Journal: International Journal of Molecular Sciences

Article Title: Hepatic FGF21 Deletion Improves Glucose Metabolism, Alters Lipogenic and Chrna4 Gene Expression, and Enhances Telomere Maintenance in Aged Female Mice

doi: 10.3390/ijms27010194

Figure Lengend Snippet: Hepatic FGF21 deletion does not alter circulating FGF21, insulin, or estrogen levels. ( A ) Basal insulin serum concentration (pg/mL). ( B ) Basal estrogen serum concentration (pg/mL). ( C ) Hepatic Fgf21 mRNA expression. ( D ) FGF21 serum levels (pg/mL). ( E ) Hepatic expression of Fgf21 receptor 4 ( Fgf21r4 ). All data are presented as mean ± SEM. ** p < 0.01 and p -values were determined by using a Student’s t -test. LoxP: n = 11; FKO: n = 9. For ELISA: LoxP: n = 10; FKO: n = 9.

Article Snippet: Serum 8-hydroxy-2-deoxyguanosine (8-OHdG), a marker of DNA oxidation, was quantified using the DNA Damage Competitive ELISA kit (EIADNAD, Thermo Fisher Scientific, Waltham, MA, USA).

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

Journal: International Journal of Molecular Sciences

Article Title: Hepatic FGF21 Deletion Improves Glucose Metabolism, Alters Lipogenic and Chrna4 Gene Expression, and Enhances Telomere Maintenance in Aged Female Mice

doi: 10.3390/ijms27010194

Figure Lengend Snippet: Gene expression changes related to lipid and glucose metabolism in the liver of FKO mice, along with serum adiponectin levels. ( A ) Relative mRNA levels of genes related to lipid metabolism: Pparα ( Peroxisome proliferator activated receptor alpha ), Hmgcs2 ( 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 2 ), Cd36 ( CD36 molecule ), Cpt1a ( Carnitine palmitoyltransferase 1a ), Chrebp ( MLX interacting protein-like ), Srebp1c ( Sterol regulatory element binding transcription factor 1 ), Fasn ( Fatty acid synthase ), Scd1 ( Stearoyl-Coenzyme A desaturase 1 ), Acox3 ( Acyl-Coenzyme A oxidase 3 ), Apoc2 ( Apolipoprotein C2 like ) and AdipoR2 ( Adiponectin Receptor 2 ). ( B ) Adiponectin serum levels (μg/mL). ( C ) Liver Triacylglycerol (TAG) levels (µmol/mg). ( D ) Total cholesterol serum levels (mg/dL). ( E ) Relative mRNA levels of genes related to gluconeogenesis: Pepck ( Phosphoenolpyruvate carboxykinase 1 ) and G6pc ( Glucose-6-phosphatase ); and glycolysis: Lpk ( Pyruvate kinase ) and Gluk ( Glucokinase ). All data are presented as mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001 and p -values were determined by using a Student’s t -test. LoxP: n = 10; FKO: n = 9. For adiponectin ELISA: LoxP: n = 8; FKO: n = 8.

Article Snippet: Serum 8-hydroxy-2-deoxyguanosine (8-OHdG), a marker of DNA oxidation, was quantified using the DNA Damage Competitive ELISA kit (EIADNAD, Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Gene Expression, Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Veterinary Science

Article Title: Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs

doi: 10.3389/fvets.2025.1702980

Figure Lengend Snippet: Nuclear DNA from canine urinary bladder organoids exposed to acrolein and assessed for DNA damage using the alkaline CometChip assay. Electrophoresed DNA was stained with SYBR™ Gold, and DNA damage was calculated as % DNA in the “comet tail” (between red and green lines) using CometAssay Analysis Software. (A) Canine urinary organoid line 3 exposed to vehicle for 6 h. (B) Canine urinary organoid line 3 exposed to 56 uM acrolein for 6 h.

