Relationship of angiotensinase and vasopressinase activities between hypothalamus, heart, and plasma in L-NAME-treated WKY and SHR.

The renin-angiotensin system (RAS), vasopressin, and nitric oxide (NO) interact to regulate blood pressure at central and peripheral level. To improve our understanding of their interaction and their relationship with the hypothalamus and the cardiovascular system, we analyzed angiotensin- and vasopressin-metabolizing activities in hypothalamus (HT), left ventricle (LV), and plasma, collected from Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) treated or not with L-NAME [N(G)-nitro-L-arginine methyl ester], which inhibits the formation of NO and over-activates the sympathetic nervous system. Previous observations in WKY suggested higher formation of Ang III and Ang IV in the HT and higher availability of Ang II in plasma after L-NAME treatment. Our current results show higher formation of Ang IV and higher metabolism of vasopressin after treatment with L-NAME in the LV of WKY rats. In SHR treated with L-NAME, there is higher availability of Ang III in the HT leading to higher release of vasopressin together with lower formation of Ang 2-10. In their LV, however, there is an increase of vasopressinase. Interestingly, while the enzymatic activities in the HT and LV of WKY rats and control SHR are poorly correlated, they are well but inversely correlated in the L-NAME treated SHR. On the other hand, no significant correlations between enzymatic activities in HT or LV and plasma were noticed. Our results suggest that eNOS inhibition in SHR induces or enhances an inverse reciprocal interaction between HT and LV involving the RAS and vasopressin, which may be mediated by the autonomic nervous system.

Effects of antihypertensive drugs on angiotensinase activities in the testis of spontaneously hypertensive rats.

Sexual dysfunction is a frequent adverse effect during antihypertensive therapy. However, the mechanisms responsible for these effects are not well understood. The renin-angiotensin system has been identified in testis where it may play a role in testicular function and be involved in the detrimental effects of antihypertensive drugs. Therefore, our objective was to compare the influence of captopril and propranolol on plasma testosterone levels and on hydrolyzing angiotensin's enzymes (angiotensinases) in the testis of spontaneously hypertensive rats (SHRs) and in control animals. Twenty-four adult male SHRs were used in this study; eight were treated with captopril in drinking water, 8 with propranolol, and 8 were controls. At the end of the 4 weeks treatment period, systolic blood pressure (SBP) was recorded, blood samples were collected, and the right testis was dissected after perfusion of the rat with saline. The soluble (Sol) and membrane-bound (MB) fractions were obtained after solubilization and ultracentrifugation. Fluorometric measurement of Sol and MB angiotensinase activities were performed using arylamide derivatives as substrates. Testosterone was measured by enzyme immunoassay. SBP decreased after captopril but did not change with propranolol treatment. Whereas captopril did not affect angiotensinase activities, highly significant reductions in Sol and MB angiotensinase activities, particularly glutamyl- and aspartyl-aminopeptidases, were observed after treatment with propranolol. Plasma testosterone decreased in captopril treated rats but propranolol had a greater effect. The present results support a general functional depression of the RAS cascade in the testis of propranolol-treated SHR, which may influence the sexual function of these animals.

Detoxification and mineralization of Acid Blue 74: study of an alternative secondary treatment to improve the enzymatic decolourization.

Many reports describe the decolourization of dyes by fungal enzymes. However, these enzymes do not contribute to dye mineralization but only to its biotransformation into less coloured or colourless molecules persisting in solution. Therefore, it is essential to analyse the identity of the metabolites produced during enzymatic treatments and its biodegradation into an appropriate system. The present work examines the decolourization/detoxification of a simulated effluent (containing Acid Blue 74) by fungal enzymes and proposes a secondary treatment using an anaerobic system to improve the enzymatic decolourization through the complete mineralization of the dye. Ligninolytic enzymes were produced by solid culture using the thermo-tolerant fungus Fomes sp. EUM1. The enzymes produced showed a high rate of decolourization (>95 % in 5 h) and were stable at elevated temperature (40 °C) and ionic strength (NaCl, 50 mM). Isatin-5-sulphonic acid was identified via (1)H-NMR as oxidation product; tests using Daphnia magna revealed the non-toxic nature of this compound. To improve the enzymatic degradation and avoid coupling reactions between the oxidation products, the effluent was subjected to an anaerobic (methanogenic) treatment, which achieved high mineralization efficiencies (>85 %). To confirm the mineralization of isatin-5-sulphonic acid, a specific degradation study, which has not been reported before, with this single compound was conducted under the same conditions; the results showed high removal efficiencies (86 %) with methane production as evidence of mineralization. These results showed the applicability of an anaerobic methanogenic system to improve the enzymatic decolourization/detoxification of Acid Blue 74 and achieve its complete mineralization.

