Trends in mortality from urologic cancers in Europe, 1970-2008.

BACKGROUND: In recent decades, there have been substantial changes in mortality from urologic cancers in Europe. OBJECTIVE: To provide updated information, we analyzed trends in mortality from cancer of the prostate, testis, bladder, and kidney in Europe from 1970 to 2008. DESIGN, SETTING, AND PARTICIPANTS: We derived data for 33 European countries from the World Health Organization database. MEASUREMENTS: We computed world-standardized mortality rates and used joinpoint regression to identify significant changes in trends. RESULTS AND LIMITATIONS: Mortality from prostate cancer has leveled off since the 1990s in countries of western and northern Europe, particularly over the last few years while it was still rising in Bulgaria, Romania, and Russia. In the European Union (EU), it reached a peak in 1995 at 15.0 per 100 000 men and declined to 12.5 per 100 000 in 2006. Mortality from testicular cancer has steadily declined in most countries in western and northern Europe since the 1970s. The declines were later and appreciably lower in central/eastern Europe. In EU, rates declined from 0.75 in 1980 to 0.32 per 100 000 men in 2006, with stronger declines up to the late 1990s and an apparent leveling off in rates thereafter. Over the last 15 years, mortality from bladder cancer has declined in most European countries in both sexes. The major exceptions were Bulgaria, Poland, and Romania. In the EU, bladder cancer mortality was stable until 1992 and declined thereafter from 7.3 to 5.5 per 100 000 men and from 1.5 to 1.2 per 100 000 women in 2006. Mortality from kidney cancer increased throughout Europe until the early 1990s and leveled off thereafter in many countries, except in a few central and eastern ones. Between 1994 and 2006, rates declined from 4.9 to 4.3 per 100 000 in EU men and from 2.1 to 1.8 per 100 000 in EU women. CONCLUSIONS: Over the last two decades, trends in urologic cancer mortality were favorable in Europe, with the exception of a few central and eastern countries.

Permian-Triassic of the Tethys: Carbon isotope sudies

Profiles of carbon isotopes were studied in marine limestones of Late Permian and Early Triassic age of the Tethyan region from 20 sections in Yugoslavia, Greece, Turkey, Armenian SSR, Iran, Pakistan, India, Nepal, and China. The Upper Permian sections continue the high positive values of 13C previously found in Upper Permian basins in NW Europe and western USA. In the more complete sections of Tethys it can now be demonstrated that the values of 13C drop from the Murgabian to the Dzhulfian Stages of the Upper Permian, then sharply to values near zero during the last two biozones of the Dorashamian. These levels of 13C sample the Tethys Sea and the world ocean, and equal values from deep-water sediments at Salamis Greece indicate that they apply to the whole water column. We hypothesize that the high values of 13C are a consequence of Late Paleozoic storage of organic carbon, and that the declines represent an episodic cessation of this organic deposition, and partial oxidation of the organic reservoir, extending over a period of several million years. The carbon isotope profile may reflect parallel complexity in the pattern of mass extinction in Late Permian time. Des profils isotopiques du carbone ont été établis dans des calcaires marins d'âge tardi-permien à  éo-triasique répartis dans 20 endroits du domaine téthysien: Yougoslavie, Grèce, Turquie, République d'Arménie, Iran, Pakistan, Inde, Népal et Chine. Les profils établis dans le Permien supérieur montrent les mêmes valeurs positives de 13C observées antérieurement dans des bassins de même âge en Europe occidentale et dans l'ouest des USA. Dans les profils les plus complets de la Téthys, il est maintenant établi que les valeurs de 13C décroissent depuis le Murgabien jusqu'au Dzhulfien (Permien supérieur) pour devenir proches de zéro dans les deux dernières biozones du Dorasharmen. Ces valeurs de 13C sont caractéristiques de la Téthys et de l'Océan mondial; elles s'appliquent à toutes les profondeurs d'eau, comme en témoignent les valeurs fournies par des sédiments de mer profonde à Salamis (Grèce). Nous formulons l'hypothèse que les hautes valeurs de 13C sont la conséquence du stockage du carbone organique au Paléozoïque supérieur et que leur décroissance traduit un arrêt épisodique de cette sédimentation organique, accompagné d'une oxydation partielle de la matière organique s'étendant sur une période de plusieurs Ma. L'influence parallèle des phénomènes d'extinction massive à le fin du Permien se refléterait également dans les profils isotopiques du carbone.

