Danger signaling through the inflammasome acts as a master switch between tolerance and sensitization.

Efficient priming of adaptive immunity depends on danger signals provided by innate immune pathways. As an example, inflammasome-mediated activation of caspase-1 and IL-1beta is crucial for the development of reactive T cells targeting sensitizers like dinitrofluorobenzene (DNFB). Surprisingly, DNFB and dinitrothiocyanobenzene provide cross-reactive Ags yet drive opposing, sensitizing vs tolerizing, T cell responses. In this study, we show that, in mice, inflammasome-signaling levels can be modulated to turn dinitrothiocyanobenzene into a sensitizer and DNFB into a tolerizer, and that it correlates with the IL-6 and IL-12 secretion levels, affecting Th1, Th17, and regulatory T cell development. Hence, our data provide the first evidence that the inflammasome can define the type of adaptive immune response elicited by an Ag, and hint at new strategies to modulate T cell responses in vivo.

Constitutively active alpha-1b adrenergic receptor mutants display different phosphorylation and internalization features.

We compared the phosphorylation and internalization properties of constitutively active alpha-1b adrenergic receptor (AR) mutants carrying mutations in two distant receptor domains, i.e., at A293 in the distal part of the third intracellular loop and at D142 of the DRY motif lying at the end of the third transmembrane domain. For the A293E and A293I mutants the levels of agonist-independent phosphorylation were 150% and 50% higher than those of the wild-type alpha-1b AR, respectively. On the other hand, for the constitutively active D142A and D142T mutants, the basal levels of phosphorylation were similar to those of the wild-type alpha-1b AR and did not appear to be further stimulated by epinephrine. Overexpression of the guanyl nucleotide binding regulatory protein-coupled receptor kinase GRK2 further increases the basal phosphorylation of the A293E mutant, but not that of D142A mutant. Both the wild-type alpha-1b AR and the A293E mutant could undergo beta-arrestin-mediated internalization. The epinephrine-induced internalization of the constitutively active A293E mutant was significantly higher than that of the wild-type alpha-1b AR. In contrast, the D142A mutant was impaired in its ability to interact with beta-arrestin and to undergo agonist-induced internalization. Interestingly, a double mutant A293E/D142A retained very high constitutive activity and regulatory properties of both the A293E and D142A receptors. These findings demonstrate that two constitutively activating mutations occurring in distant receptor domains of the alpha-1b AR have divergent effects on the regulatory properties of the receptor.

In vitro and in vivo experimental models for drug screening and development for Chagas disease

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

Exposure to Trihalomethanes through Different Water Uses and Birth Weight, Small for Gestational Age, and Preterm Delivery in Spain

BACKGROUND Evidence associating exposure to water disinfection by-products with reduced birth weight and altered duration of gestation remains inconclusive. OBJECTIVE We assessed exposure to trihalomethanes (THMs) during pregnancy through different water uses and evaluated the association with birth weight, small for gestational age (SGA), low birth weight (LBW), and preterm delivery. METHODS Mother-child cohorts set up in five Spanish areas during the years 2000-2008 contributed data on water ingestion, showering, bathing, and swimming in pools. We ascertained residential THM levels during pregnancy periods through ad hoc sampling campaigns (828 measurements) and regulatory data (264 measurements), which were modeled and combined with personal water use and uptake factors to estimate personal uptake. We defined outcomes following standard definitions and included 2,158 newborns in the analysis. RESULTS Median residential THM ranged from 5.9 μg/L (Valencia) to 114.7 μg/L (Sabadell), and speciation differed across areas. We estimated that 89% of residential chloroform and 96% of brominated THM uptakes were from showering/bathing. The estimated change of birth weight for a 10% increase in residential uptake was -0.45 g (95% confidence interval: -1.36, 0.45 g) for chloroform and 0.16 g (-1.38, 1.70 g) for brominated THMs. Overall, THMs were not associated with SGA, LBW, or preterm delivery. CONCLUSIONS Despite the high THM levels in some areas and the extensive exposure assessment, results suggest that residential THM exposure during pregnancy driven by inhalation and dermal contact routes is not associated with birth weight, SGA, LBW, or preterm delivery in Spain.

