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In vitro recordings from area postrema neurons demonstrate responsiveness to adrenomedullin.

Adrenomedullin (ADM) is a recently discovered 52-amino acid peptide that exerts potent vasodilatory effects in the periphery and influences the control of body fluid balance when injected centrally. In this study extracellular single-unit recordings were obtained from 94 AP neurons in rat brain slices. Bath application of ADM (10(-7) M) excited 47% (32 of 68) of cells tested, and these effects were found to be dose dependent from 10(-7) to 10(-9) M. Excitation was maintained during synaptic blockade in a low-Ca2+ artificial cerebrospinal fluid solution, demonstrating direct actions of ADM on these neurons. The remaining cells were either unaffected (n = 25) or inhibited (n = 11) by ADM. ADM (10(-7) M) also influenced the spontaneous activity of 9 (7 inhibited, 2 excited) of 16 neurons located in the nucleus tractus solitarii (NTS). However, these effects could be eliminated during synaptic blockade, suggesting indirect actions of the peptide on NTS neurons. These data demonstrate that a specific population of CNS neurons within the AP are directly influenced by ADM and suggest that ADM may exert its effects on the central control of fluid balance through direct actions at this circumventricular organ.

Adrenomedullin↗

Afferent renal inputs onto subfornical organ neurons responsive to angiotensin II.

Experiments were done in pentobarbital sodium-anesthetized rats to investigate the effect of electrical stimulation of afferent renal nerves (ARN) on the discharge rate of subfornical organ (SFO) neurons that responded to changes in plasma levels of angiotensin II (ANG II) and projected directly to the paraventricular nucleus of the hypothalamus (PVH). Extracellular recordings were made from 76 histologically verified single neurons in the SFO that were excited by intracarotid infusions of ANG II. Of these units, 54.8% (23 of 42) responded with excitation to ARN stimulation (mean onset latency, 125 +/- 35 ms). None of the SFO units excited by plasma ANG II were found to be inhibited by ARN stimulation. An additional 34 units in the SFO that were excited by plasma ANG II were also antidromically activated by stimulation of the PVH. Of these neurons, 17.8% (6 of 34) were also excited by stimulation of ARN. The results indicate that inputs from ARN converge onto SFO neurons that alter their discharge rate during changes in plasma concentration of ANG II and project directly to the PVH. These data suggest that ARN may play an important role in body fluid balance and circulatory regulation by modulating the activity of SFO neurons that function in the detection of blood-borne signals resulting from the decrease in extracellular fluid volume and arterial pressure and that influence the activity of hypothalamic nuclei that contain neurosecretory neurons.

Action Potentials↗

Relationship between adrenomedullin and vasopressin-aquaporin system under general anesthesia.

AIM: The roles of adrenomedullin (AM) in body fluid balance under general anesthesia were investigated. METHODS: Time course changes in plasma osmolality, AM, arginine vasopressin (AVP), and urinary aquaporin 2 (AQP2) in 17 patients undergoing abdominal surgery under general anesthesia were examined. RESULTS: Increases in plasma AM levels were observed in parallel with increases in the levels of urinary AQP2/creatinine (Cr) before induction and 90 and 180 min after initiation of anesthesia. Significant correlations between plasma AM and urinary AQP2/Cr (r = 0.62, p < 0.0001) as well as urinary AVP/Cr and AQP2/Cr (r = 0.60, p < 0.0001) were uncovered. Multivariate stepwise analysis identified plasma AM as the critical independent factor affecting urinary AQP2/Cr level. CONCLUSION: A novel correlation of AM and AQP2 which overlays an AVP-AQP2 system may play a key role in fluid homeostasis during general anesthesia.

Adrenomedullin↗

Terminations of LHRH-immunoreactive fibers in the subfornical organ of the opossum: an ultrastructural study.

Electron microscopic immunocytochemical approaches were used to analyze LHRH-containing elements in the subfornical organ of the opossum, a species in which this input to the subfornical organ is prominent. Not only were LHRH synaptic specializations easily demonstrated in the subfornical organ, forming axo-dendritic and axo-axonal contacts, but also LHRH-immunoreactive fibers contacted astrocytic end-feet on fenestrated capillaries and were found in the subependymal layer. LHRH-carrying elements in the subfornical organ may be important for relating reproductive functions to body fluid balance.

