Prospective randomized study of doxorubicin-eluting-bead embolization in the treatment of hepatocellular carcinoma: results of the PRECISION V study.

Transcatheter arterial chemoembolization (TACE) offers a survival benefit to patients with intermediate hepatocellular carcinoma (HCC). A widely accepted TACE regimen includes administration of doxorubicin-oil emulsion followed by gelatine sponge-conventional TACE. Recently, a drug-eluting bead (DC Bead) has been developed to enhance tumor drug delivery and reduce systemic availability. This randomized trial compares conventional TACE (cTACE) with TACE with DC Bead for the treatment of cirrhotic patients with HCC. Two hundred twelve patients with Child-Pugh A/B cirrhosis and large and/or multinodular, unresectable, N0, M0 HCCs were randomized to receive TACE with DC Bead loaded with doxorubicin or cTACE with doxorubicin. Randomization was stratified according to Child-Pugh status (A/B), performance status (ECOG 0/1), bilobar disease (yes/no), and prior curative treatment (yes/no). The primary endpoint was tumor response (EASL) at 6 months following independent, blinded review of MRI studies. The drug-eluting bead group showed higher rates of complete response, objective response, and disease control compared with the cTACE group (27% vs. 22%, 52% vs. 44%, and 63% vs. 52%, respectively). The hypothesis of superiority was not met (one-sided P = 0.11). However, patients with Child-Pugh B, ECOG 1, bilobar disease, and recurrent disease showed a significant increase in objective response (P = 0.038) compared to cTACE. DC Bead was associated with improved tolerability, with a significant reduction in serious liver toxicity (P < 0.001) and a significantly lower rate of doxorubicin-related side effects (P = 0.0001). TACE with DC Bead and doxorubicin is safe and effective in the treatment of HCC and offers a benefit to patients with more advanced disease.

Voriconazole-Induced Inhibition of the Fungicidal Activity of Amphotericin B in Candida Strains with Reduced Susceptibility to Voriconazole: an Effect Not Predicted by the MIC Value Alone.

An antagonistic effect of voriconazole on the fungicidal activity of sequential doses of amphotericin B has previously been demonstrated in Candida albicans strains susceptible to voriconazole. Because treatment failure and the need to switch to other antifungals are expected to occur more often in infections that are caused by resistant strains, it was of interest to study whether the antagonistic effect was still seen in Candida strains with reduced susceptibility to voriconazole. With the hypothesis that antagonism will not occur in voriconazole-resistant strains, C. albicans strains with characterized mechanisms of resistance against voriconazole, as well as Candida glabrata and Candida krusei strains with differences in their degrees of susceptibility to voriconazole were exposed to voriconazole or amphotericin B alone, to both drugs simultaneously, or to voriconazole followed by amphotericin B in an in vitro kinetic model. Amphotericin B administered alone or simultaneously with voriconazole resulted in fungicidal activity. When amphotericin B was administered after voriconazole, its activity was reduced (median reduction, 61%; range, 9 to 94%). Levels of voriconazole-dependent inhibition of amphotericin B activity differed significantly among the strains but were not correlated with the MIC values (correlation coefficient, -0.19; P = 0.65). Inhibition was found in C. albicans strains with increases in CDR1 and CDR2 expression but not in the strain with an increase in MDR1 expression. In summary, decreased susceptibility to voriconazole does not abolish voriconazole-dependent inhibition of the fungicidal activity of amphotericin B in voriconazole-resistant Candida strains. The degree of interaction could not be predicted by the MIC value alone.

The Changing Face of the Epidemiology of Tuberculosis due to Molecular Strain Typing: A Review

About one third of the world population is infected with tubercle bacilli, causing eight million new cases of tuberculosis (TB) and three million deaths each year. After years of lack of interest in the disease, World Health Organization recently declared TB a global emergency and it is clear that there is need for more efficient national TB programs and newly defined research priorities. A more complete epidemiology of tuberculosis will lead to a better identification of index cases and to a more efficient treatment of the disease. Recently, new molecular tools became available for the identification of strains of Mycobacterium tuberculosis (M. tuberculosis), allowing a better recognition of transmission routes of defined strains. Both a standardized restriction-fragment-length-polymorphism-based methodology for epidemiological studies on a large scale and deoxyribonucleic acids (DNA) amplification-based methods that allow rapid detection of outbreaks with multidrug-resistant (MDR) strains, often characterized by high mortality rates, have been developed. This review comments on the existing methods of DNA-based recognition of M. tuberculosis strains and their peculiarities. It also summarizes literature data on the application of molecular fingerprinting for detection of outbreaks of M. tuberculosis, for identification of index cases, for study of interaction between TB and infection with the human immunodeficiency virus, for analysis of the behavior of MDR strains, for a better understanding of risk factors for transmission of TB within communities and for population-based studies of TB transmission within and between countries

Individual versus standard quality of life assessment in a phase II clinical trial in mesothelioma patients: feasibility and responsiveness to clinical changes.

