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Biomedical subjects

R A Ward

Publications and source records attributed to R A Ward.

At least 37 records · Page 2Linked to original sources

Heparinization for routine hemodialysis.

Routine hemodialysis requires anticoagulation with heparin to prevent clotting in the extracorporeal circuit. Activated whole blood clotting times are used to measure the level of anticoagulation during dialysis. The magnitude of the clotting time obtained for a given level of heparin depends on the test used, and each dialysis unit must establish its own target range. The preferred dosage regimen for heparin is an initial loading dose, followed by a constant infusion. Dosage regimens based on intermittent boluses of heparin are associated with periods of under- and over-anticoagulation and require more staff attention during dialysis. Because patient responses to heparin differ markedly, the doses required to achieve suitable anticoagulation must be determined on an individual basis. The doses of heparin for an individual patient are best determined using a simple pharmacodynamic model, although they can be found empirically through trial and error. In spite of careful anticoagulation, an unsatisfactory outcome may still result. Patient-specific factors may be responsible for some of these adverse outcomes; however, in most cases they result from poor technique, including failure to deliver the prescribed dose of heparin, turbulence and foam formation in the extracorporeal circuit, and excessive access recirculation.

Anticoagulants↗

Depletion of total salivary gland protein in blood-fed Anopheles mosquitoes.

Reduction in total salivary gland protein from four anopheline vectors of human malaria, Anopheles stephensi Liston, An. albimanus Wiedmann, An. gambiae Giles, and An. freeborni Aitken, was quantified after mosquitoes blood-fed to repletion on human volunteers, hamsters or through a Baudruche artificial membrane. Total salivary gland protein from pools of six unfed mosquitoes ranged from 4.33 to 7.91 micrograms/ml. The difference between the total protein of glands from unfed and blood-fed mosquitoes for all species ranged from 1.77 to 3.12 (micrograms/ml for six pooled salivary glands. Total salivary gland protein for mosquitoes blood-fed to repletion was significantly less than that of unfed controls from the same cohort. Reduction in total salivary gland protein for An. freeborni and An stephensi blood fed to repletion on human volunteers, hamsters, and a Baudruche membrane ranged from 24 to 46%, from 43 to 56%, and from 24 to 51%, respectively. An. stephensi mosquitoes were allowed to blood feed on humans for 0 (unfed), 0.5-, 1.0-, 2.0-min time periods or to repletion (> 2-5 min). As feeding time increased, there was a significant decrease in total amount of protein in the salivary glands. This decrease was proportional over time, indicating that salivation occurred continuously from the beginning (probing) of blood feeding to withdrawal of the mosquito mouthparts at repletion. These data indicate that during blood feeding there difference between species in the salivary gland output measured as amount of protein depleted from the salivary glands and that depletion of salivary protein from the glands occurred continuously as mosquitoes fed to repletion.

Animals↗

Hemodialysis with cellulose membranes primes the neutrophil oxidative burst.

Hemodialysis with cellulose membranes causes a complement-mediated neutropenia. Changes in neutrophil function have also been reported; however, it is unclear if these changes indicate a direct effect of the membrane on neutrophils or if they are a consequence of the neutropenia. We tested the hypothesis that neutrophil oxidative burst activity is enhanced during dialysis with cellulose membranes. Resting and Staphylococcus aureus-stimulated H2O2 production were determined predialysis and in blood entering and leaving the dialyzer during the first 30 min of dialysis and in blood leaving the membrane module in a single-pass on-line model of hemodialysis. Resting H2O2 production increased slightly but significantly during the first 30 min of dialysis. Transit of neutrophils through the dialyzer caused a marked increase in stimulated H2O2 production, indicating priming of the oxidative burst. However, priming was limited to the first 5 min of dialysis before the onset of neutropenia. In contrast, stimulation and priming of H2O2 production persisted throughout 30 min of single-pass on-line perfusion. These results indicate that cellulose membranes both stimulate and prime neutrophil oxidative burst activity but that these effects are partially obscured by neutropenia.

Adult↗

Technical challenges to successful dialyzer reuse.

