Ecstasy (MDMA)-induced hyponatraemia is associated with genetic variants in CYP2D6 and COMT.

We hypothesised that genetically determined poor metabolism of 3,4-methylene dioxymetamphetamine (MDMA) due either to the presence of CYP2D6 genotypes giving absent or low CYP2D6 enzyme activity, or a COMT genotype predicting low COMT enzyme activity would be associated with a greater degree of MDMA-induced reduction in plasma sodium and osmolality than other genotypes at these genes following consumption of 'ecstasy' tablets by clubbers. Of the 48 subjects who returned to the test site post-clubbing, 30 provided samples for measurement of vasopressin (AVP), plasma sodium, urea and plasma and urine osmolality. Genotyping was performed for functional variants in CYP2D6 (n = 29) and COMT (Val158Met, n = 30). In subjects with urinary MDMA detected post-clubbing, there was a significant association between change in plasma osmolality (p = 0.009) and in plasma sodium (p = 0.012) and CYP2D6 genotypic category. Individuals with the low-activity but readily inhibitable CYP2D6 extensive metaboliser/intermediate metaboliser (EM/IM) genotype showed greater reductions in these measures than all other CYP2D6 genotypic categories. COMT low-activity genotypes (Met/Met and Val/Met) were also significantly associated with reductions in plasma osmolality (p = 0.028) and in plasma sodium (p = 0.003). On conservative Bonferroni correction for two independent genes, the CYP2D6 and COMT plasma sodium findings remain significant. The relatively high frequency of the low-activity CYP2D6 and COMT genotypes in the population warrants further attention, since consumption of free water following ingestion of MDMA in these individuals may trigger dilutational hyponatraemia and increased risk of syndrome of inappropriate antidiuretic hormone secretion.

Pharmacogenetic studies of change in cortisol on ecstasy (MDMA) consumption.

In this study we investigate the association of cytochrome P450 enzyme CYP2D6, catechol-O-methyl transferase (COMT, Val158Met) and serotonin transporter promoter (5-HTTLPR) genotypes on change in cortisol concentration following 3, 4-methylenedioxy-methamphetamine (MDMA, 'ecstasy') consumption. Forty-eight subjects (30 males, mean age 23 years), self-nominating regular clubbers provided 'in the field' pre- and post-clubbing biological samples and associated information. Of the 39 subjects who provided a post-clubbing urine sample, 21 were positive for MDMA. Plasma cortisol concentrations increased in subjects (n = 48) tested for cortisol, with changes being significantly greater in the MDMA-positive group (736.9 ± 83.2 vs. 350.9 ± 34.5 mmol/l, p = 0.001). We found a positive association between the low activity COMT genotype (Met/Met) and MDMA-induced change in cortisol and also between this and change in cortisol in the whole sample (p = 0.039, Bonferroni corrected). For CYP2D6, there was an association between genotype and change in cortisol, confined to subjects with MDMA-positive urine post-clubbing (p = 0.003, Bonferroni corrected). There was no association with 5-HTTLPR genotype. These associations suggest that chronic use of MDMA may lead to HPA axis dysregulation and that the magnitude of this may be moderated by genetic polymorphism, and warrant further investigation in a larger sample of those who consume the drug on a regular basis.

Release of vasopressin during suppression of oestrous cycle in rat by zoladex and hypovolemic challenge.

BACKGROUND: The vasopressin (AVP) response to hypovolemia has been compared in intact and chemically castrated rats. This functional ovariectomy was done to confirm the findings in surgical ovariectomy of how gonadal steroids modulate the release of AVP under hypo-volemic challenge. METHODS: Twenty female Sprague Dawley rats were checked for oestrous over two consecutive cycles. The ten control rats were given sub-cutaneous puncture only whereas the experimental were given Zoladex implant. On the fifteenth day all the rats were given intra-peitoneal injection of poly-ehylene glycol. All the rats were de-capitated after an hour. RESULTS: The uterine weight was significantly decreased in experimental group. The plasma AVP level was also significantly decreased in the experimental group. The pituitary AVP level was significantly increased in the experimental group. CONCLUSION: The chemical castration effected the AVP secretion, this proves that the sex steroids modulate the release of AVP secretion inspite of hypo-volemic challenge.

Oxytocin and cholecystokinin secretion in women with colectomy.