BACKGROUND: In patients with malignant pleural mesothelioma undergoing a multimodality therapy, treatment toxicity may outweigh the benefit of progression-free survival. The subjective experience across different treatment phases is an important clinical outcome. This study compares a standard with an individual quality of life (QoL) measure used in a multi-center phase II trial. PATIENTS AND METHODS: Sixty-one patients with stage I-III technically operable pleural mesothelioma were treated with preoperative chemotherapy, followed by pleuropneumonectomy and subsequent radiotherapy. QoL was assessed at baseline, at day 1 of cycle 3, and 1, 3 and 6 months post-surgery by using the Rotterdam Symptom Checklist (RSCL) and the Schedule for the Evaluation of Quality of Life-Direct Weighting (SEIQoL-DW), a measure that is based on five individually nominated and weighted QoL-domains. RESULTS: Completion rates were 98% (RSCL) and 92% (SEIQoL) at baseline and 98%/89% at cycle 3, respectively. Of the operated patients (N=45) RSCL and SEIQoL were available from 86%/72%, 93%/74%, and 94%/76% at months 1, 3, and 6 post-surgery. Average assessment time for the SEIQoL was 24min compared to 8min needed for the RSCL. Median changes from baseline indicate that both RSCL QoL overall score and SEIQoL index remained stable during chemotherapy with a clinically significant deterioration (change>or=8 points) 1 month after surgery (median change of -66 and -14 for RSCL and SEIQoL, respectively). RSCL QoL overall scores improved thereafter, but remained beneath baseline level until 6 months after surgery. SEIQoL scores improved to baseline-level at month 3 after surgery, but worsened again at month 6. RSCL QoL overall score and SEIQoL index were moderately correlated at baseline (r=.30; p<or=.05) and at 6-month follow-up (r=.42; p<or=.05) but not at the other time points. CONCLUSION: The SEIQoL assessment seems to be feasible within a phase II clinical trial, but may require more effort from staff. More distinctive QoL changes in accordance with clinical changes were measured with the RSCL. Our findings suggest that the two measures are not interchangeable: the RSCL is to favor when mainly information related to the course of disease- and treatment is of interest.

The effect of isolation on reproduction and growth of Pseudosuccinea columella (Pulmonata: Lymnaeidae): a snail-conditioned water experiment

A snail-conditioned water experiment was conducted in Pseudosuccinea columella to test the possible role of a chemical interaction between snails on the diminished growth and fecundity rates found for snails raised in pairs compared to those raised in complete isolation. The results permit to discard the hypothesis of an inhibition of growth and reproduction between snails due to factors released into the water.

Meta-analysis based SVM classification enables accurate detection of Alzheimer's disease across different clinical centers using FDG-PET and MRI.

The application of support vector machine classification (SVM) to combined information from magnetic resonance imaging (MRI) and [F18]fluorodeoxyglucose positron emission tomography (FDG-PET) has been shown to improve detection and differentiation of Alzheimer's disease dementia (AD) and frontotemporal lobar degeneration. To validate this approach for the most frequent dementia syndrome AD, and to test its applicability to multicenter data, we randomly extracted FDG-PET and MRI data of 28 AD patients and 28 healthy control subjects from the database provided by the Alzheimer's Disease Neuroimaging Initiative (ADNI) and compared them to data of 21 patients with AD and 13 control subjects from our own Leipzig cohort. SVM classification using combined volume-of-interest information from FDG-PET and MRI based on comprehensive quantitative meta-analyses investigating dementia syndromes revealed a higher discrimination accuracy in comparison to single modality classification. For the ADNI dataset accuracy rates of up to 88% and for the Leipzig cohort of up to 100% were obtained. Classifiers trained on the ADNI data discriminated the Leipzig cohorts with an accuracy of 91%. In conclusion, our results suggest SVM classification based on quantitative meta-analyses of multicenter data as a valid method for individual AD diagnosis. Furthermore, combining imaging information from MRI and FDG-PET might substantially improve the accuracy of AD diagnosis.