Animals↗

Hypotension and renal impairment during infusion of atrial natriuretic factor in liver cirrhosis with ascites.

Plasma immunoreactive atrial natriuretic factor (irANF) levels and the effects of alpha-human ANF (alpha-hANF) infusion were investigated in 7 patients with liver cirrhosis and ascites. Under basal conditions, supine blood pressure (BP) averaged 136/76 +/- 9/4 mm Hg (mean +/- SEM). Plasma irANF concentrations (124 +/- 33 pg/ml) were higher (p less than 0.01) than those in age-matched normal subjects (47 +/- 5 pg/ml). Plasma renin activity (PRA 5.9 +/- 2.2 ng/ml/h), aldosterone (18 +/- 7 ng/dl) and norepinephrine (NE, 66 +/- 5 ng/dl) levels were also elevated compared to the age-related normal range. Alpha-hANF infusion for 60 min at 0.036 micrograms/kg/min decreased the mean BP (-14%; p less than 0.05), increased PRA (+179%; p less than 0.05) and plasma NE (+24%; p less than 0.05). Glomerular filtration rate (GFR), effective renal plasma flow (ERPF), diuresis and natriuresis were not modified. A subsequent 60-min infusion of alpha-hANF at 0.067 micrograms/kg/min produced a marked fall in mean BP (-26%; p less than 0.001), hemoconcentration (hematocrit +6%; p less than 0.001) despite stable body fluid balance and a further increase in PRA (+350%, p less than 0.005). GFR and ERPF were severely reduced (-55 and -56%, respectively; p less than 0.001), while diuresis and natriuresis were not modified. Plasma aldosterone was unaltered during, but rose (+72%; p less than 0.01) after the cessation of alpha-hANF infusion. Variations in natriuresis during alpha-hANF infusion correlated positively with BP (r = 0.47; p less than 0.01), ERPF (r = 0.53; p less than 0.01) or GFR (r = 0.51; p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Atrial natriuretic factor in mild to moderate chronic renal failure.

The relationship between kidney function and plasma immunoreactive atrial natriuretic factor (irANF) levels as well as the effects of synthetic human ANF-(99-126) were investigated in 13 patients with mild to moderate chronic renal failure. Under basal conditions, glomerular filtration rate averaged 39 +/- 5 (SEM) ml/min/1.73 m2 and blood pressure (BP) averaged 166/107 +/- 7/2 mm Hg; 12 patients were hypertensive. Plasma irANF levels were significantly increased (98 +/- 16 vs 42 +/- 4 pg/ml in healthy control subjects; p less than 0.001) and correlated (p less than 0.05-0.005) inversely with hematocrit (r = -0.65) and positively with systolic BP (r = 0.75) or fractional sodium excretion (r = 0.75). Human ANF-(99-126) infusion for 45 minutes at 0.034 microgram/kg/min augmented (p less than 0.05-0.01) diuresis and urinary sodium, chloride, calcium, phosphate, and magnesium excretion. During the subsequent 45 minutes of human ANF-(99-126) infusion at a rate of 0.077 microgram/kg/min, diuresis and electrolyte excretion remained elevated (p less than 0.05-0.01). Glomerular filtration rate and effective renal plasma flow were not significantly modified, but filtration fraction rose progressively (p less than 0.01). Human ANF-(99-126) infusion decreased BP (p less than 0.05-0.01), produced hemoconcentration (hematocrit + 7%; p less than 0.01) without negative body fluid balance, and increased (p less than 0.01-0.001) plasma norepinephrine, insulin, and serum free fatty acids; plasma aldosterone and renin activity were unaltered during but rose after cessation of human ANF-(99-126) infusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Salt-sensitive hypertension in (mREN-2)27 transgenic rats.