Angiotensinase and vasopressinase activities in hypothalamus, plasma, and kidney after inhibition of angiotensin-converting enzyme: basis for a new working hypothesis.

Reducing angiotensin II (Ang II) production via angiotensin-converting enzyme (ACE) inhibitors is a key approach for the treatment of hypertension. However, these inhibitors may also affect other enzymes, such as angiotensinases and vasopressinase, responsible for the metabolism of other peptides also involved in blood pressure control, such as Ang 2-10, Ang III, Ang IV, and vasopressin. We analyzed the activity of these enzymes in the hypothalamus, plasma, and kidney of normotensive adult male rats after inhibition of ACE with captopril. Aspartyl- (AspAP), glutamyl- (GluAP), alanyl- (AlaAP) and cystinyl-aminopeptidase (CysAP) activities were measured fluorimetrically using arylamides as substrates. Systolic blood pressure (SBP), water intake, and urine flow were also measured. Captopril reduced SBP and increased urine flow. In the hypothalamus, GluAP and AspAP increased, without significant changes in either AlaAP or CysAP. In contrast with effects in plasma, GluAP was unaffected, AspAP decreased, while AlaAP and CysAP increased. In the kidney, enzymatic activities did not change in the cortex, but decreased in the medulla. These data suggest that after ACE inhibition, the metabolism of Ang I in hypothalamus may lead mainly to Ang 2-10 formation. In plasma, the results suggest an increased formation of Ang IV together with increased vasopressinase activity. In the kidney, there is a reduction of vasopressinase activity in the medulla, suggesting a functional reduction of vasopressin in this location. The present data suggest the existence of alternative pathways in addition to ACE inhibition that might be involved in reducing BP after captopril treatment.

The profile of fatty acids in frontal cortex of rats depends on the type of fat used in the diet and correlates with neuropeptidase activities.

The kind of fat in the diet modifies the profile of fatty acids in brain and also affects aminopeptidase activities in tissues. Although modifications in brain fatty acids, neurotransmitters, or enzymes due to dietary fat composition have been reported, no direct relationship has yet been described between specific brain fatty acid changes and neuropeptide metabolism following the fat composition of the diet. We investigated the lipid profile and some neuropeptidase activities in the frontal cortex of adult male rats after a period in which diets were supplemented with fatty acids differing in their degrees of saturation such as fish oil (rich in polyunsaturated fatty acids, PUFAs), olive oil (rich in monounsaturated fatty acids, MUFAs), and coconut oil (rich in saturated fatty acids, SAFAs). It is observed that the diet composition affects fatty acid distribution in the brain. Although there is no change of global aminopeptidase/neuropeptidase, their activities in the brain correlate positively or negatively with the dietary fat composition. It is hypothesized that fatty acid in the diet modifies membrane fluidity, peptidases tertiary structure, and therefore, the availability and function of neuropeptides. The present results support the notion that cognitive functions may be modulated depending on the type of fat used in the diet.

Hypothalamic and plasmatic angiotensin metabolism in L-NAME treated rats.

In order to study the interaction between the renin-angiotensin system (RAS) and nitric oxide (NO), we analyzed the activity of aspartyl- (AspAP), glutamyl- (GluAP), alanyl- (AlaAP), and cystinylaminopeptidase (CysAP) enzymes involved in the RAS cascade, in the hypothalamus, and plasma of normotensive adult male rats after the inhibition of NO production with the NO synthase inhibitor L-NAME (L-N (G)-nitroarginine methyl ester). L-NAME treatment produced a significant increase of systolic blood pressure (SBP). In plasma, while GluAP activity decreased significantly, suggesting a lower Ang III formation, the other aminopeptidases did not change after L-NAME treatment. In hypothalamus, the activities of AspAP and CysAP were not affected after L-NAME treatment. In contrast, GluAP and AlaAP increased significantly. These results suggested mainly a higher formation of Ang III, but also higher levels of Ang IV in the hypothalamus of L-NAME treated rats. Both peptides have hypertensive properties at central level. On the contrary, Ang III may counteract the hypertensive action of Ang II at the periphery. Therefore, the increased SBP in L-NAME treated rats may be due in part to the increased activity of GluAP and AlaAP in hypothalamus and to a decreased activity of GluAP in plasma.