Evaluating bone marrow metastasis of neuroblastoma with iodine-123-MIBG scintigraphy and MRI.

Of 10 patients with neuroblastoma who had both 123I-MIBG scintigraphy and MRI at diagnosis, four presented with bone marrow metastasis that was diagnosed by both imaging modalities and confirmed by bone marrow biopsy and smears. This report focuses on the follow up of the four patients with bone marrow metastasis. MIBG scintigraphy and MRI were concordant in two patients, a case of normalization and a case of relapse in the seventh dorsal vertebra confirmed by surgical biopsy. The last two patients presented a normalized MIBG scan for marrow infiltration after chemotherapy but persistent abnormal MRI signal of several vertebrae, suggesting marrow infiltration, up to 27 mo after the end of chemotherapy in one case. In the second patient, MRI bone marrow aspect returned to normal 4 mo after the end of chemotherapy. Bone marrow biopsy remained negative in these two MIBG-negative patients. These cases suggest that in presence of complete normalization of the MIBG scan after chemotherapy, the persistence of a hypointense signal on bone marrow on T1WI does not necessarily indicate persistence of disease but may be due to delayed normalization. Therefore, attention must be paid to the delay of signal normalization on MRI (which can be as long as more than 2 yr after the end of chemotherapy) in order to avoid false-positive interpretation.

Occam's razor in minimally invasive pituitary surgery: tailoring the endoscopic endonasal uninostril trans-sphenoidal approach to sella turcica.

BACKGROUND: Since the introduction of the endoscopic endonasal approaches in the field of skull base surgery during the last two decades, several variants of the sella turcica endoscopic surgery have been described. The aim of this study is to provide a stepwise description of one of these variants in a minimally invasive/maximally efficient perspective. METHOD: For the majority of our sella turcica pathologies, we have progressively adopted a uninostril endoscopic approach that is very conservative towards the nasal mucosa with a very limited mucosal incision, resection of the vomer and allowing an almost ad integrum sellar floor reconstruction, without compromising the efficacy and completeness of both surgical oncologic and endocrine targets. CONCLUSION: The uninostril trans-sphenoidal endoscopic endonasal approach to sella turcica is tailored to ally maximal efficiency and minimal invasiveness.

Niche, distribution and global change : modeling insights into biogeography and conservation biology Olivier Broennimann