Limitations of dialyzer reuse technology may contribute to adverse patient outcomes. Assessing changes in dialyzer performance by measuring changes in total cell volume may not be valid for all membranes and reprocessing methods. Systems which determine dialyzer performance on-line are superior. Compliance with standards for microbial contamination of reprocessing fluids is poor and improvements in water treatment system design and monitoring are needed. Technological innovation is required to enable dialyzer performance to be monitored over an expanded molecular weight range and to ensure that dialyzer reprocessing can be routinely performed with sterile, non-pyrogenic solutions.

Equipment Reuse↗

Polymorphonuclear leukocyte oxidative burst is enhanced in patients with chronic renal insufficiency.

Previous reports that polymorphonuclear leukocyte (PMN) function is impaired in hemodialysis patients do not differentiate between effects of dialysis and of uremia. The hypothesis that chronic renal insufficiency impairs PMN function was tested. Phagocytosis and oxidative burst were measured in PMN from patients with varying degrees of chronic renal insufficiency impairs PMN function was tested. Phagocytosis and oxidative burst were measured in PMN from patients with varying degrees of chronic renal insufficiency (creatinine clearance, 6 to 35 mL/min per 1.73 m2) and normal subjects. The ability of tumor necrosis factor-alpha (TNF-alpha) to prime the oxidative burst was also assessed. Phagocytosis of Staphylococcus aureus and basal H2O2 and O2- release by PMN did not differ between normal subjects and patients with chronic renal insufficiency. However, the oxidative burst stimulated by S. aureus and formyl-Met-Leu-Phe, but not phorbol myristate acetate, was significantly enhanced in PMN from patients with chronic renal insufficiency. The increase in formyl-Met-Leu-Phe-stimulated oxidative burst correlated significantly with the level of renal function. TNF-alpha significantly increased S. aureus-induced H2O2 production in normal PMN, but not in PMN from patients with chronic renal insufficiency. These data indicate that chronic renal insufficiency does not impair PMN phagocytosis and oxidative burst. To the contrary, it enhances receptor-mediated oxidative burst. The inability of TNF-alpha to further enhance the oxidative burst suggests that PMN exist in a primed state in patients with chronic renal insufficiency.

Adult↗

The effects of RU 33965 and RU 34030, two new 3-cyclopropyl carbonyl imidazobenzodiazepines, on GABAA receptor-mediated synaptic transmission in cerebellar slices in the rat.

1. Two 3-cyclopropyl carbonyl imidazobenzodiazepines, RU 33965 and RU 34030, were tested for their ability to modulate GABAA synaptic transmission in rat cerebellar slices. The action of the full benzodiazepine agonist RU 32007 and the inverse agonist Ro19-4603 were tested for comparison. 2. Extracellular recordings were made from the Purkinje cell layer of the cerebellar slices and inhibition induced by just threshold electrical stimulation of the parallel fibres was bicuculline sensitive. 3. The major effect of RU 32007 when examined at 100 nM and 1 microM was to increase the GABAA mediated inhibition in the slice. 4. In contrast the major effect of the inverse agonist Ro19-4603 was to reduce the period of inhibition. 5. RU 33965 and RU 34030 at 10 and 1 microM respectively either had little effect on GABAA mediated inhibition or decreased it slightly. 6. RU 34030, 1 microM, abolished the agonist effect of RU 32007, 1 microM. 7. The effects of RU 32007 and Ro19-4603 were abolished by the benzodiazepine antagonist flumazenil. 8. It is concluded that both RU 33965 and RU 34030 have marginal inverse agonist properties.

Animals↗

Methods for the assessment of neutrophil function during extracorporeal circulation.