Vinyl acetate degradation by Brevibacillus agri isolated from a slightly aerated methanogenic reactor.

In a previous paper, the authors showed that a slight aeration of a methanogenic reactor treating wastewater from the manufacture of polymeric resins could improve its performance, by increasing or allowing the removal of some of its contaminants, including vinyl acetate (VA). This paper reports the isolation under aerobic conditions of a VA-biodegrading axenic culture (strain C1) retrieved from the sludge of a slightly aerated methanogenic reactor at 1 mg L(-1) d(-1) of dissolved oxygen (DO). The axenic culture obtained was phenotypically (morphology, biochemical properties, VA consumption kinetics) and phylogenetically characterized. It formed white colonies with a branched and flat morphology on solid medium. The cell morphology of the isolate was bacillus with round endings and flagellate. The cells could form chains and were stained Gram-negative. The isolate required simple nutritional elements and had a growth rate of 0.024 h(-1). The phylogenetical analysis showed that the aerobic bacterium was identified as Brevibacillus agri, with 99.3% similarity. The VA consumption kinetics in the methanogenic sludge were: volumetric consumption rate (rVA) of 1.74 +/- 0.2 mg L(-1) h(-1), maximum specific consumption rate (qVAmax) of 3.98 mg g(-1) volatile suspended solids (VSS) h(-1) and affinity constant (Ks) of 457.1 mg L(-1). The same parameters in the axenic culture were 1.69 +/- 0.04 mg L(-1) (h-1), 4.09 mg g(-1) dry weight h(-1) and 421.9 mg L(-1), respectively. These results show evidence that the aerobic isolated bacterium, identified as Brevibacillus agri, carried out the VA hydrolysis in the slightly aerated methanogenic sludge, which is the limiting step in the degradation of this compound.

[Incident study of medication errors in drug use processes: prescription, transcription, validation, preparation, dispensing and administering in the hospital environment]. / Estudio de incidencia de los errores de medicación en los procesos de utilización del medicamento: prescripción, transcripción, validación, preparación, dispensación y administración en el ámbito hospitalario.

OBJECTIVE: To determine both the global Incident, and the Incident for stages of medication errors in six Catalonian hospitals, the types of error and the consequences. METHOD: A prospective design, with the global variable of the medication error. Potential errors have been excluded. The patients admitted to each hospital were studied in 2 groups of up to 300 patients and 1,500 administrations were observed. The NCCMERP taxonomy was applied. The prescription error was detected through the review of prescriptions, checking the patient, medication, adherence to protocols, interactions, contraindications, omission, duplicated therapy, doses, frequency, method, and lack of follow-up. During the transcription/validation, it was verified that the prescription matched the original order. In the dispensing process, the content of the drawers was checked, comparing it to the computer generated list, before sending out the single dose trolley. The transcription, preparation and administration were observed on the wards. The information for all the procedures was registered in a specific data sheet. There was moderate concordance amongst the inspectors (kappa = 0.525). RESULTS: 16.94 errors were detected per 100 patients-day and 0.98 errors per patient: 16 % in prescription, 27 % in transcription/validation, 48 % in dispensing, and 9 % in administration. 84.47 % were category B errors (they did not reach the patient), and < 0.5 % of the errors were harmful. The population, with an average age of 65, had a male/female ratio of 60/40. The principal therapeutic groups were: agents against peptic ulcer and GERD, antithrombotic agents, and other analgesics and antipyretics, principally in a solid oral drug form (58 %). The medications per patient-day were 5.5 and the units of medication were on average 11.21, varying greatly among the institutions. The adjustment of 10 units made the results more uniform. In all the stages, omission was the most frequent error. DISCUSSION: The different methods used and different areas of the investigations make comparisons difficult. This is evident in the harmful errors, the proportion of which is affected by the detection procedure. The number of mistakes avoided during the execution of this project demonstrates the need to improve the planning of the work systems and to establish safety measures.

