), estrogen (lane 2, panels A, B, C & D), resveratrol (lane 3, panels A & B), genistein (lane 4, panels A & B), catechin (lane 5, panels A & B), tamoxifen (lane 6, panels A & B), pterostilbene (lane 3, panels C & D), raloxifene (lane 4, panels C & D), clomiphene (lane 5, panels C & D). Panels A and C were probed with α-chicken β-actin rabbit polyclonal antibody and panels B and D were probed with α-chicken apo II rabbit polyclonal antibody.

Article Snippet: Liver cytosolic extracts (S100) were prepared as previously described [ 9 ] from livers of 6-week-old white leghorn roosters (SPAFAS, Norwich, CT) who received two intramuscular injections of the vehicle (DMSO), 5μmol/kg of 17β-estradiol or 1000 μmol/kg of the respective SERMS (Sigma Chemical Co, Mo) or a combination of 5 μmol/kg of β-estradiol and 1000 μmol/kg of each of the respective SERMs, at 3-day intervals.

Techniques: Western Blot

Journal: Theriogenology

Article Title: Estrogen-Responsive Genes Encoding Egg Yolk Proteins Vitellogenin and Apolipoprotein II in Chicken are differentially regulated by Selective Estrogen Receptor Modulators

doi: 10.1016/j.theriogenology.2015.08.015

Figure Lengend Snippet: The figure shows a chemiluminograph of a Western blot of liver cytosolic extracts from roosters who received treatment with the vehicle (lane 1, panels A, B, C & D), estrogen (lane 2, panels A, B, C & D), resveratrol (lane 3, panels A & B), genistein (lane 4, panels A & B), catechin (lane 5, panels A & B), tamoxifen (lane 6, panels A & B), pterostilbene (lane 3, panels C & D), raloxifene (lane 4, panels C & D), clomiphene (lane 5, panels C & D). Panels A and C were probed with α-chicken vitellogenin rabbit polyclonal antibody and panels B and D were probed with α-chicken β-actin rabbit polyclonal antibody.

Article Snippet: Liver cytosolic extracts (S100) were prepared as previously described [ 9 ] from livers of 6-week-old white leghorn roosters (SPAFAS, Norwich, CT) who received two intramuscular injections of the vehicle (DMSO), 5μmol/kg of 17β-estradiol or 1000 μmol/kg of the respective SERMS (Sigma Chemical Co, Mo) or a combination of 5 μmol/kg of β-estradiol and 1000 μmol/kg of each of the respective SERMs, at 3-day intervals.

Techniques: Western Blot

Journal: Theriogenology

Article Title: Estrogen-Responsive Genes Encoding Egg Yolk Proteins Vitellogenin and Apolipoprotein II in Chicken are differentially regulated by Selective Estrogen Receptor Modulators

doi: 10.1016/j.theriogenology.2015.08.015

Figure Lengend Snippet: The figure shows a chemiluminograph of a Northern blot of liver cytosolic extracts from roosters who received treatment with the vehicle (lane 1, panels A, B, C & D), estrogen (lane 2, panels A, B, C & D), resveratrol (lane 3, panels A & B), genistein (lane 4, panels A & B), catechin (lane 5, panels A & B), tamoxifen (lane 6, panels A & B), pterostilbene (lane 3, panels C & D), raloxifene (lane 4, panels C & D), clomiphene (lane 5, panels C & D). Panels A and C were probed with biotinylated antisense chicken β-actin DNA oligonucleotide probe and panels B and D were probed with biotinylated antisense chicken apo II DNA oligonucleotide probe. Panel E shows the antiestrogenic effect of the SERMs on apo II gene expression and the relative electrophoretic mobility of apo II mRNA in relation to the β-actin mRNA. Lanes 1-5 in panel E correspond to whole cell RNA from livers of roosters treated with estrogen + resveratrol (lane 1), estrogen + genistein (lane 2), estrogen (lanes 3 & 4), and estrogen + catechin (Lane 5), probed with biotinylated apo II and β-actin antisense DNA oligonucleotides.

Article Snippet: Liver cytosolic extracts (S100) were prepared as previously described [ 9 ] from livers of 6-week-old white leghorn roosters (SPAFAS, Norwich, CT) who received two intramuscular injections of the vehicle (DMSO), 5μmol/kg of 17β-estradiol or 1000 μmol/kg of the respective SERMS (Sigma Chemical Co, Mo) or a combination of 5 μmol/kg of β-estradiol and 1000 μmol/kg of each of the respective SERMs, at 3-day intervals.

Techniques: Northern Blot, Gene Expression

Journal: Theriogenology

Article Title: Estrogen-Responsive Genes Encoding Egg Yolk Proteins Vitellogenin and Apolipoprotein II in Chicken are differentially regulated by Selective Estrogen Receptor Modulators

doi: 10.1016/j.theriogenology.2015.08.015

Figure Lengend Snippet: The figure shows a chemiluminograph of a Northern blot of liver cytosolic extracts from roosters who received treatment with the vehicle (lane 1, panels A, B, C, & D), estrogen (lane 2, panels A, B, C, & D), resveratrol (lane 3, panels A, & B), genistein (lane 4, panels A, & B), catechin (lane 5, panels A, & B), tamoxifen (lane 6, panels A & B), pterostilbene (lane 3, panels C & D), raloxifene (lane 4, panels C & D), clomiphene (lane 5, panels C & D). Panels A and C were probed with biotinylated antisense chicken VG DNA oligonucleotide probe and panels B and D were probed with biotinylated antisense chicken β-actin DNA oligonucleotide probe. Panel E shows the antiestrogenic effect of the SERMs on VG gene expression and the relative electrophoretic gel mobility of the VG mRNA in relation to β-actin mRNA. Lanes 1-5 in panel E correspond to whole cell RNA from livers of roosters treated with the vehicle (lane 1), estrogen (lane 2), estrogen + resveratrol (lane 3), estrogen + genistein (lane 4), and estrogen + catechin (lane 5).

