Journal: The Journal of biological chemistry

Article Title: Lipid dependence and basic kinetics of the purified 1,2-diacylglycerol 3-glucosyltransferase from membranes of Acholeplasma laidlawii.

doi: 10.1074/jbc.272.2.929

Figure Lengend Snippet: FIG. 1. Purification of the 1,2-diacylglycerol 3-glucosyltrans- ferase in three column chromatography steps. Protein (E) and MGlcDAG synthesis activity (G) were eluted in three different column chromatography steps as described under “Materials and Methods.” A, IEC on SP-Sepharose (Pharmacia), pH 8. The solid black line is the applied NaCl concentration. B, gel filtration on Sephacryl S-100 HR (Pharmacia); one of the four best fractions from the former IEC step. C, HAC on ceramic hydroxyapatite Macro-Prep (Bio-Rad), pH 8. The solid black line is the applied KH2PO4/K2HPO4 concentration. The MGlcDAG synthesis activity was assayed as described under “Materials and Meth- ods” with 27 mM amphiphiles present (3.7 mol % DOG, 33 mol % DOPG, and 63 mol % CHAPS).

Article Snippet: Hydroxyapatite Chromatography (HAC)—A column (16 mm in diameter) packed with 6 ml of ceramic hydroxyapatite Macro-Prep (Bio-Rad) was equilibrated with HAC-buffer: 100 mM KH2PO4/K2HPO4, pH 8, 20% (v/v) glycerol, and 20 mM CHAPS.

Techniques: Purification, Column Chromatography, Activity Assay, Concentration Assay, Filtration

Journal: Nutrients

Article Title: Antimicrobial Activity of EPA and DHA against Oral Pathogenic Bacteria Using an In Vitro Multi-Species Subgingival Biofilm Model

doi: 10.3390/nu12092812

Figure Lengend Snippet: Maximum projection of images obtained by confocal laser scanning microscopy (CLSM) of the 72 h biofilms, where the growth of these biofilms was observed on the surfaces of the hydroxyapatite discs after 60 s of exposure: ( A ) to the negative control (phosphate buffer saline); ( B ) to the ethanol solution; ( C ) to the docosahexaenoic acid (DHA) extracts (100 μM concentration) and ( D ) to 0.2% chlorhexidine. Specimens were stained with the LIVE/DEAD ® BacLightTM Bacterial Viability Kit solution, containing SYTO 9 and Propidium Iodide nucleic acid stains. Cells with a compromised membrane that are considered to be dead or dying were stain red (PI), whereas cells with an intact membrane were stain green (SYTO9).

Article Snippet: Sterile calcium hydroxyapatite (HA) discs, of 7 mm of diameter and 1.8 mm (standard deviation, SD = 0.2) of thickness (Clarkson Chromatography Products, Williamsport, PA, USA), were coated with treated saliva for 4 h at 37 °C in sterile plastic tubes, and then placed in the wells of a 24-well tissue culture plate (Greiner Bio-one, Frickenhausen, Germany).

Techniques: Confocal Laser Scanning Microscopy, Negative Control, Saline, Concentration Assay, Staining, Membrane

Journal: Nutrients

Article Title: Antimicrobial Activity of EPA and DHA against Oral Pathogenic Bacteria Using an In Vitro Multi-Species Subgingival Biofilm Model

doi: 10.3390/nu12092812

Figure Lengend Snippet: Scanning electron microscopy (SEM) of biofilms with an evolution of 72 h in hydroxyapatite (HA) discs treated with the negative control: phosphate buffer saline (PBS) ( A ), with docosahexaenoic acid (DHA) at 100 µM ( B ), with EtOH ( C ) or with the positive control: 0.2% chlorhexidine (CHX)( D ). A dense bacterial population could be observed on the HA discs treated with PBS ( A ), forming discontinuous layers of bacteria bonded to the discs. Meanwhile, on the biofilms of the discs treated with DHA ( B ), a lower density of cells distributed across the surface of the disc could be seen, and some of these exhibited structural damages. Likewise, on the discs treated with EtOH ( C ) or CHX ( D ), a reduction in the bacterial density present on the surface of the disc could also be observe, although it was lower than that on the discs treated with DHA ( B ). Chains of Aggregatibacter and/or Streptococcus (blue arrow) and fusiform bacilli of the F. nucleatum genus (yellow arrow) could be identified. Magnification ( A – D ): 1500×. The samples were dried by critical points and coated with gold by sputtering.

