Journal: Microorganisms

Article Title: Nisin, a Probiotic Bacteriocin, Modulates the Inflammatory and Microbiome Changes in Female Reproductive Organs Mediated by Polymicrobial Periodontal Infection

doi: 10.3390/microorganisms12081647

Figure Lengend Snippet: Oral periodontal pathogens are present in the female upper reproductive organs, plus the heart and aorta, and nisin mitigates changes in the oral cavity. The graphs show the log copy number of the periodontal pathogens P. gingivalis ( a ), T. forsythia ( b ), T. denticola ( c ), and F. nucleatum ( d ) in the oral swabs (at baseline and infection after 8 weeks) and uterus, ovary, vagina, heart and aorta tissue. Data are represented as the mean ± SD. * indicates significant differences between marked groups ( p ≤ 0.05).

Article Snippet: In this study, Porphyromonas gingivalis (FDC 381), Treponema denticola (ATCC 35405), Tannerella forsythia (ATCC 43037), and Fusobacterium nucleatum (ATCC 10953) were cultured anaerobically (85% N 2 , 10% H 2 , 5% CO 2 ) at 37 °C in an anaerobic chamber, as previously described [ , , , ].

Techniques: Infection

Journal: Microorganisms

Article Title: Nisin, a Probiotic Bacteriocin, Modulates the Inflammatory and Microbiome Changes in Female Reproductive Organs Mediated by Polymicrobial Periodontal Infection

doi: 10.3390/microorganisms12081647

Figure Lengend Snippet: Detection frequency of the periodontal pathogens in the oral swabs, uterus, ovary, vagina, heart, and aorta. Values are expressed as the percentage (%) of total mice and with the total frequency in parenthesis (N = 6 per group).

Article Snippet: In this study, Porphyromonas gingivalis (FDC 381), Treponema denticola (ATCC 35405), Tannerella forsythia (ATCC 43037), and Fusobacterium nucleatum (ATCC 10953) were cultured anaerobically (85% N 2 , 10% H 2 , 5% CO 2 ) at 37 °C in an anaerobic chamber, as previously described [ , , , ].

Techniques:

Journal: ACS chemical biology

Article Title: Catch and Release DNA Decoys: Capture and Photochemical Dissociation of NF-κB Transcription Factors

doi: 10.1021/acschembio.6b00130

Figure Lengend Snippet: Quantitative EMSA analysis to measure equilibrium dissociation (Kd) constants for 4 with p50-p65 heterodimer and p50 and p65 homodimer proteins. Recombinant p50 (35-381) and p65 (1-306) proteins were used in this study. Mean ± SD (n = 3).

Article Snippet: For photochemical stability studies, binding reactions containing binding buffer, recombinant p50 protein (0.5 μL; 0.50 μg/μL, Enzo Life Sciences, BML-UW9885-0050, amino acids 35-381), recombinant p65 protein (0.5 μL; 0.50 μg/μL Sino Biological, 12054-H09E, amino acids 1-306), and DNase/RNase free H 2 O (to a final volume of 19 μL) were prepared in microcentrifuge tubes (0.65 mL) and incubated at room temperature for 10 mins.

Techniques: Recombinant

Journal: ACS chemical biology

Article Title: Catch and Release DNA Decoys: Capture and Photochemical Dissociation of NF-κB Transcription Factors

doi: 10.1021/acschembio.6b00130

Figure Lengend Snippet: (a) Electrophoretic mobility shift assays with 4-10. 5’-32P-labeled 4-10 incubated with p50 (35-381) and p65 (1-306) recombinant proteins displays formation of the NF-κB-CRDD complexes. NF-κB binding is observed until three abasic sites are formed (i.e., 10). Scrambled decoys 11 and 12 and mismatch decoy 13 demonstrate no observable complex formation (Supplemental Figure 2). (b) Quantitative EMSA analysis to measure equilibrium dissociation (Kd) constants for 4, 9, and 10 with recombinant p50 (35-381) and p65 (1-306) proteins. (c-d) Photolysis of CRDD 9 (350 nm light). Samples analyzed by ion-extracted LC-MS. (c) Photolytic decay curve of 9 with calculated half-life and quantum yield (R2 = 0.97). (d) Formation of abasic sites and truncation products 10 and 14-19 resulting from photolysis of 9 (P = 5’-phosphate). Dashed line denotes full-length DNA decoys containing abasic products and solid lines denote truncation products (some contain abasic sites as well). Isomers denote constitution isomers resulting from photolysis of 9 (e.g., X and _ are in different arrangements). Mean ± SD (n = 4). An LC-MS chromatogram of the photo-products from irradiation of 9 can be found in Supplementary Figure 4.

Article Snippet: For photochemical stability studies, binding reactions containing binding buffer, recombinant p50 protein (0.5 μL; 0.50 μg/μL, Enzo Life Sciences, BML-UW9885-0050, amino acids 35-381), recombinant p65 protein (0.5 μL; 0.50 μg/μL Sino Biological, 12054-H09E, amino acids 1-306), and DNase/RNase free H 2 O (to a final volume of 19 μL) were prepared in microcentrifuge tubes (0.65 mL) and incubated at room temperature for 10 mins.

Techniques: Electrophoretic Mobility Shift Assay, Labeling, Incubation, Recombinant, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Irradiation

Journal: ACS chemical biology

Article Title: Catch and Release DNA Decoys: Capture and Photochemical Dissociation of NF-κB Transcription Factors

doi: 10.1021/acschembio.6b00130

Figure Lengend Snippet: Quantitative electrophoretic mobility shift assay with CRDD 9. (a) 5’-32P-labeled 9 incubated with p50 (35-381) and p65 (1-306) recombinant proteins yields complex formation (catch, t = 0 min), which is dissociated upon photolysis with 350 nm light (release) in a time-dependent manner. Recovery of the NF-κB-9 complex can be obtained by addition of 32P-labeled 9. (b) Densitometry of EMSAs yielding half-life of NF-κB release (R2 = 0.93). Mean ± SD. (n = 3).

Article Snippet: For photochemical stability studies, binding reactions containing binding buffer, recombinant p50 protein (0.5 μL; 0.50 μg/μL, Enzo Life Sciences, BML-UW9885-0050, amino acids 35-381), recombinant p65 protein (0.5 μL; 0.50 μg/μL Sino Biological, 12054-H09E, amino acids 1-306), and DNase/RNase free H 2 O (to a final volume of 19 μL) were prepared in microcentrifuge tubes (0.65 mL) and incubated at room temperature for 10 mins.

Techniques: Electrophoretic Mobility Shift Assay, Labeling, Incubation, Recombinant