The Lausanne experience with the Wichita fusion nail for arthrodesis of the knee

Background: Arthrodesis of the knee by intramedullary fixation hasbeen reported to have a higher rate of success than external fixationor compression plating. Antegrade nailing however can lead to complicationsdue to the different diameters of the medullary canals, fracturesduring insertion, poor rotational stability, breakage of the IM-nailand insufficient compression at the fusion site.Method: This retrospective study reports all knee fusions performedby the same orthopaedic surgeon with the Wichita (Stryker) fusion nail(WFN) from 2004 to 2010. The Wichita nail is a short nail with a deviceat the knee which allows for coupling of differently sized and interlockedfemoral and tibial components and at the same time for compression.Results: We report of 18 patients with a mean follow up of 28 months(range 3-71 months). Infected TKA was the most common indicationfor arthrodesis in 9 cases. The remaining reasons included asepticfailed TKA in 3 cases, 2 patients after fracture, 1 patient with neurologicalinstability after knee dislocation, 1 patient after tumoral resectionand 1 non union after failed arthrodesis with long antegrade nail.Finally 1 patient with bilateral congenital knee dislocation operated onboth sides. As expected, patients receiving the WFN had undergonea large number of previous knee surgeries with a mean of 3.8 (range0-8) procedures per patient. The complication rate was 27% (5 of 18).Two patients had persistent pain requiring revision surgery to increasestability with plating. One case of periprosthetic fracture needed openreduction and internal fixation. 2 patients with superficial hematomawere treated one with open drainage and the other with physiotherapy.Infection was erradicated in all septic cases, we found no new infectionand the fusion rate was 100%.Conclusion: The results in these often difficult cases are satisfyingand we think that this technique is a valid alternative to the otherknown techniques of knee fusion in patients with a poor bone stockand fragile soft tissues.

Does the sun shine by p p or CNO fusion reactions?

We show that solar neutrino experiments set an upper limit of 7.8% (7.3% including the recent KamLAND measurements) to the fraction of energy that the Sun produces via the CNO fusion cycle, which is an order of magnitude improvement upon the previous limit. New experiments are required to detect CNO neutrinos corresponding to the 1.5% of the solar luminosity that the standard solar model predicts is generated by the CNO cycle.

Trans-SNARE pairing can precede a hemifusion intermediate in intracellular membrane fusion.

The question concerning whether all membranes fuse according to the same mechanism has yet to be answered satisfactorily. During fusion of model membranes or viruses, membranes dock, the outer membrane leaflets mix (termed hemifusion), and finally the fusion pore opens and the contents mix. Viral fusion proteins consist of a membrane-disturbing 'fusion peptide' and a helical bundle that pin the membranes together. Although SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complexes form helical bundles with similar topology, it is unknown whether SNARE-dependent fusion events on intracellular membranes proceed through a hemifusion state. Here we identify the first hemifusion state for SNARE-dependent fusion of native membranes, and place it into a sequence of molecular events: formation of helical bundles by SNAREs precedes hemifusion; further progression to pore opening requires additional peptides. Thus, SNARE-dependent fusion may proceed along the same pathway as viral fusion: both use a docking mechanism via helical bundles and additional peptides to destabilize the membrane and efficiently induce lipid mixing. Our results suggest that a common lipidic intermediate may underlie all fusion reactions of lipid bilayers.

Minimally invasive versus open transforaminal lumbar interbody fusion: evaluating initial experience.

The aim of this study was to compare our experience with minimally invasive transforaminal lumbar interbody fusion (MITLIF) and open midline transforaminal lumbar interbody fusion (TLIF). A total of 36 patients suffering from isthmic spondylolisthesis or degenerative disc disease were operated with either a MITLIF (n = 18) or an open TLIF technique (n = 18) with an average follow-up of 22 and 24 months, respectively. Clinical outcome was assessed using the visual analogue scale (VAS) and the Oswestry disability index (ODI). There was no difference in length of surgery between the two groups. The MITLIF group resulted in a significant reduction of blood loss and had a shorter length of hospital stay. No difference was observed in postoperative pain, initial analgesia consumption, VAS or ODI between the groups. Three pseudarthroses were observed in the MITLIF group although this was not statistically significant. A steeper learning effect was observed for the MITLIF group.

