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Biomedical subjects

P J Donaldson

Publications and source records attributed to P J Donaldson.

At least 19 recordsLinked to original sources
PubMed1995

Changes in lens connexin expression lead to increased gap junctional voltage dependence and conductance.

The differentiation of mouse lens epithelial cells into fiber cells is a useful model for studying the changes of the electrical properties of gap junction (cell-to-cell) channels that are induced by an alteration in connexin expression patterns. In this model, cuboidal lens epithelial cells differentiate into elongated fiber cells, and the expression of connexin43 (Cx43) in the epithelial cells is replaced with the production of high levels of Cx50 and Cx46 in the fiber cells. We now report a new procedure to isolate mouse lens fiber cell pairs suitable for double whole cell patch-clamp analysis. Analysis was also performed for fiberlike cell pairs differentiated from epithelial cells in culture. Voltage dependence and unitary conductance of fiber cell gap junction channels were determined and compared with the corresponding values previously measured for the channels joining lens epithelial cells and for lens connexin channels formed in Xenopus oocyte pairs. Our results support a differentiation-induced shift toward stronger gap junctional voltage dependence and larger unitary conductances in the fiber cells. Our data further reflect a balanced functional contribution of Cx50 and Cx46 in the fiber cell-to-cell channels rather than a predominance of a single connexin.

Animals↗
PubMed1991

Microelectrode studies of toad urinary bladder epithelial cells using a novel mounting method.

To optimise the conditions for recording stable membrane potentials, epithelial cells of short-circuited toad bladders were impaled via either their apical or basolateral membranes. Microelectrode impalements via the apical membrane were affected by impalement damage and were typically biphasic, consisting of an initial sharp increase in apical membrane potential (Vsc of around -26 mV), followed by a rapid depolarization of Vsc towards 0 mV in the next 10-20 s. To facilitate basolateral impalement two different methods for mounting bladders were tested. Both mounting methods yielded similar values for Vsc and Ra/Rb (the ratio of apical to basolateral membrane resistance) of around -57 mV and 5, respectively, which were larger than those recorded via the apical membrane and consistent with potential measurements from other tight epithelial tissues. Of the two basolateral mounting methods tested, the agar method gave the most stable impalements, making it possible to use amiloride and Ba2+ to assess for impalement damage. In conclusion, basolateral impalements of agar-mounted toad bladders makes this traditionally difficult tissue amenable to microelectrode studies.

Animals↗
PubMed1991

Channel reconstitution in liposomes and planar bilayers with HPLC-purified MIP26 of bovine lens.

The major intrinsic protein (MIP26) of bovine lens membranes, purified by HPLC, was incorporated into liposomes and planar bilayers. Permeability of MIP26 channels was studied in liposomes by a spectrophotometric osmotic-swelling assay, and channel electrical properties were monitored in planar bilayers following liposome fusion. Particle formation in liposomes was determined by freeze fracture. MIP26 channels were permeable to KCl and sucrose. In planar bilayers, channel-conductance transitions were observed only after addition of liposomes to both chambers and with voltages greater than +/- 20 mV. Channel open probability decreased progressively as voltage increased, and an open probability of 50% was at 60-80 mV, indicating that the channels are voltage dependent. Histograms of single-channel current amplitudes at 80 mV showed a Gaussian distribution that peaked at 10 pA (approximately 120 pS), after subtraction of 1 pA baseline current. Frequency distributions of open and closed times at 80 mV were single exponential functions with time constants of 0.13 and 1.9 sec, respectively. Open time constants ranged from 0.1 to 0.3 sec, and closed time constants ranged from 1 to 7 sec. Cs+ did not decrease conductance, but reduced mean open time from 0.2 to 0.038 sec and mean closed time from 1.5 to 0.38 sec. The increase in channel flickering with Cs+ occurred in bursts. TEA affected neither conductance nor kinetics. Channel events were also observed in Na+ solutions (zero K+). These data indicate that MIP26 channels are not K(+)-selective channels. Channel characteristics such as: permeability to molecules larger than small ions, conductance greater than 100 pS, long open and closed time constants, etc., are similar to those of gap junction channels.

Animals↗
PubMed1990

Effect of hyperosmotic challenge on basolateral membrane potential in rabbit urinary bladder.