Extremal properties of certain risk models

Cette th��se s'int��resse �� ��tudier les propri��t��s extr��males de certains mod��les de risque d'int��r��t dans diverses applications de l'assurance, de la finance et des statistiques. Cette th��se se d��veloppe selon deux axes principaux, �� savoir: Dans la premi��re partie, nous nous concentrons sur deux mod��les de risques univari��s, c'est-��- dire, un mod��le de risque de d��flation et un mod��le de risque de r��assurance. Nous ��tudions le d��veloppement des queues de distribution sous certaines conditions des risques commun��s. Les principaux r��sultats sont ainsi illustr��s par des exemples typiques et des simulations num��riques. Enfin, les r��sultats sont appliqu��s aux domaines des assurances, par exemple, les approximations de Value-at-Risk, d'esp��rance conditionnelle unilat��rale etc. La deuxi��me partie de cette th��se est consacr��e �� trois mod��les �� deux variables: Le premier mod��le concerne la censure �� deux variables des ��v��nements extr��me. Pour ce mod��le, nous proposons tout d'abord une classe d'estimateurs pour les coefficients de d��pendance et la probabilit�� des queues de distributions. Ces estimateurs sont flexibles en raison d'un param��tre de r��glage. Leurs distributions asymptotiques sont obtenues sous certaines condi��tions lentes bivari��es de second ordre. Ensuite, nous donnons quelques exemples et pr��sentons une petite ��tude de simulations de Monte Carlo, suivie par une application sur un ensemble de donn��es r��elles d'assurance. L'objectif de notre deuxi��me mod��le de risque �� deux variables est l'��tude de coefficients de d��pendance des queues de distributions obliques et asym��triques �� deux variables. Ces distri��butions obliques et asym��triques sont largement utiles dans les applications statistiques. Elles sont g��n��r��es principalement par le m��lange moyenne-variance de lois normales et le m��lange de lois normales asym��triques d'��chelles, qui distinguent la structure de d��pendance de queue comme indiqu�� par nos principaux r��sultats. Le troisi��me mod��le de risque �� deux variables concerne le rapprochement des maxima de s��ries triangulaires elliptiques obliques. Les r��sultats th��oriques sont fond��s sur certaines hypoth��ses concernant le p��rim��tre al��atoire sous-jacent des queues de distributions. -- This thesis aims to investigate the extremal properties of certain risk models of interest in vari��ous applications from insurance, finance and statistics. This thesis develops along two principal lines, namely: In the first part, we focus on two univariate risk models, i.e., deflated risk and reinsurance risk models. Therein we investigate their tail expansions under certain tail conditions of the common risks. Our main results are illustrated by some typical examples and numerical simu��lations as well. Finally, the findings are formulated into some applications in insurance fields, for instance, the approximations of Value-at-Risk, conditional tail expectations etc. The second part of this thesis is devoted to the following three bivariate models: The first model is concerned with bivariate censoring of extreme events. For this model, we first propose a class of estimators for both tail dependence coefficient and tail probability. These estimators are flexible due to a tuning parameter and their asymptotic distributions are obtained under some second order bivariate slowly varying conditions of the model. Then, we give some examples and present a small Monte Carlo simulation study followed by an application on a real-data set from insurance. The objective of our second bivariate risk model is the investigation of tail dependence coefficient of bivariate skew slash distributions. Such skew slash distributions are extensively useful in statistical applications and they are generated mainly by normal mean-variance mixture and scaled skew-normal mixture, which distinguish the tail dependence structure as shown by our principle results. The third bivariate risk model is concerned with the approximation of the component-wise maxima of skew elliptical triangular arrays. The theoretical results are based on certain tail assumptions on the underlying random radius.

The hope for an HIV vaccine based on induction of CD8+ T lymphocytes: a review

The only long-term and cost-effective solution to the human immunodeficiency virus (HIV) epidemic in the developing world is a vaccine that prevents individuals from becoming infected or, once infected, from passing the virus on to others. There is currently little hope for an AIDS vaccine. Conventional attempts to induce protective antibody and CD8+ lymphocyte responses against HIV and simian immunodeficiency virus (SIV) have failed. The enormous diversity of the virus has only recently been appreciated by vaccinologists, and our assays to determine CD8+ lymphocyte antiviral efficacy are inadequate. The central hypothesis of a CTL-based vaccine is that particularly effective CD8+ lymphocytes directed against at least five epitopes that are derived from regions under functional and structural constraints will control replication of pathogenic SIV. This would be somewhat analogous to control of virus replication by triple drug therapy or neutralizing antibodies.

Aggravation of chronic stress effects on hippocampal neurogenesis and spatial memory in LPA��� receptor knockout mice.

BACKGROUND The lysophosphatidic acid LPA��� receptor regulates plasticity and neurogenesis in the adult hippocampus. Here, we studied whether absence of the LPA��� receptor modulated the detrimental effects of chronic stress on hippocampal neurogenesis and spatial memory. METHODOLOGY/PRINCIPAL FINDINGS Male LPA���-null (NULL) and wild-type (WT) mice were assigned to control or chronic stress conditions (21 days of restraint, 3 h/day). Immunohistochemistry for bromodeoxyuridine and endogenous markers was performed to examine hippocampal cell proliferation, survival, number and maturation of young neurons, hippocampal structure and apoptosis in the hippocampus. Corticosterone levels were measured in another a separate cohort of mice. Finally, the hole-board test assessed spatial reference and working memory. Under control conditions, NULL mice showed reduced cell proliferation, a defective population of young neurons, reduced hippocampal volume and moderate spatial memory deficits. However, the primary result is that chronic stress impaired hippocampal neurogenesis in NULLs more severely than in WT mice in terms of cell proliferation; apoptosis; the number and maturation of young neurons; and both the volume and neuronal density in the granular zone. Only stressed NULLs presented hypocortisolemia. Moreover, a dramatic deficit in spatial reference memory consolidation was observed in chronically stressed NULL mice, which was in contrast to the minor effect observed in stressed WT mice. CONCLUSIONS/SIGNIFICANCE These results reveal that the absence of the LPA��� receptor aggravates the chronic stress-induced impairment to hippocampal neurogenesis and its dependent functions. Thus, modulation of the LPA��� receptor pathway may be of interest with respect to the treatment of stress-induced hippocampal pathology.

