Beyond lectures and practical courses: Teaching pharmacology using imaginative pedagogical tools.

Pharmacology has broadened its scope considerably in recent decades. Initially, it was of interest to chemists, doctors and pharmacists. In recent years, however, it has been incorporated into the teaching of biologists, molecular biologists, biotechnologists, chemical engineers and many health professionals, among others. Traditional teaching methods, such as lectures or laboratory work, have been superseded by the use of new pedagogical approaches to enable a better conceptualization and understanding of the discipline. In this article, we present several new methods that have been used in Spanish universities. Firstly, we describe a teaching network that has allowed the sharing of pedagogical innovations in Spanish universities. A European experience to improve prescribing safety is described in detail. The use of popular films and medical TV series in biomedical students shows how these audiovisual resources can be helpful in teaching pharmacology. The use of virtual worlds is detailed to introduce this new approach to teaching. The increasingly important area of the social aspects of pharmacology is also considered in two sections, one devoted to social pharmacology and the other to the use of learning based on social services to improve understanding of this important area. Finally, the use of Objective Structured Clinical Evaluation in pharmacology allows to know how this approach can help to better evaluate clinical pharmacology students. In conclusion, this article allows to know new pedagogical methods resources used in some Spanish universities that may help to improve the teaching of pharmacology.

Effectiveness of PUSH notifications from a mobile app for improving the body composition of overweight or obese women: a protocol of a three-armed randomized controlled trial.

BACKGROUND: The penetration level of mobile technology has grown exponentially and is part of our lifestyle, at all levels. The use of the smartphone has opened up a new horizon of possibilities in the treatment of health, not in vain, around 40% of existing applications are linked to the mHealth segment. Taking advantage of this circumstance to study new approaches in the treatment of obesity and prescription of physical activity is growing interest in the field of health. The primary outcome (obese adult women) will be assessed according to age, fitness status, weight, and body composition status. Data will be collected at enrollment and weekly during 6 months of intervention on dietary practices, physical activity, anthropometry, and body composition. Analysis of effect will be performed comparing the outcomes between intervention and control arms. The message delivery is in progress. METHODS: A 3-arm clinical trial was established. A series of quantitative and qualitative measures were used to evaluate the effects of self-weighing and the establishment of objectives to be reached concerning the prescription of physical activity. At the end of this pilot study, a set of appropriate measures and procedures were identified and agreed upon to determine the effectiveness of messaging in the form of PUSH technology. The results were recorded and analyzed to begin a randomized controlled trial to evaluate the effectiveness of the proposed methodology. CONCLUSIONS: The study is anticipated to establish feasibility of using PUSH notifications to evaluate whether or not an intervention of 6 months, directed by a team formed by Dietician-Nutritionist and nursing professionals, by means of an application for Smartphone and a personal consultation, improves the body composition of adult women with a fat percentage equal to or higher than 30% at the beginning of the study. TRIAL REGISTRATION: Clinical Trials ID: NCT03911583. First Submitted: April 9, 2019. Ethical oversight is provided by the Bioethical Committee of Córdoba University and registered in the platform clinicaltrials.gov. The results will be published in peer-reviewed journals and analysis data will be made public.

Changes in body composition with a hypocaloric diet combined with sedentary, moderate and high-intense physical activity: a randomized controlled trial.

BACKGROUND: There is evidence showing the effectiveness of a hypocaloric diet and the increase in physical activity on weight loss. However, the combined role of these factors, not only on weight loss but also body composition, remains unclear. The purpose of this study was to investigate the effect of a hypocaloric diet on the body composition of obese adult women throughout different degrees of physical activity during a weight loss program. METHODS: One hundred and seventeen healthy female volunteers were randomly assigned to one of the experimental groups: a control group with a low-level prescription of physical activity (1-4 METs), moderate physical activity group that performed 10.000 steps walking (5-8 METs) and intense physical activity group that trained exercises by at least 70% of VO2max three times a week (> 8 METs). All subjects followed a hypocaloric diet designed with a reduction of 500 kcal/day. Nutritional counseling was provided throughout the study period to help ensure dietary adherence. RESULTS: We found no differences in body weight compared to moderate and intense physical activity (ßstand. = - 0.138 vs. ßstand. = - 0.139). Body fat was lower in women following an intense activity (ßstand. = - 0.436) than those with moderate exercise (ßstand. = - 0.231). The high-intense activity also increased muscle mass at the end of the intervention, standing out above the moderate activity (ßstand. = 0.182 vs. ßstand. = 0.008). CONCLUSIONS: These findings indicate that a hypocaloric diet, without prescription of physical activity, is adequate to lose weight in the short term (12 weeks), but physical activity is vital to modify the body composition in women with obesity. Body fat was lower when women practiced a moderate exercise compared to hypocaloric diet only, but an intense physical activity was the most effective protocol to obtain a reduction of body fat and maintain muscle mass. TRIAL REGISTRATION: The study protocol complied with the Declaration of Helsinki for medical studies, it was approved by the bioethical committee of Córdoba University, in the Department of Health at the Regional Government of Andalusia (Act n°284, ref.4156) and retrospectively registered in clinicaltrials.gov (NCT03833791). Registered 2 January 2019.