RESUME Les évidences montrant que les changements globaux affectent la biodiversité s'accumulent. Les facteurs les plus influant dans ce processus sont les changements et destructions d'habitat, l'expansion des espèces envahissantes et l'impact des changements climatiques. Une évaluation pertinente de la réponse des espèces face à ces changements est essentielle pour proposer des mesures permettant de réduire le déclin actuel de la biodiversité. La modélisation de la répartition d'espèces basée sur la niche (NBM) est l'un des rares outils permettant cette évaluation. Néanmoins, leur application dans le contexte des changements globaux repose sur des hypothèses restrictives et demande une interprétation critique. Ce travail présente une série d'études de cas investiguant les possibilités et limitations de cette approche pour prédire l'impact des changements globaux. Deux études traitant des menaces sur les espèces rares et en danger d'extinction sont présentées. Les caractéristiques éco-géographiques de 118 plantes avec un haut degré de priorité de conservation sont revues. La prévalence des types de rareté sont analysées en relation avec leur risque d'extinction UICN. La revue souligne l'importance de la conservation à l'échelle régionale. Une évaluation de la rareté à échelle globale peut être trompeuse pour certaine espèces car elle ne tient pas en compte des différents degrés de rareté que présente une espèce à différentes échelles spatiales. La deuxième étude test une approche pour améliorer l'échantillonnage d'espèces rares en incluant des phases itératives de modélisation et d'échantillonnage sur le terrain. L'application de l'approche en biologie de la conservation (illustrée ici par le cas du chardon bleu, Eryngium alpinum), permettrait de réduire le temps et les coûts d'échantillonnage. Deux études sur l'impact des changements climatiques sur la faune et la flore africaine sont présentées. La première étude évalue la sensibilité de 227 mammifères africains face aux climatiques d'ici 2050. Elle montre qu'un nombre important d'espèces pourrait être bientôt en danger d'extinction et que les parcs nationaux africains (principalement ceux situé en milieux xériques) pourraient ne pas remplir leur mandat de protection de la biodiversité dans le futur. La seconde étude modélise l'aire de répartition en 2050 de 975 espèces de plantes endémiques du sud de l'Afrique. L'étude propose l'inclusion de méthodes améliorant la prédiction des risques liés aux changements climatiques. Elle propose également une méthode pour estimer a priori la sensibilité d'une espèce aux changements climatiques à partir de ses propriétés écologiques et des caractéristiques de son aire de répartition. Trois études illustrent l'utilisation des modèles dans l'étude des invasions biologiques. Une première étude relate l'expansion de la laitue sáuvage (Lactuca serriola) vers le nord de l'Europe en lien avec les changements du climat depuis 250 ans. La deuxième étude analyse le potentiel d'invasion de la centaurée tachetée (Centaures maculosa), une mauvaise herbe importée en Amérique du nord vers 1890. L'étude apporte la preuve qu'une espèce envahissante peut occuper une niche climatique différente après introduction sur un autre continent. Les modèles basés sur l'aire native prédisent de manière incorrecte l'entier de l'aire envahie mais permettent de prévoir les aires d'introductions potentielles. Une méthode alternative, incluant la calibration du modèle à partir des deux aires où l'espèce est présente, est proposée pour améliorer les prédictions de l'invasion en Amérique du nord. Je présente finalement une revue de la littérature sur la dynamique de la niche écologique dans le temps et l'espace. Elle synthétise les récents développements théoriques concernant le conservatisme de la niche et propose des solutions pour améliorer la pertinence des prédictions d'impact des changements climatiques et des invasions biologiques. SUMMARY Evidences are accumulating that biodiversity is facing the effects of global change. The most influential drivers of change in ecosystems are land-use change, alien species invasions and climate change impacts. Accurate projections of species' responses to these changes are needed to propose mitigation measures to slow down the on-going erosion of biodiversity. Niche-based models (NBM) currently represent one of the only tools for such projections. However, their application in the context of global changes relies on restrictive assumptions, calling for cautious interpretations. In this thesis I aim to assess the effectiveness and shortcomings of niche-based models for the study of global change impacts on biodiversity through the investigation of specific, unsolved limitations and suggestion of new approaches. Two studies investigating threats to rare and endangered plants are presented. I review the ecogeographic characteristic of 118 endangered plants with high conservation priority in Switzerland. The prevalence of rarity types among plant species is analyzed in relation to IUCN extinction risks. The review underlines the importance of regional vs. global conservation and shows that a global assessment of rarity might be misleading for some species because it can fail to account for different degrees of rarity at a variety of spatial scales. The second study tests a modeling framework including iterative steps of modeling and field surveys to improve the sampling of rare species. The approach is illustrated with a rare alpine plant, Eryngium alpinum and shows promise for complementing conservation practices and reducing sampling costs. Two studies illustrate the impacts of climate change on African taxa. The first one assesses the sensitivity of 277 mammals at African scale to climate change by 2050 in terms of species richness and turnover. It shows that a substantial number of species could be critically endangered in the future. National parks situated in xeric ecosystems are not expected to meet their mandate of protecting current species diversity in the future. The second study model the distribution in 2050 of 975 endemic plant species in southern Africa. The study proposes the inclusion of new methodological insights improving the accuracy and ecological realism of predictions of global changes studies. It also investigates the possibility to estimate a priori the sensitivity of a species to climate change from the geographical distribution and ecological proprieties of the species. Three studies illustrate the application of NBM in the study of biological invasions. The first one investigates the Northwards expansion of Lactuca serriola L. in Europe during the last 250 years in relation with climate changes. In the last two decades, the species could not track climate change due to non climatic influences. A second study analyses the potential invasion extent of spotted knapweed, a European weed first introduced into North America in the 1890s. The study provides one of the first empirical evidence that an invasive species can occupy climatically distinct niche spaces following its introduction into a new area. Models fail to predict the current full extent of the invasion, but correctly predict areas of introduction. An alternative approach, involving the calibration of models with pooled data from both ranges, is proposed to improve predictions of the extent of invasion on models based solely on the native range. I finally present a review on the dynamic nature of ecological niches in space and time. It synthesizes the recent theoretical developments to the niche conservatism issues and proposes solutions to improve confidence in NBM predictions of the impacts of climate change and species invasions on species distributions.

