Journal: Journal of dentistry

Article Title: Dental primer and adhesive containing a new antibacterial quaternary ammonium monomer dimethylaminododecyl methacrylate

doi: 10.1016/j.jdent.2013.01.004

Figure Lengend Snippet: Minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) against S. mutans. A lower MIC and MBC indicate a stronger antibacterial potency for the antibacterial agent. CHX had the lowest MIC and MBC. Regarding the two new monomers, DMADDM (dimethylaminododecyl methacrylate) with an alkyl chain length of 12 was much more potent than DMAHM (dimethylhexylamine methacrylate) with a chain length of 6. The tests were performed in triplicate (n = 3).

Article Snippet: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) The use of S. mutans (ATCC 700610, American Type Culture, Manassas, VA) was approved by the University of Maryland.

Techniques: Concentration Assay

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: CD39 gene disruption aggravates liver injury and fibrosis in Mdr2–/– mice. Mdr2–/–;CD39–/– double‐mutant mice were generated as described in Materials and Methods, and pups of both sexes phenotyped at 8 weeks of age in comparison to their Mdr2–/–;CD39+/+ and Mdr2–/–;CD39+/– littermates (males are shown). (A) Representative pictures of connective tissue (Sirius Red, left and middle) and p‐CK (right) staining demonstrate portal fibrosis and pronounced ductular reaction in Mdr2–/– mice, respectively, which is exacerbated in Mdr2–/–;CD39–/– mice. Magnification ×50 (left and right panel) and ×200 (middle panel, portal area). (B) Liver hydroxyproline content, (C) p‐CK‐positive ductal cell counts (quantified from 10 random portal HPF area/liver at ×200, n = 3/bar; * P = 0.0034, t test), and (D) serum ALT levels are significantly increased in Mdr2–/–;CD39–/– compared to Mdr2–/– mice. (E) CD39 deletion leads to an up‐regulation of Col1a1 and Tgfβ2 mRNA expression. qRT‐PCR data are shown as fold increase compared to age‐matched WT controls. Data (B‐E,G) are mean ± SEM (n = 3‐6 mice/bar). * P < 0.05 compared to Mdr2–/–;CD39+/+ controls (ANOVA followed by Dunnett's posttest). (F) Mdr2–/–;CD39–/– mice at 12 months old demonstrate severe periductular scarring that extends into liver parenchyma (sinusoidal fibrosis), while fibrotic lesions in Mdr2–/– mice are conferred to the periportal region (Sirius Red, ×200, portal area). (G) Total hepatic hydroxyproline content in Mdr2–/–;CD39–/– mice (n = 8) compared to Mdr2–/– mice (n = 12) at the age of 12 months (** P = 0.0013, t test). Abbreviations: ANOVA, analysis of variance; het, heterozygous; HPF, high‐power field; ko, knockout; p‐CK, pan‐cytokeratine; qRT‐PCR, quantitative reverse‐transcription polymerase chain reaction; Tgfβ2, transforming growth factor β2; wt, wild type.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Disruption, Mutagenesis, Generated, Comparison, Staining, Expressing, Quantitative RT-PCR, Knock-Out, Reverse Transcription, Polymerase Chain Reaction

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: Loss of CD39 is associated with CD8+ T‐cell infiltration in the liver. (A) FACS analysis of liver‐infiltrating lymphocytes isolated from 8‐week‐old male mice. Representative dot plots of CD3‐gated hepatic lymphocytes (average values shown) and mean ± SEM for CD8+ T‐cell quantification (gated on CD3) in Mdr2–/– (n = 3) versus Mdr2–/–;CD39–/– (n = 5). (B,C) qRT‐PCR analysis of T‐cell subsets and T‐cell‐related tropism markers shown as fold relative to Mdr2–/–. (D) Immunohistochemical staining of liver for MAdCAM‐1 and CCL25. Values are presented as mean ± SEM. * P < 0.05 for Mdr2–/– versus Mdr2–/–;CD39–/– ( t test). Abbreviations: FACS, fluorescence‐activated cell sorting; qRT‐PCR, quantitative reverse‐transcription polymerase chain reaction.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Isolation, Quantitative RT-PCR, Immunohistochemical staining, Staining, Fluorescence, FACS, Reverse Transcription, Polymerase Chain Reaction