The (mREN-2)27 transgenic model of hypertension was developed to investigate the effect of genetic over activity of angiotensin II systems as a contributing factor in the development of arterial hypertension. In this model, transgene-positive rats demonstrate elevated renin-angiotensin system activity not only in the circulatory system but also in adrenal gland, reproductive organs, and brain. Since evidence indicates that angiotensin peptides and osmotic stimuli interact synergistically to produce exaggerated behavioral, endocrine, and cardiovascular effects, we examined the effect of salt consumption on arterial pressure, plasma vasopressin, and body fluid balance in male (mREN-2)27 transgene-positive and -negative rats. Four days of drinking 2% NaCl increased mean arterial pressure from 165 +/- 10 to 199 +/- 7 mm Hg in transgene-positive rats. In contrast, transgene-negative rats showed no change in arterial pressure (126 +/- 5 to 128 +/- 3 mm Hg). Plasma vasopressin levels were significantly elevated only in transgene-positive rats, whereas pituitary levels of vasopressin were significantly lower in transgene-positive rats compared with transgene-negative controls (18 +/- 3 and 118 +/- 14 ng, respectively). Although transgene-positive rats consumed significantly more 2% NaCl than did transgene-negative rats, during this period 24-hour sodium balance did not differ between the groups. Since fluid and electrolyte balance is similar between the two groups of rats, the data suggest that transgene-positive rats may be more sensitive to the effects of increased NaCl intake in terms of both endocrine and cardiovascular responses.

Animals↗

Blood levels and renal effects of atrial natriuretic peptide in normal man.

Since mammalian atria were recently found to contain vasoactive and natriuretic peptides, we investigated the following in normal humans: plasma human atrial natriuretic peptide concentrations, effective renal plasma flow (ERPF), glomerular filtration rate (GFR), urinary water and electrolyte excretion, blood pressure (BP), and catecholamine, antidiuretic hormone (ADH), angiotensin II, and aldosterone levels before, during, and after intravenous administration of the newly synthetized alpha-human atrial natriuretic peptide (alpha hANP). In 10 subjects alpha hANP given as an initial bolus of 50 micrograms followed by a 45-min maintenance infusion at 6.25 micrograms/min increased plasma alpha hANP from 58 +/- 12 to 625 +/- 87 (mean +/- SEM) pg/ml; caused an acute fall in diastolic BP (-12%, P less than 0.001) and a hemoconcentration (hematocrit +7%, P less than 0.01) not fully explained by a negative body fluid balance; increased GFR (+15%, P less than 0.05) despite unchanged or decreased ERPF (filtration fraction +37%, P less than 0.001); augmented (P less than 0.05- less than 0.001) urinary chloride (+317%), sodium (+224%), calcium (+158%), magnesium (+110%), phosphate excretion (+88%), and free water clearance (from -0.76 to +2.23 ml/min, P less than 0.001) with only little change in potassium excretion; and increased plasma norepinephrine (P less than 0.001) while plasma and urinary epinephrine and dopamine, and plasma ADH, angiotensin II, and aldosterone levels were unchanged. The magnitude and pattern of electrolyte and water excretion during alpha hANP infusion could not be accounted for by increased GFR alone. Therefore, in normal man, endogenous alpha hANP seems to circulate in blood. alpha hANP can cause a BP reduction and hemoconcentration which occur, at least in part, independently of diuresis and are accompanied by sympathetic activation. An increase in GFR that occurs in the presence of unchanged or even decreased total renal blood flow is an important but not sole mechanism of natriuresis and diuresis induced by alpha hANP in man.

Atrial Natriuretic Factor↗

Cardiotonic steroids: potential endogenous sodium pump ligands with diverse function.

The highly conserved cardiotonic steroid (CS) binding site present on the ubiquitous membrane sodium pump, sodium, potassium-ATPase, appears to have been conserved by no force other than its capacity to bind CS: a family that includes plant-derived cardiac glycosides and putative endogenous vertebrate counterparts. Binding of ligand is inhibited by increased extracellular potassium. This implies functional coordination because inhibition of the sodium pump would be counterproductive when extracellular potassium is elevated. The interesting biology of the CS binding site continues to stimulate investigations into the identity of endogenous ligands, their role as pump regulators at the cellular level, and as mediators of body fluid balance and blood pressure regulation. In addition to inhibition of sodium and potassium transport, there is considerable recent evidence suggesting that the sodium pump may act as a cell signaling receptor activated by CS binding and responding by coordination of intracellular signaling pathways that can be dependent on and also independent of the reduction in transmembrane ion flux resulting directly from pump inhibition. This signaling may influence cell survival, growth, and differentiation. Recent insight into the biology of pump regulation by CS is reviewed.