Use of granules and free salts for Fe and Zn fortification of leafy vegetables: Improvements in trace element bioaccessibility and fulfillment of Dietary Reference Intakes.

BACKGROUND: Leafy vegetables represent an excellent dietary source of trace elements such as Fe and Zn. Nevertheless, Fe and Zn bioaccessibility can lessen due to a high concentration of anti-nutritional compounds. The encapsulation of Fe and Zn salts as granules could be used to fortify these leafy vegetables. METHOD: Three leafy vegetables, spinach, Swiss chard and Ethiopian mustard were fortified with iron sulfate and zinc sulfate as granules and free salts in order to test the improvements in the bioaccessibility and fulfillments of DRIs. Fe and Zn granules were prepared in a fluidized bed granulator. A probabilistic analysis was performed, using experimental data, to assess bioaccessible intake and fulfillments of DRIs in European populations. RESULTS: Fe contents ranged between 4.8 mg/100 g of Ethiopian mustard to 157.4 mg/100 g of spinach. Fe and Zn bioaccessibility percentages were low for Swiss chard and spinach without fortification. Fortification with granules improved Fe bioaccessibility of these latter vegetables (196 and 223 mg/100 g). Zn contents in samples without fortification ranged between 2.3 mg/100 g for Ethiopian mustard and 7.4 mg/100 g for spinach. Zn fortification as granules improved Zn bioaccessibility for the three vegetables studied. Thus, Zn bioccessible concentrations ranged between 17.4 and 108 mg/100 g for the solubility assay and between 5.9 and 31.1 mg/100 g for the dialyzability assay. Besides, the probability analysis showed that fortification had a better performance in meeting DRIs for those populations with higher consumption levels of leafy vegetables. CONCLUSIONS: The probability analysis demonstrated that fortification can be a suitable strategy to meet DRIs for both trace elements, which was especially remarkable for Fe. Fortification with granule was more effective in most the cases, although for Ethiopian mustard, free salt of Fe showed a better performance.

Bioaccessibility of glucosinolates, isothiocyanates and inorganic micronutrients in cruciferous vegetables through INFOGEST static in vitro digestion model.

Green tissues and seeds from cruciferous vegetables growing in conventional and ecological conditions (Brassica carinata; Brassica rapa; Eruca vesicaria and Sinapis alba) were analyzed to determine their contents of glucosinolates, isotihiocyanates (ITCs) and inorganic micronutrients (Ca, Cr, Cu, Fe, Mn, Ni, Se and Zn), and the bioaccessibility of these compounds. Regarding total contents and bioaccessibility values of these compounds, no clear difference was found between the organic and conventional systems. Glucosinolates bioaccessibility present in green tissues were high, with values around 60-78%. In additon, it was quantified in bioaccessible fraction ITCs concentrations such as Allyl - ITC; 3 - Buten - 1 - yl - ITC and 4 - Penten - 1 - yl - ITC. Trace elements bioaccessibility in green tissues was also high for Ca (2.26-7.66 mg/g), Cu (0.60-2.78 µg/g), Se (9.93-74.71 µg/Kg) and Zn (12.98-20.15 µg/g). By contrast, the bioaccessibility of glucosinolates and trace elements in cruciferous seeds was extremely low. With the exception of Cu, these bioaccessibility percentages did not exceed 1% in most cases.

Quantification and in vitro bioaccessibility of glucosinolates and trace elements in Brassicaceae leafy vegetables.