PAX6 aniridia and interhemispheric brain anomalies.

PURPOSE: To report the clinical and genetic study of patients with autosomal dominant aniridia. METHODS: We studied ten patients with aniridia from three families of Egyptian origin. All patients underwent full ophthalmologic, general and neurological examination, and blood drawing. Cerebral magnetic resonance imaging was performed in the index case of each family. Genomic DNA was prepared from venous leukocytes, and direct sequencing of all the exons and intron-exon junctions of the Paired Box gene 6 (PAX6) was performed after PCR amplification. Phenotype description, including ophthalmic and cerebral anomalies, mutation detection in PAX6 and phenotype-genotype correlation was acquired. RESULTS: Common features observed in the three families included absence of iris tissue, corneal pannus with different degrees of severity, and foveal hypoplasia with severely reduced visual acuity. In Families 2 and 3, additional findings, such as lens dislocation, lens opacities or polar cataract, and glaucoma, were observed. We identified two novel (c.170-174delTGGGC [p.L57fs17] and c.475delC [p.R159fs47]) and one known (c.718C>T [p.R240X]) PAX6 mutations in the affected members of the three families. Systemic and neurological examination was normal in all ten affected patients. Cerebral magnetic resonance imaging showed absence of the pineal gland in all three index patients. Severe hypoplasia of the brain anterior commissure was associated with the p.L57fs17 mutation, absence of the posterior commissure with p.R159fs47, and optic chiasma atrophy and almost complete agenesis of the corpus callosum with p.R240X. CONCLUSIONS: We identified two novel PAX6 mutations in families with severe aniridia. In addition to common phenotype of aniridia and despite normal neurological examination, absence of the pineal gland and interhemispheric brain anomalies were observed in all three index patients. The heterogeneity of PAX6 mutations and brain anomalies are highlighted. This report emphasizes the association between aniridia and brain anomalies with or without functional impact, such as neurodevelopment delay or auditory dysfunction.

Regulation of free corticosterone and CBG capacity under different environmental conditions in altricial nestlings.

The concentration of circulating glucocorticoids is regulated in response to environmental and endogenous conditions. Total circulating corticosterone, the main glucocorticoid in birds, consists of a fraction which is bound to corticosterone-binding globulins (CBG) and a free fraction. There is increasing evidence that the environment modulates free corticosterone levels through varying the concentration of CBG, but experimental evidence is lacking. To test the hypothesis that the regulation of chronic stress in response to endogenous and environmental conditions involves variation in both corticosterone release and CBG capacity, we performed an experiment with barn owl (Tyto alba) nestlings in two different years with pronounced differences in environmental conditions and in nestlings experimentally fed ad libitum. In half of the individuals we implanted a corticosterone-releasing pellet to artificially increase corticosterone levels and in the other half we implanted a placebo pellet. We then repeatedly collected blood samples to measure the change in total and free corticosterone levels as well as CBG capacity. The increase in circulating total corticosterone after artificial corticosterone administration varied with environmental conditions and with the food regime of the nestlings. The highest total corticosterone levels were found in nestlings growing up in poor environmental conditions and the lowest in ad libitum fed nestlings. CBG was highest in the year with poor environmental conditions, so that, contrary to total corticosterone, free corticosterone levels were low under poor environmental conditions. When nestlings were fed ad libitum total corticosterone, CBG and free corticosterone did not increase when administering corticosterone. These results suggest that depending on the individual history an animal experienced during development the HPA-axis is regulated differently.

Orexin/hypocretin (Orx/Hcrt) transmission and drug-seeking behavior: is the paraventricular nucleus of the thalamus (PVT) part of the drug seeking circuitry?