Use of topical negative pressure with a lipidocolloid dressing: results of a clinical evaluation.

OBJECTIVE: To evaluate the protection and acceptability of Urgotul wound dressing in the local management of acute or chronic wounds receiving topical negative pressure (TNP) therapy. METHOD: This was a prospective multicentre non-comparative open-label trial. At each dressing change the investigating physician clinically evaluated and photographed the wound. Planimetric measurement was undertaken and wound depth was assessed at the start and end of the treatment. Follow-up was undertaken until deemed clinically unnecessary by the investigator. RESULTS: Sixty-six patients were included (42 acute wounds and 24 chronic wounds) and followed up for an average of 17 days. Dressing changes were deemed entirely painless in 52% of cases (compared with 18% at baseline) and pain between two consecutive dressing changes was absent in 66% of cases (34% at baseline). Removal of the TNP-interface dressing combination was considered'very easy' or 'easy' in 94% of cases and adherence to the wound was recorded as 'absent' in 88%. On average, the dressings were changed every 3.8 +/- 1.1 days (all wounds were considered), and wound area and depth were reduced by 19% and 54% respectively by the end of the follow-up period. CONCLUSION: Use of the interface dressing in combination with TNP substantially reduced the pain caused by dressing changes. It therefore makes more acceptable the use of this technique, which aims to optimise the management of wounds that are sometimes considered to be in a therapeutic impasse.

Handedness and gender influence blood pressure in young healthy men and women: A pilot study.

OBJECTIVE: The type and level of sex steroids influence blood pressure (BP). It has been suggested that functional brain asymmetries may be influenced by sex hormones. In addition, there are inter-arm differences in BP not yet related with handedness. In this study, we hypothesize a possible association between sex hormones, handedness, and inter-arm differences in blood pressure. METHODS: To analyze this hypothesis, we measured BP in the left and right arm of the left and right handed adult young men and women in menstrual and ovulatory phase and calculated their mean arterial pressure (MAP). RESULTS: Significant differences depending on sex, arm, handedness or phase of the cycle were observed. MAP was mostly higher in men than in women. Remarkably, in women, the highest levels were observed in the left handed in menstrual phase. Interestingly, the level of handedness correlated negatively with MAP measured in the left arm of right-handed women in the ovulatory phase but positively with the MAP measured in the right arm of right-handed women in the menstrual phase. CONCLUSIONS: These results may reflect an asymmetrical modulatory influence of sex hormones in BP control.

Angiotensinase activity is asymmetrically distributed in the amygdala, hippocampus and prefrontal cortex of the rat.

There are important asymmetries in brain functions such as emotional processing and stress response in humans and animals. Knowledge of the bilateral distribution of brain neurotransmitters is important to appropriately understand its functions. Some peptides such as those included in the renin-angiotensin system (RAS) and cholecystokinin (CCK) are related to modulation of behavior and stress. However, although angiotensin AT1 and CCK type 2 receptors were found in adult rat brain, there are no studies of their bilateral distribution in stress-related areas. The function of angiotensin peptides is depending on the action of several aminopeptidases (AP) called angiotensinases, some of them being also involved in the metabolism of CCK. We have studied the bilateral distribution of soluble (SOL) and membrane-bound (MEM) alanyl- (AlaAP), cystinyl- (CysAP), glutamyl- (GluAP) and aspartyl- (AspAP) AP activities in stress-related areas such as amygdala, hippocampus and medial prefrontal cortex of adult male rats in resting conditions. These enzymes are involved in the metabolism of angiotensins (AlaAP, CysAP, GluAP, AspAP) and CCK (GluAP, AspAP). In the amygdala, all the activities studied showed a right predominance with a significant difference ranging from 30% for SOL CysAP to 125% for SOL GluAP. In the hippocampus, there was a left predominance for SOL AlaAP, SOL and MEM CysAP and MEM AspAP activities (100, 80, 300 and 100% higher, respectively). In contrast, GluAP predominated remarkably in the right hippocampus (eight-fold for SOL and three-fold for MEM). In the prefrontal cortex, SOL and MEM CysAP and SOL AspAP predominated in the left hemisphere (40, 100 and 40% higher, respectively). These results demonstrated a heterogeneous bilateral pattern of angiotensinase activities in motivation and stress-related areas. This may reflect an uneven asymmetrical distribution of their endogenous substrates depending on the brain location and consequently, it would be also a reflect of the asymmetries in the functions they are involved in.