In the rabbit urinary bladder, serosal hyperosmotic challenge (SHOC) with either 33 mM NaCl or 66 mM mannitol caused basolateral membrane potential (Vbl) to initially depolarize from -52.6 +/- 1.6 to -48.4 +/- 1.4 mV, followed by a recovery of Vbl to -57.5 +/- 1.3 mV after 13.7 +/- 1.0 min. The voltage recovery was dependent on both serosal HCO3- and Cl-, and in the absence of both, Vbl depolarized to -11.6 +/- 1.5 mV and the ratio of apical-to-basolateral resistance (Ra/Rbl) decreased from 21.0 +/- 3.4 to 8.3 +/- 3.1. This decrease in Ra/Rbl and consequent depolarization of Vbl is caused by a decrease in basolateral K+ conductance. Replacement of serosal Cl- with NO3- or SCN- followed by SHOC caused a sustained depolarization of Vbl to -32.5 +/- 4.4 and -40.9 +/- 0.9 mV, respectively. However, when Br- was used to replace Cl-, voltage recovery occurred but was slowed (24.0 +/- 2.7 min) and reduced in magnitude (-47.5 +/- 3.5 mV). Addition of amiloride (1 mM) or niflumic acid (100 microM), but not bumetanide (1 microM), to the serosal bathing solution inhibited voltage recovery causing Vbl to depolarize to -36.3 +/- 2.6 and -41.5 +/- 4.5 mV, respectively. Serosal addition of ouabain after SHOC caused Vbl to depolarize by 10.8 +/- 0.9 mV in 2 min. We speculate that the SHOC-induced initial depolarization of Vbl is a loss of Ba2(+)-sensitive K+ conductance caused by cell shrinkage. The subsequent repolarization/hyperpolarization of Vbl is caused by an enhanced basolateral membrane Na+ pump current and a reappearance of the Ba2(+)-sensitive K+ conductance. The parallel operation of Na(+)-H+ and Cl(-)-HCO3- exchanges will then supply Na+ for the pump current and, via cellular accumulation of Na+, K+, and Cl-, might result in a partial recovery of cell volume and thus Ba2(+)-sensitive K+ conductance.

Amiloride↗
PubMed1990

Vectorial secretion of a kallikrein-like enzyme by cultured renal cells. I. General properties.

Urinary kallikreins are proteolytic enzymes known to be secreted by distal nephron tubules. In this study, we demonstrate (using the chromogenic tripeptide substrate S 2266) that the renal cell line A6 from Xenopus laevis secretes a kallikrein-like enzyme. Secretion is present only when the cells are grown on filters, and enzyme is secreted only into the apical membrane bathing solution. Enzyme secretion consists of two components, one soybean trypsin inhibitor (SBTI) sensitive (SSBTI) and the other insensitive to SBTI (ISBTI). Both enzymes were inhibited by aprotinin, a kallikrein-like enzyme inhibitor. Using a bioassay, only the ISBTI enzyme produced a hypotensive effect on blood pressure and is thus a kallikrein-like enzyme. The apical membrane of cells grown on filters contains both enzyme species, whereas the basolateral membrane contains only the ISBTI (kallikrein-like) enzyme. Both enzymes were present in the apical membrane of cells grown on plastic. Initiation of enzyme secretion occurred after the cells formed electrically tight monolayers and the increase in membrane activity always preceded enzyme secretion. Using an irreversible inhibitor of the apical membrane-bound enzymes, the turnover rate for the SSBTI and ISBTI enzymes (cells on filters) was 3 and 7 h, respectively. Because the recovery of enzyme secretion was proportional to the recovery of membrane-bound enzyme activities, this suggests that enzyme secretion is due to the release of membrane-bound enzyme.

Animals↗
PubMed1989

Effects of serosal anion composition on the permeability properties of rabbit urinary bladder.

This study describes the effects of serosal Cl- and HCO3- substitution on transepithelial Na+ transport and basolateral membrane properties of the rabbit urinary bladder. Replacement of Cl- with NO3-, SCN-, and Br- had no effect on transepithelial Na+ transport or the basolateral membrane potential (Vbl). However, gluconate, isethionate, and cyclamate (anions that were shown previously to be not as permeable as Cl- through the basolateral membrane anion channel), decreased transepithelial Na+ transport and depolarized Vbl. Replacement of HCO3- also produced a decrease in transepithelial Na+ transport and a depolarization of Vbl. Utilizing conventional and K+-specific microelectrodes, we found the depolarization to be due to a reduction in basolateral K+ conductance. This depolarization was reversible only when Cl- was returned to the serosal solution, the normally permeant anion NO3- being unable to affect repolarization, suggesting that both the K+ and Cl- conductance are reduced during depolarization. A lag period of some 4 min preceded the repolarization of Vbl. The Na+-H+ exchange blocker amiloride prolonged the lag phase associated with repolarization, whereas niflumic acid, a Cl-(-)HCO3- exchange blocker (in red blood cells) reduced the magnitude of Vbl repolarization. Because of the possible involvement of the exchangers it is believed that the lag phase represents a volume-dependent and/or pH-dependent reactivation of the basolateral membrane conductances.

Amiloride↗
PubMed1987

The beta-2 microglobulin urine-to-serum ratio: an early marker of renal dysfunction in primary hyperparathyroidism.

Human beta-2 microglobulin (beta-2 M) is released into body fluids as a result of cell turnover, excreted by the kidney, and catabolized in the proximal tubule. Urinary excretion rates constitute a sensitive index of renal tubular function. The beta-2 M urine-to-serum ratio was measured in 25 patients with primary hyperparathyroidism (15 preoperative and 10 treated conservatively) in addition to 20 age- and sex-matched control subjects. The ratio was found to be significantly higher in both the operative and the conservatively managed groups compared to controls (p less than 0.05, Mann-Whitney U test). After surgical excision of a single parathyroid adenoma in the 15 operative cases, the beta-2 M urine-to-serum ratio fell to normal limits. These preliminary findings indicate that the urine-to-serum beta-2 M ratio may be of value in detecting change in renal function in asymptomatic patients with hyperparathyroidism. Further studies are indicated to establish whether these subtle changes are associated with long-term morbidity.

