Journal: International Journal of General Medicine

Article Title: Cardiac ß-adrenoceptor expression is markedly depressed in Ossabaw swine model of cardiometabolic risk

doi: 10.2147/ijgm.s18175

Figure Lengend Snippet: Figure 2 Using real-time PCR, no significant changes in mRNA of β1- and β2-ARs were detected in left ventricle obtained from high-fat-fed Ossabaw swine when compared with controls. (A) Melt curve graph for SYBR®-β1- and β2-ARs and β-actin. (B) PCR amp/cycle graph for β1- and β2-ARs and β-actin. Abbreviations: CF, curve fit; AR, adrenoceptor; PCR, polymerase chain reaction; RFU, relative fluorescence units; CT, mean cycle threshold value.

Article Snippet: Real-time PCR reactions were performed for β 1 -, β 2 -ARs, and β-actin in triplicate with a custom-designed SYBR® Green mix (2.4 μL of 25 mM MgCl 2 , 5 μL of 1:10,000 dilution SYBR Green I, Molecular Probes, Inc, Eugene, OR) and 5 μL of 1 nM fluorescein calibration dye (1 mM/L in dimethyl sulfoxide, Bio-Rad) in 50 μL of total reaction using Taq DNA polymerase (Promega).8,17 Primers were designed using the Genomatix Software Suite program (Müchen, Germany) based on sequences published in the National Center for Biotechnology Information GenBank database.18 Gene-specific primers were used: for β 1 -ARs, sense 5′-GACCGAAAGCAGGTGAACTC-3′ and antisense 5′-CTCCCATCCCTTCCCTAGTC-3′ (242 bp product; accession number AF042454); for β 2 -ARs, sense 5′-GCCATCGACTGCTATCACAA-3′ and antisense 5′-GGTTTGGGGAGTGGAATCTT-3′ (193 bp product; accession number U53185); for β-actin, primers were designed based on published sequences in the GenBank database (sense 5′-ACGTGGACATCAGGAAGGAC-3′ and antisense 5′-ACATCTGCTGGAAGGTGGAC-3′; accession number U07786).

Techniques: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Fluorescence

Journal: Biological Research

Article Title: Exposure to polystyrene nanoparticles leads to dysfunction in DNA repair mechanisms in Caco-2 cells

doi: 10.1186/s40659-025-00629-y

Figure Lengend Snippet: Identification of SSBs and DSBs in Caco-2 cells after PNPs exposure (100 µg/mL, 24 h) with alkaline comet assay. 50 µM etoposide (2 h incubation) was used as a positive control. A : Representative comets with a comet tail (fragmented DNA) and a head (intact DNA), visualized under 40x immersion oil, confocal microscope Leica SP5 with SYBR SAFE™ staining using fluorescein filter with maximum excitation/emission 496 nm/522 nm). B : Quantification of comet assay images was performed using OpenComet v1.3.1 software for % of DNA in the head and % of DNA in the tail (box plots). Results are based on the analysis of at least 50 comets per replicate ( n = 3). P-values were considered significant: * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001 and **** p ≤ 0.0001

Article Snippet: The expression of genes was assessed in two Caco-2 cell lines (untreated vs. nanoparticle PNPs-100 μg/mL treated) using a PrimePCR assay with a custom-designed SYBR plate (Bio-Rad), following the manufacturer’s protocol.

Techniques: Alkaline Single Cell Gel Electrophoresis, Incubation, Positive Control, Microscopy, Staining, Single Cell Gel Electrophoresis, Software