Serum pyroglutamyl aminopeptidase activity: a promising novel biomarker candidate for liver cirrhosis.

OBJECTIVE: As a reflect of tissue damage, serum aminopeptidases have been proposed as biomarkers of various diseases. In order to search new serologic markers for liver cirrhosis we conducted a preliminary study in which we analyzed a broad range of aminopeptidase activities in serum of controls and patients diagnosed with pancreatitis, hepatitis, and liver cirrhosis without distinction among the etiological type or the degree of severity of each condition. METHODS: Alanyl-, arginyl-, glutamyl-, cystinyl- pyroglutamyl-, and aspartyl-aminopeptidase activities were analyzed fluorometrically, using aminoacyl-ß-naphthylamides as substrates. In addition, various parameters, such as alanine transaminase, aspartate transaminase, alkaline phosphatase, total bilirubin, direct bilirubin, and gamma glutamyl transpeptidase were assayed as routine laboratory test for liver function. RESULTS: Compared with control group, alanyl- and arginyl-aminopeptidase activities increased nonspecifically in pancreatitis, hepatitis and liver cirrhosis, glutamyl- and cystinyl-aminopeptidases did not differ between groups and pyroglutamyl-aminopeptidase demonstrated that while pancreatitis and hepatitis did not differ between them and with controls, this activity decreased selectively in liver cirrhosis compared with all the rest of groups (p<0.001 vs. control and p<0.01 vs. pancreatitis and hepatitis). Aspartyl-aminopeptidase also decreased significantly (p<0.05) in liver cirrhosis compared with controls. Routine parameters for liver function test increased, as expected, in the three pathologies analyzed. CONCLUSIONS: Despite the heterogeneous composition of the three patient groups, the specific reduction of the levels of pyroglutamyl-aminopeptidase activity in serum of liver cirrhosis patients might be considered as a potential candidate to be included in a combination of markers for the diagnosis of this disease.

Influence of thyroid disorders on the kidney expression and plasma activity of aminopeptidase A.

OBJECTIVE: Thyroid disorders may affect blood pressure and renal function modifying factors of the plasmatic and kidney renin-angiotensin system such as aminopeptidase A (AP A) that metabolizes angiotensin II to angiotensin III. We investigated the expression of AP A in the kidney, as well as its enzymatic activity in the plasma of euthyroid, hyperthyroid, and hypothyroid adult male rats. METHODS: Hyperthyroidism was induced by daily subcutaneous injections of tetraiodothyronine. Hypothyroid rats were obtained by administration of methimazole in drinking water. Expression of AP A was determined by Western blot analysis. Plasma AP A activity was measured fluorometrically using glutamyl-ß-naphthylamide as substrate. RESULTS: While hyperthyroid rats exhibited lower levels of plasma AP A activity than controls, the kidney of hyperthyroid animals expressed significantly higher AP A than controls and hypothyroid animals. CONCLUSIONS: A discrepancy between the high expression of AP A in kidney of hyperthyroid rats and the low activity of AP A measured in plasma and kidney of hyperthyroid animals was found. The posttranslational influence of environmental biochemical factors may be in part responsible for that divergence.

Rapid mineralisation of the Organic Fraction of Municipal Solid Waste.

The effect of pH, C/N ratio, addition of a microbial consortium (MC) and temperature upon mineralisation of Organic Fraction of Municipal Solid Waste (OFMSW) was studied; mineralisation was measured through the CO2 production rate and total CO2 formation. Through this process up to 432.9mg of CO2g(-1) initial dry matter (IDM) after 2days of treatment was obtained. It was found that under a slightly acidic pH (5-6) and C/N of 30, the mineralisation process was accelerated. Moreover, temperature (27-50°C) had no effect on the total CO2 produced. The highest CO2 production rate (5.28d(-1)) was observed at 27°C, C/N ratio of 30 and 8% of microbial consortium; it is at least 3.52 times higher than that reported (1.5d(-1)). The highest release of reducing sugars was determined at 50°C, possibly due to an increase in hydrolytic enzymes. Results suggest the potential use of rapid mineralisation of OFMSW for further friendly environmental processes.

Strain-responsive regions in the platelet-derived growth factor-A gene promoter.

