Margarida Caramona

Improving the well-being of elderly patients via community pharmacy-based provision of pharmaceutical care: a multicentre study in seven European countries.

ObjectiveThis study aimed to measure the outcomes of a harmonised, structured pharmaceutical care programme provided to elderly patients (≥65 years of age) by community pharmacists in a multicentre international study performed in 7 European countries.Design and settingThe study was a randomised, controlled, longitudinal, clinical trial with repeated measures performed over an 18-month period. A total of 104 intervention and 86 control pharmacy sites participated in the research and 1290 intervention patients and 1164 control patients were recruited into the study.Main outcome measures and resultsA general decline in health-related quality of life over time was observed in the pooled data; however, significant improvements were achieved in patients involved in the pharmaceutical care programme in some countries. Intervention patients reported better control of their medical conditions as a result of the study and cost savings associated with pharmaceutical care provision were observed in most countries. The new structured service was well accepted by intervention patients and patient satisfaction with the services improved during the study. The pharmacists involved in providing pharmaceutical care had a positive opinion on the new approach, as did the majority of general practitioners surveyed. The positive effects appear to have been achieved via social and psychosocial aspects of the intervention, such as the increased support provided by community pharmacists, rather than via biomedical mechanisms.ConclusionsThis study is the first large-scale, multicentre study to investigate the effects of pharmaceutical care provision by community pharmacists to elderly patients. Future research methodology and implementation will be informed by the experience gained from this challenging trial.

Differential roles of hydrogen peroxide and superoxide in mediating IL-1-induced NF-KB activation and iNOS expression in bovine articular chondrocytes

Our previous studies showed that reactive oxygen species (ROS) are required for the pro‐inflammatory cytokine interleukin‐1β (IL‐1) to induce the activity of the Nuclear transcription Factor‐kappaB (NF‐κB) and the expression of the inducible isoform of the nitric oxide synthase (iNOS) in bovine articular chondrocytes. This study aimed at elucidating the role of hydrogen peroxide (H2O2) and the superoxide radical, two major ROS, in mediating those IL‐1‐induced responses. The results obtained show that chondrocytes produce both H2O2 and superoxide radical in response to IL‐1. Treatment of the chondrocyte cultures with H2O2 alone did not induce NF‐κB activation or iNOS expression. Addition of H2O2 simultaneously with IL‐1 did neither enhance nor inhibit NF‐κB activation and iNOS expression, relatively to treatment with IL‐1 alone. Accordingly, treatment with catalase did not inhibit those IL‐1‐induced responses. Treatment with superoxide dismutase, however, effectively prevented IL‐1‐induced IκB‐α degradation and iNOS expression. Taken together, the results obtained indicate that superoxide mediates IL‐1‐induced IκB‐α degradation and the consequent NF‐κB activation and iNOS expression in chondrocytes, whereas H2O2 does not seem to participate in those IL‐1‐induced responses. In conclusion, the present study identifies the superoxide radical as the ROS involved in mediating the IL‐1‐induced signaling pathway that leads to NF‐κB activation and to the expression of NF‐κB‐dependent genes in bovine articular chondrocytes.

Lamotrigine analysis in blood and brain by high-performance liquid chromatography

A reversed-phase high-performance liquid chromatography assay was developed and validated to determine plasma and brain lamotrigine concentrations allowing pharmacokinetic-pharmacodynamic studies of this new antiepileptic drug in patients and laboratory animals. Lamotrigine and its internal standard were extracted, under alkaline conditions, from plasma and brain homogenate, into ethyl acetate; brain proteins were previously precipitated with trichloroacetic acid. The method was linear between 0.1 and 15.0 mg/l for plasma, with a detection limit of 0.008 mg/l, and between 0.1 and 5.0 mg/l for brain homogenate, with a detection limit of 0.023 mg/l. The method proved to be simple, useful and appropriate, not only for clinical and experimental research, but also for routine monitoring of lamotrigine concentrations in patients.