Many different assays and test systems have been used to study the effects of biocompatibility on neutrophil function in haemodialysis patients. This review examines the most common and recommends three assays that are widely available, amenable to standardization, and minimally susceptible to artefacts. The recommended assays are neutrophil count, plasma concentrations of the elastase- alpha 1-proteinase inhibitor complex (a measure of neutrophil degranulation), and, the neutrophil oxidative burst response as measured by the reduction of ferricytochrome-c by superoxide or the oxidation of dihydrorhodamine 123 by hydrogen peroxide. This review also discusses the advantages and disadvantages of using whole blood or isolated cell assays and the choice of in-vitro, on-line, or clinical dialysis test systems. While whole blood assays are associated with fewer artefacts than those using isolated cells, specialized equipment is needed to study neutrophils and some questions concerning localization of defects in neutrophil function can only be addressed in isolated cells. For these reasons a role for isolated cell assays continues. For similar reasons there remains a role for in-vitro and on-line test systems, even though clinical dialysis studies are preferred whenever possible.

Blood Specimen Collection↗

Phagocytic cell function as an index of biocompatibility.

This review describes the physiology and biochemistry of phagocytic cells and examines the rationale for using their function to assess biocompatibility. Haemodialysis modulates phagocyte adhesion receptors to a degree dependent on the level of complement activation by the membrane, and this modulation leads to a proportional change in the number of circulating phagocytes because of pulmonary sequestration of activated cells. However, it is not clear that other acute changes in phagocyte function are directly attributable to the nature of the membrane; they may merely reflect changes in the population of circulating cells resulting from pulmonary sequestration. While haemodialysis is associated with stimulation of monocyte cytokine production, the relative contributions to this phenomenon of the membrane material, its mass transfer properties, and the microbiological quality of the dialysate are incompletely understood. A lack of well-designed studies and difficulty in separating the effects of uraemia from those of dialysis make it impossible to know if chronic changes in phagocyte function are a consequence of the choice of dialysis membrane. Thus, while changes in the number of circulating phagocytes and their expression of adhesion receptors are useful indices of biocompatibility, a clear association has yet to be shown between membrane properties and changes in other measures of phagocytes function.

Cytokines↗

Biocompatibility of a new high-permeability modified cellulose membrane for haemodialysis.

The biocompatibility and solute permeability characteristics of a high-permeability modified cellulose membrane (Hemophan-HP) (He-HP) were compared with those of two synthetic membranes (poly(ethylene-co-vinyl alcohol) (EVAL) and poly(acrylonitrile-co-sodium methallyl sulphonate) (AN69)) and Cuprophan in a multicentre, four-way cross-over clinical trial. Cuprophan membranes caused significant complement activation, leukopenia, and granulocyte elastase release. He-HP membranes demonstrated a lesser effect, which was similar to that observed for the EVAL membrane, although less than that seen with the AN69 membrane. A similar order for the four membranes was seen for their effect on platelets. Cuprophan membranes provided superior small-molecule removal to the other three membranes. In contrast, Cuprophan was essentially impermeable to beta 2-microglobulin, whereas He-HP, EVAL, and AN69 allowed the removal of 60-90 mg of beta 2-microglobulin per treatment. However, a decrease in the plasma concentration of beta 2-microglobulin was observed only with the AN69 membrane, most probably as a result of the ability of that membrane to adsorb proteins. Our results demonstrate that high-permeability membranes of comparable biocompatibility to some synthetic membranes can be fabricated from cellulose derivatives.

Adult↗

Prevention of blood loss in dialysers with DEAE-cellulose membranes does not require increased doses of heparin.

Binding of heparin to DEAE-cellulose membranes may reduce bioavailable heparin, thus increasing the amount of heparin needed for anticoagulation during dialysis. To test this hypothesis, blood loss and coagulation were evaluated during dialysis with DEAE-cellulose and polysulphone membranes. The heparin dose required to effect a given increase in the baseline recalcified activated clotting time (RACT) was determined using a pharmacokinetic model. Blood remaining in the dialyser post-dialysis (RBV) was measured by red cell lysis and haemoglobinometry. Plasma thrombin-antithrombin III complex (TAT) was used to assess activation of the coagulation system. RBV and changes in TAT were determined in two crossover studies. Firstly, DEAE-cellulose membranes were used at doses of heparin calculated to increase baseline RACT by 12.5% and 25%. Secondly, DEAE-cellulose and polysulphone membranes were compared at a heparin dose calculated to increase baseline RACT by 15%. RBV for DEAE-cellulose membranes was independent of the dose of heparin and did not differ from that found for polysulphone membranes. TAT concentrations increased during dialysis; however, there was no difference between the two membranes. These results show that DEAE-cellulose membranes do not require increased heparin to avoid dialyser-associated blood loss.