Intra- and intersexual selection and their potential consequences on fitness-relevant traits in several fish species

R��sum�� : Les m��canismes de s��lection sexuelle, en particulier la comp��tition entre m��les (s��lection inter-sexuelle) et le choix des femelles (s��lection intra-sexuelle), peuvent fortement influencer le succ��s reproducteur d'un individu, c'est-��-dire son nombre de descendants. On observe ainsi que les m��les dominants et les m��les ��laborant des caract��res sexuels secondaires marqu��s ont un succ��s reproducteur ��lev��. Toutefois, le succ��s reproducteur ne suffit pas pour garantir une contribution g��n��tique ��lev��e, parce que la fitness d��pend ��galement de la performance des descendants (c'est-��-dire de leur survie et de leur propre succ��s reproducteur). Si cette performance d��pend en partie des g��nes paternels, les males ont un avantage certain �� signaler leur qualit�� aux femelles afin d'atteindre des taux de reproduction ��lev��. Ce m��canisme de signalisation est connu sous le nom de 'good genes hypothesis', toutefois tr��s peu d'��tudes ont clairement d��montr�� le lien entre la qualit�� g��n��tique des individus et la signalisation. De plus, la performance des descendants peut aussi d��pendre des effets g��n��tiques de compatibilit�� entre m��les et femelles ('compatible genes'). C'est-��-dire que certains all��les paternels n'apporteraient un avantage aux descendants qu'en combinaison avec certains all��les maternels. Nous avons d��termin��, durant la p��riode de reproduction, le statut de dominance des m��les pour deux esp��ces de poissons d'eau douce : la truite (Salmo trotta) et le vairon (Phoxinus phoxinus), puis nous avons ��valu�� la relation entre le succ��s reproducteur et le statut de dominance et/ou la quantit�� de signalisation des caract��res sexuels secondaires. Nous avons ��galement f��cond��s artificiellement des oeufs de truites et de cor��gones (Coregonus palaea), en croisant chaque m��le avec chaque femelle (full-factorial breeding design). Ce type de design autorise la quantification pr��cise des effets g��n��tiques et permet de s��parer les effets de 'good genes' et de 'compatible genes'. Cela a ��t�� fait sous diff��rentes intensit��s de stress bact��rien, ainsi que dans des conditions naturelles, et nous avons pu ainsi tester si certains indicateurs de qualit�� g��n��tique des m��les ('good genes') ��taient li��s a) �� la dominance et/ou b) �� l'expression des caract��res sexuels secondaires des m��les comme l'intensit�� m��lanique ou la taille des tubercules sexuels. En outre, nous cherchons �� savoir si la survie des descendants est li��e �� certaines combinaison des g��nes du complexe d'histocompatibilit�� majeur (MHC) et/ou �� la parent�� g��n��tique des parents, les deux traits ��tant soup��onn��s d'avoir des influences g��n��tique de compatibilit�� (`compatible genes') �� la performance des descendants. Nous avons constat�� que la dominance des m��les est directement li��e �� la taille et au poids des m��les (truites, vairons), mais ��galement aux caract��res sexuels secondaires (tubercules). De plus, les m��les vairons dominant ont eu un succ��s de f��condation plus ��lev��s que les m��les subordonn��s. Nous montrons que les truites et cor��gones m��les diff��rent dans leur qualit�� g��n��tique, qui a ��t�� mesur��e av��c la survie embryonnaire, le temps avant l'��closion et enfin la croissance juv��nile. Contrairement aux pr��dictions, la dominance (ou les traits indicatifs de dominance) n'��tait li��e �� la qualit�� g��n��tique, dans aucun des traitements, et ne fonctionne donc pas comme indicateur de qualit��. Par contre, la qualit�� g��n��tique ��tait li��e aux caract��res sexuels secondaires, particuli��rement par la teinte m��lanique chez les truites. Les embryons de truites issus de p��res sombres survivaient mieux que ceux issus de p��res clairs dans des environnements difficiles, de plus leur croissance ��tait plus ��lev��e lors de leur premi��re ann��e dans des conditions naturelles. La taille des juv��niles lors de leur premi��re ann��e est un trait important li�� au succ��s dans la comp��tition pour des ressources telles qu'abri ou nourriture. De plus, les femelles truites peuvent augmenter la survie de leurs descendants en choisissant des m��les selon leur type de MHC ou selon leur degr�� de parent��. En outre, chez les cor��gones, la morphologie des tubercules sexuels ne semble pas signaler la qualit�� g��n��tique. Nous avons ��galement remarqu�� que l'exposition �� des pathog��nes non-l��taux pouvait influencer la performance des alevins �� court et long terme, probablement en affaiblissant leur syst��me immunitaire. Cette th��se montre que les m��les diff��rent dans leur qualit�� g��n��tique et que diff��rents m��canismes de s��lection inter- ou intra-sexuelle (par exemple la pr��f��rence pour des m��les sombres, pour des g��notypes MHC ou pour des couples avec degr�� de parent�� basse) pouvait avoir un effet positif sur la qualit�� des descendants, bien que cet effet g��n��tique pouvait changer au cours du temps et entre diff��rents environnements. Contrairement �� nos attentes, le r��sultat de la comp��tition intra-sexuelle (la hi��rarchie de dominance entre