Comment on: Deuterium nuclear fusion at room temperature: A pertinent inequality on barrier penetration

In a recent letter, Rosen claims to justify ...

Recent misconceptions about the "database search problem": a probabilistic analysis using Bayesian networks

This paper analyses and discusses arguments that emerge from a recent discussion about the proper assessment of the evidential value of correspondences observed between the characteristics of a crime stain and those of a sample from a suspect when (i) this latter individual is found as a result of a database search and (ii) remaining database members are excluded as potential sources (because of different analytical characteristics). Using a graphical probability approach (i.e., Bayesian networks), the paper here intends to clarify that there is no need to (i) introduce a correction factor equal to the size of the searched database (i.e., to reduce a likelihood ratio), nor to (ii) adopt a propositional level not directly related to the suspect matching the crime stain (i.e., a proposition of the kind 'some person in (outside) the database is the source of the crime stain' rather than 'the suspect (some other person) is the source of the crime stain'). The present research thus confirms existing literature on the topic that has repeatedly demonstrated that the latter two requirements (i) and (ii) should not be a cause of concern.

The role of the vacuolar H+ - ATPase in membrane fusion

RésuméLa H+-ATPase vacuolaire (V-ATPase) est un complexe enzymatique composé de deux secteurs multimériques (VQ et Vi) dont l'association dans la cellule est réversible. Le secteur intramembranaire de la V-ATPase (V0) interagit physiquement avec des protéines SNARE et stimule la fusion homotypique des vacuoles de la levure (lysosomes), la sécrétion de neurotransmetteurs et d'insuline, la fusion entre phagosome et lysosome ainsi que la sécrétion des corps multivésiculaires par un mécanisme inconnu. Dans cette étude j'ai identifié des résidues d'acides amines situés dans des sous-unités de V0 impliqués dans le mécanisme de fusion des vacuoles mais non essentiels pour l'acidification vacuolaire par la V-ATPase. j'ai utilisé un protocole de mutagenèse aléatoire pour produire des libraries de mutants des sous unités de V0. Ces libraries ont été analysées in vivo afin d'identifier des alleles qui permettent la translocation des protons mais produisent une vacuole fragmentée, phénotype indiquant un défaut dans la fusion membranaire. Les vacuoles des mutants ont été isolées et caractéisées en utilisant une grande variété d'outils biochimiques pour déterminer précisément l'impact des différentes mutations sur l'accomplissement d'événements clés du processus de fusion.J'ai identifié des mutations associées à des défauts spécifiques de la fusion dans plusieurs sous-unités de V0. Dans les protéolipides c, c' et c" ces mutations se concentrent dans la partie cytosolique des domaines transmembranaires. Elles renforcent les associations entre les secteurs de la V-ATPase et entre V0 et les SNAREs. Dans la fusion vacuolaire ces mutations permettent la formation de complexes SNAREs en trans mais inhibent l'induction de la fusion. Par contre, la deletion de la sous- unité d influence les étapes de la fusion qui précèdent la formation des complexes trans-SNAREs. Mes résultats démontrent que V0 joue des rôles différents dans plusieurs étapes de la fusion et que ces fonctions sont liées au système des SNAREs. Ils différencient génétiquement les activités de V0 dans la translocation des protons et dans la fusion et identifient de nombreux résidus importants pour la fusion vacuolaire. De plus, compte tenu de la grande conservation de sequence des protéolipides chez les eukaryotes les mutations identifiées dans cette l'étude apportent de nouvelles informations pour analyser la fonction de V0 dans des organismes multicellulaires pour lesquels la function catalytique de la V-ATPase est essentielle à la survie.Résumé pour le large publicLe transport de protéines et de membranes est important pour maintenir la fonction des organelles dans la cellule. Il s'excerce au niveau des vesicules. La fusion membranaire est un processus élémentaire de ce transport. Pour fusionner deux membranes, il faut la coordination de deux activités: le rapprochement et la déstabiiization des deux membranes. La collaboration d'un ensemble de proteins conservés chez les eukaryotes, est nécessaire pour catalyser ces activités. Les proteins SNAREs sont les protagonistes