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Biomedical subjects

H Pontoppidan

Publications and source records attributed to H Pontoppidan.

6 recordsLinked to original sources

Direct dye binding--a quantitative assay for solid-phase immobilized protein.

A direct dye-binding procedure was established for the quantification of protein after its immobilization on a solid phase, using IgG and BSA as model proteins. The assay, which in the range 0-5 mg protein/ml gel correlates well with indirect protein determination by A280 as well as determination of protein hydrolyzed from the gel, is based on a modified Bradford dye-binding assay. As the protein coupled to the gel binds the dye, a decrease in A465 of the supernatant is measured. Three solid supports commonly used for protein immobilization (Sepharose, Sephadex, Sephacryl) were found to be compatible with the dye-binding assay while nonspecific dye binding was found to HEMA gels. Protein was coupled to Sephacryl S-1000 using three different activation methods (aldehyde, hydrazine, and adipic acid dihydrazide). Artifactual dye-binding was not observed using any of the three different "linkers." The assay is easily carried out and represents a useful tool, e.g., when optimizing procedures for protein immobilization.

Coloring Agents

Hemodynamic responses to mechanical ventilation with PEEP: the effect of hypervolemia.

The hemodynamic effects of prolonged mechanical ventilation with positive end-expiratory pressure (PEEP), with and without blood volume augmentation, were studied in 18 beagles anesthetized with halothane (0.7 per cent end-tidal). Addition of 12 cm H2O PEEP during mechanical ventilation in normavolemic dogs was associated with reductions of transmural cardiac filling pressures, cardiac index and stroke index to 50 per cent of control values. Circulatory adaptation did not occur. Filling pressures and flow remained unchanged during the ensuing 8 hours when PEEP was maintained. They returned to control levels when PEEP was discontinued, except for the transmural right ventricular end-diastolic pressure, which remained elevated above control levels. Systemic vascular resistance was unchanged, but pulmonary vascular resistance doubled upon addition of PEEP. Following autologous whole blood transfusion (25 ml/kg) during mechanical ventilation with PEEP, cardiac index returned to, and remained at, control levels. After PEEP was discontinued, cardiac index increased acutely and remained elevated for the remainder of the study period (as long as 7 hours). Comparable transfusion during mechanical ventilation without PEEP elevated cardiac index only transiently. Right atrial, pulmonary capillary wedge, and right and left ventricular end-diastolic pressures showed marked increases relative to atmospheric with PEEP and after transfusion. Calculated transmural pressures demonstrated clear reductions with application of PEEP, followed by increases to control levels with transfusion and further increases to above control when PEEP was discontinued. Study of ventricular function curves revealed that changes in filling pressures and not to changes in ventricular contractility. Transmural pulmonary arterial diastolic pressure rose throughout the 12 hours of study, despite return of pulmonary vascular resistance to control level with removal of PEEP. Thus, acute decreases in cardiac filling pressure, cardiac index, and stroke index persist consequent to application of PEEP, and circulatory adaptation does not occur. The apparent hemodynamic deterioration may be reversed by blood volume augmentation, but when PEEP is discontinued, hypervolemia with consequent increases in filling pressures and a move along a ventricular function curve will occur. Changes in cardiac index will depend upon the overall state of right and left ventricular contractility.

Animals

Extracorporeal perfusion for acute respiratory failure: recent experience with the spiral coil membrane lung.

Selection criteria, clinical data, and physiological measurements obtained during five extracorporeal membrane lung perfusions for acute respiratory insufficiency are presented. Four patients died and 1 survived. A new technique of femoral artery cannulation to allow aortic arch perfusion is described. When properly monitored, this route provides improved oxygen delivery to the brain during venoarterial (VA) perfusion. The importance of monitoring the equivalent of carotid artery Po2 during VA perfusion is emphasized. Recognition of the effects of high cardiac output in limiting the quality of extracorporeal perfusion, plus the use of hypothermia to reduce output, are stressed. We have confirmed that perfusion can be accomplished with small quantities of heparin, so that bleeding is reduced, but thrombocytopenia and occasional hemorrhage continue to be persistent problems.

Acute Disease

Effect of mechanical ventilation with end-inspiratory pause on blood-gas exchange.

The effects of end-inspiratory pause (EIP) on gas exchange were measured in 10 adult patients with acute respiratory insufficiency while maintained on mechanical ventilation. Four inspiratory patterns were studied with a constant tidal volume (10 to 15 ml/kg body weight), respiratory rate (9 to 12 breaths/min), FIO2 (0.5) and end-expiratory pressure. Inspiratory flow rate (V insp) and EIP time were varied to produce a control pattern (V insp = 60 L/min, EIP = 0), 2 EIP patterns of 0.6 and 1.2 seconds with a similar V insp and a "slow" flow pattern (V insp = 30 L/min) without EIP. The control pattern was applied before and after each study period. Arterial oxygenation was unchanged with both EIP and "slow" flow patterns when compared to control. Dead-space ventilation (VD/VT) and Paco2 were significantly decreased (p less than 0.01) as EIP was increased from 0 to 1.2 seconds, but remained unchanged with slow inspiratory flow. Thus, EIP improved the efficiency of ventilation with no apparent improvement in oxygenation in patients with acute respiratory insufficiency.

Adult