m��les) n'��tait pas li�� �� la qualit�� g��n��tique individuelle ('good genes'). Dans ce sens, ce travail permet ��galement de contribuer �� l'explication du fait que la s��lection sexuelle, de par sa forte s��lection directionnelle, ne conduit pas �� la diminution de la variance g��n��tique, mais plut��t �� la maintenance du polymorphisme g��n��tique. Summary : Sexual selection mechanisms, especially male-male competition (inteasexual selection) and female mate choice (inteasexual selection), can strongly influence individual mating success, often resulting in dominant males and males with elaborate secondary sexual characters having higher fertilisation success. However, siring a high number of offspring alone does not guarantee high individual fitness, as fitness does also strongly depend on offspring performance (i.e. survival, fecundity). If this superiority in offspring performance depends on paternally inherited genes, the fathers are expected to signal this potential indirect benefit to females in order to attain high mating rates. This mechanism is also known as the 'good genes' hypothesis of sexual selection but until now most studies failed to conclusively show the relation of an individual genetic quality and its potential signalling traits. Further, offspring performance could also depend on compatible gene effects. These are alleles that increase offspring performance only in combination with other specific alleles. We first determined male dominance status from intrasexual competition during mating season for brown trout (Salmo trutta) and European minnows (Phoxinus phoxinus). For minnows we additionally checked if dominance and/or secondary sexual traits were linked to fertilisation success. Further, we artificially fertilised brown trout and alpine whitefish (Coregonus palaea) eggs, following full factorial breeding designs, enabling to properly measure `good gene' and `compatible gene' effects on offspring performance. This was done under different intensities of natural stressors, as well as under natural conditions. This procedure allowed us to test if the obtained male genetic quality measures (good genes effects) were indicated by a) dominance or lay traits linked to dominance and/or by b) secondary sexual characteristics such as melanin-based male skin darkness or breeding tubercles. Further, we investigated if offspring survival was linked to the MHC (major histocompatibility complex) gene combinations and/or to the parental genetic relatedness, as both traits were shown to have 'compatible gene' effects that may influence offspring performance. We found that male dominance in intrasexual competition was positively linked to body size, body weight (brown trout, minnows) but also to elaborate secondary sexual characteristics (breeding tubercles in minnows). Further, dominant minnow males did have an increased fertilisation success compared to subordinate ones. We show that brown trout and whitefish males do usually differ in their genetic quality, which was measured as embryo survival, hatching timing and finally as juvenile growth. Contrary to prediction male dominance or dominance indicating traits do not function as a quality signal as they were not linked to genetic quality. This result was constant when measuring genetic quality under different levels of natural stressors and under natural conditions (brown trout). On the other hand genetic quality seemed to be indicated by secondary sexual characteristics, specifically by melanin-based skin darkness in brown trout as brown trout embryos sired by darker fathers had increased survival rates when raised under harsh conditions and. they grew larger as juveniles after one year of growth in a natural stream, which is an important trait influencing success of juveniles in competition for hidings, food and other resources. Furthermore, brown trout females may increase the survival of their embryos when choosing males according to their MHC genotypes or to the general genetic relatedness between themselves and their potential mates. In whitefish on the other hand breeding tubercle morphology did not seem to signal genetic quality. Eventually, we saw that anon-lethal exposure to pathogens might influence short term and long term offspring performance probably by weakening an exposed individual's immune system. This thesis shows that males usually differ in their genetic quality and that different inter- or intrasexual selection mechanisms (e.g. mate selection favouring dark males, preference for MHC genotype combinations or for unrelated mates) may have strong positive effects on genetically dependent offspring performance but that such genet��c effects can change over time and environments. In contrast to our a priori expectations, the outcome of intrasexual selection, namely male dominance hierarchies, with dominant males often having high fertilisation success, was not linked to individual genetic quality (`good genes'). In this sense the present thesis may also be a helpful contribution to understand why sexual selection does not lead to rapid loss of genetic variation by strong directional selection but could even lead to the maintenance of genetic variation in natural populations.