Proliferation of cultured neonatal vascular smooth muscle (VSM) cells is enhanced by exposure to cyclic mechanical strain, in part through autocrine action of secreted platelet-derived growth factor (PDGF). We examined transcription factors and DNA response elements that may participate in the induction of PDGF-A gene transcription by mechanical strain. PDGF-A mRNA increased gradually over 4 to 24 hours exposure to cyclic (1 Hz) strain. This was due, at least in part, to increased transcription since a full length (890 bp) PDGF-A promoter reporter construct was induced 3.5-fold in transfected VSM cells exposed to strain for 24 hours. A series of PDGF-A promoter truncation reporter constructs was used to identify potential regions of the promoter involved in regulation by strain. Strain-responsive regions were found between -262 bp and -92 bp and between -92 bp and -41 bp of the promoter. Since these regions are GC-rich and contain response elements for Egr-1 and Sp-1, we examined expression of these transcription factors in response to strain. mRNA for both factors increased over 0.5 to 4 hours of strain, while protein expression for both increased gradually over a 24 hours period. Gel shift assays with a probe specific for Egr-1 demonstrated at least 1 prominent new shifted band after 4 to 12 hours exposure to strain. An Sp-1 probe demonstrated constitutive shifted bands that did not change in response to strain. Thus, GC-rich regions in the proximal 92 bp of the PDGF-A promoter contain mechanical strain-responsive elements that bind Egr-1 and possibly Sp-1.

Electrophysiological study of the effects of D1 and D2 dopamine antagonists on the interaction of converging inputs from the sensory-motor cortex and substantia nigra neurons in the rat.

The interaction of stimulation of the cerebral cortex and of the substantia nigra on the activity of neostriatal neurons was investigated in urethane-anesthetized rats. Neurons of the dorsal striatum were activated by single pulse stimulation of the sensory-motor cortex. The effects of nigral conditioning stimulation on this excitatory response of striatal neurons to cortical stimulation were studied in a series of parametric experiments in which the length of the train of pulses and the intensity of the nigral stimulation were varied. One and five pulses of nigral conditioning stimulation had little or no effect. Ten pulses of nigral conditioning stimulation reduced the excitatory response, the magnitude of the reduction being greater with higher current intensities. In another series of experiments, the effects of dopaminergic receptor antagonists on the interaction of cortical and nigral inputs to striatal neurons were studied. Sulpiride, a D2 antagonist, reversed the attenuating effects of nigral conditioning stimulation on the excitatory response of striatal neurons to cortical stimulation, whereas SCH 23390 a D1 antagonist, had no effect. The present findings support the hypothesis that the nigrostriatal dopaminergic pathway modulates the excitatory response of striatal neurons to cortical stimulation by means of dopamine D2 receptors.

Modification of nitric oxide synthase activity and neuronal response in rat striatum by melatonin and kynurenine derivatives.

Tryptophan is mainly metabolized in the brain through methoxyindole and kynurenine pathways. The methoxyindole pathway produces (among other compounds) melatonin, which displays inhibitory effects on human and animal central nervous systems, including a significant attenuation of excitatory, glutamate-mediated responses. The kynurenine pathway produces kynurenines that interact with brain glutamate-mediated responses. Nitric oxide (NO) increases glutamate release, and melatonin and kynurenines may act via modification of NO synthesis. In the present study, the effects of melatonin and four synthetic kynurenines were studied on the activity of rat striatal nitric oxide synthase (NOS) and on the response of rat striatal neurons to sensorimotor cortex (SMCx) stimulation, a glutamate-mediated response. Melatonin inhibited both NOS activity and the striatal glutamate response, and these effects were dose-related. Compound A (2-acetamide-4-(3-methoxyphenyl)-4-oxobutyric acid) did not inhibit NOS activity but inhibited the striatal response similarly to melatonin. Compound B (2-acetamide-4-(2-amino-5-methoxyphenyl)-4-oxobutyric acid) was more potent than melatonin in inhibiting both NOS activity and the striatal response. Compound C (2-butyramide-4-(3-methoxyphenyl)-4-oxobutyric acid) did not change NOS activity, but increased the striatal response. Compound D (2-butyramide-4-(2-amino-5-methoxyphenyl)-4-oxobutyric acid) showed potent inhibitory effects on both NOS activity and striatal glutamate-mediated response. A structure-related effect of the kynurenine derivatives was observed, and those with an amino group in position 2 of the benzenic ring had more potent effects than melatonin itself in inhibiting striatal NOS activity and the response of striatal neurons to SMCx.

