Effects of Mediterranean diet in patients with recurring colds and frequent complications.

INTRODUCTION: In recent years, traditional diets enriched with fresh plant-based foods have been gradually abandoned, increasing the consumption of animal foods and highly processed food. The aim of this study was to assess the effects of a nutritional intervention with a Traditional Mediterranean Diet in patients with recurring colds (RC) and frequent inflammatory complications (IC). METHODS: Prospective before-after comparison study of 63 girls and 65 boys aged 1-5 years were included over a year in the nutritional programme "Learning to eat from the Mediterranean". We studied clinical and therapeutic variables and various anthropometric parameters. RESULTS: All the studied indicators (number of catarrhal episodes CB, degree of intensity, emergency and hospital admissions) showed a positive and statistically significant evolution, evidenced from the first weeks of starting treatment, until the end of the year, after which 53.9% of patients had no CB, 25% had only one, and 16.4% had two episodes, compared to the 4.64 episodes on average in the previous year. Antibiotic use decreased by 87.4%, from 3.85±1.27 times/patient/year to 0.49±0.79 (p<0.001). Symptomatic treatment decreased by 56.7%, from 7.03±2.76 to 3.05±1.69 (p<0.001). The satisfaction of the families was very high. The Kidmed index, which assesses the quality of the Mediterranean Diet, increased from 7.8 to 10.9 points. CONCLUSION: The adoption of a Traditional Mediterranean Diet could be a major contribution to the improvement of patients with recurring colds and frequent inflammatory complications.

Cocaine facilitates protein synthesis-dependent LTP: the role of metabotropic glutamate receptors.

Cocaine addiction alters synaptic plasticity in many brain areas involved in learning and memory processes, including the hippocampus. Long-term potentiation (LTP) is one of the best studied examples of hippocampal synaptic plasticity and it is considered as one of the molecular basis of learning and memory. We previously demonstrated that in the presence of cocaine, a long lasting form of hippocampal LTP is induced by a single pulse of high frequency stimulation, which in normal conditions evokes only an early form of LTP. In this study, we further explore the molecular basis of this modulation of synaptic plasticity by cocaine. By performing pharmacological experiments on hippocampal slices, we were able to show that cocaine converts early LTP to a form of LTP dependent on protein synthesis, probably through the cAMP-dependent protein kinase and extracellular signal-regulated kinase signaling cascades. We also found that metabotropic glutamate receptors are involved in this phenomenon. These studies further clarify the molecular machinery used by cocaine to alter synaptic plasticity and modulate learning and memory processes.

Differential regulation of CDK5 and c-Fos expression by morphine in the brain of Lewis and Fischer 344 rat strains.

The aim of this study was to comparatively study cyclin-dependent kinase 5 (CDK5) and c-Fos regulation by morphine in the brains of Lewis and Fischer 344 (F344) rats, which are known to differ in their behavioral sensitivities to several drugs of abuse. Two hours after an acute i.p. administration of morphine (10 mg kg(-1)) or saline (control), the animals were perfused and their brains prepared for immunohistochemistry. The number of CDK5 immunoreactive cells was significantly higher in the nucleus accumbens (NAC), the locus coeruleus (LC) and the nucleus tractus solitarius (NTS) of saline-injected F344 rats than in those of the Lewis rats. Morphine upregulated CDK5 with a varying pattern depending on the strain and brain area. The effect of the opioid was more marked in the NTS of the Lewis rats and the NAC of the F344 rats. Immunostaining of c-Fos was very low or absent in the control animals and was consistently up-regulated by morphine, especially in the LC and NTS of the F344 rats and the NAC of the Lewis rats. We propose that the acute morphine regulation of CDK5 expression in the NAC may predict the rate of drug intake and/or extinction of drug seeking, while the pattern of c-Fos activation may be more related to the differential acquisition of morphine-seeking behaviors.

Undernutrition upregulates fumarate hydratase in the rat nucleus accumbens.