Diphenyleneiodonium inhibits NF-kappaB activation and iNOS expression induced by IL-1beta: involvement of reactive oxygen species

AIMS: In this work, we studied the mechanisms by which diphenyleneiodonium chloride (DPI) inhibits nitric oxide (NO) synthesis induced by the proinflammatory cytokine interleukin-1beta (IL-1) in bovine articular chondrocytes. To achieve this, we evaluated the ability of DPI to inhibit the expression and activity of the inducible isoform of the NO synthase (iNOS) induced by IL-1. We also studied the ability of DPI to prevent IL-1-induced NF-kappaB activation and reactive oxygen species (ROS) production. RESULTS: Northern and Western blot analysis, respectively, showed that DPI dose-dependently inhibited IL-1-induced iNOS mRNA and protein synthesis in primary cultures of bovine articular chondrocytes. DPI effectively inhibited NO production (IC50=0.03+/-0.004 microM), as evaluated by the method of Griess. Nuclear factor-kappa B (NF-kappaB) activation, as evaluated by electrophoretic mobility shift assay, was inhibited by DPI (1-10 microM) in a dose-dependent manner. IL-1-induced ROS production, as evaluated by measurement of dichlorofluorescein fluorescence, was inhibited by DPI at concentrations that also prevented NF-kappaB activation and iNOS expression. CONCLUSIONS: DPI inhibits IL-1-induced NO production in chondrocytes by two distinct mechanisms: (i) by inhibiting NOS activity, and (ii) by preventing iNOS expression through the blockade of NF-kappaB activation. These results also support the involvement of reactive oxygen species in IL-1-induced NF-kappaB activation and expression of NF-kappaB-dependent genes, such as iNOS.

Intra- and extraneuronal metabolism of 5-hydroxytryptamine in the isolated saphenous vein of the dog

SummaryThe uptake and metabolism of 3H-5-hydroxytryptamine (5-HT) in the isolated saphenous vein of the dog was studied, as well as the sensitivity to 5-HT and its termination of action (as determined by oil immersion experiments).After exposure of the vein to 0.23 μmol/15-HT for 30 min, about one half of the 5-HT removed was accumulated in the tissue, the other half appearing in the form of metabolites. Cocaine, chlorimipramine or denervation reduced the accumulation, but not the metabolism, of 5-HT; hydrocortisone reduced both accumulation and metabolism. Hydrocortisone plus cocaine (or phenoxybenzamine alone) caused a greater reduction of accumulation and metabolism than hydrocortisone or cocaine alone. Pargyline caused an increase in accumulation and virtually abolished metabolism; amezinium also abolished metabolism, but reduced accumulation (the same effects as caused by cocaine plus pargyline).Cocaine (12 μmol/l) caused supersensitivity to 5-HT and a prolongation of the half-time for relaxation in oil; hydrocortisone (40 μmol/l) had no effect on sensitivity, but prolonged somewhat the half-time for relaxation in oil, showing supra-additive effects with cocaine.5-HT, taken up into adrenergic nerve terminals, was released by electrical stimulation; no appreciable reduction in fractional release occurred during repeated stimulations. Exposure to cocaine prior to incubation with 5-HT prevented the neuronal uptake of 5-HT.Compartmental analysis, done with the help of washout curves of strips pretreated with pargyline and loaded with 3H-5-HT, shows that 3H-5-HT distributed into 3 compartments plus a bound fraction. The latter (and probably compartment III) is of neuronal nature; compartments II and I appear to be of extraneuronal and extracellular nature, respectively.Strips from animals pretreated with reserpine and exposed to pargyline and cocaine were used for the study of the kinetics of the extraneuronal uptake of 5-HT. A saturable and a non-saturable component were found to be present. The saturable component of this uptake had a Km of 391 μmol/l and a Vmax of 45 nmol · g−1 · min−1; the k value of the non-saturable component was 0.022 · min−1.It is concluded that 5-HT behaves in a very similar way to biogenic catecholamines: it is taken up into adrenergic nerve terminals by a cocaine-sensitive mechanism and partially incorporated into vesicles, from where it may be released by electrical stimulation, thus behaving as a co-transmitter; it is also subject to uptake into extraneuronal cells, through corticosteroid-sensitive and corticosteroid-resistant mechanisms. After uptake of any kind, metabolism by monoamine oxidase of type A may occur.

