Presynaptic autoinhibition of the electrically evoked dopamine release studied in the rat olfactory tubercle by in vivo electrochemistry.

Evoked dopamine release was monitored in vivo from the olfactory tubercle of anaesthetized rats by differential pulse amperometry combined with carbon fibre electrodes which, in most cases, were electrochemically treated. Dopamine release was evoked by electrical stimulation of the ascending dopaminergic pathway. The dopamine release evoked by burst stimulation (20 s with a mean frequency of 6 Hz) was dose-dependently decreased by D,L-apomorphine (25-800 micrograms/kg, s.c.) or by quinpirole (50 micrograms/kg, s.c.) while the opposite effect was observed with haloperidol (12.5 micrograms/kg-0.5 mg/kg, s.c.) or with D,L-sulpiride (2-200 mg/kg, s.c.). Neither the D1 agonist SKF 38393 (10 mg/kg, s.c.) nor the D1 antagonist SCH 23390 (0.5 mg/kg, s.c.) affected the evoked dopamine release. Moreover, sulpiride competitively antagonized the effects of apomorphine. The relative amplitude of the apomorphine inhibition was inversely correlated with the stimulation frequency (6 or 9 Hz). The increase induced either by haloperidol or by sulpiride was positively related to the stimulation frequency (from 3 to 9 Hz) and reached a stable value (+700% of the pre-drug-evoked dopamine release) with higher frequencies (from 9 to 20 Hz). This increase also depended on the duration of the stimulation: both single-train (10 pulses) or burst stimulations for 20 s, whose frequency inside the trains was in both cases 14 Hz, evoked a dopamine release which was minimally affected by sulpiride or haloperidol. In conclusion, in physiological conditions the amplitude of the impulse flow-dependent dopamine release is regulated by the extrasynaptic extracellular dopamine concentration which varies from 10 to 100 nM. This presynaptic autoinhibition is mediated by autoreceptors of the D2 type and is involved in the nonlinear relationship between impulse flow and dopamine release.

Hemorrhage-Induced Activations of Adrenocorticotropin Release and Catecholamine Metabolism in the Ventrolateral Medulla are Differently Affected by Glucocorticoid Feedback.

We have compared the effects of increasing doses of dexamethasone on the hemorrhage-induced stimulation of the corticotropic axis and the metabolism of the catecholamines of the A1 group in the ventrolateral medulla. Adrenocorticotropin was measured in sequential samples of plasma while the metabolism of the catecholamines was recorded by in vivo electrochemistry in urethane-anesthetized rats. Combined intracerebroventricular injection of specific adrenergic blockers (α(1) -antagonist, prazosin and ß-antagonist, propranolol) prevented the stimulation of the adrenocorticotropin release by hemorrhage. Pretreatment with dexamethasone (1 mg/kg sc) fully blocked the hemorrhage-induced adrenocorticotropin release but did not affect the concomitant stimulation of the catecholamine metabolism in A1 cells. The latter was partially decreased only with the highest dose (10 mg/kg sc). While a central catecholaminergic input appears to be necessary for the hemorrhage-induced stimulation of the corticotropic axis, it does not seem to play a significant role in the feedback regulation by glucocorticoids.

Comparison of the effects of dDAVP and AVP on the sodium transport in the frog skin.

The effect of 1-deamino-8-D-arginine-vasopressin, dDAVP, the synthetic analogue of vasopressin, upon the active sodium transport across the frog skin was studied using standard microelectrode technique and compared with the effect of synthetic arginine-vasopressin, AVP. dDAVP applied to the basolateral side of the epithelium stimulated the active sodium transport as reflected by the increase of short-circuit current, Isc, and transepithelial electrical potential difference, Voc. Potential difference across both the apical, Vo, and the basolateral, Vi, cell membranes decreased. The driving force of transepithelial sodium transport, ENa, did not change. The transepithelial electrical resistance, Rt, ohmic resistance of the active sodium transport, RNa, and apical cell membrane resistance, Ro, rapidly decreased, while the resistance of the basolateral cell membrane, Ri, and the resistance of the shunt pathway, Rs, remained unchanged. It is concluded that dDAVP primarily increases sodium permeability of the apical cell membrane which subsequently stimulates sodium pump activity. This action is similar to that of AVP.

