Rectangular quartz cell

All spectroscopic experiments were conducted in 50 mM, pH 7.4 Tris-HCl buffer containing 100 mM sodium chloride. CD and UV absorption spectra were recorded on a JASCO J-715 spectropolarimeter at 25 ± 0.2 °C. Temperature control was provided by a Peltier thermostat equipped with magnetic stirring. CD/UV titration experiments were performed in a rectangular quartz cell of 1 cm optical path length (Hellma, USA). Each spectrum represents the average of two scans obtained by collecting data at a scan speed of 200 nm/min. Absorption spectra were obtained by conversion of the high voltage (HT) values of the photomultiplier tube of the CD equipment into absorbance units. CD and UV curves of surfen-CDX and CDX-surfen mixtures were corrected by blank buffer or buffer solutions of CDXs, respectively. JASCO CD spectropolarimeters record CD data as ellipticity ('') in units of millidegrees (mdeg).

Spectroscopic experiments were conducted in ultra-pure deionized water (18.2 mΩ) or in 50 mM, pH 7.0/7.4 phosphate buffer (80/140 mM NaCl) using a rectangular quartz cell of 1 cm optical path length (Hellma, USA). CD and absorption spectra were recorded on a JASCO J-715 spectropolarimeter at 25 ± 0.2 °C and represent the average of three scans obtained by collecting data at a scan speed of 100 nm/min. Temperature control was provided by a Peltier thermostat equipped with magnetic stirring. Absorption spectra were obtained by conversion of the high voltage (HT) values of the photomultiplier tube of the CD equipment into absorbance units. CD and UV curves of drug-GAG mixtures were corrected by blank water or buffer solution. JASCO CD spectropolarimeters record CD data as ellipticity ('Θ') in units of millidegrees (mdeg). The quantity of 'Θ' is converted to molar circular dichroic absorption coefficient (∆ε in M -1 cm -1 ) using the equation ∆ε = Θ/(33982cl), where, 'c' is the molar concentration of the ligand (mol/L), and 'l' is the optical pathlength expressed in cm.

UV absorption spectra were recorded on a JASCO J-715 spectropolarimeter at 25 ± 0.2 °C.
Temperature control was provided by a Peltier thermostat equipped with magnetic stirring.
Each spectrum represents the average of two scans obtained by collecting data at a scan speed of 200 nm/min. 2.3 mM stock solution of RX-207 was prepared using ultra-pure water and a small volume of ethanol. 25 or 30 L stock solution of the ligand was added to 1.8 mL ultra-pure water in a rectangular quartz cell of 1 cm optical path length (Hellma, USA). Drug samples obtained by this way were subsequently titrated by small aliquots of concentrated aqueous solution of heparin and chondroitin 6-sulfate. Due to their poly-disperse nature, glycosaminoglycan (GAG) concentrations were expressed using the molecular weight of the average repeating disaccharide units: 665 and 503 g/mol for heparin and chondroitin 6sulfate, respectively. Absorption curves of drug-GAG mixtures were corrected by subtracting the spectra of blank water solution. Thioglycollate induced peritonitis that results in a massive neutrophil infiltrate into the peritoneum [16] , is a frequently used animal model to study acute inflammation [17] . Oral administration of RX-205 and RX-207 (50 mg/kg) inhibited neutrophil migration into the peritoneal cavity by 38% and 37%, respectively (Figure 2).