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Biomedical subjects

H E Edwards

Publications and source records attributed to H E Edwards.

29 records · Page 2Linked to original sources

The effects of autoclaving on the physical properties and biological activity of parenteral heparin preparations.

Heparin ampoules have been autoclaved at 115 degrees C, 121 degrees C, 126 degrees C and 130 degrees C for time intervals up to 50 min and the biological potency and physicochemical integrity of the preparations assessed. The anticoagulant activity, determined with the APTT assay, did not change significantly for the autoclaved samples but a decrease was observed using the BP assay. Autoclaving was also associated with a depolymerization process, as confirmed by high performance liquid chromatography, and the formation of 'species' with an absorption in the UV spectrum. However, autoclaving had no detrimental effect on the integrity of the anionic sites on heparin. It is proposed that Maillard-type reactions are responsible for these observations and the results indicate that autoclaving could be used to sterilize parenteral heparin solutions.

Animals↗

Quantitative analysis of a polysulfated xylan (SP54) in urine using gas-liquid chromatography.

A sensitive analytical method for the quantitation of a polysulfated xylan (SP54) in urine has been developed. SP54 and urinary glycosaminoglycans have been isolated from urine using cetylpyridinium chloride. This method removes all glycosaminoglycans with molecular weights less than 3000 Da. Following isolation, SP54 and urinary glycosaminoglycans have been selectively hydrolyzed under conditions (0.5 M HCl/105 degrees C/30 min) which produce an efficient yield of xylose from SP54 but not from the glycosaminoglycans. Xylose derived from SP54 has subsequently been determined using gas-liquid chromatography. Levels of SP54 down to 10 micrograms/ml have been determined using this technique.

Chromatography, Gas↗

The determination of the heparin neutralising capacity of protamine using acridine orange fluorescence.

A new method to estimate the heparin neutralising capacity of protamine has been investigated. The technique has been applied to two commercial heparin preparations tested against the W.H.O. 1st International Reference preparation of protamine. The technique is based on the fluorescence of acridine orange dye and the selective binding of heparin with protamine. A titration procedure has been devised and the two variations of the assay are compared and evaluated. The titration is rapid, accurate and convenient.

Acridine Orange↗

An investigation of calcium ion binding to heparins.

Using the line charge model of Manning and, where appropriate, a form of the empirical corrections to his limiting laws proposed by Wells, we have investigated the ion condensation phenomenon for four pure calcium heparin preparations and similar Ca-heparin/CaCl2 mixtures in aqueous solution. This has been expressed in terms of the single ion activity coefficient of the total calcium counter ion present. The results found for the single counter ion activity coefficient of pure Ca-heparin agree moderately well with the limiting law of Manning for a pure divalent counter ion polyanion salt in dilute solution. The results for Ca-heparin/CaCl2 mixtures when corrected for small ion-small ion interactions give a surprisingly good fit to the theoretical counter ion activity coefficient for a divalent counter ion/divalent cation simple electrolyte system, for values of X (X = the polyanion/co-ion ratio) up to 4, with slight deviations for values of X > 4. In the mixtures with the largest excess of polyelectrolyte, the single ion activity coefficients of the Ca2+ ion are in excess of the value predicted by Manning's theory. Ion exchange of sodium heparins to give the Ca-heparin samples used in the work did not appear to cause regular changes in the anticoagulant activity.

Blood Coagulation↗

Analysis of glycosaminoglycans in urine by using acridine orange fluorescence.

The fluorescence technique described here utilizes the electrostatic interaction between the polyanionic sites of glycosaminoglycans and the cationic dye Acridine Orange to analyse urinary glycosaminoglycans from patients suffering from mucopolysaccharidoses. The basis of the titration is the decrease in the fluorescence of free Acridine Orange that occurs when it is bound to polyanions. The effect of the presence of possible interfering materials such as salt, proteins and trace materials in urine was evaluated. This fluorescence technique is technically simple.

Acridine Orange↗

Effects of ionizing radiations on human costal cartilage and exploration of the procedures to protect the tissue from radiation damage.

The chemical changes produced following 60Co-gamma-irradiation of human costal cartilage have been monitored using "critical electrolyte concentration" (CEC) measurements utilizing Alcian Blue--magnesium chloride and Toluidine Blue--sodium chloride systems. The decrease in the CEC with radiation can be related to the decrease in hexose and hexosamine contents of the cartilage. Such changes arise as a result of degradation of the glycosaminoglycan component of the tissue. The state of the collagen fibres after irradiation has been examined using the Van Giesson stain and by biochemical assay of the hydroxyproline moeity. Energy transfer methods have been utilized to protect the glycosaminoglycan component of the cartilage from radiation damage.

Aged↗

Effects of 60Co gamma-irradiation on chondromucoprotein.

Pulse radiolysis and 60Co gamma-radiolysis have been used to study the effects of ionizing radiations on the protein-polysaccharide complex, chondromucoprotein. Both hydroxyl radicals and hydrated electrons readily react with the complex, with second-order rate-constants of 4-55 X 10(8)M-1s-1, respectively. Depolymerization of the carbohydrate component, which is initiated by reaction of OH, readily occurs after irradiation in aqueous solution, as is evidenced by an increase in the formation of reducing-end groups and a decrease in the viscosity of the solution. The ability of the anionic carbohydrate component to bind cationic counter-ions also decreases.

Animals↗

The study of rous sarcoma virus-transformed baby hamster kidney cells using fluorescent probes.

The fluorescent probes pyrene, pyrene butyric acid and N-phenyl 1-naphthylamine have been used to investigate the changes that accompany in vitro transformation of a baby hamster kidney cell line using Rous sarcoma virus. The fluorescent probes which reside in the membrane were used to compare the changes in microviscosity and polarity of the membranes of normal cells with two transformed cell lines. The spectrofluorimetric data indicate that following transformation the probe N-phenyl 1-naphthylamine resides in a more polar environment. However, using the probe pyrene, the yield of excimer indicates decreased mobility of this probe in the membrane of transformed cells. The data also indicate differences between the two transformed cell lines. Laser photolysis was used to study the lifetime of the pyrene probes and the quenching of the pyrene fluorescence in the membrane by several different quenching molecules. The data indicate differences between the three cell lines and suggest that transformation decreases movement within the membrane.

1-Naphthylamine↗