BACKGROUND: Cholecystokinin (CCK) concentrations in plasma have been shown to be significantly higher in colectomised subjects compared to healthy controls. This has been ascribed to reduced inhibition of CCK release from colon. In an earlier study CCK in all but one woman who was colectomised, induced release of oxytocin, a peptide present throughout the gastrointestinal (GI) tract. The aim of this study was thus to examine if colectomised women had a different oxytocin response to CCK compared to healthy controls. METHODS: Eleven women, mean age 34.4 +/- 2.3 years, who had undergone colectomy because of ulcerative colitis or constipation were studied. Eleven age-matched healthy women served as controls. All subjects were fasted overnight and given 0.2 microg/kg body weight of CCK-8 i.v. in the morning. Samples were taken ten minutes and immediately before the injection, and 10, 20, 30, 45, 60, 90 and 120 min afterwards. Plasma was collected for measurement of CCK and oxytocin concentrations. RESULTS: The basal oxytocin and CCK concentrations in plasma were similar in the two groups. Intravenous injection of CCK increased the release of oxytocin from 1.31 +/- 0.12 and 1.64 +/- 0.19 pmol/l to 2.82 +/- 0.35 and 3.26 +/- 0.50 pmol/l in controls and colectomised women, respectively (p < 0.001). Given the short half-life of CCK-8 in plasma, the increased concentration following injection could not be demonstrated in the controls. On the other hand, in colectomised women, an increase of CCK in plasma was observed for up to 20 minutes after the injection, concentrations increasing from 1.00 +/- 0.21 to a maximum of 1.81 +/- 0.26 pmol/l (p < 0.002). CONCLUSION: CCK stimulates the release of oxytocin in women. There is no difference in plasma concentrations between colectomised and controls. However, colectomy seems to reduce the metabolic clearance of CCK. The hyperCCKemia in patients who had undergone colectomy is consequently not only dependent on CCK release, but may also depend on reduced clearance.

The soya isoflavone content of rat diet can increase anxiety and stress hormone release in the male rat.

RATIONALE: Most commercial rodent diets are formulated with soya protein and therefore contain soya isoflavones. Isoflavones form one of the main classes of phytoestrogens and have been found to exert both oestrogenic and anti-oestrogenic effects on the central nervous system. The effects have not been limited to reproductive behaviour, but include effects on learning and anxiety and actions on the hypothalamo-pituitary axis. It is therefore possible that the soya content of diet could have significant effects on brain and behaviour and be an important source of between-laboratory variability. OBJECTIVES: To determine whether behaviour in two animal tests of anxiety, and stress hormone production, would differ between rats that were fed a diet which was free of soya isoflavones and other phytoestrogens (iso-free) and those that were fed a diet which contained 150 microg/g of the isoflavones genistein and daidzein (iso-150). This controlled diet has an isoflavone concentration similar to that in the maintenance diet routinely used in our institution. METHODS: Male rats were randomly allocated to the iso-free and iso-150 diets and their body weights and food and water consumption were recorded for 14 days. They were then maintained on the same diets, but housed singly for 4 days, before testing in the social interaction and elevated plus-maze tests of anxiety. Corticosterone concentrations in both dietary groups were determined under basal conditions and after the stress of the two tests of anxiety. Vasopressin and oxytocin concentrations were determined after brief handling stress. RESULTS: The groups did not differ in food or water intake, body weight or oxytocin concentrations. Compared with the rats fed the iso-free diet, the rats fed the iso-150 diet spent significantly less time in active social interaction and made a significantly lower percentage of entries onto the open arms of the plus-maze, indicating anxiogenic effects in both animal tests. The groups did not differ in their basal corticosterone concentrations, but the iso-150 group had significantly elevated stress-induced corticosterone concentrations. Stress-induced plasma vasopressin concentrations were also significantly elevated in the iso-150 diet group compared with the iso-free rats. CONCLUSIONS: Major changes in behavioural measures of anxiety and in stress hormones can result from the soya isoflavone content of rat diet. These changes are as striking as those seen following drug administration and could form an important source of variation between laboratories.

The effect of exogenous melatonin on stimulated neurohypophysial hormone release in man.

OBJECTIVE: Melatonin is known to have a dose-dependent effect on basal neurohypophysial hormone release in man, low physiological doses being stimulatory and high doses inhibitory. We have performed a study to determine if this is true also for stimulated release. DESIGN: This was a double-blind randomized crossover study with subjects being given oral doses of melatonin (0.5 mg or 5.0 mg) or placebo, before infusion of hypertonic saline or exercise. SUBJECTS: Studies were performed on a total of 24 male subjects aged 19-23 years, who were entrained to a normal light-dark cycle and who refrained from taking heavy exercise, alcohol and smoking for 24 h prior to study. MEASUREMENTS: Plasma vasopressin, oxytocin, sodium and osmolality and packed cell volume were measured in control samples and at regular intervals after the administration of melatonin and the challenge tests. Blood pressure and urine flow, sodium potassium and osmolality were determined during hypertonic saline infusion and blood pressure and pulse rate during exercise. RESULTS: Plasma vasopressin increased during hypertonic saline infusion and exercise, the responses being attenuated by 5.0 mg melatonin and augmented with 0.5 mg melatonin. Although exercise had no effect on oxytocin release, there was a small but statistically significant (P < 0.05) increase in plasma oxytocin after hypertonic saline administration, which was inhibited by 5.0 mg and augmented by 0.5 mg melatonin. CONCLUSIONS: Melatonin modulates the neurohypophysial response to different stimuli, low doses enhancing the response. This effect could contribute to the night-time increase in circulating concentrations of the hormones.