Adult↗

Origin and fate of fetuin-containing neurons in the developing neocortex of the fetal sheep.

The development of the neocortex has previously been extensively studied in carnivores (cat and ferret), rodents (rat and mouse) and primates (monkey and human). In these species, it has been shown that the initial population of cells migrating from the ventricular zone forms the primordial plexiform layer. This is subsequently split into marginal zone and subplate zone by the insertion of later-migrating cells into the primordial plexiform layer, to form the cortical plate proper. Many of the cells derived from the split primordial plexiform layer are transient. The neurons of the subplate zone are found in the deeper part of layer VI, and white matter deep to layer VI in the more mature cortex; most of these neurons disappear by adulthood. [3H]-thymidine labelling in the present study has shown a similar pattern of neocortical development in Artiodactyla (sheep). In addition it has been shown that the previously described staining of subplate and cortical plate cells for the fetal protein fetuin indicates that fetuin is a useful marker for a proportion of this transient population of neurons and defines its extent in neocortical development more clearly. Dividing cells were labelled by a single intra-amniotic injection of [3H]-thymidine at E26 to E35 (birth is at E150). The brains were subsequently examined at E40 or E80 for [3H]-thymidine labelling and fetuin staining by a combination of autoradiography and immunocytochemistry. The earliest generated neocortical cells detected in this study (E26) were found in two layers by E40, the outer marginal zone and inner subplate zone. Neurons of the marginal zone were generated up to E28; those of the early subplate zone were generated up to E31. The cortical plate proper was generated by cells "born" on E32 and later. This sequence is similar to that described in other species, especially the cat. A proportion of the early-generated neurons in the marginal zone, subplate zone and early cortical plate stained for fetuin. By E80 these earliest-generated, fetuin-positive cells were found in the white matter deep to the forming neocortical layers and in layer VI. In adult brains no fetuin-positive neurons could be identified in the neocortex, and neurons had almost entirely disappeared from the white matter. The fetal glycoprotein fetuin seems to be specifically associated with a population of cells that has the same developmental history as the transient marginal zone and subplate neurons described in other species.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Effects of RU33368, a low affinity ligand for neuronal benzodiazepine receptors, on rodent behaviours and GABA-mediated synaptic transmission in rat cerebellar slices.

1. The effects of the low affinity benzodiazepine receptor ligand RU33368 were studied on rodent behaviours and on GABA-mediated synaptic transmission in rat cerebellar slices. 2. RU33368 inhibited stress induced ultrasounds in rat pups without inducing marked muscle relaxation. RU33368 also enhanced operant responding in rats that had been suppressed by mild footshock. These effects of RU33368 in these two models of anxiety were both blocked by the benzodiazepine antagonist Ro15-1788 (flumazonil). 3. In cerebellar slices RU33368 enhanced stimulus-induced synaptic inhibition of Purkinje layer cells with a minimal effective concentration in the order of 1 microM. The classical benzodiazepine agonist RU32007 was approx. 10 times more potent. This action of RU33368 was blocked by Ro15-1788. 4. The minimal effective concentration of RU33368 fully blocked the effect of RU32007 in 2 of 4 cells tested and partially antagonized it in a third cell. 5. These data suggest that RU33368 is a partial agonist at benzodiazepine receptors and this, at least in part, explains its non-sedative anxiolytic behavioural profile.

Animals↗

Characterization of thromboxane and prostacyclin effects on pulmonary vascular resistance.