Leaf samples from five Brassicaceae species (Brassica carinata, Brassica oleracea, Brassica rapa, Eruca vesicaria and Sinapis alba) were analyzed to determine their contents of glucosinolates and trace elements, and the bioaccessibility of these compounds. Considerable variability in the total contents and glucosinolate profiles was observed in the Brassicaceae species, with the total amounts ranging from 8.5 µmol/g dw in Brassica oleracea to 32.9 µmol/g dw in Sinapis alba. Bioaccessibilities of the predominant glucosinolates were moderate, ranging from 13.1% for glucoraphanin to 43.2% for gluconapin, which is particularly relevant as they have been implicated in a variety of anti-carcinogenic mechanisms. Trace element concentrations were: Se (28-160 µg/Kg dw); Cr (0.31-4.03 µg/g dw); Ni (0.19-1.53 µg/g dw); Fe (8.6-18.8 µg/g dw); Zn (20.8-41.5 µg/g dw); Ca (6.2-15.2 mg/g dw). Brassicaceae leaves were also moderate dietary sources of Se, Ni, Zn and Ca.

Selenium and cadmium in bioaccessible fraction of organic weaning food: Risk assessment and influence of dietary components.

BACKGROUND: The tendency of some sectors of the population to consume organic food has also come to include baby food. Nevertheless, it is necessary to develop studies to support the true nutritional and toxicological value of these products, making special emphasis in several trace elements. To our knowledge, no studies have been conducted on this type of organic food. METHODS: Weaning foods with different formulations categorized as organic were analyzed to determine Se and Cd contents as well as its bioaccesibility. The analyses were conducted by electro thermal atomic absorption spectroscopy (ET - AAS) after the treatment of the samples with acid mineralization. Besides, macronutrient analyses (protein, fat and dietary fiber) were also developed. Finally, a novelty statistic approach such as @Risk was used to evaluate contributions to DRI or PTWI of Se and Cd derived for consumption of these weaning foods. RESULTS: Se content ranged between 2.44-15.4 µg Kg 1. Samples with meat ingredients showed the highest Se contents, while weaning foods consisting of fruits or vegetables presented the lowest concentrations. Se bioccessible concentration ranged between 1.90-4.35 µg Kg-1 with a greater uniformity amongst analyzed samples. Regarding Cd, concentrations of this heavy metal ranged between 1.23 and 3.64 µg Kg-1. Furthermore, Cd bioaccessibility of organic weaning foods ranged between 0.17 and 1.38 µg Kg-1. The solubility of all samples studied was around 20% from the initial Cd concentration. A negative statistical correlation between fat content - Cd bioaccesible (p < 0.05; r = - 0.756) and Cd content - Se bioaccesible (p < 0.05; r = - 0.777) were also found. CONCLUSIONS: Cd concentrations are considerably lower than those reported in weaning formulas which were not categorized as organic. On the other hand, the analysed organic jars did not represent a significant source of Se. The probabilistic assessment developed, showed that contributions to DRI of Se for infants 1-3 years old by consumption of these weaning foods, are excessively low (15% at best).

Complications associated with Brucella melitensis infection: a study of 530 cases.

We carried out a prospective study of 530 patients older than 14 years of age with brucellosis. We describe the incidence and clinical features of the focal forms of the disease, analyzing some of the possible factors associated with their appearance. One hundred sixty-nine patients (31.9%) had a focal form or complication. Osteoarticular complications were the most frequent, totaling 113 cases (66%), followed by genitourinary with 18 cases (5.1% of males), hepatic (2.5%), neurologic (1.7%), and heart (1.5%). Nine patients (1.7%) had more than 1 complication. In a multivariate analysis, diagnostic delay greater than 30 days (OR 2.0), ESR > 40 mm/hr (OR 1.9), and levels of alpha-2 globulin > 7.5 g/L (OR 6.8) were statistically significant independent variables associated with the presence of focal forms. Twenty-five patients with complications (14.8%) required surgical treatment. The relapse rate was 3.6% for those patients without complications and 4.1% for patients with focal forms (p > 0.05). However, when therapeutic failure, relapses, and mortality were considered together, the risk of an unfavorable evolution was significantly greater in patients with focal forms (10.6% versus 3.6% in patients without complications; OR 1.9, 95% CI 1.4-7.1, p < 0.005). Given the worse prognosis, knowledge and early diagnosis of the focal forms of B. melitensis infection is especially important.