Previous comparative studies of fumarate hydratase (FH) protein density revealed that the enzyme was overexpressed in the striatum of rodents that are less influenced by rewarding stimuli, from cocaine to food. Therefore, we recently proposed FH as a potential striatal biomarker of brain reward deficiency and addiction vulnerability. This work has been focused to investigate FH activity in the Nucleus Accumbens (NAc) of undernourished rats, taking into account that malnutrition has been related to increased responsiveness to food and drug reward. To this end, we have studied adult female Wistar rats severely food restricted from the 16th day of intrauterine life until adulthood. Animals were sacrificed to dissect the NAc and obtain mitochondrial and cytosolic fractions after homogenisation and centrifugation. FH activity was measured by conversion of malate to fumarate, and protein levels were compared by Western blot analysis when fractions showed differences in activity. Undernutrition did not change cytosolic FH activity but led to a marked increase of mitochondrial FH activity (72 %) and protein content (50 %) in the NAc. This change was in the opposite direction that one would predict if it was related to addiction vulnerability of some kind, but strongly suggests that mitochondrial FH needs to be at some optimal level for normal reward responsiveness.

Genetic differences in hippocampal synaptic plasticity.

Synaptic plasticity is considered a physiological substrate for learning and memory [Lynch MA (2004) Long-term potentiation and memory. Physiol Rev 84:87-136] that contributes to maladaptive learning in drug addiction [Schoenbaum G, Roesch MR, Stalnaker TA (2006) Orbitofrontal cortex, decision-making and drug addiction. Trends Neurosci 29:116-124]. Many studies have revealed that drug addiction has a strong hereditary component [Kosten TA, Ambrosio E (2002) HPA axis function and drug addictive behaviors: insights from studies with Lewis and Fischer 344 inbred rats. Psychoneuroendocrinology 27:35-69; Uhl GR (2004) Molecular genetic underpinnings of human substance abuse vulnerability: likely contributions to understanding addiction as a mnemonic process. Neuropharmacology 47 (Suppl 1):140-147], however the contribution of the genetic background to drug-induced changes in synaptic plasticity has been scarcely studied. The present study reports on an analysis of long-term potentiation (LTP) and depotentiation in Lewis (LEW) and Fischer-344 (F344) rats, two inbred rat strains that show different proneness to drugs of abuse and are considered an experimental model of genetic vulnerability to addiction [Kosten TA, Ambrosio E (2002) HPA axis function and drug addictive behaviors: insights from studies with Lewis and Fischer 344 inbred rats. Psychoneuroendocrinology 27:35-69]. The induction of saturated-LTP was similar in LEW and F344 rats treated with saline or cocaine. However, only slices from LEW saline-treated rats showed the reversal of LTP; thus, the depotentiation of saturated-LTP was not observed in cocaine-injected LEW rats and in F344 animals (treated either with cocaine or saline). These results suggest significant differences in hippocampal synaptic plasticity between Lewis and Fischer 344 rats.

Capillary electrophoresis for caffeine and pyroglutamate determination in coffees study of the in vivo effect on learning and locomotor activity in mice.

In a preliminary study pyroglutamate showed to be over 10 times increased in some lyophilised coffees with respect to brewed or filtered coffees, and probably that increase is related to some stage of the industrial process. Pyroglutamate is known to have a number of remarkable cognitive enhancing effects, which could be also related to the properties of coffee traditionally associated to caffeine. Pyroglutamate improves memory and learning and has anti-anxiety effects in rats. Therefore, a method has been developed and validated for the simultaneous determination of caffeine and pyroglutamate in coffee by capillary electrophoresis. Separation conditions employed MECK conditions with 50 mM borate buffer at pH 9.5 with 130 mM SDS. The applied potential was 10 kV and detection was performed at 200 nm. Afterwards, 10 soluble coffees from the market were measured and caffeine and pyroglutamate levels were compared. Those coffees with higher pyroglutamate with or without caffeine were preliminarily tested for sedative/stimulant properties and cognition enhancing effects in mice. The most relevant finding was a partial reversal of scopolamine-induced amnesia in the passive avoidance paradigm after oral administration of one coffee.

Histamine H3 receptor: a potential drug target for the treatment of central nervous system disorders.

Histamine H(3) receptors were first described in the eighties but finally cloned four years ago. They are G-protein coupled, mostly presynaptic, and are involved in the control of the synthesis and/or release of different neurotransmitters both in the central nervous system and the periphery. The availabiliy of specific ligands has permitted the study of potential therapeutic applications of either stimulating or blocking the function of these receptors. There is experimental evidence that drugs targeted at histamine H(3) receptors could be beneficial for neurodegenerative diseases such as Alzheimer and Parkinson's disease, epilepsy, drug abuse and several affective, appetite and sleeping disorders, among others. This review presents recent advances in this field.