The effects of chemical sympathectomy on dopamine, noradrenaline and adrenaline content in some peripheral tissues.

1 Dopamine, noradrenaline (NA) and adrenaline (Ad) depletion by 6‐hydroxydopamine (6‐OHDA) and pargyline plus 6‐OHDA was investigated in the cat left ventricle, mesenteric and renal arteries, renal cortex, renal medulla and adrenal medulla. Catecholamine concentrations in plasma were also analyzed in these two experimental conditions. 2 6‐OHDA alone or in combination with pargyline induced parallel decreases of NA and dopamine contents in the left ventricle. In the main trunk and proximal branches of the mesenteric artery and renal artery 6‐OHDA selectively reduced NA without a parallel decrease in dopamine content. Previous treatment with pargyline abolished this selectivity. 3 In the kidney of control animals, dopamine content was greater than could be attributed to its presence only in noradrenergic neurones. In the renal cortex 6‐OHDA reduced significantly dopamine and NA contents, and in the renal medulla only NA levels were decreased by this drug. Pargyline plus 6‐OHDA did not deplete the NA content either in the renal cortex or in the renal medulla, and only reduced significantly the dopamine content in the renal cortex. 4 NA concentrations in plasma were increased by pargyline plus 6‐OHDA whilst Ad remained unaffected. In the adrenal medulla only NA content was reduced either by 6‐OHDA or pargyline plus 6‐OHDA. 5 The present findings suggest a NA‐independent dopamine pool in both segments of the mesenteric artery and renal artery but not in the left ventricle.

Effects of Methylene Blue on the Uptake, Release and Metabolism of Noradrenaline in Mesenteric Arterial Vessels

Abstract— Methylene blue (3, 10 and 30 μM) increased the spontaneous outflow of endogenous dopamine and noradrenaline from sympathetic nerves supplying the dog mesenteric artery and drastically reduced the formation of endogenous dihydroxyphenylglycol (DOPEG). In addition, it decreased the accumulation of [3H]noradrenaline in the tissue, reduced the formation of [3H]DOPEG and [3H]normetanephrine, without altering the formation of [3H]dihydroxymandelic acid. In tissue homogenates of the same blood vessel, methylene blue 30 and 100 μM produced a significant reduction in the deamination of 5‐hydroxytryptamine (5‐HT), β‐phenylethylamine (β‐PEA) and tyramine. Methylene blue increased the accumulation of [3H]isoprenaline in the tissue, and markedly reduced the formation of [3H]O‐methylisoprenaline ([3H]OMI). These results show that methylene blue alters the storage and disposition of the adrenergic transmitter.

Evidence for an extraneuronal location of monoamine oxidase in renal tissues

Summary(1) Homogenates of renal cortex and renal medulla of control and 6-hydroxydopamine-denervated cat kidneys were prepared. (2) Monoamine oxidase (MAO) activity was determined with [3H]-5-hydroxytryptamine ([3H]-5HT) and [14C]-β-phenylethylamine ([14C]-β-PEA) as preferential substrates for MAO-A and MAO-B, respectively. (3) The endogenous dopamine and noradrenaline tissue contents of control and chemicallydenervated kidneys were compared with the MAO activities. (4) The results show that a 70% depletion of monoamine content by chemical denervation resulted only in a 23% reduction of MAO-A activity in the renal cortex, whereas MAO-13 was unaffected either in the cortical or the medullary zones; in the renal medulla MAO-A activity was not changed by denervation. Most of the MAO activity in the cat kidney is of the B type (74%) and is located in the renal cortex.