Article Snippet: Liver cytosolic extracts (S100) were prepared as previously described [ 9 ] from livers of 6-week-old white leghorn roosters (SPAFAS, Norwich, CT) who received two intramuscular injections of the vehicle (DMSO), 5μmol/kg of 17β-estradiol or 1000 μmol/kg of the respective SERMS (Sigma Chemical Co, Mo) or a combination of 5 μmol/kg of β-estradiol and 1000 μmol/kg of each of the respective SERMs, at 3-day intervals.

Techniques: Northern Blot, Gene Expression

Journal: Frontiers in Veterinary Science

Article Title: A monoclonal antibody-based sandwich ELISA for measuring canine Thymidine kinase 1 protein and its role as biomarker in canine lymphoma

doi: 10.3389/fvets.2023.1243853

Figure Lengend Snippet: Dot blot results using recombinant enzymes from canine (rCTK1), feline (rFTK1), equine (rETK1) and human (rHTK1) at concentration ranging from 0.6–20 ng. The membranes were developed with (A) the monoclonal antibody produced against the active site: Mab-1 (3 μg/mL) and (B) the monoclonal antibody produced against the C terminal region: Mab-2 (3 μg/mL). (C) Western blot experiments result with 2 μg/mL of Mab-1. (D) Western blot results with 2 μg/mL of Mab-2. Immunoprecipitation results with recombinant canine TK1 5 and 2.5 ng and sera from dogs with different malignancies diluted 10x and incubated at 4°C and room temperature in (E) with 4 μg/mL of Mab-1, in (F) with 4 μg/mL of Mab- 2. The results are presented as the percentage of TK1 protein bound to the magnetic beads coated with anti-mouse IgG.

Article Snippet: Recombinant dog and human TK1 (10, 5 and 0.5 ng) as well as cytosolic extract from human CEM cells positive and negative for TK1 , containing 25 μg of protein, were diluted in the denaturing sample buffer, boiled, and loaded onto a 12% polyacrylamide gel.

Techniques: Dot Blot, Recombinant, Concentration Assay, Produced, Western Blot, Immunoprecipitation, Incubation, Magnetic Beads

Journal: Frontiers in Veterinary Science

Article Title: A monoclonal antibody-based sandwich ELISA for measuring canine Thymidine kinase 1 protein and its role as biomarker in canine lymphoma

doi: 10.3389/fvets.2023.1243853

Figure Lengend Snippet: (A) Log STK1 protein concentrations measured in sera from healthy dogs ( n = 124) and dogs with lymphoma ( n = 93), the lines represent the median values (B) . The distribution of Log STK1 activity values in sera from healthy dogs ( n = 124) and in sera from dogs with lymphoma ( n = 93), error bars represent the median values. (C) Receiver operating characteristic (ROC) curves of the STK1 protein concentrations (blue line) and TK1 activity values (green line) in sera from dogs with tumors and healthy dogs.

Article Snippet: Recombinant dog and human TK1 (10, 5 and 0.5 ng) as well as cytosolic extract from human CEM cells positive and negative for TK1 , containing 25 μg of protein, were diluted in the denaturing sample buffer, boiled, and loaded onto a 12% polyacrylamide gel.

Techniques: Activity Assay

Journal: Frontiers in Veterinary Science

Article Title: A monoclonal antibody-based sandwich ELISA for measuring canine Thymidine kinase 1 protein and its role as biomarker in canine lymphoma

doi: 10.3389/fvets.2023.1243853

Figure Lengend Snippet: (A) Log STK1 protein concentration in sera from healthy dogs ( n = 124), dogs with T-cell lymphoma ( n = 25) and B-cell lymphoma ( n = 32). (B) Log STK1 activity levels in healthy dogs, dogs with T-cell lymphoma and B-cell lymphoma. (C) ROC curves of STK1 protein values (blue line) and TK1 activity values (green line) to differentiate between dogs with T-cell lymphoma and B-cell lymphoma. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Recombinant dog and human TK1 (10, 5 and 0.5 ng) as well as cytosolic extract from human CEM cells positive and negative for TK1 , containing 25 μg of protein, were diluted in the denaturing sample buffer, boiled, and loaded onto a 12% polyacrylamide gel.

Techniques: Protein Concentration, Activity Assay

Journal: Environmental Health Perspectives

Article Title: Sulfated Metabolites of Polychlorinated Biphenyls Are High-Affinity Ligands for the Thyroid Hormone Transport Protein Transthyretin

doi: 10.1289/ehp.1206198

Figure Lengend Snippet: OH-PCBs are substrates for human SULT1A1. The sulfation of 2´OH-PCB 3, 3´OH-PCB 3, 4´OH-PCB 3, 4´OH-PCB 11, and 4´OH-PCB 12 is catalyzed by a cytosolic extract of E. coli expressing recombinant hSULT1A1. * p < 0.05, and ** p < 0.001 compared with 0 µM OH-PCB

Article Snippet: Cytosolic extract of E. coli expressing recombinant hSULT1A1 (cytosolic protein concentration of 10 mg/mL) (Xenotech, Lenexa, KS) was used without further purification.

Techniques: Expressing, Recombinant