Adrenal Cortex↗

Short-term and long-term inhibition of endogenous atrial natriuretic peptide in dogs with early-stage heart failure.

Early-stage heart failure (HF) is characterized by an increase in circulating atrial natriuretic peptide (ANP) without activation of the renin-angiotensin-aldosterone system (RAAS) or body fluid retention. To test the hypothesis that elevated endogenous ANP suppresses the RAAS, maintains body fluid balance, and regulates vascular tone in early-stage HF, we assessed the effects of short-term and long-term inhibition of ANP on cardiorenal and neurohormonal functions. Short-term antagonism was produced by bolus administration (3 mg/kg) of HS-142-1, an antagonist of guanylate-cyclase coupled ANP receptors, and long-term antagonism was produced by continuous infusion (1 mg/kg per h) of HS-142-1 for 8 h to dogs with early-stage HF induced by rapid ventricular pacing (270 beats/min, 8 days). In this experimentally produced HF, plasma ANP was significantly increased relative to the pre-pacing value, but not plasma renin activity (PRA) or plasma aldosterone level. HS-142-1 significantly suppressed plasma and urinary guanosine 3',5'-cyclic monophosphate (cGMP) levels, markers of endogenous ANP activity, in both experiments. Although mean arterial pressure and cardiac output did not change significantly, pulmonary capillary wedge pressure and right atrial pressure were elevated in both experiments. While short-term inhibition of ANP did not change PRA and aldosterone levels, long-term inhibition significantly increased these hormonal levels, resulting in decreases in urine flow rate, urinary sodium excretion rate, glomerular filtration rate, and renal plasma flow. These findings suggest that endogenous ANP plays a critical role in regulating venovascular tone, inhibiting activation of RAAS, and maintaining renal functions in early-stage HF.

Animals↗

Long-term anaesthesia using inhalatory isoflurane in different strains of mice-the haemodynamic effects.

The aim of this study was to establish a simple and safe method of anaesthesia for intravital microcirculatory observations in small laboratory animals. The usefulness of isoflurane inhalation anaesthesia has been investigated in different strains of mice commonly used in experimental medicine. These were the hairless (hr/hr, n = 12), the BALB/c (n = 12) and the nude mouse (nu/nu, n = 3). Anaesthesia was maintained by mask inhalation of isoflurane vaporized at concentrations of up to 4% in the induction phase, at 1.5% during acute surgical procedures and at 0.8-1.3% during prolonged experimental observations. Isoflurane was vapoured in a N(2)O/O(2) mixture and saturated with 32-36% F(i)O(2). During observations the body temperature was kept constant at 37 degrees C. The tail artery was cannulated for monitoring of mean arterial blood pressure (MAP) and heart rate (HR). To maintain the body fluid balance, isotonic saline was administered at a constant rate of 0.2 ml/h. Arterial blood samples were drawn for blood-gas analysis at the end of the experiments. All animals survived the anaesthesia protocol lasting between 3 and 6.5 h. During isoflurane inhalation, no breathing complications or changes in systemic circulatory parameters were observed. Mean values of MAP and HR were 79+/- 3 mmHg and 486+/- 13 min(-1), respectively, over the entire observation period. A moderate acidosis was recorded in animals under isoflurane anaesthesia, with alterations of arterial blood pH, p(a)O(2) and pCO(2) values (7.29+/- 0.06, 130+/- 19 mmHg and 35.6+/- 4.7 mmHg, respectively). In conclusion, inhalation anaesthesia with isoflurane is useful for experimental studies in the mouse due to (1) the simplicity of administration of the anaesthetic, (2) the rapid induction of anaesthesia, (3) easy control of the depth of anaesthesia, (4) the low percentage of complications, and (5) stable MAP and HR during observations lasting several hours. The proposed technique is especially suitable for observations of the microcirculation under intravital fluorescence microscopy.

Anesthesia↗

Historical perspective of the renin-angiotensin system.