Complexos de inclusão de indometacina com hidroxipropil-beta-ciclodextrina: estudos de dissolução e coeficiente de partição

Indomethacin, a non steroidal anti-inflamatory, is practically water insoluble. Hydroxypropyl-b-cyclodextrin grants better solubility characteristics to included drugs. Indomethacins complexation protects from hydrolysis, enhancing solubility and dissolution. The aim is to study the influence of complexation methods, freeze and spray-dryer, on indomethacin dissolution and partition coefficient. Dissolution results show that freeze-dried inclusion complex has a better time needed to dissolve 50% and 90% - 1 and 1.8 minutes then other products. The same happens with percentage of indomethacin dissolved at 5, 30 and 60 minutes - 92.6±1.8%; 98.9±1.2% e 100.01±0.1% and also with dissolution efficiency and dissolution profile. Partition coefficient results with the complexes obtained by both methods corroborate the theory that there are several intervenient strengths on this process and not only drugs free fraction that regulates transport to organic phase, reinforcing the environmental pH significance. With phosphate buffer pH 7.0, variations on transport grade by cyclodextrin addition are very small, showing no significant changes on log P*. With phosphate buffer pH 5.5, although not significant, variations are slightly higher. We can conclude that complexation enhances dissolution capacities of lipophylic drugs without changing the characteristics that give them a good ability for membrane diffusion.

Factores associados à hipertensão arterial nos utentes de farmácias em Portugal

OBJETIVO: Estimar a prevalencia, tratamento e controlo da hipertensao e identificar factores associados em utentes de farmacias comunitarias. METODOS: Estudo transversal com 1.042 utentes de 40 a 65 anos em 60 farmacias comunitarias de Portugal Continental entre outubro de 2005 e janeiro de 2006. Os dados foram obtidos pela aplicacao de questionario e medicao de parâmetros biologicos. Foram realizadas tres regressoes logisticas sequenciais para verificar associacao entre as variaveis. RESULTADOS: A idade media foi de 53,7 anos e a razao homem/mulher foi 0,68. A prevalencia da hipertensao arterial foi de 54,8%. Cerca de 70% dos hipertensos encontravam-se sob tratamento anti-hipertensivo e, destes, 47,7% estavam controlados. A hipertensao esteve positivamente associada a idade mais elevada, sexo masculino, ser casado, apresentar indice de massa corporal e nivel de colesterol total mais alto, ser diabetico, ter doenca cardiovascular pessoal ou familiar precoce e reportar mais consultas medicas por ano. A hipertensao tratada mostrou-se positivamente associada a ser mulher, nao casado, ser diabetico, viver numa area urbana e reportar mais de tres consultas medicas por ano. Nos hipertensos tratados, estar controlado foi positivamente associado a ter comportamento aderente a terapeutica anti-hipertensiva (auto-reporte), percepcionar o efeito desta medicacao e ser de baixo risco cardiovascular. Os modelos preditivos apresentaram areas sob as respectivas curvas ROC entre 0,72 e 0,78, com capacidade discriminatoria aceitavel. CONCLUSOES: A prevalencia da hipertensao foi elevada, mas similar a encontrada em outros estudos realizados em Portugal. A proporcao de doentes tratados foi satisfatoria, em contraste com o nivel insuficiente de controlo.OBJECTIVE To estimate the prevalence, treatment and control of hypertension, and to identify factors associated in community pharmacy users. METHODS A cross-sectional study was conducted with 1,042 pharmacy users, aged between 40 and 65 years, in 60 community pharmacies of continental Portugal, between October 2005 and January 2006. Data were obtained with the application of a questionnaire and measurement of biological parameters. A total of three sequential logistic regressions were performed to verify an association among variables. RESULTS Mean age was 53.7 years and the male/female ratio was 0.68. Prevalence of arterial hypertension was 54.8%. Approximately 70% of hypertensive individuals were undergoing antihypertensive treatment and, of these, 47.7% were controlled. Hypertension was positively associated with older age, male sex, being married, higher body mass index and higher total cholesterol level, being a diabetic, having a family or personal history of premature cardiovascular disease, and reporting more medical visits per year. When treated, hypertension was found to be positively associated with the female sex, not being married, being a diabetic, living in an urban area, and reporting more than three medical visits per year. In hypertensive users who were treated, being controlled was positively associated with self-reporting adherent behavior towards antihypertensive treatment, perceiving the effect of these drugs and having a low cardiovascular risk. The predictive models showed areas under the respective ROC curves between 0.72 and 0.78, with an acceptable discriminatory power. CONCLUSIONS The prevalence of hypertension was high, although similar to that found in other studies conducted in Portugal. The proportion of hypertensive individuals under treatment was satisfactory, in contrast to an insufficient level of control.