Muscarinic receptor subtypes in human and rat colon smooth muscle.

Muscarinic receptor subtypes in human and rat colon smooth muscle homogenates were characterized with [3H]N-methylscopolamine ([3H]NMS) by ligand binding studies. [3H]NMS saturation experiments show the existence of a homogeneous population of non-interacting binding sites with similar affinity (KD values of 1.38 +/- 0.20 nM in human colon smooth muscle and 1.48 +/- 0.47 nM in rat colon smooth muscle) and with Hill slopes close to unity in both samples of tissue. However, a significant (P less than 0.01) increase in muscarinic receptor density (Bmax) is found in human colon (29.9 +/- 2.9 fmol/mg protein) compared with rat colon (17.2 +/- 1.5 fmol/mg protein). Inhibition of [3H]NMS binding by non-labelled compounds shows the following order in human colon: atropine greater than AF-DX 116 greater than pirenzepine. Whereas in rat colon the rank order obtained is atropine greater than pirenzepine greater than AF-DX 116. Atropine and pirenzepine bind to a homogeneous population of binding sites, although pirenzepine shows higher affinity to bind to the sites present in rat colon (Ki = 1.08 +/- 0.08 microM) than those in human colon (Ki = 1.74 +/- 0.02 microM) (P less than 0.05). Similarly, IC50 values obtained in AF-DX 116 competition experiments were significantly different (P less than 0.01) in human colon (IC50 = 1.69 +/- 0.37 microM) than in rat colon (IC50 = 3.78 +/- 0.75 microM). Unlike atropine and pirenzepine, the inhibition of [3H]NMS binding by AF-DX 116 did not yield a simple mass-action binding curve (nH less than 1, P less than 0.01) suggesting the presence of more than one subtype of muscarinic receptor in both species. Computer analysis of these curves with a two binding site model suggests the presence of two populations of receptor. The apparent Ki1 value for the high affinity binding site is 0.49 +/- 0.07 microM for human colon smooth muscle and 0.33 +/- 0.05 microM for rat colon smooth muscle. The apparent Ki2 for the low affinity binding site is 8.01 +/- 1.0 microM for human samples and 6.07 +/- 1.1 microM for rat samples. These values are close enough to suggest that the first subtype of muscarinic receptor may be considered cardiac (M2) and the second subtype glandular (M3). The relative densities of the receptor subtypes are significantly different for both species. Human colon samples show the major densities of subtype M2, 22.62 +/- 1.11 fmol/mg protein, this represents 75.66 +/- 3.73% of the total receptors.(ABSTRACT TRUNCATED AT 400 WORDS)

Characterization of muscarinic receptors in salivary and lacrimal glands of the rat.

The binding characteristics of muscarinic receptors in rat salivary and lacrimal glands were studied by means of radioligand binding techniques. In competition experiments against [3H]N-methylscopolamine, classical muscarinic antagonists ipratropium bromide, N-methylscopolamine and N-methylatropine exhibited very similar KD values in all the glands and their binding behavior was well described by a one binding site model (nH congruent to 1). The novel cardioselective antimuscarinic compound, AF-DX 116, displayed an equally low affinity in all the tissues examined. Pirenzepine and dicyclomine, two other selective muscarinic antagonists, showed a similar behaviour in all but the sublingual gland, where their binding profile indicated the presence of a heterogeneous receptor population (nH = 0.74 and 0.84, respectively). Histological studies of the sublingual-submandibular glandular complex demonstrated the presence of ganglionic structures mainly located in the hilum of the sublingual-submandibular glandular complex connected with the sublingual gland. Binding studies carried out with pirenzepine on the hilum and on a synaptosomal preparation from this region again revealed the presence of two populations of muscarinic receptors with KD values of 22-25 and 270-463 nM. These results are best explained by the presence of M1 and M2 receptors located on neuronal and glandular structures.

Characterization of muscarinic receptors in human submandibular salivary glands.

We studied the binding of the muscarinic antagonist [3H]N-methyl-scopolamine [( 3H]NMS) in order to characterize muscarinic receptors located in human submandibular salivary glands obtained from intrasurgical biopsy. [3H]NMS bound with a Kd value of 1.56 nM to a single class of muscarinic receptors (Bmax 37.3 fmol/mg protein) since pirenzepine exhibited a homogeneous binding profile.