The orexin/hypocretin (Orx/Hcrt) system has long been considered to regulate a wide range of physiological processes, including feeding, energy metabolism, and arousal. More recently, concordant observations have demonstrated an important role for these peptides in the reinforcing properties of most drugs of abuse. Orx/Hcrt neurons arise in the lateral hypothalamus (LH) and project to all brain structures implicated in the regulation of arousal, stress, and reward. Although Orx/Hcrt neurons have been shown to massively project to the paraventricular nucleus of the thalamus (PVT), only recent evidence suggested that the PVT may be a key relay of Orx/Hcrt-coded reward-related communication between the LH and both the ventral and dorsal striatum. While this thalamic region was not thought to be part of the "drug addiction circuitry," an increasing amount of evidence demonstrated that the PVT-particularly PVT Orx/Hcrt transmission-was implicated in the modulation of reward function in general and several aspects of drug-directed behaviors in particular. The present review discusses recent findings that suggest that maladaptive recruitment of PVT Orx/Hcrt signaling by drugs of abuse may promote persistent compulsive drug-seeking behavior following a period of protracted abstinence and as such may represent a relevant target for understanding the long-term vulnerability to drug relapse after withdrawal.

Targeting anticancer therapy : clinical and preclinical approaches

AbstractCancer treatment has shifted from cytotoxic and nonspecific chemotherapy to chronic treatment with targeted molecular therapies. These new classes of drugs directed against cancer-specific molecules and signaling pathways, act at a particular level of the tumor cell development. However, in both types of therapeutic approaches (standard cytotoxic chemotherapy and targeted signal transduction inhibitions), toxicity and side effects can occur. The aim of this thesis was to investigate various approaches to improve the activity and tolerability of cancer treatment, in a clinical setting, a) by molecular targeting through the use of tyrosine kinase inhibitors (TKIs), whose dosage can be adapted to each patient according to plasma levels, and, b) in a preclinical model, by tissue targeting with locoregional administration of cytotoxic chemotherapy to increase drug exposure in the target tissue while reducing systemic toxicity of the treatment.A comprehensive program for the Therapeutic Drug Monitoring (TDM) of the new class of targeted anticancer drugs of TKIs in patient's blood has been therefore initiated comprising the setting up, validation and clinical application of a multiplex assay by liquid chromatography coupled to tandem mass spectrometry of TKIs in plasma from cancer patients. Information on drugs exposure may be clinically useful for an optimal follow-up of patients' anticancer treatment, especially in case of less than optimal clinical response, occurrence of adverse drug reaction effects and the numerous risks of drug-drug interactions. In this context, better knowledge of the potential drug interactions between TKIs and widely prescribed co- medications is of critical importance for clinicians, to improve their daily care of cancer patients. For one of the first TKI imatinib, TDM interpretation is nowadays based on total plasma concentrations but, only the unbound (free) form is likely to enter cell to exert its pharmacological action. Pharmacokinetic analysis of the total and free plasma level of imatinib measured simultaneously in patients have allowed to refine and validate a population pharmacokinetic model integrating factors influencing in patients the exposure of pharmacological active species. The equation developed from this model may be used for extrapolating free imatinib plasma concentration based on the total plasma levels that are currently measured in TDM from patients. Finally, the specific influence of Pglycoprotein on the intracellular disposition of TKIs has been studies in cell systems using the siRNA silencing approach.Another approach to enhance the selectivity of anticancer treatment may be achieved by the loco-regional administration of a cytostatic agent to the target organ while sparing non- affected tissues. Isolated lung perfusion (ILP) was designed for the treatment of loco-regional malignancies of the