Differences of mu-opioid receptors between Lewis and F344 rats.

F344 and Lewis rats show different responses to opioids in several experimental paradigms. In this study we have used the specific mu-opioid agonist DAMGO to find out if these differences could be attributed to heterogeneity of mu-opioid receptors. The density of [H3]DAMGO binding sites was similar in the brain cortex and spinal cord of both strains, but DAMGO affinity for mu-opioid receptors was higher in F344 tissues. Moreover, a parallel study of the effects of DAMGO on electrically-evoked twitches of isolated vasa deferentia revealed that this drug was also more effective in F344 preparations. These results suggest that mu-opioid receptors of F344 rats are more sensitive to pharmacological stimulation in vitro, which could be related to a higher drug affinity.

Biological activity of extracts from Catalpa bignonioides Walt. (Bignoniaceae).

Catalpa bignonioides Walt. (Bignoniaceae) is a species that belongs to a tropical family but has been introduced in many countries as ornamental. Although this plant is consumed by indigenous cultures of South America for medical uses, experimental studies of the biological properties of Catalpa bignonioides are lacking. The aim of this work was to study the biological activity of crude extracts from either pods, seeds or leaves of Catalpa bignonioides which were collected in Spain. Ethyl ether, butanolic and aqueous fractions of the pod extract were also prepared and studied. We have examined the antimicrobial activity against five bacteria and one yeast, the cytotoxic activity against HepG2 cells and the anti-inflammatory and antinociceptive effects in rodents. A preliminary phytochemical analysis of the extracts and fractions was also conducted. Results showed no antimicrobial or antitumoral effects, but prominent anti-inflammatory and antinociceptive actions of the extracts. These last activities may be a result of the presence of either of saponins, sterols or phenols, mainly found in the leaves and pods of the plants.

Effects of yohimbine on the antinociceptive and place conditioning effects of opioid agonists in rodents.

1. The pharmacological modulation of opioid actions by drugs acting on heterologous mechanisms could be useful to overcome some of the main problems associated with the use of opiate agonists. Based on previous findings on the interactions between yohimbine and opioid drugs, we have further studied the effects of yohimbine on the antinociceptive and positive-negative reinforcing effects of morphine (mu opioid receptor-preferring agonist), U-50,488 (kappa agonist) and SNC80 (delta agonist). 2. Pretreatment with yohimbine completely blocked the antinociception provided by the three opioid agonists in the mouse tail-immersion test. 3. A similar blockade of SNC80 and U-50,488-induced antinociception was observed with yohimbine in the mouse hot plate test at the same doses. In this paradigm, the effect of the kappa agonist was very slight and the actions of yohimbine rather variable. 4. In place conditioning experiments with SD (Sprague -- Dawley) male rats, yohimbine alone was inactive but it limited the preference induced by morphine and SNC80 and the aversive effect of U-50,488. Impaired novelty preference was also observed with the combination of yohimbine and U-50,488. 5. It is concluded that yohimbine tends to limit opioid antinociception and the addictive potential of mu and delta opioid agonists. More selective drugs could help to understand the mechanisms involved in these actions.

Tecoma sambucifolia: anti-inflammatory and antinociceptive activities, and 'in vitro' toxicity of extracts of the 'huarumo' of peruvian incas.

Aqueous and alcoholic extracts of pods and flowers of Tecoma sambucifolia H.B.K. (Bignoniaceae) ('huarumo') were analysed to determine their anti-inflammatory activity (carrageenan-induced edema test), antinociceptive activity (acetic acid writhing test) and 'in vitro' toxicity in Chinese hamster ovary cells, human hepatome cells and human larynx epidermal carcinoma cells. The cytotoxic effects of both extracts were evaluated by two endpoint systems: neutral red uptake assay and tetrazolium assay. The results showed that all extracts have anti-inflammatory and antinociceptive activity, but the highest potency is that of the alcoholic extracts. There were significant differences in cytotoxicity between extracts and among the response of cells to them. The highest cytotoxicity was noted with the alcoholic extract, and the human hepatome cell line was the most sensitive, especially to the alcoholic extract of flowers. The aqueous pod extract appeared to have the best pharmaco-toxicological profile, since it provided a significant reduction of both pain and inflammation together with the lowest cytotoxicity.