Cholecystokinin stimulation leads to increased oxytocin secretion in women.

OBJECTIVE: To find out if cholecystokinin (CCK) stimulates the secretion of oxytocin in humans, and if there are any differences in secretion between healthy women and those with normal-transit constipation. DESIGN: Prospective open study. SETTING: Teaching hospital, Sweden. SUBJECTS: 8 healthy female volunteers and 6 women with chronic refractory normal-transit constipation. INTERVENTIONS: Subjects were fasted before experiments. On one day they were given emulsified corn oil and another an intravenous injection of 1 Ivy dog unit (IDU) CCK/kg body weight. Blood samples were taken before each experiment at 10 minutes and at the time the experiments started. Blood samples were also taken after each experiment at 10, 20, 30, 45, 60, 90 and 120 minutes. MAIN OUTCOME MEASURES: Concentrations of CCK and oxytocin. RESULTS: Ingestion of corn oil significantly increased the plasma concentration of CCK in both groups (healthy women p = 0.03 and constipated women p = 0.008). Injection of CCK also led as expected to hypercholecystokininaemia in both groups (p = 0.008 and p = 0.03, respectively). The corn oil increased oxytocin secretion in both groups (p = 0.02 and 0.03, respectively) and exogenous CCK increased the secretion still further (p = 0.008 and 0.03, respectively). CONCLUSIONS: Both corn oil and injection of CCK led to an increased CCK concentration in plasma. Oxytocin was secreted in response to endogenous as well as exogenous CCK stimulation. There was no difference between healthy and constipated women in either parameter analysed.

The effect of 3,4-methylenedioxymethamphetamine (MDMA, 'ecstasy') and its metabolites on neurohypophysial hormone release from the isolated rat hypothalamus.

Methylenedioxymethamphetamine (MDMA, 'ecstasy'), widely used as a recreational drug, can produce hyponatraemia. The possibility that this could result from stimulation of vasopressin by MDMA or one of its metabolites has been investigated in vitro. Release of both oxytocin and vasopressin from isolated hypothalami obtained from male Wistar rats was determined under basal conditions and following potassium (40 mM) stimulation. The results were compared with those obtained for basal and stimulated release in the presence of MDMA or metabolites in the dose range 1 microM to 100 pM (n=5 - 8) using Student's t-test with Dunnett's correction for multiple comparisons. All compounds tested affected neurohypophysial hormone release, HMMA (4-hydroxy-3-methoxymethamphetamine) and DHA (3,4-dihydroxyamphetamine) being more active than MDMA, and DHMA (3,4-dihydroxymethamphetamine) being the least active. The effect on vasopressin release was greater than that on oxytocin. In the presence of HMMA the ratio test:control for basal release increased for vasopressin from 1.1+/-0.16 to 2.7+/-0.44 (s.e.m., P<0.05) at 10 nM and for oxytocin from 1.0+/-0.05 to 1.6+/-0.12 in the same hypothalami. For MDMA the ratio increased to 1.5+/-0.27 for vasopressin and to 1.28+/-0.04 for oxytocin for 10 nM. MDMA and its metabolites can stimulate both oxytocin and vasopressin release in vitro, the response being dose dependent for each drug with HMMA being the most potent.

Renal response to arginine vasopressin during the oestrous cycle in the rat: comparison of glucose and saline infusion using physiological doses of vasopressin.

The renal response to arginine vasopressin in the rat has been shown to depend on reproductive status. However there is no consensus as to when the kidney is most responsive. The varying results could depend on the protocol and the dose of hormone used. A study has been performed, with physiological doses of vasopressin, comparing the responses during infusion of hypotonic saline and glucose. After an equilibration period of 150 min, conscious rats were infused on each of the four days of the oestrous cycle with either isotonic saline (0.077 M) or 0.14 M glucose for a control period of 45 min. Vasopressin was then infused at 10-40 fmol min(-1) for 1 h, followed by a recovery period of 90 min. Timed urine samples were collected for determination of volume, sodium concentration and osmolality. During the control period urine flow was greatest at oestrus and dioestrus day 2 and sodium excretion on dioestrus day 2 irrespective of the infusate. Vasopressin concentrations achieved lay within the physiological range and no difference was observed between the different days for a given dose. Infusion of vasopressin in both saline and glucose produced a dose-dependent antidiuresis, the greatest responses being seen of pro-oestrus and dioestrus day 2. It was only with the highest rate of infusion that a significant increase in sodium excretion was seen on each day of the cycle and the greatest responses were seen on pro-oestrus and dioestrus day 1 for both infusates. Thus the kidney shows the greatest response to physiological doses of vasopressin at pro-oestrus and dioestrus day 1 irrespective of the infusate employed.