Determination of trace concentrations of dissolved nitric oxide in a biological buffer.

A new real-time method for measuring a trace concentration of nitric oxide (NO) in a complex matrix routinely used in pharmacological studies of its bioactivity is described. NO was quantified as a gas by chemiluminescence after extraction from a continuous liquid sample flow with a limit of detection of 0.042 nmol dm(-3) at a signal to noise ratio of 3. Theories to calculate the concentration of NO in the liquid sample flow from a direct measurement of NO in the extraction carrier gas are presented. The efficiency of extraction is determined by a stopflow experiment. An example is presented of the measurement of the steady-state concentrations of NO in Krebs-bicarbonate buffer at pH 7.4 and 37 degrees C when its liquid surface is sequentially exposed to gases containing various concentrations of NO in O2 plus CO2.

Circulating nitrite anions are a directly acting vasodilator and are donors for nitric oxide.

Inhaled nitric oxide (NO) is a pulmonary vasodilator, but also acts systemically, causing negative cardiac inotropic effects and a fall in systemic vascular resistance. Circulating metabolites of NO are presumed to be responsible. We questioned the role of nitrite anions and the manner in which they might contribute to these effects. Nitrite and nitrate anions coexist in blood, while circulating levels of dissolved NO are very low. Nitrate anions are not biologically active, but nitrite anions may have a biological role through the release of NO. In vitro, at 37 degrees C and in aerated Krebs bicarbonate solution, the steady-state concentration of dissolved NO was proportional to the concentration of NO in the gas. Nanomolar concentrations of dissolved NO coexisted with micromolar concentrations of nitrite anions. The idea of an equilibrium between the two in solution was also supported by the observed release of NO from nitrite anions in the absence of gas. With rings of precontracted pig pulmonary arteries (prostaglandin F(2alpha); 10 micromol/l), the steady-state concentration of dissolved NO causing 50% relaxation (EC(50)) was 0.84+/-0.25 nmol/l, corresponding to a gaseous concentration of 2.2 p.p.m. The EC(50) of nitrite was 4.5+/-0.7 micromol/l, a concentration normally found in plasma. The estimated concentration of dissolved NO derived from this nitrite was 4.5 pmol/l, some 100 times lower than would be needed to cause relaxation. The rate of exhalation of NO was increased and pulmonary vascular resistance was reduced by the addition of nitrite solution to the perfusate of isolated perfused and ventilated pig lungs, but only when millimolar concentrations were achieved. Thus circulating nitrite anions are a direct vasodilator, only being a carrier of effective amounts of "free" NO at higher than physiological concentrations.

Effect of varying alveolar oxygen partial pressure on diffusing capacity for nitric oxide and carbon monoxide, membrane diffusing capacity and lung capillary blood volume.

1. To examine the effect of varying oxygen partial pressure (PAO2) on nitric oxide (DLNO) and carbon monoxide (DLCO) diffusing capacity (transfer factor), 10 subjects performed combined DLCO/DLNO measurements with the inspired mixture made up with three different oxygen concentrations (25%, 18% and 15%) to give PAO2 values of 12-20 kPa. 2. A novel method is described for calculating membrane diffusing capacity (DM) and pulmonary capillary volume (Qc) from DLNO and DLCO. 3. The mean DMCO was 52.89 mmol min-1 kPa-1 and Qc was 0.056 litre. Reducing PAO2 from 20 to 12 kPa resulted in an increase in DLCO = -0.124 (O2%) + 11.67 (P less than 0.001) and a fall in DLNO = 0.538 (O2%) + 32.01 (P less than 0.001) and a fall in DLNO/DLCO = 0.107 (O2%) + 2.52 (P less than 0.001). DM (P = 0.59) and Qc (P = 0.64) also tended to fall with falling PAO2. 4. It appears more likely that the minor reduction in DLNO that we have observed with falling PAO2 is due to diffusion rather than reaction limitation.

A simultaneous single breath measurement of pulmonary diffusing capacity with nitric oxide and carbon monoxide.

Pulmonary diffusing capacity (DL) for carbon monoxide (CO) and nitric oxide (NO) were simultaneously measured in man using the single breath method, by adding 4O ppm of NO to the inspired gas and analysing the expirate for NO by a chemiluminescent method. The mean ratio of DLNO to DLCO in thirteen subjects was 4.3 (SD 0.3), mean DLNO = 49 mmol.min-1.kPa-1 (SD 10) and mean DLCO = 11 mmol.min-1.kPa-1 (SD 2). An increase in alveolar oxygen concentration from a mean of 18 to 68% in five subjects was associated with a 54% fall in DLCO but no change in DLNO. A reduction of lung volume from total lung capacity (TLC) (mean of 7 l) to a mean volume of 3.9 l in five subjects caused a fall in both DLNO (by 34%) and DLCO (by 8%). With 175 watts cycle exercise in three subjects the DLCO rose by 45% and DLNO by 25%. Since NO reacts much faster with haemoglobin than CO, DLNO should be influenced much less by reaction with haemoglobin, and perhaps represents a better index for the diffusing capacity of the alveolar-capillary membrane (Dm) than DLCO.

Pulmonary oedema in meningococcal meningitis.

Two cases of meningococcal meningitis complicated by pulmonary oedema are described. The pulmonary arterial wedge pressure was raised in the one case studied. Profound sympathetic over-activity may be the cause of the pulmonary oedema occurring in this situation. If this is so, adrenergic blockade would appear to be a rational approach to therapy.