principaux dans la fusion membranaire. Néanmoins, d'autres protéines, comme des Rab-GTPases et des chaperonnes, sont nécessaires pour permettre ce phénomène de fusion. Toutes ces protéines sont temporairement associées avec les SNAREs et leur fonction dans la fusion membranaire est souvent directement liée à leur activité dans cette association. Le secteur transmembranaire V0 de la V-ATPase rnteragit avec des SNAREs et est essentiel pour la fusion dans une variété de systèmes modèles comme la mouche, la souris et la levure. Le secteur V0 est composé de six protéines différentes. Avec te secteur Va, qui réside dans le cytosol, il forme la V-ATPase dont la fonction principale est l'acidification des organelles par translocation des protons à travers la membrane par un mécanisme ressemblant à celui d'une pompe. V0joue un role dans la fusion membranaire, indépendamment de son activité catalytique liée au pompage des protons, et ce rôle est encore largement méconnu à ce jour. Le but de ma thèse était de mieux comprendre l'implication de V0 dans ce contexte.Pour étudier des activités liées à la V-ATPase, la levure est un excellent modèle d'étude car elle survie à une inactivation de l'enzyme alors que le meme traitement serait léthal pour des organismes multicellulaires. Dans ma thèse j'ai utilisé la fusion homotypique de la vacuole de levure comme système modèle pour étudier le rôle de V0 dans la fusion. J'ai muté des gènes qui encodent des sous- unités de V0 et les ai introduit dans des souches privées des gènes respectifs. Dans les librairies de souches portant différentes versions de ces gènes j'ai cherché des clones exprimant une V-ATPase intacte et fonctionnelle mais qui possèdent une vacuole fragmentée. Le plus souvent, une vacuole fragmentée indique un défaut dans la fusion vacuolaire. Dans les trois types de protéolipides qui composent un cylindre dans le secteur V0, j'ai trouvé des clones avec une vacuole fragmentée. Après avoir isolé les mutations responsable de ce type de morphologie vacuolaire, j'ai isolé les vacuoles de ces clones pour étudier leur activités dans différentes étapes de la fusion vacuolaire. Les résultats de ces analyses mettent en évidence une implication de V0 dans plusieurs étapes de la fusion vacuolaire. Certaines mutations sélectionnées dans mon étude inhibent une étape précoce de la fusion qui inclue la dissociation des complexes SNARE, tandis que d'autres mutations inhibent une étape tardive du processus de fusion qui inclue la transmission d'une force disruptive dans la membrane.AbstractThe membrane-integral V0 sector of the vacuolar H+-ATPase (V-ATPase) interacts with SNARE proteins. V0 stimulates fusion between yeast vacuoles (lysosomes) (Peters et al., 2001b), secretion of neurotransmitters and insulin (Hiesinger et al., 2005a, Sun-Wada et al., 2006a), phagosome-lysosome fusion (Peri and Nusslein-Volhard, 2008) and secretion of multivesicular bodies (Liegeois et al., 2006b) by a yet unknown mechanism. In my thesis, I identified sites in V0 subunits that are involved in yeast vacuole fusion but dispensable for the proton pumping by the V-ATPase. I randomly mutagenized V0 subunits and screened in vivo for mutant alleles that support proton pumping but cause fragmented vacuoles, a phenotype indicative of a fusion defect. Mutant vacuoles were isolated and analyzed in a cell-free system, allowing assay of key events in fusion, such as trans-SNARE pairing, lipid transition and fusion pore opening (Reese et al., 2005b).Mutants with selective fusion defects were found in several V0 subunits. In the proteolipids c, c' and c", critical mutations are concentated in the cytosolic half of the transmembrane domains. These mutations rendered the V-ATPase holoenzyme more stable and modulated V0-SNARE associations. In vacuole fusion critical proteolipid mutations permitted trans-SNARE pairing but impeded the induction of lipid flow between the membranes. Deletion of subunit d, by contrast, influenced early stages of fusion that precede trans-SNARE pairing. My results show that V0 acts in several steps of the fusion process and that its function is intimately connected to the SNARE system. They genetically separate the proton pump and fusion activities of V0 and identify numerous critical residues. Given the high sequence conservation of proteolipids in eukaryotic life, the identified mutations may be helpful in analyzing the fusion function of V0 also in mammalian cells, where V- ATPase pump function is essential for survival.