Microelectrode study of insulin effect on apical and basolateral cell membrane of frog skin: comparison with the effect of 1-deamino-8-D-arginine-vasopressin (dDAVP).

The standard microelectrode technique was used to electrophysiologically characterize the effect of insulin on both apical and basolateral cell membranes of the frog skin, a model Na-transporting epithelium. Insulin applied to the basolateral side of the epithelium stimulated the sodium transport as shown by both increased short-circuit current, Isc, (P less than 0.02) and transepithelial potential difference, Voc, (P less than 0.002). Potential difference across the apical cell membrane, Vo, decreased (P less than 0.002) as did the apical cell membrane resistance, Ro, (P less than 0.05). The driving force of sodium ions, ENa, increased after insulin (P less than 0.005). These findings confirm that insulin acts both to increase the apical cell membrane permeability for ions and to stimulate the sodium pump in the basolateral membrane. The effects of insulin were compared with those of a vasopressin analog (dDAVP), known to stimulate transepithelial sodium transport by increasing the permeability of the apical cell membrane for sodium ions. dDAVP applied at the height of insulin effect further stimulated transepithelial transport, but insulin applied at the height of dDAVP action did not. It is concluded that the direct stimulation of the sodium pump by insulin may not represent a decisive component in the stimulation of transepithelial transport across the frog skin. A more potent stimulus for sodium transport is obviously the increased permeability of the apical membrane for ions.

Effects of lysine-vasopressin (LVP) and 1-deamino-8-D-arginine-vasopressin (dDAVP) upon electrical potential, short-circuit current and transepithelial D.C. resistance of the frog skin.

The synthetic analogue of vasopressin, 1-deamino-8-D-arginine-vasopressin (dDAVP), possesses a protracted antidiuretic activity while having practically no pressoric activity as compared to arginine-vasopressin (AVP) or lysine-vasopressin (LVP). The effects of LVP and dDAVP were studied on the frog skin (Rana temporaria) sodium transport as reflected by the short-circuit current (SCC) level, on an Ussing apparatus. The application two different equimolar doses of LVP or dDAVP (approx. 9.4 X 10(-8) mol X l-1 and 18.8 X 10(-8) mol X l-1 to the inner surface of the skin resulted in identical maximal increases of sodium transport. However, the maximum transport stimulation after the application of dDAVP was delayed by about 30 min as compared to the stimulation by LVP (P less than 0.01). In addition, a protracted recovery of SCC towards its original levels was observed in experiments with dDAVP application after the hormone removal (P less than 0.01). It is concluded that dDAVP stimulates Na+ transport through the frog skin despite its lacking pressoric activity. Thus, the natriferic activity of vasopressin is related to its antidiuretic rather than pressoric activity. Maximum increase in the sodium transport following dDAVP application was delayed and more protracted as compared to the effect of LVP.

Stimulation of the sodium transport across the frog skin by three N-terminally extended arginine-vasopressins.

The standard Ussing method was used to electrophysiologically characterize the effects of three analogs of arginine-vasopressin (AVP) on the frog skin, a model Na-transporting epithelium. The analogs tested were N-terminally extended Arg8-vasopressins: Ala-AVP, Ser-Ala-AVP and Thr-Ser-Ala-AVP; synthetic Arg8-AVP was used as the reference agent. The vasopressins were applied to the basolateral side of the frog skin in concentrations ranging between 10(-8) to 10(-5) mol.l-1. All the three analogs increased both the short-circuit current (Isc) and the open-circuit transepithelial potential (Voc), and decreased the transepithelial d.c. resistance (Rt) similarly as did synthetic Arg8-AVP. The results show that N-terminal extension of the Arg8-AVP did not alter the natriferic properties of AVP.

Suppression of natriferic activity of the vasopressin molecule by modifications in positions 1, 2 and 4.