lung but clinical results have been so far disappointing. It has been shown in a preclinical model in rats that ILP with the cytotoxic agent doxorubicin alone allows a high drug uptake in lung tissue, and a low systemic toxicity, but was characterized by a high spatial tissular heterogeneity in drug exposure and doxorubicin uptake in tumor was comparatively smaller than in normal lung tissue. Photodynamic therapy (PDT) is a new approach for the treatment of superficial tumors, and implies the application of a sensitizer activated by a laser light at a specific wavelength, that disrupts endothelial barrier of tumor vessels to increase locally the distribution of cytostatics into the tumor tissue. PDT pre-treatment before intravenous administration of liposomal doxorubicin was indeed shown to selectively increase drug uptake in tumors in a rat model of sarcoma tumors to the lung.RésuméLe traitement de certains cancers s'est progressivement transformé et est passé de la chimiothérapie, cytotoxique et non spécifique, au traitement chronique des patients avec des thérapies moléculaires ciblées. Ces médicaments ont une action ciblée en interférant à un niveau spécifique du développement de la cellule tumorale. Dans les deux types d'approches thérapeutiques (chimiothérapie cytotoxique et traitements ciblés), on est confronté à la présence de toxicité et aux effets secondaires du traitement anticancéreux. Le but de cette thèse a donc été d'étudier diverses approches visant à améliorer l'efficacité et la tolérabilité du traitement anticancéreux, a) dans le cadre d'une recherche clinique, par le ciblage moléculaire grâce aux inhibiteurs de tyrosines kinases (TKIs) dont la posologie est adaptée à chaque patient, et b) dans un modèle préclinique, par le ciblage tissulaire grâce à l'administration locorégionale de chimiothérapie cytotoxique, afin d'augmenter l'exposition dans le tissu cible et de réduire la toxicité systémique du traitement.Un programme de recherche sur le suivi thérapeutique (Therapeutic Drug Monitoring, TDM) des inhibiteurs de tyrosine kinases a été ainsi mis en place et a impliqué le développement, la validation et l'application clinique d'une méthode multiplex par chromatographie liquide couplée à la spectrométrie de masse en tandem des TKIs chez les patients souffrant de cancer. L'information fournie par le TDM sur l'exposition des patients aux traitements ciblés est cliniquement utile et est susceptible d'optimiser la dose administrée, notamment dans les cas où la réponse clinique au traitement des patients est sous-optimale, en présence d'effets secondaires du traitement ciblé, ou lorsque des risques d'interactions médicamenteuses sont suspectés. Dans ce contexte, l'étude des interactions entre les TKIs et les co-médications couramment associées est utile pour les cliniciens en charge d'améliorer au jour le jour la prise en charge du traitement anticancéreux. Pour le premier TKI imatinib, l'interprétation TDM est actuellement basée sur la mesure des concentrations plasmatiques totales alors que seule la fraction libre (médicament non lié aux protéines plasmatiques circulantes) est susceptible de pénétrer dans la cellule pour exercer son action pharmacologique. L'analyse pharmacocinétique des taux plasmatiques totaux et libres d'imatinib mesurés simultanément chez les patients a permis d'affiner et de valider un modèle de pharmacocinétique de population qui intègre les facteurs influençant l'exposition à la fraction de médicament pharmacologiquement active. L'équation développée à partir de ce modèle permet d'extrapoler les concentrations libres d'imatinib à partir des concentrations plasmatiques totales qui sont actuellement mesurées lors du TDM des patients. Finalement, l'influence de la P-glycoprotéine sur la disposition cellulaire des TKIs a été étudiée dans un modèle cellulaire utilisant l'approche par la technologie du siRNA permettant de bloquer sélectivement l'expression du gène de cette protéine d'efflux des médicaments.Une autre approche pour augmenter la sélectivité du traitement anticancéreux consiste en une administration loco-régionale d'un agent cytostatique directement au sein de l'organe cible tout en préservant les tissus sains. La perfusion isolée du poumon (ILP) a été conçue pour le traitement loco-régional des cancers affectant les tissus pulmonaires mais les résultats cliniques ont