Pharmacological characterization and distribution of muscarinic receptors in human placental syncytiotrophoblast brush-border and basal plasma membranes.

Based on the existence of choline acetyltransferase and acetylcholine in human placenta, we have investigated the presence of muscarinic acetylcholine receptors in brush-border and basal plasma membranes from human term placenta. Radioligand binding assay, using [3H]N-methyl-scopolamine as tracer, showed the existence of acetylcholine muscarinic receptors in brush-border (Kd 0.28 +/- 0.04 nM; Bmax 9.4 +/- 1.6 fmol/mg protein) and basal plasma membranes (Kd 0.24 +/- 0.05 nM; Bmax 34.3 +/- 6.3 fmol/mg protein). In order to perform a pharmacological characterization of these receptors, competition binding experiments were carried out using the muscarinic receptor antagonists pirenzepine, (11(2-diethyl-amino)methyl)-1-piperidinylacetyl-5-11-dihydro-6H-py rido(14) benzodiazepine (AF-DX 116), himbacine, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), dicyclomine and hexahydro-sila-difenidol (HHSD). The results obtained showed that the muscarinic receptors in brush-border and basal plasma membranes belong to different subtypes. In brush-border membranes, the receptor found match in terms of affinity for the antagonists with the muscarinic M1 receptor subtype (Ki pirenzepine, 13.6 +/- 8.2 nM; Ki AF-DX 116, 1680 +/- 271 nM; Ki himbacine, 212 +/- 6.5 nM; Ki 4-DAMP. 1.5 +/- 0.4 nM; Ki dicyclomine, 5.1 +/- 0.8 nM; Ki HHSD, 34.3 +/- 7.3 nM), whereas the receptor in basal plasma membrane seems to be of the muscarinic M2 receptor subtype (Ki pirenzepine, 202 +/- 48 nM; Ki AF-DX 116, 124 +/- 60 nM; Ki himbacine, 20.6 +/- 4.8 nM; Ki 4-DAMP, 4.5 +/- 1.2 nM; Ki dicyclomine, 54.6 +/- 22 nM; Ki HHSD, 89.2 +/- 15.8 nM). The results obtained show the existence of muscarinic acetylcholine receptors in brush-border and basal plasma membranes from human term placenta with a different distribution pattern in terms of number of receptors and distribution of different subtypes. The functional significance of these findings is as yet unknown, but these receptors probably mediate different functions as they belong to different subtypes and are coupled to different second messengers.

Characterization of muscarinic receptor subtypes in human tissues.

The affinities of selective, pirenzepine and AF-DX 116, and classical, N-methylscopolamine and atropine, muscarinic cholinergic receptor antagonists were investigated in displacement binding experiments with [3H]Pirenzepine and [3H]N-methylscopolamine in membranes from human autoptic tissues (forebrain, cerebellum, atria, ventricle and submaxillary salivary glands). Affinity estimates of N-methylscopolamine and atropine indicated a non-selective profile. Pirenzepine showed differentiation between the M1 neuronal receptor of the forebrain and the receptors in other tissues while AF-DX 116 clearly discriminated between muscarinic receptors of heart and glands. The results in human tissues confirm the previously described selectivity profiles of pirenzepine and AF-DX 116 in rat tissues. These findings thus reveal the presence also in man of three distinct muscarinic receptor subtypes: the neuronal M1, the cardiac M2 and the glandular M3.

Effect of S-adenosyl methionine on muscarinic receptors in young rats.

In the present study we have investigated the effect of a chronic administration of S-Adenosyl Methionine (SAM) on muscarinic receptor subtypes in young rat forebrain, cerebellum, heart and lacrimal gland. Saturation binding experiments were performed using 3H-N-methylscopolamine (3H-NMS) to label the total population of muscarinic receptors in plasma membranes from forebrain, cerebellum, heart and lacrimal gland. 3H-Pirenzepine (3H-Pz) was used to label the M1 subtype in plasma membranes from forebrain. The results obtained in cerebellum, heart and lacrimal gland show no changes in the affinity (Kd) nor in the number of receptors (Bmax) of the treated versus control groups. Saturation experiments in forebrain show an increase in the number of receptors of the treated versus control groups when using 3H-NMS (Bmax 2117 +/- 63 versus 1643 +/- 104 fmol/mg protein) without changes in the affinity. Saturation experiments with 3H-Pz, show an increase in the number of M1 receptors in the treated group with no changes in the affinity (Bmax 421 +/- 16 versus 225 +/- 19 fmol/mg protein). From our results, we conclude that SAM increase the number of receptors in forebrain and this increase is mainly due to changes in the number of M1 receptor subtypes.