Study of drug tolerance mechanisms and of the calcineurin pathway in the opportunistic pathogen "Candida albicans"

ABSTRACT :Azole antifungal drugs possess fungistatic activity in Candida albicans making this human pathogen tolerant to these agents. The conversion of azoles into fungicidal agents is of interest since their fungistatic properties increase the ability of C. albicans to develop drug resistance. In C. albicans, the phosphatase calcineurin (calcineurin) is essential for antifungal drug tolerance. Up to now, the only known target of calcineurin is Crzl, which is a transcription factor (TF) involved in responses to ionic stress. Thus, most of the components of the calcineurin signaling remain to be identified in C. albicans.In this work, the calcineurin pathway was investigated in order to i) characterize the role of calcineurin in the biology of C. albicans, ii) identify putative targets of calcineurin and iii) characterize the phenomenon of tolerance to antifungal drugs. Towards these aims, four different approaches were used.First, using C. albicans microarrays, an attempt was made to identify a set of calcineurindependent genes (CDGs). Since CDGs were highly dependent upon the external stimulus used to activate calcineurin (Ca2+ or terbinafine), this stimulus bias was bypassed by the construction of strains expressing a truncated autoactive form of calcineurin (Cmp1tr) in a doxycyclinedependent manner. The characterization of Cmpltr was undertaken and results showed that it mimicked awild-type activated calcineurin for all tested phenotypes (i.e. Cnbl-dependence, inhibition by FK506, phosphatase 2B activity, ability to dephosphorylate Crzl and to regulate Crz1-and calcineurin-dependent genes, role in antifungal drug tolerance and susceptibility, role in colony formation on Spider agar). Cmp1tr was therefore considered as a valid tool to study the calcineurin signaling pathway. In silico analysis of CDGs allowed the identification of i) a significant overlap between CDGs and genes regulated by the Cyrl signal��ng pathway, ii) putative interactions between calcineurin activation and cell wall reorganization and phospholipid transport, iii) a putative interacti��n between calcineurin and the regulation of translation and iv) a putative relation between calcineurin and proteasome regulation. Further in silico analyses of the promoters of Crz1-independent CDGs were performed to identify TFs (other than Crz1) that were likely to regulate CDGs and therefore to be a direct target of calcineurin. The analyses revealed that Rpn4 and Mnl1 were TFs likely to be regulated by calcineurin.Second, in order to better characterize azole tolerance, an attempt was made to i) confirm the role of Hsp90 in fluconazole tolerance with a doxycycline-dependent Hsp90 expression system and ii) assess its calcineurin-dependence. Hsp90 was found to be significantly involved in fluconazole tolerance. However, results were not in agreement with the hypothesis that Hsp90 mediates fluconazole tolerance by the only downstream effector calcineurin. Rather Hsp90 is interacting with numerous components for fluconazole tolerance.Third, a collection of C. albicans TFs mutants were screened for loss of tolerance to terbinafine and fluconazole in order to identify TFs involved in antifungal drug tolerance. Out of the 265 TFs mutants screened, only the upc2Δ/Δ mutant showed a loss of fluconazole and terbinafine tolerance. Interestingly, no relation between Upc2 and calcineurin activity was found. These results suggested that the tolerance to antifungal drugs must not be only considered as a calcineurin-dependent phenomenon in C. albicans.Fourth, using FRCS analyses, an attempt was made to identify putative signs of programmed cell death (PCD) in calcineurin mutant cells upon loss of tolerance to terbinafine. A high proportion of cells died from both RO5-dependent (which is a sign of PCD) and ROS-independent (which is a sign of loss of homeostasis) processes in the calcineurin mutant. While these results suggest that calcineurin represses both loss of homeostasis and PCD, the role of calcineurin in PCD is still an open question.In conclusion, this work allowed i) the identification of several putative calcineurin targets, ii) the discovery of several links between calcineurin and signaling pathways and important biological processes and iii) the identification of novel components of calcineurin-independent mechanisms that participate in tolerance to antifungal drugs in C. albicans.R��SUME :Les azoles sont des antifongiques qui pr��sentent une activit�� fongistatique contre Candida albicans et rendent cette levure tol��rante �� ces agents. La conversion des azoles en agents fongicides est d'int��r��ts car leurs propri��t��s fongistatiques