été jusqu'à ce jour décevants. Dans des modèles précliniques chez le rat, il a pu être démontré que l'ILP avec la doxorubicine, un agent cytotoxique, administré seul, permet une exposition élevée au niveau du tissu pulmonaire, et une faible toxicité systémique. Toutefois, cette technique est caractérisée par une importante variabilité de la distribution dans les tissus pulmonaires et une pénétration du médicament au sein de la tumeur comparativement plus faible que dans les tissus sains.La thérapie photodynamique (PDT) est une nouvelle approche pour le traitement des tumeurs superficielles, qui consiste en l'application d'un agent sensibilisateur activé par une lumière laser de longueur d'onde spécifique, qui perturbe l'intégrité physiologique de la barrière endothéliale des vaisseaux alimentant la tumeur et permet d'augmenter localement la pénétration des agents cytostatiques.Nos études ont montré qu'un pré-traitement par PDT permet d'augmenter sélectivement l'absorption de doxorubicine dans les tumeurs lors d'administration i.v. de doxorubicine liposomale dans un modèle de sarcome de poumons de rongeurs.Résumé large publicDepuis une dizaine d'année, le traitement de certains cancers s'est progressivement transformé et les patients qui devaient jusqu'alors subir des chimiothérapies, toxiques et non spécifiques, peuvent maintenant bénéficier de traitements chroniques avec des thérapies ciblées. Avec les deux types d'approches thérapeutiques, on reste cependant confronté à la toxicité et aux effets secondaires du traitement.Le but de cette thèse a été d'étudier chez les patients et dans des modèles précliniques les diverses approches visant à améliorer l'activité et la tolérance des traitements à travers un meilleur ciblage de la thérapie anticancéreuse. Cet effort de recherche nous a conduits à nous intéresser à l'optimisation du traitement par les inhibiteurs de tyrosines kinases (TKIs), une nouvelle génération d'agents anticancéreux ciblés agissant sélectivement sur les cellules tumorales, en particulier chez les patients souffrant de leucémie myéloïde chronique et de tumeurs stromales gastro-intestinales. L'activité clinique ainsi que la toxicité de ces TKIs paraissent dépendre non pas de la dose de médicament administrée, mais de la quantité de médicaments circulant dans le sang auxquelles les tumeurs cancéreuses sont exposées et qui varient beaucoup d'un patient à l'autre. A cet effet, nous avons développé une méthode par chromatographie couplée à la spectrométrie de masse pour mesurer chez les patients les taux de médicaments de la classe des TKIs dans la perspective de piloter le traitement par une approche de suivi thérapeutique (Therapeutic Drug Monitoring, TDM). Le TDM repose sur la mesure de la quantité de médicament dans le sang d'un patient dans le but d'adapter individuellement la posologie la plus appropriée: des quantités insuffisantes de médicament dans le sang peuvent conduire à un échec thérapeutique alors qu'un taux sanguin excessif peut entraîner des manifestations toxiques.Dans une seconde partie préclinique, nous nous sommes concentrés sur l'optimisation de la chimiothérapie loco-régionale dans un modèle de sarcome du poumon chez le rat, afin d'augmenter l'exposition dans la tumeur tout en réduisant la toxicité dans les tissus non affectés.La perfusion isolée du poumon (ILP) permet d'administrer un médicament anticancéreux cytotoxique comme la doxorubicine, sélectivement au niveau le tissu pulmonaire où sont généralement localisées les métastases de sarcome. L'administration par ILP de doxorubicine, toxique pour le coeur, a permis une forte accumulation des médicaments dans le poumon, tout en épargnant le coeur. Il a été malheureusement constaté que la doxorubicine ne pénètre que faiblement dans la tumeur sarcomateuse, témoignant des réponses cliniques décevantes observées avec cette approche en clinique. Nous avons ainsi étudié l'impact sur la pénétration tumorale de l'association d'une chimiothérapie cytotoxique avec la thérapie photodynamique (PDT) qui consiste en l'irradiation spécifique du tissu-cible cancéreux, après l'administration d'un agent photosensibilisateur. Dans ce modèle animal, nous avons observé qu'un traitement par PDT permet effectivement d'augmenter de façon sélective l'accumulation de doxorubicine dans les tumeurs lors d'administration intraveineuse de médicament.