Chirurgie endoscopique endonasale des tumeurs de la base du crane : historique, état de l'art et perspectives d'avenir [Shifting paradigm in skull base surgery: Roots, current state of the art and future trends of endonasal endoscopic approaches].

During the last two decades, endoscopic endonasal approach has completed the minimally invasive skull base surgery armamentarium. Endoscopic endonasal skull base surgery (EESBS) was initially developed in the field of pituitary adenomas, and gained an increasing place for the treatment of a wide variety of skull base pathologies, extending on the midline from crista galli process to the occipitocervical junction and laterally to the parasellar areas and petroclival apex. Until now, most studies are retrospective and lack sufficient methodological quality to confirm whether the endoscopic endonasal pituitary surgery has better results than the microsurgical trans-sphenoidal classical approach. The impressions of the expert teams show a trend toward better results for some pituitary adenomas with the endoscopic endonasal route, in terms of gross total resection rate and probably more comfortable postoperative course for the patient. Excepting intra- and suprasellar pituitary adenomas, EESBS seems useful for selected lesions extending onto the cavernous sinus and Meckel's cave but also for clival pathologies. Nevertheless, this infatuation toward endoscopic endonasal approaches has to be balanced with the critical issue of cerebrospinal fluid leaks, which constitutes actually the main limit of this approach. Through their experience and a review of the literature, the authors aim to present the state of the art of this approach as well as its limits.

Recent trends in colorectal cancer mortality in Europe.

Colorectal cancer mortality has been declining over the last two decades in Europe, particularly in women, the trends being, however, different across countries and age groups. We updated to 2007 colorectal cancer mortality trends in Europe using data from the World Health Organization (WHO). Rates were analyzed for the overall population and separately in young, middle-age and elderly populations. In the European Union (EU), between 1997 and 2007 mortality from colorectal cancer declined by around 2% per year, from 19.7 to 17.4/100,000 men (world standardized rates) and from 12.5 to 10.5/100,000 women. Persisting favorable trends were observed in countries of western and northern Europe, while there were more recent declines in several countries of eastern Europe, including the Czech Republic, Hungary and Slovakia particularly in women (but not Romania and the Russian Federation). In 2007, a substantial excess in colorectal cancer mortality was still observed in Slovakia, Hungary, Croatia, the Czech Republic and Slovenia in men (rates over 25/100,000), and in Hungary, Norway, Denmark and Slovakia in women (rates over 14/100,000). Colorectal mortality trends were more favorable in the young (30-49 years) from most European countries, with a decline of ∼2% per year since the early 1990s in both men and women from the EU. The recent decreases in colorectal mortality rates in several European countries are likely due to improvements in (early) diagnosis and treatment, with a consequent higher survival from the disease. Interventions to further reduce colorectal cancer burden are, however, still warranted, particularly in eastern European countries.