Mutual control of membrane fission and fusion proteins.

Membrane fusion and fission are antagonistic reactions controlled by different proteins. Dynamins promote membrane fission by GTP-driven changes of conformation and polymerization state, while SNAREs fuse membranes by forming complexes between t- and v-SNAREs from apposed vesicles. Here, we describe a role of the dynamin-like GTPase Vps1p in fusion of yeast vacuoles. Vps1p forms polymers that couple several t-SNAREs together. At the onset of fusion, the SNARE-activating ATPase Sec18p/NSF and the t-SNARE depolymerize Vps1p and release it from the membrane. This activity is independent of the SNARE coactivator Sec17p/alpha-SNAP and of the v-SNARE. Vps1p release liberates the t-SNAREs for initiating fusion and at the same time disrupts fission activity. We propose that reciprocal control between fusion and fission components exists, which may prevent futile cycles of fission and fusion.

Mechanism of reduction of virus release and cell-cell fusion in persistent canine distemper virus infection.

Canine distemper virus (CDV), a mobillivirus related to measles virus causes a chronic progressive demyelinating disease, associated with persistence of the virus in the central nervous system (CNS). CNS persistence of morbilliviruses has been associated with cell-to-cell spread, thereby limiting immune detection. The mechanism of cell-to-cell spread remains uncertain. In the present study we studied viral spread comparing a cytolytic (non-persistent) and a persistent CDV strain in cell cultures. Cytolytic CDV spread in a compact concentric manner with extensive cell fusion and destruction of the monolayer. Persistent CDV exhibited a heterogeneous cell-to-cell pattern of spread without cell fusion and 100-fold reduction of infectious viral titers in supernatants as compared to the cytolytic strain. Ultrastructurally, low infectious titers correlated with limited budding of persistent CDV as compared to the cytolytic strain, which shed large numbers of viral particles. The pattern of heterogeneous cell-to-cell viral spread can be explained by low production of infectious viral particles in only few areas of the cell membrane. In this way persistent CDV only spreads to a small proportion of the cells surrounding an infected one. Our studies suggest that both cell-to-cell spread and limited production of infectious virus are related to reduced expression of fusogenic complexes in the cell membrane. Such complexes consist of a synergistic configuration of the attachment (H) and fusion (F) proteins on the cell surface. F und H proteins exhibited a marked degree of colocalization in cytolytic CDV infection but not in persistent CDV as seen by confocal laser microscopy. In addition, analysis of CDV F protein expression using vaccinia constructs of both strains revealed an additional large fraction of uncleaved fusion protein in the persistent strain. This suggests that the paucity of active fusion complexes is due to restricted intracellular processing of the viral fusion protein.

Bayesian estimation of speciation and extinction from incomplete fossil occurrence data.