Preweanling naltrindole administration differentially affects clonidine induced antinociception and plasma adrenaline levels in male and female neonatal rats.

1. The influence of a chronic treatment with the delta-selective opioid antagonist naltrindole (1 mg kg-1) during the preweanling period (daily injections from birth to postnatal day 19), on the antinociceptive and sympatholytic effects of the alpha2-adrenergic agonist clonidine in male and female rats of 20 and 25 days of age was investigated. 2. Nociception was assessed using the tail immersion test (water at 50 degrees C) and plasma levels of adrenaline were measured by high-performance liquid chromatography. 3. The dose of clonidine (1.5 mg kg-1) and the time point at which nociceptive responses were recorded (30 min after the administration of the drug) were chosen on the basis of dose-response (0.5, 1, 1.5 and 2 mg kg-1) and time-response (5, 10, 15, 30 and 60 min) curves which were previously carried out in naive control neonatal rats. 4. In females, the functional blockade of the delta-receptor by neonatal naltrindole treatment did not modify the sympatholytic effect of clonidine but prevented clonidine induced antinociception. Conversely, in males naltrindole treatment allowed the appearance of clonidine antinociception and the sympatholytic effect of clonidine. 5. The results indicate that the delta-receptor is involved in the modulation of antinociceptive and sympatholytic responses to clonidine in neonatal rats and suggest the existence of sex differences in the interactions between delta-opioid and alpha2-adrenergic receptors.

Effects of histamine H3 receptor ligands in experimental models of anxiety and depression.

Histamine H3 receptor ligands have been proposed to be of potential therapeutic interest for the treatment of different central nervous system disorders; however, the psychopharmacological properties of these drugs have not been studied extensively. In this work, we investigated the possible involvement of histamine H3 receptor function in experimental models of anxiety (elevated plus-maze) and depression (forced swimming test). Male Sprague-Dawley rats were treated i.p. with the histamine H3 receptor agonist R-alpha-methylhistamine (10 mg/kg) or the histamine H3 receptor antagonist thioperamide (0.2, 2 and 10 mg/kg) and 30 min afterwards the time spent in the open arms of an elevated plus-maze was registered for 5 min. The immobility time of male OF1 mice in the forced swimming test was recorded for 6 min, 1 h after the i.p. administration of R-alpha-methylhistamine (10 and 20 mg/kg), thioperamide (0.2, 2, 10 and 20 mg/kg) or another histamine H3 receptor antagonist, clobenpropit (5 mg/kg). The locomotor activity of mice was checked in parallel by means of an activity meter. Both saline controls and active drug controls were used in all the paradigms. Neither thioperamide nor R-alpha-methylhistamine significantly changed animal behaviour in the elevated plus-maze. R-alpha-methylhistamine and the higher dose of thioperamide assayed (20 mg/kg) were also inactive in the forced swimming test. By contrast, thioperamide (0.2-10 mg/kg) dose-dependently decreased immobility, the effect being significant at 10 mg/kg (33% reduction of immobility); clobenpropit produced an effect qualitatively similar (24% reduction of immobility). None of these histamine H3 receptor antagonists affected locomotor activity. These preliminary results suggest that the histamine H3 receptor blockade could be devoid of anxiolytic potential but have antidepressant effects. Besides, the stimulation of these receptors does not seem to be followed by changes in the behavioural parameters studied.

Histamine H3 receptor desensitization in the guinea-pig ileum.

Histamine H3 receptor ligands are usually tested in guinea-pig intestine preparations. A possible desensitization of agonist-induced twitch inhibition was studied in longitudinal muscle-myenteric plexus from ileal segments. A cumulative concentration-response curve for R-alpha-methylhistamine was made; when a second curve was made 30 min afterwards, a marked decrease of pD2 and a more modest decrease of Emax were observed without changes in tissue sensitivity to electrical stimulation or morphine inhibition. At 120 min, pD2 and Emax were not different from those for the first curve. Receptor desensitization seems homologous and reversible and could interfere with repetitive testing of histamine H3 receptor ligands.

Effect of oral yohimbine on withdrawal jumping behaviour of morphine-dependent mice.