Combination of lentivector immunization and low-dose chemotherapy or PD-1/PD-L1 blocking primes self-reactive T cells and induces anti-tumor immunity.

In the last two decades, anti-cancer vaccines have yielded disappointing clinical results despite the fact that high numbers of self/tumor-specific T cells can be elicited in immunized patients. Understanding the reasons behind this lack of efficacy is critical in order to design better treatment regimes. Recombinant lentivectors (rLVs) have been successfully used to induce antigen-specific T cells to foreign or mutated tumor antigens. Here, we show that rLV expressing a murine nonmutated self/tumor antigen efficiently primes large numbers of self/tumor-specific CD8(+) T cells. In spite of the large number of tumor-specific T cells, however, no anti-tumor activity could be measured in a therapeutic setting, in mice vaccinated with rLV. Accumulating evidence shows that, in the presence of malignancies, inhibition of T-cell activity may predominate overstimulation. Analysis of tumor-infiltrating lymphocytes revealed that specific anti-tumor CD8(+) T cells fail to produce cytokines and express high levels of inhibitory receptors such as programmed death (PD)-1. Association of active immunization with chemotherapy or antibodies that block inhibitory pathways often leads to better anti-tumor effects. We show here that combining rLV vaccination with either cyclophosphamide or PD-1 and PD-L1 blocking antibodies enhances rLV vaccination efficacy and improves anti-tumor immunity.

Phenotypic plasticity in salmonid embryos : characterizing pathogen-induced stress effects on heritable variation in traits and reaction norms

