Journal: Pain

Article Title: Downregulation of adenosine and adenosine A1 receptor contributes to neuropathic pain in resiniferatoxin neuropathy

doi: 10.1097/j.pain.0000000000001246

Figure Lengend Snippet: Changes in prostatic acid phosphatase (PAP) ectonucleotidase activity and alterations of adenosine hydrolysis in resiniferatoxin (RTX) neuropathy. (A–C) Prostatic acid phosphatase ectonucleotidase activity was assessed through enzyme histochemistry using adenosine monophosphate (AMP, 3 mM) as a substrate. The reaction product was developed with 1% sodium sulfide and appeared dark brown in the dorsal root ganglion (DRG) sections of the vehicle (A) and day 7 after RTX neuropathy (RTX group) (B). (C) The graph shows AMP(+) neuronal densities in the vehicle (open bar, n = 5) and RTX (filled bar, n = 5) groups according to (A and B). (D–G) High-performance liquid chromatography (HPLC) was conducted to assess the change in the contents of adenosine and AMP, which were extracted from DRG. (D) This graph shows the HPLC chromatograms of the eluted standards (upper panel; AMP, 6.5 minutes; adenosine, 10.2 minutes), the vehicle group (middle panel), and the RTX group (bottom panel). Number designation on the chromatograms: 1 for AMP and 2 for adenosine. (E and F) These figures provide verification of the peaks for AMP (E) and adenosine (F) samples extracted from DRG tissues with respect to a spectral library. The absorbance spectra of the adenosine and AMP samples (pink line in E and F) were the same as the absorbance spectra of the adenosine standard (similarity index = 0.99) and the AMP standard (similarity index = 1.00) (black line in E and F). (G) A significant difference in adenosine signaling molecules was present between the vehicle (open bar, n = 6) and RTX groups (filled bar, n = 5) (ie, decreased adenosine and increased AMP). Elution time scale, minutes. * P < 0.05, ** P < 0.01. Bar, 50 µm.

Article Snippet: A Brownlee SPP HPLC column (C18, 2.7 µm, 4.6 × 150 mm; PerkinElmer, Waltham, MA) protected with a guard cartilage (C18, 2.7 µm, 4.6 × 5 mm; PerkinElmer) was used in this assay.

Techniques: Activity Assay, High Performance Liquid Chromatography

Journal: Scientific Reports

Article Title: Burkholderia pseudomallei biofilm resists Acanthamoeba sp. grazing and produces 8- O -4′-diferulic acid, a superoxide scavenging metabolite after passage through the amoeba

doi: 10.1038/s41598-023-43824-1

Figure Lengend Snippet: Schematic flow chart of the co-cultivation of two different B. pseudomallei biofilm phenotypes and Acanthamoeba sp. Adhesion and intracellular-survival assays at MOI 100 using non-encapsulated biofilm cells were performed. Burkholderia pseudomallei were passaged through Acanthamoeba sp. up to three times and were then collected for metabolomic analysis using ultra-high-performance liquid chromatography-electrospray ionization-quadruple time-of-flight mass spectrometry in parallel with observations of colony morphology on Ashdown’s agar. In addition, preformed 24-h and 48-h B. pseudomallei biofilm was cocultured with Acanthamoeba sp. to monitor the biofilm structure and biofilm biomass using confocal laser scanning microscopy. Amoeba cells were counted using a hemocytometer.

Article Snippet: In brief, the aqueous phase extracts of samples were analysed on a reverse-phase liquid chromatography platform using a Bruker intensity HPLC C18 (2.1 × 100 mm, 2 µm column).

Techniques: High Performance Liquid Chromatography, Mass Spectrometry, Confocal Laser Scanning Microscopy