Article Snippet: Sterile calcium hydroxyapatite (HA) discs, of 7 mm of diameter and 1.8 mm (standard deviation, SD = 0.2) of thickness (Clarkson Chromatography Products, Williamsport, PA, USA), were coated with treated saliva for 4 h at 37 °C in sterile plastic tubes, and then placed in the wells of a 24-well tissue culture plate (Greiner Bio-one, Frickenhausen, Germany).

Techniques: Electron Microscopy, Negative Control, Saline, Positive Control, Bacteria

Journal: Nutrients

Article Title: Antimicrobial Activity of EPA and DHA against Oral Pathogenic Bacteria Using an In Vitro Multi-Species Subgingival Biofilm Model

doi: 10.3390/nu12092812

Figure Lengend Snippet: Maximum projection of images obtained by confocal laser scanning microscopy (CLSM) of the 72 h biofilms, where the growth of these biofilms was observed on the surfaces of the hydroxyapatite discs, stained with LIVE/DEAD ® BacLightTM Bacterial Viability Kit, after 60 s of exposure: ( A ) to the negative control (phosphate buffer saline); ( B ) to the ethanol solution; ( C ) to the docosahexaenoic acid (EPA) extracts (100 μM concentration) and ( D ) to 0.2% chlorhexidine. Specimens were stained with the LIVE/DEAD ® BacLightTM Bacterial Viability Kit solution, containing SYTO 9 and Propidium Iodide nucleic acid stains. Cells with a compromised membrane that are considered to be dead or dying were stained red (PI), whereas cells with an intact membrane were stained green (SYTO9).

Article Snippet: Sterile calcium hydroxyapatite (HA) discs, of 7 mm of diameter and 1.8 mm (standard deviation, SD = 0.2) of thickness (Clarkson Chromatography Products, Williamsport, PA, USA), were coated with treated saliva for 4 h at 37 °C in sterile plastic tubes, and then placed in the wells of a 24-well tissue culture plate (Greiner Bio-one, Frickenhausen, Germany).

Techniques: Confocal Laser Scanning Microscopy, Staining, Negative Control, Saline, Concentration Assay, Membrane

Journal: Nutrients

Article Title: Antimicrobial Activity of EPA and DHA against Oral Pathogenic Bacteria Using an In Vitro Multi-Species Subgingival Biofilm Model

doi: 10.3390/nu12092812

Figure Lengend Snippet: Scanning electron microscopy (SEM) of biofilms with an evolution of 72 h in hydroxyapatite (HA) discs treated with the negative control: phosphate buffer saline (PBS) ( A ), with docosahexaenoic acid (EPA) at 100 µM ( B ), with ethanol ( C ) or with the positive control: 0.2% chlorhexidine (CHX)( D ). A dense bacterial population could be observed on the HA discs treated with PBS ( A ), forming discontinuous layers of bacteria bonded to the discs. Meanwhile, on the biofilms of the discs treated with EPA ( B ), a lower density of cells distributed across the surface of the disc could be seen, and some of these exhibited structural damages. Likewise, on the discs treated with EtOH ( C ) or CHX ( D ), a reduction could also be observed in the bacterial density present on the surface of the disc, although this reduction was slighter than that on the discs treated with EPA ( B ). Chains of Aggregatibacter and/or Streptococcus (blue arrow) and fusiform bacilli of the F. nucleatum genus (yellow arrow) could be identified. Magnification ( A – D ): 1500×. The samples were dried by critical points and coated with gold by sputtering.

Article Snippet: Sterile calcium hydroxyapatite (HA) discs, of 7 mm of diameter and 1.8 mm (standard deviation, SD = 0.2) of thickness (Clarkson Chromatography Products, Williamsport, PA, USA), were coated with treated saliva for 4 h at 37 °C in sterile plastic tubes, and then placed in the wells of a 24-well tissue culture plate (Greiner Bio-one, Frickenhausen, Germany).

Techniques: Electron Microscopy, Negative Control, Saline, Positive Control, Bacteria