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: CD8+ T‐cell depletion ameliorates liver injury and has beneficial effects on fibrosis‐related gene expression. (A) Immunofluorescent staining of CD8+ (red) and bile ducts visualized by pan‐CK (green) in liver sections from male Mdr2–/–;CD39–/– mice 3 days after administration of anti‐CD8 monoclonal antibody or the respective isotype control (original magnification, ×200). (B) Flow cytometry analysis of CD8 expression in splenocytes isolated from Mdr2–/–;CD39–/– mice treated for 3 days with anti‐CD8 or isotype control. Cells were gated on CD3+ subsets. (C) CD8+ T‐cell depletion resulted in a significant decrease in serum ALT and ALP. (D) CD8+ T‐cell depletion led to a profibrolytic shift in gene expression with significantly reduced TIMP‐1 expression and increased MMP‐8 and MMP‐13 expression. Data are mean ± SEM (n = 3‐5 mice/bar). * P < 0.05 compared to isotype‐treated control mice ( t test). Abbreviations: CK, cytokeratine; IgG, immunoglobulin G; Tgfβ2, transforming growth factor β2; TIMP‐1, tissue inhibitor of metalloproteinase 1.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Gene Expression, Staining, Control, Flow Cytometry, Expressing, Isolation

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: Intestinal bacteria promote sclerosing cholangitis in Mdr2–/– mice. (A) Experimental design of experimental colitis model in 6‐week‐old male FVB.Mdr2–/– mice. After 7 days of DSS administration, colitis was assessed, the DSS‐supplemented water was replaced by normal drinking water, and livers assessed 7 days later. Hematoxylin and eosin staining of colons (×200) and Sirius Red staining of liver specimens from Mdr2–/– mice receiving drinking water (left), 3% DSS (middle), or 5% DSS (right) for 7 days. Magnification ×50 (middle panel) and ×200 (lower panel, portal area). (B) Hydroxyproline content in livers from control and DSS‐treated Mdr2–/– mice. (C) Hepatic expression of Col1a1 and Tgfβ2. (D) Schematic outline for the antibiotic treatment in male Mdr2–/– mice. (E) Hepatic hydroxyproline content and (F) representative connective tissue stain (Sirius Red, ×50) in livers from 8‐week‐old Mdr2–/– mice after 4 weeks of antibiotic treatment with polymyxin B (100 mg/kg) + neomycin (220 mg/kg). Data are shown as mean ± SEM (n = 4‐5 mice/bar). * P < 0.05 compared to age‐matched Mdr2–/– controls (analysis of variance followed by Dunnett's posttest). Abbreviations: DSS, dextran sulfate sodium; FVB, Friend virus B‐type; Tgfβ2, transforming growth factor β2.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Bacteria, Staining, Control, Expressing, Virus

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: Colonic ATP administration to Mdr2–/– mice recapitulates the liver phenotype of Mdr2–/–;CD39–/– mice. Stable ATP agonist (αβ‐ATP, 1 mg/kg/day) or vehicle (RPMI medium) was rectally administered into 5‐week‐old male Mdr2–/– mice daily for 6 days. (A) Double immunofluorescence for CD8+ (red) and biliary marker p‐CK (green) showed remarkable CD8+ T‐cell (arrowheads) infiltration in portal areas in Mdr2–/– mice challenged with αβ‐ATP (original magnification, ×200). (B) CD8+ T cells were quantified by counting 10 randomly selected high‐power fields and by measuring hepatic CD8 mRNA expression. (C) Elevation of serum ALT and ALP in mice treated with αβ‐ATP. (D) mRNA expression of Col1a1, Tgfb2, as well as the gut‐homing markers Itgb7, Ccr9 (n.s.). Data are mean ± SEM (n = 3 mice/bar). * P < 0.05 compared to vehicle‐treated Mdr2–/– ( t test). Abbreviations: HPF, high‐power field; Itgb7, integrin b7; n.s., not significant; p‐CK, pan‐cytokeratine; RPMI, Roswell Park Memorial Institute; Tgfb2, transforming growth factor β2.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Immunofluorescence, Marker, Expressing