favorisent le d��veloppement de r��sistance aux drogues chez C. albicans. La calcineurine (calcineurin) est une phosphatase essentielle pour la tol��rance aux antifongiques chez C. albicans. La seule cible connue de la calcineurin est Crz1, un facteur de transcription (FT) impliqu�� dans la r��ponse aux stress ionique. Ainsi, la plupart des constituants de la voie de signalisation de la calcineurin restent encore �� ��tre identifi��s chez C. albicans.Dans ce travail de th��se, la voie de signalisation de la calcineurin a ��t�� ��tudi��e de sorte �� i) caract��riser le r��le de la calcineurin dans la biologie de C. albicans, ii) identifier de nouvelles cibles de la calcineurin et iii) caract��riser le ph��nom��ne de tol��rance aux antifongiques. A ce propos, quatre approches ont ��t�� entreprises.Premi��rement, des puces �� ADN de C. albicans ont ��t�� utilis��es afin d'identifier les g��nes d��pendants de la calcineurin (GDCs). Les GDCs ��tant ��troitement d��pendants du stimulus utilis�� pour activer la calcineurin, le biais ��stimulus�� a ��t�� ��vit�� via la construction d'une souche exprimant une forme tronqu��e et autoactive de la calcineurin (Cmp1tr), en pr��sence de doxycycline. La caract��risation de Cmp1tr a ��t�� entreprise et les r��sultats ont montr�� qu'elle mimait une calcineurin sauvage et activ��e pour la plupart des ph��notypes test��s (i.e. d��pendance �� Cnb1, inhibition par le FK506, activit�� phosphatase 2B, d��phosphorylation de Crz1 et r��gulation de g��nes d��pendant de la calcineurin, r��le dans la tol��rance et la susceptibilit�� aux antifongiques, r��le dans la formation des colonies sur milieu Spider). Cmp1tr a donc ��t�� consid��r�� comme un outil pertinent pour l'��tude de la voie de signalisation de la calcineurin. Les analyses in silico des GDCs ont permis l'identification i) d'un chevauchement entre les GDCs ��t les g��nes r��gul��s par la voie de signalisation de Cyrl, ii) d'une interaction entre la calcineurin et la r��organisation de la paroi cellulaire ainsi que le transport des phospholipides, iii) d'une interaction entre calcineurin et la r��gulation de la traduction et iv) une relation entre la calcineurin et la r��gulation du prot��asome. De plus, une analyse in silico des promoteurs des GDCs avec une r��gulation ind��pendante de Crz1 a permis d'identifier deux FTs qui pourraient ��tre des cibles directes de la calcineurin, Rpn4 et Mnll.Deuxi��mement, afin de caract��riser la tol��rance aux azoles, il a ��t�� entrepris i) de confirmer le r��le de Hsp90 dans la tol��rance au fluconazole en utilisant un syst��me d'expression d��pendant de la doxycycline et ii) de caract��riser sa d��pendance �� la calcineurin. Hsp90 a ��t�� montr�� impliqu�� dans la tol��rance aux azoles. Cependant, les r��sultats n'ont pas corrobor�� une hypoth��se expliquant le r��le d'Hsp90 dans la tol��rance aux antifongiques par son unique. interaction avec la calcineurin. Il a ��t�� propos�� que le r��le d'Hsp90 dans la tol��rance aux antifongiques soit d�� �� ces multiples interactions avec le prot��ome de C. albicans plut��t que par son interaction avec un partenaire unique.Troisi��mement, une collection de mutant pour des FTs de C. albicans a ��t�� cribl��e pour une perte de tol��rance au fluconazole ou �� la terbinafine, de sorte �� identifier les FTs impliqu��s dans la tol��rance aux antifongiques. Sur les 265 FTs pass��s au crible, seul le mutant upc2Δ/Δ a montr�� une perte de tol��rance au fluconazole et �� la terbinafine. Aucune relation n'a ��t�� trouv��e entre la calcineurin et l'activit�� d'Upc2. Ces r��sultats sugg��rent que la perte de tol��rance aux antifongiques ne doit pas ��tre consid��r��e comme un ph��nom��ne exclusivement li�� �� la voie de signalisation de la calcineurin.Quatri��mement, en utilisant la cytom��trie de flux, la pr��sence de signes de mort cellulaire programm��e (MCP) a ��t�� recherch��e lors de la perte de tol��rance du mutant calcineurin incub�� avec de la terbinafine. Une grande proportion de cellules mortes incluant ou non une production de ROS (un signe de MCP) a ��t�� d��tect��e dans le mutant calcineurin. Ces r��sultats pr��liminaires sugg��rent que la calcineurin r��prime autant la perte d'hom��ostasie qu'elle r��gule l'entr��e en MCP. Cependant d'autres analyses sont n��cessaires pour d��montrer clairement le r��le de la calcineurin dans la r��gulation de la MCP.En conclusion, ce travail de th��se a permis i) l'identification de plusieurs cibles possibles de la calcineurine, ii) la d��couverte de plusieurs interactions entre la calcineurine et d'autres voies de signalisation et processus biologiques importants et iii) de d��montrer la pr��sence de voies ind��pendantes de la calcineurine impliqu��es dans la tol��rance aux antifongiques chez C. albicans.