Acute administration of the alpha-2 adrenoceptor agonist clonidine and chronic administration of the alpha2 antagonist yohimbine both inhibit opioid withdrawal signs in experimental models of dependence and also in clinical studies with opiate abusers. There are exceptions to this general rule: restlessness or self-reported abstinence in humans and withdrawal-induced escape behaviour in rodents are resistant to inhibition by acute clonidine. We have explored the effect of the alpha-2 antagonist yohimbine on morphine withdrawal-induced escape behaviour in a mouse model that we have proposed to differentiate between the urge to escape (number of jumps) and non-specific sedative/motor actions (height of jumps). Morphine dependence was induced by s.c. administration of a sustained-release preparation (1 g/kg). Naloxone (1 mg/kg) was injected to precipitate withdrawal jumping 72 hours after morphine injection. Co-treatment with yohimbine dissolved in the tap water (70 mg/l) decreased the number of jumps upon naloxone challenge, an effect which did not seem to be related with a sedative or toxic effect of the drug. This result confirms previous data and suggests that yohimbine could prevent the development of opioid dependence being active to decrease withdrawal-induced escape behaviour. The mechanisms of this action are discussed.

Cardiovascular parameters of heroin-withdrawn addicts treated with guanfacine.

Blood pressure and heart rate were monitored in heroin addicts detoxified with guanfacine in an outpatient program. Retrospective analysis of the cardiovascular parameters from the addicts who failed this detoxification during the first 5 days of treatment (they consumed heroin within this period) showed that these patients experienced less hypotension and bradycardia when treated with the alpha 2-agonist than the group of successful patients. Therefore, the relapse to opiate use could be predicted, at least in some patients, by a minimal cardiovascular reaction to guanfacine. Opiate use was followed by significant increases in blood pressure and heart rate, as had happened in previous studies with clonidine-treated addicts. This finding further confirms a similarity between alpha 2-agonists adrenergic and opiates with regard to their cardiovascular effects.

Effect of yohimbine on the development of morphine dependence in the rat: lack of involvement of cortical beta-adrenoceptor modifications.

The alpha-2 receptor antagonist yohimbine has been previously shown to prevent the development of morphine dependence in a rat behavioral model. This study was directed to clarify the mechanism of this interaction, which is presently unknown. Since upregulation of cortical beta-adrenoceptors has been suggested to be involved in morphine withdrawal, we have tested the possible correlation between receptor density and withdrawal behaviors in the presence of yohimbine. Sprague-Dawley male rats received a s.c. suspension of morphine (300 mg/kg) or the vehicle. Animals received saline or yohimbine (4 mg/kg, IP) 24, 28, 48 and 52 h after morphine and finally naloxone (1 mg/kg i.p) at 72 h; the subsequent signs of withdrawal (mainly wet-dog shakes and escape attempts) were recorded and the cerebral cortex dissected to study [3H]-CGP 12177 binding. Morphine-treated animals displayed a marked withdrawal behavior together with beta-adrenoceptor upregulation; nevertheless, these effects were not correlated. As expected, yohimbine prevented morphine withdrawal behavior but did not reverse the beta-adrenoceptor upregulation induced by the opiate. These results confirm previous evidence against the involvement of beta-adrenoceptor upregulation on morphine withdrawal behaviors and also permit to discard beta-adrenoceptor regulation as the neurochemical basis of the antiwithdrawal effect of yohimbine. The possible contribution of some other neurochemical effects of yohimbine are discussed to explain the inhibition of morphine dependence by that drug.

Social deprivation stress induces adaptative changes of opioid mechanisms in the rat tail artery.

Brief (7-14 days) social deprivation stress has been found to increase blood pressure in Wistar rats, an effect dependent on activation of opioid function. The role of central opioids in this and other responses to stress has been repeatedly determined, but the possible involvement of modifications of peripheral opioid mechanisms is poorly understood. To further increase this knowledge, we have examined the opioid sensitivity of tail arteries taken from social deprived Wistar rats by studying the effect of beta-endorphin and DADLE "in vitro". Both opioids inhibited the electrically-induced constriction of the preparations in a dose-dependent manner, but these actions were significantly attenuated after 7-14 days of social deprivation. When the rats were isolated for 30-35 days, the hypertensive response was still present but the arteries from group-housed and isolated animals no longer showed differential sensitivity to opioids. This difference with respect to 7-14 days of isolation could be related to age-dependent changes of opioid function, which were observed among group-housed animals. The results suggest that social deprivation stress induces an adaptation of the tail arteries to the opioid effects on contractility. It is suggested that this endogenous adaptation could be contributing to the hypertensive response observed after social deprivation.