The temporal dynamics of species diversity are shaped by variations in the rates of speciation and extinction, and there is a long history of inferring these rates using first and last appearances of taxa in the fossil record. Understanding diversity dynamics critically depends on unbiased estimates of the unobserved times of speciation and extinction for all lineages, but the inference of these parameters is challenging due to the complex nature of the available data. Here, we present a new probabilistic framework to jointly estimate species-specific times of speciation and extinction and the rates of the underlying birth-death process based on the fossil record. The rates are allowed to vary through time independently of each other, and the probability of preservation and sampling is explicitly incorporated in the model to estimate the true lifespan of each lineage. We implement a Bayesian algorithm to assess the presence of rate shifts by exploring alternative diversification models. Tests on a range of simulated data sets reveal the accuracy and robustness of our approach against violations of the underlying assumptions and various degrees of data incompleteness. Finally, we demonstrate the application of our method with the diversification of the mammal family Rhinocerotidae and reveal a complex history of repeated and independent temporal shifts of both speciation and extinction rates, leading to the expansion and subsequent decline of the group. The estimated parameters of the birth-death process implemented here are directly comparable with those obtained from dated molecular phylogenies. Thus, our model represents a step towards integrating phylogenetic and fossil information to infer macroevolutionary processes.

Cointegrase, a naturally occurring, truncated form of IS21 transposase, catalyzes replicon fusion rather than simple insertion of IS21.

The bacterial insertion sequence IS21 contains two genes, istA and istB, which are organized as an operon. IS21 spontaneously forms tandem repeats designated (IS21)2. Plasmids carrying (IS21)2 react efficiently with other replicons, producing cointegrates via a cut-and-paste mechanism. Here we show that transposition of a single IS21 element (simple insertion) and cointegrate formation involving (IS21)2 result from two distinct non-replicative pathways, which are essentially due to two differentiated IstA proteins, transposase and cointegrase. In Escherichia coli, transposase was characterized as the full-length, 46 kDa product of the istA gene, whereas the 45 kDa cointegrase was expressed, in-frame, from a natural internal translation start of istA. The istB gene, which could be experimentally disconnected from istA, provided a helper protein that strongly stimulated the transposase and cointegrase-driven reactions. Site-directed mutagenesis was used to express either cointegrase or transposase from the istA gene. Cointegrase promoted replicon fusion at high frequencies by acting on IS21 ends which were linked by 2, 3, or 4 bp junction sequences in (IS21)2. By contrast, cointegrase poorly catalyzed simple insertion of IS21 elements. Transposase had intermediate, uniform activity in both pathways. The ability of transposase to synapse two widely spaced IS21 ends may reside in the eight N-terminal amino acid residues which are absent from cointegrase. Given the 2 or 3 bp spacing in naturally occurring IS21 tandems and the specialization of cointegrase, the fulminant spread of IS21 via cointegration can now be understood.

MBIS: multivariate Bayesian image segmentation tool.

We present MBIS (Multivariate Bayesian Image Segmentation tool), a clustering tool based on the mixture of multivariate normal distributions model. MBIS supports multichannel bias field correction based on a B-spline model. A second methodological novelty is the inclusion of graph-cuts optimization for the stationary anisotropic hidden Markov random field model. Along with MBIS, we release an evaluation framework that contains three different experiments on multi-site data. We first validate the accuracy of segmentation and the estimated bias field for each channel. MBIS outperforms a widely used segmentation tool in a cross-comparison evaluation. The second experiment demonstrates the robustness of results on atlas-free segmentation of two image sets from scan-rescan protocols on 21 healthy subjects. Multivariate segmentation is more replicable than the monospectral counterpart on T1-weighted images. Finally, we provide a third experiment to illustrate how MBIS can be used in a large-scale study of tissue volume change with increasing age in 584 healthy subjects. This last result is meaningful as multivariate segmentation performs robustly without the need for prior knowledge.