Researchers continue to be fascinated with the renin-angiotensin system (RAS) more than 100 yr after its discovery because of its powerful role in controlling sodium balance, body fluid volumes, and arterial pressure. Development of drugs that block different components of this system has led to powerful treatments for hypertension, heart failure, diabetes, and other diseases. Molecular approaches to studying this system offer new possibilities for better understanding the physiology and pathophysiology of the RAS, and for developing new therapeutic paradigms. Our challenge in the future will be to effectively utilize the technological advances that are taking place in virtually all areas of science, including the RAS, and to translate them into a better understanding of the pathophysiology and treatment of human diseases.

Angiotensin-Converting Enzyme Inhibitors↗

Relationship between Doppler transmitral flow velocity pattern and plasma atrial and brain natriuretic peptide concentrations in anuric patients on maintenance hemodialysis.

Plasma atrial (ANP) and brain (BNP) natriuretic peptide levels were compared to determine if transmitral flow velocity pattern is an instantaneous marker of body fluid balance in anuric patients on hemodialysis (HD). We measured plasma ANP and BNP levels and performed Doppler echocardiography in 38 anuric patients before and after HD. Patients with valvular disease, left ventricular systolic dysfunction having a fractional shortening < 0.3, arrhythmia, or left ventricular hypertrophy were excluded. The relationships between plasma ANP or BNP levels and the transmitral flow velocity pattern were evaluated. We also determined if the magnitude of the decrease in plasma ANP level was related to that in the early peak of transmitral flow velocity (peak E). The mean age of the subjects was 61.1 +/- 9.7 years. The ANP level of 213.6 +/- 146.1 pg/mL was related to peak E of 61 +/- 15 cm/s before HD (R = 0.504, P < 0.001), but not after HD. Plasma ANP level was not related to peak late transmitral flow velocity (peak A) or peak E/peak A before or after HD. BNP level was not related to the transmitral flow velocity pattern. The magnitude of decrease in hANP level during HD was significantly related to that in peak E (R = 0.342, P < 0.05). Before HD, peak E was related to the plasma ANP level, reflecting volume overload. Change in peak E showed a weak relationship with that of plasma ANP level in the same HD patient. The measurement of peak E during a HD session may potentially enable the assessment of hydration status during HD.

Aged↗

Irritable bowel syndrome: current concepts and future trends.

About five per cent of the adult population each year will see their doctor with complaints that are finally characterised as irritable bowel syndrome (IBS). The complaints are constipation (perhaps alternating with diarrhoea), abdominal pain (dull or colicky), abdominal distension, abdominal rumbling and flatulence. The diagnosis of IBS implies that a relevant examination has precluded any organic disease. The etiology is unknown and the syndrome probably does not represent a disease entity. It is therefore difficult, if not impossible, to produce a definite rationale of treatment. However, several aspects of the pathogenesis of the individual symptoms of IBS are well known: 1) chronic constipation is most likely due to fibre-depleted diet, psychological factors, local organic disorders (e.g., anal fissures, hemorrhoids, diverticulosis) and disturbance of the body fluid balance (e.g., high consumption of diuretic compounds such as coffee and tea); 2) pain is related to spasms and motility disturbances causing increased intraluminal pressure; 3) meteorism is not due to an increased amount of intestinal gas, but "air traps" and segmental accumulation of gas seem to occur. Furthermore, psychopathological factors and perhaps also food intolerance may play an etiological role. At present the rationale of treatment in IBS is: 1) management of constipation, 2) ease of spasms, 3) reduction of surface tension of intestinal contents, 4) ease of mental stress.

Abdomen↗

Orexin-A depolarizes dissociated rat area postrema neurons through activation of a nonselective cationic conductance.

The area postrema (AP) is involved in the regulation of body fluid balance, feeding behavior, and cardiovascular function. Orexin (ORX)-A is a 33 aa peptide that regulates energy metabolism and sympathetic and cardiovascular actions. ORX immunoreactive axons and their varicose terminals have been found in AP. In this study, whole-cell, current- or voltage-clamp recordings were obtained from 108 dissociated rat AP neurons. The mean resting membrane potential of these neurons (n = 48) was -59.24 +/- 0.87 mV, the mean input resistance was 3.57 +/- 0.22 G(Omega), and the action potential amplitude of these cells was always >90 mV. Current-clamp studies showed bath application of ORX-A depolarized the majority of AP neurons tested (68.8%; 33 of 48), whereas small proportions of cells were either hyperpolarized (16.7%; 8 of 48) or unaffected (14.6%; 7 of 48). These depolarizing effects were found to be concentration dependent from 10(-8) to 10(-11) m. We then examined the contributions of specific ionic conductances to the ORX-A-induced excitation of AP neurons through whole-cell, voltage-clamp studies. Our results demonstrate that in contrast to previous studies on other neuronal populations, ORX-A did not affect net whole-cell potassium currents in AP neurons. Slow depolarizing voltage ramps, however, revealed that ORX-A enhanced a nonselective cationic conductance in AP neurons, effects which would explain the depolarizing effects of the peptide. These data demonstrate that AP neurons are directly influenced by ORX-A and suggest that ORX-A may exert its effects on the central control of feeding behavior and cardiovascular function through direct actions in AP.

