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Biomedical subjects

A J Pope

Publications and source records attributed to A J Pope.

50 records · Page 3Linked to original sources

The photodynamic effect of a pulsed dye laser on human bladder carcinoma cells in vitro.

The photodynamic effect of a pulsed flashlamp pumped dye laser on cultured human bladder carcinoma cells was studied. MGH-U1 cells were incubated for 1 h in dihaematoporphyrin either (DHE) and then exposed to green laser light (504 nm, 20 Hz) for varying laser power densities (50-100 mW/cm2 and exposure times (2-15 s), representing incident pulse energy fluences of 2.5-5 mJ/cm2 and energy densities of 0.1-1.5 J/cm2. The cell survival was measured by clonogenic assay and controls exposed to either laser light alone or DHE in the dark showed no cytotoxicity. Sensitised cells were killed by energy densities of less than 1 J/cm2 (LD90 = 0.54 J/cm2). This demonstrates the probable effectiveness of a pulsed dye laser for photodynamic therapy provided that pulse fluence are below the saturation threshold of the photosensitiser (10 mJ/cm2).

Carcinoma, Transitional Cell↗

Characterization of Cl- transport in vacuolar membrane vesicles using a Cl(-)-sensitive fluorescent probe: reaction kinetic models for voltage- and concentration-dependence of Cl- flux.

The effects of Cl- concentration and membrane potential (delta psi) on Cl- influx in isolated vesicles of vacuolar membrane (tonoplast) from red beet (Beta vulgaris L.) storage tissue have been characterized using the Cl(-)-sensitive fluorescent probe, 6-methoxy-1-(3-sulfonatopropyl)quinolinium (SPQ). The initial rate of Cl- transport into the vesicles was enhanced both by the imposition of a positive delta psi and by increases in extravesicular Cl- concentration. The kinetic mechanism underlying these responses was investigated by examining the accuracy with which the data could be described by several transport models. A model based on constant field theory yielded a poor description of the data, but satisfactory fits were generated by pseudo-two-state reaction kinetic models based on classical carrier schemes. Fits were equally good when it was assumed that charge translocation accompanied Cl- entry, or when charge was carried by the unloaded transport system, as long as only a single charge is translocated in each carrier cycle. Expansion of the models to three states enabled description of the Cl- concentration dependence of transport by changes in a single, voltage insensitive rate constant which is tentatively identified with Cl- binding at the external surface of the membrane. The derived value of the dissociation constant between Cl- and the transport system is estimated at between 30 and 52 mM.

Biological Transport↗

Dissipation of pH Gradients in Tonoplast Vesicles and Liposomes by Mixtures of Acridine Orange and Anions: Implications for the Use of Acridine Orange as a pH Probe.

Acridine orange altered the response to anions of both ATP and in-organic pyrophosphate-dependent pH gradient formation in tonoplast vesicles isolated from oat (Avena sativa L.) roots and red beet (Beta vulgaris L.) storage tissue. When used as a fluorescent pH probe in the presence of I(-), ClO(3) (-), NO(3) (-), Br(-), or SCN(-), acridine orange reported lower pH gradients than either quinacrine or [(14)C]methylamine. Acridine orange, but not quinacrine, reduced [(14)C]methylamine accumulation when NO(3) (-) was present indicating that the effect was due to a real decrease in the size of the pH gradient, not a misreporting of the gradient by acridine orange. Other experiments indicated that acridine orange and NO(3) (-) increased the rate of pH gradient collapse both in tonoplast vesicles and in liposomes of phosphatidylcholine and that the effect in tonoplast vesicles was greater at 24 degrees C than at 12 degrees C. It is suggested that acridine orange and certain anions increase the permeability of membranes to H(+), possibly because protonated acridine orange and the anions form a lipophilic ion pair within the vesicle which diffuses across the membrane thus discharging the pH gradient. The results are discussed in relation to the use of acridine orange as a pH probe. It is concluded that the recently published evidence for a NO(3) (-)/H(+) symport involved in the export of NO(3) (-) from the vacuole is probably an artefact caused by acridine orange.

Journal Article↗

Directional Ca2+ effect on stimulation of mucin secretion from chicken trachea in vitro.