Article Snippet: To produce a single-cell suspension for use in in vitro DNA damage assessments, the organoids were dissociated using TrypLETM Express (ThermoFisher; cat. #12604013) for 10 min and passed through 40-micron filters to remove clumps.

Techniques: Staining, Software

Journal: Frontiers in Veterinary Science

Article Title: Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs

doi: 10.3389/fvets.2025.1702980

Figure Lengend Snippet: Urothelial genotoxic thresholds for acrolein in dogs. (A) Genotoxicity of acrolein in canine urinary organoids, measured as DNA strand breaks using the alkaline CometChip assay. ** p ≤ 0.001 compared to vehicle. (B) Genotoxicity of acrolein as direct DNA strand breaks plus oxidative DNA base modifications using the CometChip assay with the addition of the enzyme Fpg. * p = 0.018; ** p ≤ 0.0002. Data shown are from three individual canine urinary organoids, each assayed in triplicate on two occasions. (C) Urinary concentrations of the stable urinary acrolein metabolite, 3-HPMA (3-hydroxypropylmercapturic acid) previously measured in the urine of pet dogs with and without urothelial carcinoma (UC) . The dotted line indicates a 20 uM threshold for combined DNA damage from acrolein in vitro , with no significant difference between the number of cases and controls reaching this threshold ( p ≥ 0.99).

Article Snippet: To produce a single-cell suspension for use in in vitro DNA damage assessments, the organoids were dissociated using TrypLETM Express (ThermoFisher; cat. #12604013) for 10 min and passed through 40-micron filters to remove clumps.

Techniques: In Vitro

Journal: Frontiers in Veterinary Science

Article Title: Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs

doi: 10.3389/fvets.2025.1702980

Figure Lengend Snippet: Genotoxicity of inorganic arsenic in canine urinary organoids. (A) Genotoxicity of sodium arsenite measured as direct DNA strand breaks using the alkaline CometChip assay. * p = 0.038; *** p < 0.0001. (B) Genotoxicity of sodium arsenite as direct DNA strand breaks plus oxidative DNA base modifications using the CometChip assay with the addition of the enzyme Fpg. *** p < 0.0001. Data shown are from three separate urinary organoids, each assayed on two occasions in triplicate. (C) Total urinary concentrations of inorganic arsenic species previously measured in pet dogs without and without urothelial carcinoma (UC) . No cases or controls reached 20 uM of total inorganic arsenic.

Article Snippet: To produce a single-cell suspension for use in in vitro DNA damage assessments, the organoids were dissociated using TrypLETM Express (ThermoFisher; cat. #12604013) for 10 min and passed through 40-micron filters to remove clumps.

Techniques:

Journal: Frontiers in Veterinary Science

Article Title: Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs

doi: 10.3389/fvets.2025.1702980

Figure Lengend Snippet: Urothelial genotoxicity of the aromatic amine 2,6-dimethylaniline (2,6-DMA) in dogs. (A) DNA strand breaks from 2,6-DMA in canine urinary organoids, measured using the alkaline CometChip assay. Data shown are from three separate urinary organoids, each assayed on two occasions in triplicate (concentrations between 0.1 and 100 uM could only be obtained in one set of organoids). * p = 0.04, *** p ≤ 0.0002. (B) DNA strand breaks plus oxidative DNA base modifications from 2,6-DMA in canine urinary organoids, measured using the CometChip assay with the addition of the enzyme Fpg. * p = 0.03, ** p = 0.0008, *** p < 0.0001. (C) Urinary 2,6-DMA previously measured in the urine of pet dogs without and without urothelial carcinoma (UC) . The dotted line indicates 0.01 uM; 3 of 37 UC cases and none of 37 unaffected controls reached this urinary 2,6-DMA concentration.

Article Snippet: To produce a single-cell suspension for use in in vitro DNA damage assessments, the organoids were dissociated using TrypLETM Express (ThermoFisher; cat. #12604013) for 10 min and passed through 40-micron filters to remove clumps.

Techniques: Concentration Assay