The capacity of five synthetic analogs of [8-arginine] vasopressin (AVP) to stimulate frog skin sodium transport (natriferic activity) was characterized electrophysiologically using the method of short-circuit current, and compared to that of synthetic AVP. The analogs used were [8-arginine] vasopressins modified in positions 1 and 2: [1-(1-mercapto-4-tert-butylcyclohexaneacetic acid)] AVP (I); [1-(1-mercapto-4-methylcyclohexaneacetic acid)] AVP (II); [1-(1-mercapto-4-methylcyclohexaneacetic acid)-2-O-methyltyrosine] AVP (III); and in position 4: [1-(1-mercaptocyclohexaneacetic acid)-4-arginine] AVP (IV); [1-(2-mercaptopropionic acid)-4-arginine] AVP (V). The addition of synthetic vasopressins I, II and V to the frog skin resulted in a weaker stimulation of the skin sodium transport, measured as the level of the short-circuit current (Isc), as compared to that induced by synthetic AVP. In relation to natriferic activity, analogs III and IV did not change the electrical parameters of the skin. It is concluded that introduction of cyclic structure at the beta-carbon in position 1 of the vasopressin molecule decreased its natriferic activity by about 70%. The same reduction of the activity was caused by the replacement of the glutamine residue in position 4 with arginine, and deamination in position 1. Cyclic structure bound in position 1 together with methylation of tyrosine in position 2 resulted in a full suppression of natriferic activity. Similarly, introduction of cyclic group in position 1 in combination with substitution of glutamine in position 4 with arginine totally abolished natriferic activity.

Hydroosmotic activities of arginine-vasopressins modified either in positions 1, 2 and 4 or at N-terminal extensions.

Vasopressin and its synthetic analogs were studied for their effect on transepithelial water flux in frog urinary bladder. As compared with AVP, 1-deamino-8-D-arginine vasopressin (dDAVP) was about 40 times less effective in stimulating osmotic water flow. The vasopressin analogs obtained by modification in positions 1 and 2 were: [1-(1-mercapto-4-tert-butylcyclohexaneacetic acid)] AVP (I); [1-(1-mercapto-4-methylcyclohexaneacetic acid)]AVP (II); [1-(1-mercapto-4-methylcyclohexaneacetic acid)-2-O-methyltyrosine]AVP (III); and those modified in position 4 were: [1-(1-mercaptocyclohexaneacetic acid)-4-arginine] AVP (IV); [1-(2-mercaptopropionic acid)-4-arginine]AVP (V). Any of the above analogs did not influence basal, but antagonized vasopressin-stimulated water flux. N-terminally extended analogs of AVP: Ala-AVP (VI); Ser-Ala-AVP (VII) and Thr-Ser-Ala-AVP (VIII) stimulated osmotic water flux to the same extent in concentration 200 times higher as that of AVP. We conclude from these studies that vasopressin analogs (I-V) competitively antagonize vasopressin-stimulated hydroosmotic activity in frog urinary bladder probably at the epithelial vasotocin V1 and/or V2 receptor site. N-terminal extension of the vasopressin molecule did not influence the capacity of AVP to induce V2 receptor-mediated action, even when used at higher concentrations.

Serum free carnitine in medium chain acyl-CoA dehydrogenase deficiency.

Medium chain acyl-CoA dehydrogenase (MCAD) deficiency is the most common disorder of fatty acid beta-oxidation and presents acutely with hypoglycemia, or a Reye-like illness with low free carnitine, often provoked by an infection or an excessive period of fasting. After acute attack these children are for the most time asymptomatic and may have normal plasma free carnitine concentrations. We observed a regularity in time course of serum free carnitine concentration during two attacks of Reye-like illness in patient with MCAD deficiency. Molecular investigation confirmed that the patient was homozygote for A985G mutation. Free carnitine was measured by enzymatic UV-test. First attack of severe hypoglycemia and Reye-like symptoms started at the age of 15 months and the second at the age of 25 months. In both episodes, treatment with intravenous glucose was given immediately, but without carnitine supplementation. Between the attacks patient was on a normal diet. In both attacks, low serum free carnitine concentration from the time of acute attack continually decreased for up to 8-13 days and then normalized at about 25 days after attack. We think that the time course of serum free carnitine may help in knowledge about carnitine depletion in MCAD deficiency. This is the first observation of this pattern during an acute attack and needs to be confirmed by other patients with MCAD deficiency. (Fig. 2, Ref. 7.).