Calcium-dependent effects of melatonin inhibition of glutamatergic response in rat striatum.

The effects of melatonin, amlodipine, diltiazem (L-type Ca2+ channel blockers) and omega-conotoxin (N-type Ca2+ channel blocker) on the glutamate-dependent excitatory response of striatal neurones to sensory-motor cortex stimulation was studied in a total of 111 neurones. Iontophoresis of melatonin produced a significant attenuation of the excitatory response in 85.2% of the neurones with a latency period of 2 min. Iontophoresis of either L- or N-type Ca2+ channel blocker also produced a significant attenuation of the excitatory response in more than 50% of the recorded neurones without significant latency. The simultaneous iontophoresis of melatonin + amlodipine or melatonin + diltiazem did not increase the attenuation produced by melatonin alone. However, the attenuation of the excitatory response was significantly higher after ejecting melatonin + omega-conotoxin than after ejecting melatonin alone. The melatonin-Ca2+ relationship was further supported by iontophoresis of the Ca2+ ionophore A-23187, which suppressed the inhibitory effect of either melatonin or Ca2+ antagonists. In addition, in synaptosomes prepared from rat striatum, melatonin produced a decrease in the Ca2+ influx measured by Fura-2AM fluorescence. Binding experiments with [3H]MK-801 in membrane preparations from rat striatum showed that melatonin did not compete with the MK-801 binding sites themselves although, in the presence of Mg2+, melatonin increased the affinity of MK-801. The results suggest that decreased Ca2+ influx is involved in the inhibitory effects of melatonin on the glutamatergic activity of rat striatum.

Opioid peptides and self-stimulation of the medial prefrontal cortex in the rat.

The possible involvement of opioid peptides as part of the neurochemical substrates of self-stimulation (SS) in the medial prefrontal cortex (MPC) of the rat was investigated in two different groups of rats bilaterally implanted with monopolar electrodes in the MPC. In the first group, morphine (5, 10, and 20 micrograms) and an enkephalin analogue (BW 180) (5, 10, 20 and 40 micrograms) were injected through cannulae implanted into the lateral ventricles (IV). In the second group, naloxone (0.04, 0.4, and 1.6 micrograms) and morphine (5, 10 and 20 micrograms) were injected through cannulae implanted into the MPC, 1.5 mm above the tip of the stimulating electrodes. In the first group, spontaneous motor activity (SMA) was measured as a control for non-specific effects (sedation or motor dysfunction). In the second group SS, contralateral to the microinjected side, served as control. SS and SMA were were measured 1 and 2 h postinjection. One hour after IV injection of morphine SS was not affected, although SMA was decreased. Two hours postinjection, on the contrary, SS was increased while SMA remained decreased. Similar effects were found with IV microinjections of BW 180. Naloxone, intraperitoneally injected, reversed all these effects. Naloxone or morphine injected intracerebrally (MPC) produced no changes in SS either in the injected or in the contralateral side, which served as control. The present results suggest that the effects found with IV injections of opioids on SS of the MPC are indirect (through activation of other brain areas) and not mediated by a direct action on the neurochemical substrates underlying this behaviour in the MPC.

Melatonin-dopamine interaction in the striatal projection area of sensorimotor cortex in the rat.

The excitatory response to motor cortex stimulation of 201 striatal neurones was recorded electrophysiologically to test the effects of melatonin (aMT) and/or D1 and D2 antagonists. Iontophoresis of aMT attenuated the excitatory response in 68.5% of neurones, with a latency of 2-4 min and enhanced the excitatory response in 11.9% of the neurones; 19.6% showed no change in response. Iontophoresis of sulpiride (D2 antagonist) produced an immediate increase in the excitatory response in 62.8% of neurones, an attenuation in 2.3% and no change in the response of 34.9%. The ejection of sulpiride counteracted the aMT-dependent inhibition of the excitatory response of striatal neurones. SCH-23390 (D1 antagonist) iontophoresis had no significant effect. The results show that the same striatal units may be driven by aMT and D2 receptors. However, the significant difference in the latency of the responses suggests that the effects of these two substances are mediated by different receptor/intracellular messengers.