Les pressions écologiques peuvent varier tant en nature qu'en intensité dans le temps et l'espace. C'est pourquoi, un phénotype unique ne peut pas forcément conférer la meilleure valeur sélective. La plasticité phénotypique peut être un moyen de s'accommoder de cette situation, en augmentant globalement la tolérance aux changements environnementaux. Comme pour tout trait de caractère, une variation génétique doit persister pour qu'évoluent les traits plastiques dans une population donnée. Cependant, les pressions extérieures peuvent affecter l'héritabilité, et la direction de ces changements peut dépendre du caractère en question, de l'espèce mais aussi du type de stress. Dans la présente thèse, nous avons cherché à élucider les effets des pressions pathogéniques sur les phénotypes et la génétique quantitative de plusieurs traits plastiques chez les embryons de deux salmonidés, la palée (Coregonus palaea), et la truite de rivière (Salmo trutta). Les salmonidés se prêtent à de telles études du fait de leur extraordinaire variabilité morphologique, comportementale et des traits d'histoire de vie. Par ailleurs, avec le déclin des salmonidés dans le monde, il est important de savoir combien la variabilité génétique persiste dans les normes de réaction afin d'aider à prédire leur capacité à répondre aux changements de leur milieu. Nous avons observé qu'une augmentation de la croissance des communautés microbiennes symbiotiques entraînait une mortalité accrue et une éclosion précoce chez la palée, et dévoilait la variance génétique additive pour ces deux caractères (Chapitres 1-2). Bien qu'aucune variation génétique n'ait été trouvée pour les normes de réaction, nous avons observé une variabilité de la plasticité d'éclosion. Néanmoins, on a trouvé que les temps d'éclosion étaient corrélés entre les environnements, ce qui pourrait limiter l'évolution de la norme de réaction. Le temps d'éclosion des embryons est lié à la taille des géniteurs mâles, ce qui indique des effets pléiotropiques. Dans le Chapitre 3, nous avons montré qu'une interaction triple entre la souche bactérienne {Pseudomonas fluorescens}, l'état de dévelopement de l'hôte ainsi que ses gènes ont une influence sur la mortalité, le temps d'éclosion et la taille des alevins de la palée. Nous avons démontré qu'une variation génétique subsistait généralement dans les normes de réaction des temps d'éclosion, mais rarement pour la taille des alevins, et jamais pour la mortalité. Dans le même temps, nous avons exhibé que des corrélations entre environnements dépendaient des caractères phénotypiques, mais contrairement au Chapitre 2, nous n'avons pas trouvé de preuve de corrélations transgénérationnelles. Le Chapitre 4 complète le chapitre précédent, en se plaçant du point de vue moléculaire, et décrit comment le traitement d'embryons avec P. fluorescens s'est traduit par une régulation négative d'expression du CMH-I indépendemment de la souche bactérienne. Nous avons non seulement trouvé une variation génétique des caractères phénotypiques moyens, mais aussi de la plasticité. Les deux derniers chapitres traitent de l'investigation, chez la truite de rivière, des différences spécifiques entre populations pour des normes de réaction induites par les pathogènes. Dans le Chapitre 5, nous avons illustré que le métissage entre des populations génétiquement distinctes n'affectait en rien la hauteur ou la forme des normes de réaction d'un trait précoce d'histoire de vie suite au traitement pathogénique. De surcroît, en dépit de l'éclosion tardive et de la réduction de la taille des alevins, le traitement n'a pas modifié la variation héritable des traits de caractère. D'autre part, dans le Chapitre 6, nous avons démontré que le traitement d'embryons avec des stimuli contenus dans l'eau de conspécifiques infectés a entraîné des réponses propre à chaque population en terme de temps d'éclosion ; néanmoins, nous avons observé peu de variabilité génétique des normes de réaction pour ce temps d'éclosion au sein des populations. - Ecological stressors can vary in type and intensity over space and time, and as such, a single phenotype may not confer the highest fitness. Phenotypic plasticity can act as a means to accommodate this situation, increasing overall tolerance to environmental change. As with any trait, for plastic traits to evolve in a population, genetic variation must persist. However, environmental stress can alter trait heritability, and the direction of this shift can be trait, species, and stressor-dependent. In this thesis, we sought to understand the effects of pathogen stressors on the phenotypes and genetic architecture of several plastic traits in the embryos of two salmonids, the whitefish (Coregonus palaea), and the brown trout (Salmo trutta). Salmonids lend themselves to such studies because their extraordinary variability in morphological, behavioral, and life-history traits. Also, with declines in salmonids worldwide, knowing how much genetic variability persists in reaction norms may help predict their ability to respond to environmental change. We found that increasing growth of symbiotic microbial communities increased mortality and induced hatching in whitefish, and released additive genetic variance for both traits (Chapters 1-2). While no genetic variation was found for survival reaction norms, we did find variability in hatching plasticity. Nevertheless, hatching time was correlated across environments, which could constrain evolution of the reaction norm. Hatching time in the induced environment was also correlated to sire size, indicating pleiotropic effects. In Chapter 3 we report that a three-way interaction between bacterial strain (Pseudomonas fluorescens), host developmental stage, and host genetics impacted mortality, hatching time, and hatchling size in whitefish. We also showed that genetic variation generally persisted in hatching age reaction norms, but rarely for hatchling length, and never for mortality. At the same time, we demonstrated that cross-environmental correlations were trait-dependent, and unlike Chapter 2, we found no evidence of cross-generational correlations. Chapter 4 expands on the previous chapter, moving to the molecular level, and describes how treatment of embryos with P. fluorescens resulted in strain-independent downregulation of MHC class I. Genetic variation was evident not only in trait means, but also in plasticity. In the last two chapters, we investigated population level differences in pathogen- induced reaction norms in brown trout. In Chapter 5, we found that interbreeding between genetically distinct populations did not affect the elevation or shapes of the reaction norms of early life-history traits after pathogen challenge. Moreover, despite delaying hatching and reducing larval length, treatment produced no discernable shifts in heritable variation in traits. On the other hand, in Chapter 6, we found that treatment of embryos with water-borne cues from infected conspecifics elicited population-specific responses in terms of hatching time; however, we found little evidence of genetic variability in hatching reaction norms within populations. We have made considerable progress in understanding how pathogen stressors affect various early life-history traits in salmonid embryos. We have demonstrated that the effect of a particular stressor on heritable variation in these traits can vary according to the trait and species under consideration, in addition to the developmental stage of the host. Moreover, we found evidence of genetic variability in some, but not all reaction norms in whitefish and brown trout.