Humans↗
PubMed1985

The effect of oral contraceptive formulation and field-workers: a cautionary tale.

This paper examines oral contraceptive continuation rates and reported side effects in a large scale clinical research project conducted in Sri Lanka. The project evaluated standard- and low-dose pills used with and without vitamin supplements. Differences in continuation rates and perception of side effects are used to illustrate the impact that family planning field-workers can have on women's use of oral contraceptives.

Attitude↗
PubMed1984

Intracellular ionic activities in the EDL muscle of the mouse.

The intracellular ionic concentrations of sodium, potassium and chloride in the mouse EDL muscle were measured by chemical analysis using inulin as the extracellular marker. Cellular concentrations of 157 +/- 8, 38 +/- 3, 44 +/- 5 mmol kg-1 intracellular water were estimated for potassium, sodium and chloride respectively. The resting membrane potential was measured by a conventional microelectrode filled with 3 mol KCl and found to be -76 +/- 0.5 mV. Ion-selective microelectrodes were used to measure the intracellular ionic activities of potassium, sodium and chloride. The activities measured were 117 +/- 5, 16 +/- 2, 5 +/- 0.1 mmol l-1 for potassium, sodium and chloride respectively. Apparent activity coefficients for the intracellular ions were calculated. The observed discrepancies between the extracellular activity coefficient and the calculated apparent intracellular activity coefficients for sodium and chloride might be explained in terms of the binding to cellular macromolecules and/or the compartmentalisation of these ions. Potassium appears uniformly distributed throughout the cellular water. Intracellular chloride activity was similar to that predicted by the Donnan distribution and it is concluded, therefore, that chloride is distributed at electrochemical equilibrium.

Animals↗
PubMed1984

The impact of prenatal care on birth weight. Evidence from an international data set.

The impact of prenatal visits on birth weight in six countries is studied using data from an international collaborative maternity care monitoring project. The analyses presented pertain only to those women having singleton live births and whose pregnancies went to term. Results of the analyses indicate that around the world the number of prenatal visits is significantly related to birth weight. Controlling for other variables known to influence birth weight does not change the relationship. Further, contrary to previous research, there are no significant interactions with other variables.

Birth Weight↗
PubMed1983

Metronidazole v. cefoxitin in severe appendicitis--a trial to compare a single intraoperative dose of two antibiotics given intravenously.

In severe appendicitis, the effect of a single intravenous dose of metronidazole (500 mg) was compared with cefoxitin (1 g). The antibiotics were given by random allocation once the diagnosis had been established at operation. In the metronidazole group, 5 out of 48 patients developed a wound infection whilst in hospital compared with 13 out of 48 in the cefoxitin group (P = 0.036). However, 57% of wound infections became apparent after the patient went home and the overall infection rates were similar. The mean length of postoperative hospital stay was 5.0 days in the metronidazole group and 6.8 days in the cefoxitin group (P = 0.052), but of those who did develop a wound infection, the length of stay was almost double in the cefoxitin group. Anaerobic organisms were cultured from the wound in 7 out of 15 patients who received cefoxitin but in none of 5 patients in the metronidazole group. Whilst metronidazole only delayed the discharge of pus from the wound, it did seem to reduce the severity of infection. Cefoxitin appeared to be less effective, given as a single intravenous injection at a dose of 1 g.

Adult↗
PubMed1977

Medical education in Thailand: an alternate perspective.

This research is an attempt to provide greater understanding of two previously described medical education programmes in Thailand by placing them in historical perspective. One is a PhD programme to train medical school teachers and the other is a community health programme for medical students. The underying assumptions of the programmes are specified and through the use of archival material it is shown that in some respects the programmes are quite similar to earlier efforts to create western medical education programmes in Thailand. The analysis focuses on four major concerns of the programmes: creating small numbers of less well trained personnel; distributing medical resources to unserved rural areas; dealing with the special health needs of Thailand in the curricula; and self-consciously creating a professional elite. The discussion points up the need for historical data in evaluating the efficacy of medical education programmes.

Community Medicine↗
PubMed1976

Foreign intervention in medical education: a case study of the Rockefeller Foundation's involvement in a Thai medical school.

A case study of the process of foreign intervention in medical education in the developing world is presented. Material collected from the Rockefeller Foundation Archives on a Foundation program in Thailand is used to analyze the conditions under which foreign agencies and their personnel intervene in the development of medical professionals in the Third World and to study the problems that may occur as a result of such intervention. The importance of value consensus and the competitive advantage foreigners have in the marketing of professional models are highlighted as reasons for the diffusion of Western models of medical education throughout the developing world.

Community Health Services↗