Journal: Hepatology Communications

Article Title: The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis

doi: 10.1002/hep4.1084

Figure Lengend Snippet: Extracellular ATP promotes RA‐dependent imprinting of gut‐homing receptor α4β7 on naive CD8+ T cells. (A) Frozen tissue sections of intestine from male Mdr2–/– mice were double stained for CD39 (green) and CD11c (red) using DAPI for nuclei visualization. Photographs in the upper panel (magnification ×200) show CD39‐positive cells in the mucosa, villi, and Peyer's patch. CD11c‐positive cells were detected in villi and the Peyer's patch. CD11c cells expressing CD39 were found in the Peyer's patch and in the villi. The lower panel at a higher magnification (×400) clearly demonstrates that a subset of DC is CD11c/CD39 double positive. (B) Flow cytometry analysis of α4β7 expression in naive CD8+ T cells cultured for 4 days with CD3/CD28 beads in the presence of DMSO, 50 nM RA, or 50 nM RA + 250 μM ATP. Representative dot plots of CD45+ gated cells are shown with numbers indicating the percentage of cells positive for CD8 and α4β7; in parentheses is the mean fluorescence intensity for α4β7 in CD8+ cells. Results are shown from three different experiments (n = 9). Bar graph: α4β7 expression (MFI) in CD45+/CD8+ cells stimulated with 50 nM RA with or without ATP was normalized to DMSO‐stimulated cells (mean ± SD). *** P < 0.001. Abbreviations: DAPI, 4′,6‐diamidino‐2‐phenylindole; DC, dendritic cell; DMSO, dimethyl sulfoxide; MFI, mean fluorescence intensity; PP, Peyer's patch.

Article Snippet: A minimum of 19 male Mdr2 mutation carriers were produced in each generation, and their genome screened using microsatellite marker differences (simple sequence length polymorphisms) at Jackson Laboratory ( https://www.jax.org/jax-mice-and-services/breeding-and-rederivation-services/speed-congenic ) for the percentage of C57Bl/6J background.

Techniques: Staining, Expressing, Flow Cytometry, Cell Culture, Fluorescence

Journal: Journal of Lasers in Medical Sciences

Article Title: Photodynamic Therapy With Propolis: Antibacterial Effects on Staphylococcus aureus, Streptococcus mutans and Escherichia coli Analysed by Atomic Force Microscopy

doi: 10.34172/jlms.2020.S17

Figure Lengend Snippet: The Minimum Inhibitory Concentration of the Photosensitizer Towards the Tested Bacteria.

Article Snippet: Preparation of Microorganisms and Determination of Minimum Inhibitory Concentration Bacterial strains ( S. aureus ATCC 29213, E. coli ATCC 25922 and S. mutans ATCC 25175) were previously cultured in a bacteriological incubator according to the protocol suggested by the Clinical and Laboratory Standards Institute in 2015.

Techniques: Concentration Assay, Bacteria

Journal: Journal of Lasers in Medical Sciences

Article Title: Photodynamic Therapy With Propolis: Antibacterial Effects on Staphylococcus aureus, Streptococcus mutans and Escherichia coli Analysed by Atomic Force Microscopy

doi: 10.34172/jlms.2020.S17

Figure Lengend Snippet: The minimum inhibitory concentration of the photosensitizing agents against the bacteria after laser irradiation. Results are expressed in mg/mL.

Article Snippet: Preparation of Microorganisms and Determination of Minimum Inhibitory Concentration Bacterial strains ( S. aureus ATCC 29213, E. coli ATCC 25922 and S. mutans ATCC 25175) were previously cultured in a bacteriological incubator according to the protocol suggested by the Clinical and Laboratory Standards Institute in 2015.

Techniques: Concentration Assay, Bacteria, Irradiation