[IgG subclasses in children with recurrent respiratory diseases]. / Podtriedy IgG u detí s recidivujúcimi respiracnými chorobami.

The authors examined IgG sub-classes, using the ELISA method, in 35 patients with relapsing respiratory diseases and 27 children of a control group aged 3-15 years. In children with relapsing respiratory diseases without deficiency of the main immunoglobulin classes they found a statistically significantly reduced value in sub-class IgG2 (P < or = 0.01). The changes found in other sub-classes were not statistically significant. The authors draw attention to the fact that deficiency in IgG sub-classes should be considered in case of repeated respiratory infections caused by encapsulated microorganisms--H. influenzae, Str. pneumoniae--and also when reduced levels of IgG and IgA are found. However, even normal levels do not rule out deficiency.

Decreased free water excretion after indomethacin in the absence of antidiuretic hormone in saline loaded hypophysectomized Wistar and hydrapaenic Brattleboro rats.

Indomethacin (40 mg kg-1 in a single dose) was applied by stomach tube to inhibit the synthesis of prostaglandins during extracellular fluid volume expansion with isotonic saline in normal and acutely (2 h) hypophysectomized Wistar rats and hydropaenic diabetes insipidus Brattleboro rats. In the normal rats indomethacin reduced urine output due to decreased free water excretion. In the rats deprived of antidiuretic hormone (ADH) by acute hypophysectomy or due to hereditary eror in ADH synthesis indomethacin also decreased the urine output and clearance of osmotically free water. It is concluded that also other mechanisms besides the established competition between ADH and prostaglandins for ADH receptors may be responsible for the indomethacin effect on the osmotic movement of water in the distal nephron.

Failure to demonstrate a natriuretic effect of neurophysin in rats.

Partly purified rat neurophysin was tested for its possible natriuretic activity in acutely hypophysectomized saline-loaded rats (hypophysectomized 2 h prior to saline infusion). Such animals were unable to excrete the saline load. When partly purified rat neurophysin was injected i. p. in a dose of 40 microgram per animal (which corresponds to its content in one rat posterior pituitary) before saline loading, the sodium excretion was not different from that found in acutely hypophysectomized animals, but it was significantly lower than in normal saline-loaded rats. It was concluded that any apparent natriuretic effect of rat neurophysin could not be established by the present experiments under the experimental conditions used.

The effect of small doses of antidiuretic hormone on renal excretion of sodium and water in saline loaded acutely hypophysectomized rats.

Small doses of vasopressin (200 micronU min-1 kg-1) did not influence the loss of ability of acutely hypophysectomized rats to react by natriuresis to the extracellular fluid volume expansion with saline, inspite of restoring their arterial blood pressure to normal. It is concluded that the level of antidiuretic hormone efficient enough to concentrate urine is not a decisive factor in the homeostatic mechanism promoting the 'volume' natriuresis.

Failure to demonstrate natriuretic activity in the posterior pituitary after immunoneutralization of the vasopressin content.

The attempt to demonstrate the presence of a natriuretic substance in the posterior pituitary after immunoneutralization of the AVP content failed. Rats infused i.v. AVP-immunoneutralized posterior pituitary extract did not respond by natriuresis to a saline infusion, in contrast to those infused untreated posterior pituitary extract. Thus, vasopressin seems to be the natriuretic substance in the posterior pituitary extract.

Cholesterol and triacylglycerols in human breast milk before and after nursing.

Cholesterol and triacylglycerols concentrations were estimated in human breast milk taken before and after nursing at the period of 1 week, 3 months, and 6 months after delivery. In all postfeeding samples significantly higher concentration of cholesterol and triacylglycerols was found as compared with the appropriate samples obtained before the feeding. In cholesterol concentration such difference was most remarkable in the milk of mothers who were breast feeding their infants over 6 months. The highest cholesterol concentrations were found during the first week after the delivery, whereas the highest concentrations of triglycerides were found 6 months after the delivery. The elevations of the lipid levels were not associated with changes of osmolality and/or electrolytes (Na+, K+, Ca2+) during the nursing.