Osmoregulation of vasopressin secretion and thirst in cyclical oedema.

Osmoregulation of vasopressin release and thirst was studied in the mid-follicular and mid-luteal phases of the menstrual cycle of five patients with cyclical oedema defined by peripheral oedema and weight gain (greater than 3.0 kg) manifest in two consecutive luteal phases. Results are compared to those already obtained in eight healthy women. In the patients, basal plasma osmolality in the mid-luteal phase was significantly lower than in the mid-follicular periods (patients, 283 +/- 1, 287 +/- 1 mOsmol/kg, respectively, mean +/- SEM, P less than 0.05; controls, 282 +/- 1, 286 +/- 1 mOsmol/kg, respectively, P less than 0.05). Plasma osmolality (pOsm) and plasma arginine vasopressin (pAVP) were measured during hypertonic (850 mmol/l) saline infusion in both phases of the cycle; linear regression analyses of these data gave the following mean regression equations, (i) mid-follicular, pAVP = 0.55 (pOsm - 285), r = 0.94 and (ii) mid-luteal, pAVP = 0.42 (pOsm - 281), r = 0.93. The abscissal intercept was significantly different (P less than 0.025). Osmotic threshold for severe thirst onset was lower in the mid-luteal phase compared to the mid-follicular value (296 +/- 1, 299 +/- 1 mOsmol/kg, respectively, P less than 0.01). Basal data and results of thirst onset and theoretical threshold for vasopressin release in response to osmotic stimulation obtained in the patients were similar to healthy control women. We conclude that osmoregulation in cyclical oedema is normal.

Acute suppression of plasma vasopressin and thirst after drinking in hypernatremic humans.

Drinking rapidly abolishes thirst and vasopressin secretion in dehydrated humans before major changes in plasma osmolality are observed. We studied the effects of drinking on plasma vasopressin and thirst in seven healthy volunteers rendered hypernatremic by the infusion of hypertonic (855 mmol/l) sodium chloride solution. Thirst was measured on a visual analogue scale (0-10 cm). Infusion of hypertonic saline caused linear increases in plasma osmolality (289 +/- 1 to 306 +/- 1 mosmol/kg, mean +/- SE, P less than 0.001), plasma vasopressin (0.6 +/- 0.2 to 6.4 +/- 1.9 pmol/l, P less than 0.001), and thirst (1.4 +/- 0.4 to 7.4 +/- 0.5 cm, P less than 0.001). Water was allowed 15 min after cessation of the infusion, and within 5 min of drinking both plasma vasopressin and thirst were significantly lower than postinfusion. After 20 min of drinking, plasma vasopressin had fallen from 6.5 +/- 0.9 to 1.3 +/- 0.3 pmol/l (P less than 0.001) and thirst from 7.7 +/- 0.5 to 1.0 +/- 0.2 cm (P less than 0.001) despite no significant change in plasma osmolality (306 +/- 0.9 to 304 +/- 0.8 mosmol/kg, P = 0.17), and the drinking of 1,200 +/- 60 ml of water, over 85% of the mean cumulative water intake in the 30-min drinking period. Control studies in the same subjects showed comparable rises in plasma vasopressin, plasma osmolality, and thirst during hypertonic saline infusion but no fall in any of these parameters during an equivalent 30-min period after the infusions, during which water was withheld.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of enzyme inhibitors of pregnancy associated oxytocinase: application to the measurement of plasma immunoreactive oxytocin during human labour.

The presence of the human placental enzyme, oxytocinase, in blood samples taken during pregnancy causes major methological problems in the radioimmunoassay for plasma oxytocin. Inadequate inhibition of the enzyme activity may lead to spuriously high or low values of plasma oxytocin. This study systematically investigates a variety of enzyme inhibitors. The optimum inhibitory system was obtained by the addition of 10 microliters of cold 125 mmol/l 1.10 phenanthrolene and 1 mol/l EDTA per ml of whole blood into the syringe. Complete enzyme inhibition was maintained for up to 60 min, during which time the lithium heparinized plasma samples were extracted by the Florisil method. Following extraction there was no enzyme activity in the extract residue. Concentrations of phenanthrolene and EDTA necessary to eliminate enzyme activity were 50- and 10-fold greater, respectively, than in any previously reported method. Recovery of synthetic oxytocin added to pregnancy plasma with inhibitors was 80% or higher, over the concentration range 1-100 pmol/l. Extract residue could be stored at -20 degrees C for up to 7 weeks. Dilutions of pregnancy plasma extracts ran parallel to the oxytocin standard curve. Studies on plasma concentrations of oxytocin (OT) during the first stage of labour in 6 patients showed that 3 had pulsatile plasma OT, peak values ranging from 4-10 pmol/l in phase with uterine concentrations, but 2 who had regular uterine activity had no episodic changes in plasma OT. One patient with hypocontractile labour had low non-fluctuating plasma OT.

Interference in the cytochemical assay of plasma vasopressin by a circulating factor induced by salt-loading in man.

Infusion of hypertonic saline into six normal volunteers caused an increase in plasma osmolality from 286.8 +/- 1.7 (mean +/- S.E.M.) to 307.6 +/- 2.6 mosmol/kg (P less than 0.001), a 7.1% increase in estimated blood volume, a rise in plasma immunoreactive arginine vasopressin (AVP) concentrations from 1.3 +/- 0.2 to 12.7 +/- 3.6 pmol/l (P less than 0.001) but no change in plasma AVP concentrations (2.1 +/- 0.9 and 1.9 +/- 1.3 pmol/l) as measured by a cytochemical technique based on the ability of AVP to stimulate rat renal medullary Na+/K+-ATPase activity. Addition of synthetic AVP to plasma obtained before, during and after hypertonic saline infusion also failed to stimulate Na+/K+-ATPase activity. The results suggest that infusion of hypertonic saline interfered with the cytochemical assay for AVP by inhibiting AVP-stimulated medullary Na+/K+-ATPase activity. We conclude that the use of this cytochemical method to detect plasma AVP has severe limitations under these experimental conditions.

Solid phase radioimmunoassay for direct measurement of human plasma oxytocin.

Synthetic oxytocin (OT) conjugated to bovine thyroglobulin by the carbodiimide reaction was injected into rabbits to raise a high titre, specific OT antiserum which was then coupled to microcrystalline cellulose activated by cyanogen bromide. High affinity of the coupled antiserum was defined by Scatchard analysis, Keq = 7.1 X 10(11)1/mol. Cross-reactivity studies revealed little binding of antiserum to analogues of OT. Iodination was performed by the Chloramine T method, giving specific activity of 125I-OT, range 1.1 - 1.7 X 10(3) Ci/mmol. After incubation for 40 hours under disequilibrium conditions, specific and non-specific bindings were 10.6 +/- 2.7% and 0.2 +/- 0.1% (n=15), respectively. Displacement of 50% 125I-OT occurred with 2.9 pg OT/tube. Coefficients of variation of standard OT concentrations (0.03 - 16 pg/tube) were less than 5%. Limit of detection was 2 pg OT/ml plasma. Recovery of synthetic OT added to non-pregnant plasma was 81.8% (n=34) at 20 pg/ml and 97.4% (n=32) at 100 pg/ml. Two patients, 17 and 18 weeks post-partum, had increases in plasma OT from less than 2 pg/ml to 18.3 and 16.0 pg/ml after 6 and 4 minutes breast feeding infants, respectively. We conclude that this solid phase OT radioimmunoassay is quick, relatively sensitive and reliable, and does not require prior extraction of plasma samples.