Differences in the effects of extended-release aspirin and plain-formulated aspirin on prostanoids and nitric oxide in healthy volunteers.

This study was designed to evaluate the effects of extended-release aspirin on platelet aggregation and the production of prostanoids and nitric oxide. The participants in this double blind, randomized and crossover study were 20 healthy volunteers. Interventions were 150 mg of plain-formulated aspirin (PFASA) and 150 mg of extended-release aspirin (ERASA). Blood samples were collected before and 10, 20, 60, 120, 240, 480 and 1440 min after the first dose; 3, 7 and 14 days after daily administration and 24 h after the last dose. The main measures were platelet aggregometry, thromboxane B2, 6-keto-prostaglandin (PG) F1alpha and nitric oxide in each control. Platelet aggregation was inhibited by 50% with ERASA, and by 77% with PFASA. No differences were found in chronic treatment. Thromboxane B2 was inhibited more by the latter (51-67%), but 90% inhibition was observed in both groups after 3 days. The levels of 6-keto-PGF1alpha was reduced by 20% with ERASA and by 58% with PFASA. Nitric oxide production increased in both groups, but after 24 h, and 7-14 days, elevated concentrations of nitric oxide were found only in the ERASA. The antiplatelet effects of ERASA provide pharmacological advantages (greater prostacyclin synthesis and prolonged increase in nitric oxide production) over those provided by the plain formulation.

Diagnostic usefulness of percutaneous liver biopsy in HIV-infected patients with fever of unknown origin.

OBJECTIVES: to determine the value of percutaneous liver biopsy (PLB) in the diagnosis of fever of unknown origin (FUO) in HIV-infected patients and establish a prediction model for its usefulness to enable diagnosis of FUO in these patients to be standardized. METHODS: a total of 58 HIV-infected patients who underwent PLB for the evaluation of FUO were studied at 'Carlos Haya' Hospital in Malaga, Spain. The patients were classified into three groups, according to the results of the PLB: (a) diagnostic PLB (when a definitive diagnosis was obtained); (b) helpful PLB (the tissue sample showed suggestive, but not definitive, findings); and (c) normal or non-specific PLB (no contribution to diagnosis, the findings being normal or irrelevant). Multivariate analysis was made to establish a prediction model for the diagnostic usefulness of PLB, calculating the positive (PPV) and negative (NPV) predictive values. RESULTS: PLB was carried out in 58 HIV-infected patients during diagnosis of FUO. Risk factors for HIV infection included intravenous drug use (72.4%), homosexual or bisexual activities (12.1%), and heterosexual transmission (15.5%). Fifty-two out of 58 patients (89.6%) had previous AIDS-defining illnesses. The mean CD4 lymphocyte count +/-SD was 56.4+/-80.9/mm3. The mean duration of fever was 43 days. Diagnosis could be established in 51 (87.9%) patients, with tuberculosis (50%) and leishmaniasis (20.7%) being the most common. The PLB was diagnostic in 25 cases (43.1%), helpful in 13 (22.4%), and normal or non-specific in the remaining 20 (34.5%). Biopsy-associated complications occurred in two cases. The presence of hepatomegaly or splenomegaly were the most useful factors in predicting the usefulness of the PLB, with a PPV of 86.1% and NPV of 68.2%. In patients with tuberculosis, an increased alkaline phosphatase and hepatomegaly had a PPV of 86.4% and a NPV of 71.4%. CONCLUSIONS: PLB is a useful technique for the diagnosis of FUO in HIV-infected persons. Early PLB should be considered in those patients with hepatosplenomegaly and increased alkaline phosphatase levels.

Circadian rhythm in muscarinic receptor subtypes in rat forebrain.