Although thromboxane and prostacyclin (PGI2) have long been described as major controllers of pulmonary vascular resistance, little has been reported on the characteristics of the interactions between the two arachidonic acid products. The current study uses segmental vascular resistance and compliance measurements to evaluate the actions of thromboxane and PGI2 in isolated blood-perfused rat lung. The thromboxane analogue U-46619 increases pulmonary vascular resistance by increasing only small artery resistance and decreases pulmonary vascular compliance in the middle compartment. Among the vascular effects of U-46619 are a maximum increase in resistance (RmaxU-46619) of 60.3 +/- 15.6 cmH2O.l-1.min.100 g-1 and a concentration required for 50% of maximum increase (K0.5,U-46619) of 1.60 +/- 0.85 nM for small artery resistance, a minimum vascular compliance (CminU-46619) of -0.93 +/- 0.58 cmH2O, and a K0.5,U-46619 of 1.10 +/- 1.60 nM for middle compartment compliance. Similar results were obtained for total resistance and total compliance. The effects of PGI2 on thromboxane-induced resistance and compliance changes were evaluated using K0.5,PGI2, RmaxPGI2, and CmaxPGI2 at each dose of thromboxane. PGI2 was more effective in reversing the thromboxane constriction at higher concentrations of thromboxane. These data show that the absolute concentration of PGI2 and thromboxane and not a simple ratio of thromboxane to PGI2 determines vascular tone.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Evaluation of prostaglandin F2 alpha and prostacyclin interactions in the isolated perfused rat lung.

To characterize the interactions between prostaglandin F2 alpha and prostacyclin in controlling tone in the pulmonary circulation, isolated rat lungs were ventilated, perfused with blood, and subjected to challenge by prostaglandin F2 alpha in increasing doses. The pulmonary resistance was evaluated using occlusion techniques that separate the resistance into segments of large and small arteries and veins. The total vascular compliance was evaluated using outflow occlusion. Resistance increased after prostaglandin F2 alpha, and this resistance change was primarily in the small artery segment. The maximum resistance increase by prostaglandin F2 alpha (Rmax,PGF2 alpha), calculated from the Michaelis-Menton equation, was 16.6 +/- 3.6 cmH2O.l-1.min.100 g-1 for total vascular resistance with a concentration required to produce 50% Rmax (K0.5) of 5.26 +/- 3.57 nM. The Rmax,PGF2 alpha for small artery resistance was 13.5 +/- 2.4 cmH2O.l-1.min.100 g-1 with a K0.5 of 2.35 +/- 1.57 nM. The vascular compliance decreased during vasoconstriction by prostaglandin F2 alpha, and the maximum decrease in compliance (Cmin,PGF2 alpha) was -0.43 +/- 0.12 ml/cmH2O with a K0.5 of 2.84 +/- 2.99 nM. At each dose of prostaglandin F2 alpha, prostacyclin was administered in increasing doses to reverse the vasoconstriction caused by prostaglandin F2 alpha. For each concentration of prostaglandin F2 alpha, prostacyclin almost completely reversed the resistance increases and approximately one-half the compliance decrease. The maximum change in vascular resistance or compliance produced by prostacyclin was dependent on the dose of prostaglandin F2 alpha; yet the K0.5 for prostacyclin was within the picomolar range for all doses of prostaglandin F2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Modulation of GABA-mediated inhibition in rat cerebellar slices by benzodiazepine receptor ligands.

1. Extracellular recordings were made from the Purkinje cell layer of rat cerebellar slices. Compounds were perfused over the slice and bipolar stimulating electrodes placed in the external layer of the slice close to the recording electrode. 2. Stimulus-evoked inhibition of Purkinje layer cell activity was sensitive to bicuculline methiodide and picrotoxin, suggesting it was gamma-aminobutyric acid (GABA) mediated. The benzodiazepine ligands RU 32007 and Ro 19-0528 reversibly increased the period of inhibition, as did pentobarbital. This benzodiazepine effect was antagonised by Ro 15-1788. 3. Five inverse agonists all reduced the period of stimulated inhibition and this effect was reversed by Ro 15-1788, suggesting the involvement of benzodiazepine receptors. 4. It is concluded that this system provides a convenient physiological and possibly quantitative model for studying the action of benzodiazepine receptor ligands.

Animals↗