Parts, places, and perspectives : a theory of spatial relations based an mereotopology and convexity

This thesis suggests to carry on the philosophical work begun in Casati's and Varzi's seminal book Parts and Places, by extending their general reflections on the basic formal structure of spatial representation beyond mereotopology and absolute location to the question of perspectives and perspective-dependent spatial relations. We show how, on the basis of a conceptual analysis of such notions as perspective and direction, a mereotopological theory with convexity can express perspectival spatial relations in a strictly qualitative framework. We start by introducing a particular mereotopological theory, AKGEMT, and argue that it constitutes an adequate core for a theory of spatial relations. Two features of AKGEMT are of particular importance: AKGEMT is an extensional mereotopology, implying that sameness of proper parts is a sufficient and necessary condition for identity, and it allows for (lower- dimensional) boundary elements in its domain of quantification. We then discuss an extension of AKGEMT, AKGEMTS, which results from the addition of a binary segment operator whose interpretation is that of a straight line segment between mereotopological points. Based on existing axiom systems in standard point-set topology, we propose an axiomatic characterisation of the segment operator and show that it is strong enough to sustain complex properties of a convexity predicate and a convex hull operator. We compare our segment-based characterisation of the convex hull to Cohn et al.'s axioms for the convex hull operator, arguing that our notion of convexity is significantly stronger. The discussion of AKGEMTS defines the background theory of spatial representation on which the developments in the second part of this thesis are built. The second part deals with perspectival spatial relations in two-dimensional space, i.e., such relations as those expressed by 'in front of, 'behind', 'to the left/right of, etc., and develops a qualitative formalism for perspectival relations within the framework of AKGEMTS. Two main claims are defended in part 2: That perspectival relations in two-dimensional space are four- place relations of the kind R(x, y, z, w), to be read as x is i?-related to y as z looks at w; and that these four-place structures can be satisfactorily expressed within the qualitative theory AKGEMTS. To defend these two claims, we start by arguing for a unified account of perspectival relations, thus rejecting the traditional distinction between 'relative' and 'intrinsic' perspectival relations. We present a formal theory of perspectival relations in the framework of AKGEMTS, deploying the idea that perspectival relations in two-dimensional space are four-place relations, having a locational and a perspectival part and show how this four-place structure leads to a unified framework of perspectival relations. Finally, we present a philosophical motivation to the idea that perspectival relations are four-place, cashing out the thesis that perspectives are vectorial properties and argue that vectorial properties are relations between spatial entities. Using Fine's notion of "qua objects" for an analysis of points of view, we show at last how our four-place approach to perspectival relations compares to more traditional understandings.

On-line desorption of dried blood spot: A novel approach for the direct LC/MS analysis of micro-whole blood samples.

The aim of this work is to present a new concept, called on-line desorption of dried blood spots (on-line DBS), allowing the direct analysis of a dried blood spot coupled to liquid chromatography mass spectrometry device (LC/MS). The system is based on an inox cell which can receive a blood sample (10 microL) previously spotted on a filter paper. The cell is then integrated into LC/MS system where the analytes are desorbed out of the paper towards a column switching system ensuring the purification and separation of the compounds before their detection on a single quadrupole MS coupled to atmospheric pressure chemical ionisation (APCI) source. The described procedure implies that no pretreatment is necessary in spite the analysis is based on whole blood sample. To ensure the applicability of the concept, saquinavir, imipramine, and verapamil were chosen. Despite the use of a small sampling volume and a single quadrupole detector, on-line DBS allowed the analyses of these three compounds over their therapeutic concentrations from 50 to 500 ng/mL for imipramine and verapamil and from 100 to 1000 ng/mL for saquinavir. Moreover, the method showed good repeatability with relative standard deviation (RSD) lower than 15% based on two levels of concentration (low and high). Function responses were found to be linear over the therapeutic concentration for each compound and were used to determine the concentrations of real patient samples for saquinavir. Comparison of the founded values with those of a validated method used routinely in a reference laboratory showed a good correlation between the two methods. Moreover, good selectivity was observed ensuring that no endogenous or chemical components interfered with the quantitation of the analytes. This work demonstrates the feasibility and applicability of the on-line DBS procedure for bioanalysis.