The binding of different ligands to muscarinic receptors in the central nervous system is regulated by several factors. Among these are the administration of drugs, disease, ontogeny or aging. Studies carried out in rat brains have demonstrated changes in the density of the muscarinic receptors at different times of the day. These changes might be related to variations in the circadian rhythms. In this work we have studied the binding of the [3H]-N-methyl-escopolamine, the agonist carbachol and the antagonist pirenzepine to muscarinic receptors in rat forebrains at 10.00, 14.00, 18.00, 22.00, 02.00 and 06.00 hr. We have observed changes in the density of muscarinic receptors but not changes in affinity to the radioligand. The Bmax values obtained by saturation studies were maximum at 14.00 hr and minimum at 02.00 hr (P less than 0.05 Mann-Whitney's test). Inhibition studies in the presence of the non-selective agonist carbachol and the selective antagonist pirenzepine, at the same time-points, did not show statistically significant changes in the Bmax values. These data indicate that changes in the Bmax values are only observed in the total population of muscarinic receptors and are not due to modifications in the subtypes of muscarinic receptors nor to the different affinity states of agonist binding.

M1 and M3 muscarinic receptor subtypes in rat forebrain.

At least three pharmacologically different muscarinic receptor subtypes (M1, M2 and M3) have been identified in rat brain. While M1 and M2 subtypes can be directly labelled by selective ligands (3H-pirenzepine and 3H-AF-DX 116, respectively), there are no truly selective ligands for the M3 subtype. In the present study, we have investigated a possible method of studying the pharmacological M3 subtype in rat forebrain using the non-selective labelled antagonist 3H-N-methyl-scopolamine (3H-NMS) in the presence of unlabelled pirenzepine to protect the M1 subtype. The results obtained in kinetic experiments using 3H-NMS in presence of 30.10(-9) M unlabelled pirenzepine (Kon 1.2.10(-8) M-1 m-1, Koff 4.7.10(-2) m-1 and Kd 0.4.10(-9) M) are compatible with the studies carried out in rat pancreatic islets and submaxillary gland which contain predominantly the M3 subtype. We have also performed inhibition experiments with the selective antagonist AF-DX 116. Due to the small proportion of M2 receptors present in rat forebrain, this drug is able to discriminate between M1 and non M1 non M2 receptor subtypes in competition experiments with 3H-NMS versus AF-DX 116 (Ki values 0.28.10(-6) M and 4.3.10(-6) M, respectively). When the competition experiments were performed using 3H-NMS in presence of 30.10(-9) M unlabelled pirenzepine, the Ki value obtained was 3.8.10(-6) M, very close to the value obtained for the non M1 non M2 receptor in competition experiments with 3H-NMS versus AF-DX 116 and in excellent agreement with the affinity of this drug for the glandular M3 subtype. All these data suggest that the approach using the non-selective antagonist 3H-N-methyl-scopolamine in presence of unlabelled pirenzepine allows the study of the pharmacological M3 subtype in rat forebrain.

Influence of dietary factors on calcium bioavailability: a brief review.

There are several factors that affected calcium bioavailability, such as physiological and dietary factors. These dietary factors help to achieve an appropiate status of calcium for a correct bone mineralization. In this pathway, recently some compounds present in milk that seem improve calcium absorption such as lactose and certain caseinophosphopeptides formed during digestion of caseins have been studied. On the other hand, the possible inhibitatory effect of fiber has been also studied, without conclusive results between in vitro and in vivo studies and the role of phytic acid on impairs calcium bioavailability could be prevented by using fructo-oligosaccharides, which cannot be digested in the small intestine and arrive practically intact to the colon, where are fermented. Finally, calcium fortification must be executed by suitable compounds with high bioavailability, better technological properties, and a correct calcium:phosphorus ratio. For that reason, the objective of the present article is to review the influence of all these conditional factors on calcium bioavailability.

[Generic drugs: good or bad? Physician's knowledge of generic drugs and prescribing habits]. / Genéricos: ¿buenos o malos? Conocimientos y actitudes de los médicos ante los medicamentos genéricos.

In this article we analyze the responses of 1220 Spanish physicians who participated in a survery about generic drugs. A previously validated questionnaire was sent to physicians through the Spanish Medical Councils of the different provinces. Four items were analyzed: what doctors know about generic drugs (knowledge); physicians' prescribing habits concerning these drugs (attitude and professional competence); how prescription of generic drugs effects pharmaceutical costs amd, finally, what doctors believe a generic drug should be. The influence of physician-related variables (age, type of contract, specialty, workload, etc.) on prescribing of generic drugs was also analyzed. In view of the results, we believe that to rationalize expenditure through and appropriate policy on generic drugs Spanish health authorities should offer more and better training and information (clear and independent) about what generic drugs are.