Expression and localization of PPARs in the rat ovary during follicular development and the periovulatory period.

PPARs are a family of nuclear hormone receptors involved in various processes that could influence ovarian function. We investigated the cellular localization and expression of PPARs during follicular development in ovarian tissue collected from rats 0, 6, 12, 24, and 48 h post-PMSG. A second group of animals received human CG (hCG) 48 h post-PMSG. Their ovaries were removed 0, 4, 8, 12, and 24 h post-hCG to study the periovulatory period. mRNAs corresponding to the PPAR isotypes (alpha, delta, and gamma) were localized by in situ hybridization. Changes in the levels of mRNA for the PPARs were determined by ribonuclease protection assays. PPAR gamma mRNA was localized primarily to granulosa cells, and levels of expression did not change during follicular development. Four hours post-hCG, levels of mRNA for PPAR gamma decreased (P < 0.05) but not uniformly in all follicles. At 24 h post-hCG, levels of PPAR gamma mRNA were reduced 64%, but some follicles maintained high expression. In contrast, mRNAs for PPAR alpha and delta were located primarily in theca and stroma, and their levels did not change during the intervals studied. To investigate the physiologic significance of PPAR gamma in the ovary, granulosa cells from PMSG-primed rats were cultured for 48 h with prostaglandin J(2) (PGJ(2)) and ciglitazone, PPAR gamma activators. Both compounds increased progesterone and E2 secretion (P < 0.05). These data suggest that PPAR gamma is involved in follicular development, has a negative influence on the luteinization of granulosa cells, and/or regulates the periovulatory shift in steroid production. The more general and steady expression of PPARs alpha and delta indicate that they may play a role in basal ovarian function.

The role of hypocretin in driving arousal and goal-oriented behaviors.

The hypocretins (Hcrts), also called orexins, are two neuropeptides secreted by a few thousand neurons restricted to the lateral hypothalamus. The Hcrt peptides bind to two receptors located in nuclei associated with diverse cognitive and physiological functions. Experimental evidence has demonstrated that the physiological roles of hypocretins extend far beyond its initial role in food consumption and has emerged as a key system in the fields of sleep disorders and drug addiction. Here, we discuss recent evidence demonstrating a key role of hypocretin in the motivation for reward seeking in general, and drug taking in particular, and we delineate a physiological framework for this peptidergic system in orchestrating the appropriate levels of alertness required for the elaboration and the execution of goal-oriented behaviors. We propose a general role for hypocretins in mediating arousal, especially when an organism must respond to unexpected stressors and environmental challenges, which serve to shape survival behaviors. We also discuss the limit of the current experimental paradigms to address the question of how a system normally involved in the regulation of vigilance states and hyperarousal may promote a pathological state that elicits compulsive craving and relapse to drug seeking.

Additive functions in boolean models of gene regulatory network modules.

Gene-on-gene regulations are key components of every living organism. Dynamical abstract models of genetic regulatory networks help explain the genome's evolvability and robustness. These properties can be attributed to the structural topology of the graph formed by genes, as vertices, and regulatory interactions, as edges. Moreover, the actual gene interaction of each gene is believed to play a key role in the stability of the structure. With advances in biology, some effort was deployed to develop update functions in Boolean models that include recent knowledge. We combine real-life gene interaction networks with novel update functions in a Boolean model. We use two sub-networks of biological organisms, the yeast cell-cycle and the mouse embryonic stem cell, as topological support for our system. On these structures, we substitute the original random update functions by a novel threshold-based dynamic function in which the promoting and repressing effect of each interaction is considered. We use a third real-life regulatory network, along with its inferred Boolean update functions to validate the proposed update function. Results of this validation hint to increased biological plausibility of the threshold-based function. To investigate the dynamical behavior of this new model, we visualized the phase transition between order and chaos into the critical regime using Derrida plots. We complement the qualitative nature of Derrida plots with an alternative measure, the criticality distance, that also allows to discriminate between regimes in a quantitative way. Simulation on both real-life genetic regulatory networks show that there exists a set of parameters that allows the systems to operate in the critical region. This new model includes experimentally derived biological information and recent discoveries, which makes it potentially useful to guide experimental research. The update function confers additional realism to the model, while reducing the complexity and solution space, thus making it easier to investigate.

Unison or cacophony: postgraduate training in pathology in Europe.