Sepsis: time to reconsider the concept.

OBJECTIVE: To discuss the difficulty in using the concept of sepsis for clinical trials and propose new ways for designing future trials for severe infections. DESIGN: Short position statement. METHODS AND MAIN RESEARCH: Using a thorough evaluation of the recent literature in the field of severe sepsis and septic shock, the authors challenge the concept of sepsis as used in the past two decades and propose new ideas for designing future trials in this setting. The two main proposals are first to use a systematic assessment of the targeted inflammatory mediators when the study intends to counteract or replace those mediators (e.g., anti-tumor necrosis factor-alpha, activated protein C) and, second, to select more homogeneous populations, coming back to "precise infectious diseases," such as severe community-acquired pneumonia, severe peritonitis, or meningitis. CONCLUSIONS: The concept of sepsis has been useful to help clinicians to suspect and detect severe infections. Due to a considerable heterogeneity in the patients and type of infections included in the trials performed in the last two decades, it has not been useful in demonstrating the efficacy of new compounds. The authors propose a dramatic change in the design of future trials dealing with severe infections.

Quantify this! Report on a round table discussion on quantitative mass spectrometry in proteomics.

Following the success of the first round table in 2001, the Swiss Proteomic Society has organized two additional specific events during its last two meetings: a proteomic application exercise in 2002 and a round table in 2003. Such events have as their main objective to bring together, around a challenging topic in mass spectrometry, two groups of specialists, those who develop and commercialize mass spectrometry equipment and software, and expert MS users for peptidomics and proteomics studies. The first round table (Geneva, 2001) entitled "Challenges in Mass Spectrometry" was supported by brief oral presentations that stressed critical questions in the field of MS development or applications (Stöcklin and Binz, Proteomics 2002, 2, 825-827). Topics such as (i) direct analysis of complex biological samples, (ii) status and perspectives for MS investigations of noncovalent peptide-ligant interactions; (iii) is it more appropriate to have complementary instruments rather than a universal equipment, (iv) standardization and improvement of the MS signals for protein identification, (v) what would be the new generation of equipment and finally (vi) how to keep hardware and software adapted to MS up-to-date and accessible to all. For the SPS'02 meeting (Lausanne, 2002), a full session alternative event "Proteomic Application Exercise" was proposed. Two different samples were prepared and sent to the different participants: 100 micro g of snake venom (a complex mixture of peptides and proteins) and 10-20 micro g of almost pure recombinant polypeptide derived from the shrimp Penaeus vannamei carrying an heterogeneous post-translational modification (PTM). Among the 15 participants that received the samples blind, eight returned results and most of them were asked to present their results emphasizing the strategy, the manpower and the instrumentation used during the congress (Binz et. al., Proteomics 2003, 3, 1562-1566). It appeared that for the snake venom extract, the quality of the results was not particularly dependant on the strategy used, as all approaches allowed Lication of identification of a certain number of protein families. The genus of the snake was identified in most cases, but the species was ambiguous. Surprisingly, the precise identification of the recombinant almost pure polypeptides appeared to be much more complicated than expected as only one group reported the full sequence. Finally the SPS'03 meeting reported here included a round table on the difficult and challenging task of "Quantification by Mass Spectrometry", a discussion sustained by four selected oral presentations on the use of stable isotopes, electrospray ionization versus matrix-assisted laser desorption/ionization approaches to quantify peptides and proteins in biological fluids, the handling of differential two-dimensional liquid chromatography tandem mass spectrometry data resulting from high throughput experiments, and the quantitative analysis of PTMs. During these three events at the SPS meetings, the impressive quality and quantity of exchanges between the developers and providers of mass spectrometry equipment and software, expert users and the audience, were a key element for the success of these fruitful events and will have definitively paved the way for future round tables and challenging exercises at SPS meetings.