Animals↗

Enhanced food and water intake in renin transgenic rats.

In short term experiments angiotensin II (Ang II) is a potent stimulant of thirst, however it is not known whether prolonged activation of the renin-angiotensin system is associated with chronic alteration of water or food intake. Renin transgenic rats TGRmRen(2)27 (TGR) exhibit significant elevation of AngII in the brain regions involved in regulation of body fluid balance. The purpose of the present study was to find out whether TGR rats manifest also different water (WI) and food (FI) intake and renal excretory functions in comparison to their parent Sprague Dawley (SD) strain. To this end 24 h WI and FI as well as urine excretion (Vu) and urinary outputs of solutes (Cosm), sodium (UNaV) and potassium (UKV) were compared under baseline conditions in 16 TGR and 15 SD rats having free access to water and food. In 15 TGR and 17 SD rats effect of 24 h dehydration on water intake was investigated. Under baseline conditions TGR rats consumed significantly greater amount of food and water than SD rats. Vu, UNaV and UKV were not significantly different in both strains. Cumulative water intakes in SD and TGR rats subjected to 24 h dehydration did not differ. The results reveal that under baseline conditions TGR rats manifest greater food and water intakes than SD rats whereas stimulation of thirst by water deprivation is similar in both strains. The results suggest that the ingestive behavior may be chronically altered by upregulation of the renin-angiotensin system.

Animals↗

[Unexpected properties of angiotensin mediated by the AT2 receptor: possible therapeutic implications].

The renin-angiotensin-aldosterone system plays an important role in the regulation of electrolyte balance, body fluid volume and blood pressure. As yet, angiotensin II has been ascribed actions mediated by first type of receptors (AT1) such as increased blood pressure, antinatriuretic effect, cell proliferation. Since several years studies have been conducted on the role of second type receptor (AT2) through which angiotensin manifests its effects opposing those resulting from stimulation of type 1 receptor. They include: release of bradykinin and nitric oxide, vasodilation, natriuretic and antiproliferative effects. Blockade of the function of this receptor causes excessive reaction induced by action exerted on AT1 receptor.

Angiotensin-Converting Enzyme Inhibitors↗

[Effects of female sex hormones on atrial natriuretic factor gene expression in rats].

To clarify the effects of female sex hormones on ANF gene expression, emasculated and intact female rats were subcutaneously injected with estradiol (E2), progesterone (P) and olive oil solvent (O) respectively, once a day for 7 days. The relative rANF-mRNA contents of rat atria were measured by molecular hybridization, and rANF-cDNA was labeled with alpha-32P as probe. The groups and hormone doses per 100 g body weight were: (1) sham, 0.2 ml O; (2) rats were bilaterally ovariectomized(OVX), 0.2 ml O; (3) OVX, 80 ng E2; (4) OVX, 80 micrograms E2; (5) OVX, 0.2 mg P; (6) OVX, 2 mg P; (7) OVX, 80 ng E2, 0.2 mg P. The results showed that the rANF-mRNA of group (2) was the lowest, it was 75% of that in group (1), while the rANF-mRNA levels in groups (3)-(7) were 1.1, 2.0, 1.9, 2.3 and 2.1 folds of that in group (2) respectively. The results revealed that E2 and P increased ANF gene expression. Their effects were associated with the dosage. The two may have synergistic actions. Physiological amount of E2 and P may maintain suitable level of rANF-mRNA. This experiment suggested that female sex hormone may have dual effects in body fluid balance.

Animals↗