With the free movement of people in the European Union, medical mobility has increased significantly. This is notably the case for disciplines for which shortage of well-trained staff has occurred. Pathology is among those specialties and effectively the discipline is confronted with a striking increase in mobility among trainees and qualified specialists. The presumption underlying unlimited mobility is that the competencies of the medical specialists in the European countries are more or less equal, including significant similarities in the postgraduate training programs. In order to assess whether reality corresponds with this presumption, we conducted a survey of the content and practice requirements of the curricula in the EU and affiliated countries. The results indicate a striking heterogeneity in the training program content and practice requirements. To name a few elements: duration of the training program varied between 4 and 6 years; the number of autopsies required varied between none at all and 300; the number of biopsies required varied between none at all and 15,000. We conclude that harmonization of training outcomes in Europe is a goal that needs to be pursued. This will be difficult to reach through harmonization of training programs, as these are co-determined by political, cultural, and administrative factors, difficult to influence. Harmonization might be attained by defining the general and specific competencies at the end of training and subsequent testing them through a test to which all trainees in Europe are subjected.

Moderate-Vigorous Physical Activity across Body Mass Index in Females: Moderating Effect of Endocannabinoids and Temperament.

BACKGROUND Endocannabinoids and temperament traits have been linked to both physical activity and body mass index (BMI) however no study has explored how these factors interact in females. The aims of this cross-sectional study were to 1) examine differences among distinct BMI groups on daytime physical activity and time spent in moderate-vigorous physical activity (MVPA), temperament traits and plasma endocannabinoid concentrations; and 2) explore the association and interaction between MVPA, temperament, endocannabinoids and BMI. METHODS Physical activity was measured with the wrist-worn accelerometer Actiwatch AW7, in a sample of 189 female participants (43 morbid obese, 30 obese, and 116 healthy-weight controls). The Temperament and Character Inventory-Revised questionnaire was used to assess personality traits. BMI was calculated by bioelectrical impedance analysis via the TANITA digital scale. Blood analyses were conducted to measure levels of endocannabinoids and endocannabinoid-related compounds. Path-analysis was performed to examine the association between predictive variables and MVPA. RESULTS Obese groups showed lower MVPA and dysfunctional temperament traits compared to healthy-weight controls. Plasma concentrations of 2-arachidonoylglyceryl (2-AG) were greater in obese groups. Path-analysis identified a direct effect between greater MVPA and low BMI (b = -0.13, p = .039) and high MVPA levels were associated with elevated anandamide (AEA) levels (b = 0.16, p = .049) and N-oleylethanolamide (OEA) levels (b = 0.22, p = .004), as well as high Novelty seeking (b = 0.18, p<.001) and low Harm avoidance (b = -0.16, p<.001). CONCLUSIONS Obese individuals showed a distinct temperament profile and circulating endocannabinoids compared to controls. Temperament and endocannabinoids may act as moderators of the low MVPA in obesity.

Modalities and future prospects of gene therapy in heart transplantation.

Heart transplantation is the treatment of choice for many patients with end-stage heart failure. Its success, however, is limited by organ shortage, side effects of immunosuppressive drugs, and chronic rejection. Gene therapy is conceptually appealing for applications in transplantation, as the donor organ is genetically manipulated ex vivo before transplantation. Localised expression of immunomodulatory genes aims to create a state of immune privilege within the graft, which could eliminate the need for systemic immunosuppression. In this review, recent advances in the development of gene therapy in heart transplantation are discussed. Studies in animal models have demonstrated that genetic modification of the donor heart with immunomodulatory genes attenuates ischaemia-reperfusion injury and rejection. Alternatively, bone marrow-derived cells genetically engineered with donor-type major histocompatibility complex (MHC) class I or II promote donor-specific hyporesponsiveness. Genetic engineering of naïve T cells or dendritic cells may induce regulatory T cells and regulatory dendritic cells. Despite encouraging results in animal models, however, clinical gene therapy trials in heart transplantation have not yet been started. The best vector and gene to be delivered remain to be identified. Pre-clinical studies in non-human primates are needed. Nonetheless, the potential of gene therapy as an adjunct therapy in transplantation is essentially intact.

Sunshine Advisory, November 2001

A Bulletin on Iowa Open Meetings and Public Records Laws By Attorney General Tom Miller