Chicken tracheal mucosa in vitro transported and incorporated radioactive precursors into mucins, which were secreted at a steady rate into the tracheal lumen. Secretion of mucins labelled with (35)S and (3)H after pulse-labelling of the mucosal layer with Na(2) (35)SO(4) and d-[1-(3)H]glucosamine as precursors was an energy-dependent process, as it was strongly inhibited by the action of respiratory-chain inhibitors, an uncoupler of oxidative phosphorylation, a metabolic blocker and a temperature shift from 41 degrees C to 5 degrees C. On the other hand, both cholinergic and parasympathomimetic agents considerably increased the secretion of dual-radiolabelled mucins when applied on the submucosal side of the trachea. The effect of Ca(2+) was directional, since only high submucosal (3.6 or 18mm) or low luminal (zero or 0.18mm) Ca(2+) massively enhanced the secretion of radiolabelled mucin compared with the mucin output measured under physiological Ca(2+) conditions (1.8mm). Whereas application of ionophore A23187 on either side of the trachea significantly increased mucin output, its presence in the appropriate tracheal compartment and under appropriate Ca(2+) conditions further accentuated the output of radiolabelled mucins. Addition of acetylcholine under appropriate conditions also had an additive effect on the Ca(2+)-stimulated secretion of mucins. Ca(2+) stimulation of mucin secretion appears to be dependent on the metabolic integrity of the mucosal cells. Mucins secreted in response to high submucosal and low luminal [Ca(2+)] appear to consist of a number of different types of glycoproteins, as judged from their ion-exchange-chromatographic behaviour.

Animals↗

Factors influencing the viscous properties of chicken tracheal mucins.

1. Reduced viscosities, in water, of different types of mucin, such as fibrillar, gelatinous and soluble phase, separated from chicken tracheal secretions were measured. 2. H-bond breaking agents caused a significant decrease in the reduced viscosity of these mucins, but thiol-reagents alone did not have any effect. 3. Papain and Pronase did not cause any decrease in the reduced viscosity of these mucins. Neuraminidase decreased the reduced viscosity of soluble phase mucin by 50% by removing about 30% of its N-acetylneuraminic acid but had no effect on fibrillar and gelatinous mucins. Sulphatase neither removed any sulphate ester groups nor decreased the reduced viscosity. Due to some nonspecific intermolecular interaction, mixtures of mucins and enzymes or ovalbumin exhibited elevated reduced viscosities. 4. Ionic strength of the solutions appeared to decrease the reduced viscosity of these mucins. Increasing concentrations of Ca2+ in solutions of ionic strength of approx. 0.1 caused significant decrease in the reduced viscosity, but had no such effect in solutions of ionic strength of more than 0.1. 5. N-Acetylneuraminic acid and sulphate ester residues were 46.6 +/- 0.2, 43.4 +/- 0.6, 27.9 +/- 3.3 mg/g and 66.0 +/- 2.0, 34.2 +/- 3.3, 2.5 +/- 0.8 mg/g for fibrillar, gelatinous and soluble phase mucins, respectively. There appeared to be a good correlation between viscosity and N-acetylneuraminic acid contents among mucins of low reduced viscosities and between viscosity and sulphate ester residues among mucins of high reduced viscosities.

Animals↗

Physicochemical properties of avian tracheal mucus.

Dual-radiolabelled avian tracheal secretions were obtained by giving Na235SO4 and D-[1-3H]glucosamine simultaneously into the lumen of the trachea in preparations in vitro. These secretions comprised fibrillar, gelatinous and soluble-phase mucins. These were eluted as single components in the non-retarded fractions from Bio-Gel A-15m. Although no evidence of the presence of subunit structure was found, chemical and radiolabelling analyses showed a high degree of internal inhomogeneity among the three types of mucins. The differences among these mucins could be attributed to the chemical nature of their constituent glycoproteins. Glycoprotein fractions separated by ion-exchange chromatography were found to contain sulphate and N-acetylneuraminic acid residues in differing amounts. The overall acidic properties appeared to be correlated with ester sulphate content. A close similarity in the carbohydrate composition and a reciprocal relationship between the total ester sulphate residue contents and 35S- and 3H-labelling suggested that, in addition to stepwise glycosylation and sulphation, some pre-existing sulphated oligosaccharides might have been utilized for the synthesis of acidic glycoproteins.

Animals↗