Atomic force microscopy imaging of living cells

Over the last two decades, Atomic Force Microscopy (AFM) has emerged as the tool of choice to image living organisms in a near-physiological environment. Whereas fluorescence microscopy techniques allow labeling and tracking of components inside cells and the observation of dynamic processes, AFM is mainly a surface technique that can be operated on a wide range of substrates including biological samples. AFM enables extraction of topographical, mechanical and chemical information from these samples.

Postmortem chemistry update part II.

As a continuation of "Postmortem Chemistry Update Part I," Part II deals with molecules linked to liver and cardiac functions, alcohol intake and alcohol misuse, myocardial ischemia, inflammation, sepsis, anaphylaxis, and hormonal disturbances. A very important array of new material concerning these situations had appeared in the forensic literature over the last two decades. Some molecules, such as procalcitonin and C-reactive protein, are currently researched in cases of suspected sepsis and inflammation, whereas many other analytes are not integrated into routine casework. As in part I, a literature review concerning a large panel of molecules of forensic interest is presented, as well as the results of our own observations, where possible.

Monocarboxylate transporters: new players in body weight regulation.

Over the last two decades, several genes have been identified that appear to play a role in the regulation of energy homeostasis and body weight. For a small subset of them, a reduction or an absence of expression confers a resistance to the development of obesity. Recently, a knockin mouse for a member of the monocarboxylate transporter (MCT) family, MCT1, was demonstrated to exhibit a typical phenotype of resistance to diet-induced obesity and a protection from its associated metabolic perturbations. Such findings point out at MCTs as putatively new therapeutic targets in the context of obesity. Here, we will review what is known about MCTs and their possible metabolic roles in different organs and tissues. Based on the description of the phenotype of the MCT1 knockin mouse, we will also provide some insights about their putative roles in weight gain regulation.

Investigation of resins suitable for the preparation of biological sample for 3-D electron microscopy.

In the last two decades, the third-dimension has become a focus of attention in electron microscopy to better understand the interactions within subcellular compartments. Initially, transmission electron tomography (TEM tomography) was introduced to image the cell volume in semi-thin sections (∼500nm). With the introduction of the focused ion beam scanning electron microscope, a new tool, FIB-SEM tomography, became available to image much larger volumes. During TEM tomography and FIB-SEM tomography, the resin section is exposed to a high electron/ion dose such that the stability of the resin embedded biological sample becomes an important issue. The shrinkage of a resin section in each dimension, especially in depth, is a well-known phenomenon. To ensure the dimensional integrity of the final volume of the cell, it is important to assess the properties of the different resins and determine the formulation which has the best stability in the electron/ion beam. Here, eight different resin formulations were examined. The effects of radiation damage were evaluated after different times of TEM irradiation. To get additional information on mass-loss and the physical properties of the resins (stiffness and adhesion), the topography of the irradiated areas was analysed with atomic force microscopy (AFM). Further, the behaviour of the resins was analysed after ion milling of the surface of the sample with different ion currents. In conclusion, two resin formulations, Hard Plus and the mixture of Durcupan/Epon, emerged that were considerably less affected and reasonably stable in the electron/ion beam and thus suitable for the 3-D investigation of biological samples.