Medical geography and phthisic immunity in the high altitudes: The origins of a Therapeutic hypothesis

Medical geography expanded considerably in the 19 th century. Its expansion was aided by a Neo-Hippocratic trend in medical thinking, progress in statistics and hygiene, and an overall vision of geography formulated early in the century by French and German geographers inspired by Alexander von Humboldt. By tracing out the process that prompted certain �� doctor-geographers �� to put forth the hypothesis of immunity phthisis in elevated regions, this article seeks to show how various trends in medical geography led to the establishment of the �� altitude cure �� as a treatment for tuberculosis.

Satisfaction of patients hospitalised in psychiatric hospitals: a randomised comparison of two psychiatric-specific and one generic satisfaction questionnaires.

BACKGROUND: While there is interest in measuring the satisfaction of patients discharged from psychiatric hospitals, it might be important to determine whether surveys of psychiatric patients should employ generic or psychiatry-specific instruments. The aim of this study was to compare two psychiatric-specific and one generic questionnaires assessing patients' satisfaction after a hospitalisation in a psychiatric hospital. METHODS: We randomised adult patients discharged from two Swiss psychiatric university hospitals between April and September 2004, to receive one of three instruments: the Saphora-Psy questionnaire, the Perceptions of Care survey questionnaire or the Picker Institute questionnaire for acute care hospitals. In addition to the comparison of response rates, completion time, mean number of missing items and mean ceiling effect, we targeted our comparison on patients and asked them to answer ten evaluation questions about the questionnaire they had just completed. RESULTS: 728 out of 1550 eligible patients (47%) participated in the study. Across questionnaires, response rates were similar (Saphora-Psy: 48.5%, Perceptions of Care: 49.9%, Picker: 43.4%; P = 0.08), average completion time was lowest for the Perceptions of Care questionnaire (minutes: Saphora-Psy: 17.7, Perceptions of Care: 13.7, Picker: 17.5; P = 0.005), the Saphora-Psy questionnaire had the largest mean proportion of missing responses (Saphora-Psy: 7.1%, Perceptions of Care: 2.8%, Picker: 4.0%; P < 0.001) and the Perceptions of Care questionnaire showed the highest ceiling effect (Saphora-Psy: 17.1%, Perceptions of Care: 41.9%, Picker: 36.3%; P < 0.001). There were no differences in the patients' evaluation of the questionnaires. CONCLUSION: Despite differences in the intended target population, content, lay-out and length of questionnaires, none appeared to be obviously better based on our comparison. All three presented advantages and drawbacks and could be used for the satisfaction evaluation of psychiatric inpatients. However, if comparison across medical services or hospitals is desired, using a generic questionnaire might be advantageous.

Efficacy of prescribed injectable diacetylmorphine in the Andalusian trial: Bayesian analysis of responders and non-responders according to a multi domain outcome index.

BACKGROUND The objective of this research was to evaluate data from a randomized clinical trial that tested injectable diacetylmorphine (DAM) and oral methadone (MMT) for substitution treatment, using a multi-domain dichotomous index, with a Bayesian approach. METHODS Sixty two long-term, socially-excluded heroin injectors, not benefiting from available treatments were randomized to receive either DAM or MMT for 9 months in Granada, Spain. Completers were 44 and data at the end of the study period was obtained for 50. Participants were determined to be responders or non responders using a multi-domain outcome index accounting for their physical and mental health and psychosocial integration, used in a previous trial. Data was analyzed with Bayesian methods, using information from a similar study conducted in The Netherlands to select a priori distributions. On adding the data from the present study to update the a priori information, the distribution of the difference in response rates were obtained and used to build credibility intervals and relevant probability computations. RESULTS In the experimental group (n = 27), the rate of responders to treatment was 70.4% (95% CI 53.287.6), and in the control group (n = 23), it was 34.8% (95% CI 15.354.3). The probability of success in the experimental group using the a posteriori distributions was higher after a proper sensitivity analysis. Almost the whole distribution of the rates difference (the one for diacetylmorphine minus methadone) was located to the right of the zero, indicating the superiority of the experimental treatment. CONCLUSION The present analysis suggests a clinical superiority of injectable diacetylmorphine compared to oral methadone in the treatment of severely affected heroin injectors not benefiting sufficiently from the available treatments. TRIAL REGISTRATION Current Controlled Trials ISRCTN52023186.

Cell size distribution in a random tessellation of space governed by the Kolmogorov-Johnson-Mehl-Avrami model: Grain size distribution in crystallization

The space subdivision in cells resulting from a process of random nucleation and growth is a subject of interest in many scientific fields. In this paper, we deduce the expected value and variance of these distributions while assuming that the space subdivision process is in accordance with the premises of the Kolmogorov-Johnson-Mehl-Avrami model. We have not imposed restrictions on the time dependency of nucleation and growth rates. We have also developed an approximate analytical cell size probability density function. Finally, we have applied our approach to the distributions resulting from solid phase crystallization under isochronal heating conditions