Événements et déformations tardi-métamorphiques dans le segment Ossola-Ticino (Val Vigezzo-Centovalli, Italie-Suisse)

RESUME: Une zone tectonique large et complexe, connue sous le nom de ligne des Centovalli, traverse le secteur des Alpes Centrales compris entre Domodossola et Locarno. Cette région, formée par le Val Vigezzo et la vallée des Centovalli, constitue la terminaison méridionale du dôme Lepontin et représente une portion de la zone des racines des nappes alpines. Elle fait partie d’une grande et complexe zone de cisaillement, en partie associée à des phénomènes hydrothermaux d’âge alpin (<20 Ma), qui comprend le système tectonique Insubrien et celui du Simplon. Le Val Vigezzo et les Centovalli constituent un vrai carrefour entre les principaux accidents tectoniques des Alpes ainsi qu'une zone de juxtaposition du socle Sudalpin avec la zone des racines de l’Austroalpin et du Pennique. Les phases de déformation et les structures géologiques qui peuvent être étudiées s'étalent sur une période comprise entre environ 35 Ma et l'actuel. L’étude détaillée de terrain a mis en évidence la présence de nombreuses roches et structures de déformation de type ductile et cassant tels que des mylonites, des cataclasites, des pseudotachylites, des kakirites, des failles minéralisées, des gouges de faille et des plis. Sur le terrain on a pu distinguer au moins quatre générations de plis liés aux différentes phases de déformation. Le nombre et la complexité de ces structures indiquent une histoire très compliquée, selon plusieurs étapes distinctes, parfois liées, voire même superposées. Une partie de ces structures de déformation affectent aussi les dépôts sédimentaires d’âge quaternaire, notamment des limons et des sables lacustres. Ces sédiments constituent les restes d'un bassin lacustre attribué à l'époque interglaciaire Riss/Würm (éemien, 67.000-120.000 ans) et ils affleurent dans la partie centrale de la zone étudiée, à l'Est de la plaine de Santa Maria Maggiore. Ces sédiments montrent en leur sein toute une série de structures de déformation tels que des plans de faille inverses, des structures conjuguées de raccourcissement et des véritables plis. Ces failles et ces plis représenteraient les évidences de surface d’une déformation probablement active en époque quaternaire. Une autre formation rocheuse a retenu tout notre attention; il s'agit d'un corps de brèches péridotitiques monogéniques qui affleure en discontinuité le long du versant méridional et le long du fond de la vallée Vigezzo sur environ 20 km. Ces brèches se posent indifféremment sur le socle (unités Finero, Orselina) ou sur les sédiments lacustres. Elles sont traversées par des plans de failles qui développent des véritables stries de faille et des gouges de faille; l’orientation de ces plans est la même que celle affectant les failles à gouges du socle. La genèse de cette brèche est liée à l'altération et au modelage glacier (rock-glaciers) d'une brèche tectonique originelle qui borde la partie externe du Corps de Finero. Les structures de déformation de cette brèche, pareillement à celles des sédiments lacustres, ont été considérées comme les évidences de surface d'une tectonique quaternaire active dans la région. La dernière phase de déformation cassante qui affecte cette région peut donc être considérée comme active en époque quaternaire. Une vue d’ensemble de la région étudiée nous permet de reconnaître à l’échelle régionale une zone de cisaillement complexe orientée E-W, parallèlement à l’axe de la vallée Centovalli-Val Vigezzo. Les données de terrain, indiquent que cette zone de cisaillement débute sous conditions ductiles et évolue en plusieurs étapes jusqu’à des conditions de failles cassantes de surface. La reconstruction de l'évolution géodynamique de la région a permis de définir trois étapes distinctes qui marquent le passage, de ce secteur de socle cristallin, de conditions P-T profondes à des conditions de surface. Dans ce contexte, on a reconnu trois phases principales de déformation à l’échelle régionale qui caractérisent ces trois étapes. La phase la plus ancienne est constituée par des mylonites en faciès amphibolite, associées à des mouvements de cisaillement dextre, qui sont ensuite remplacés par des mylonites en faciès schistes verts et des plis rétrovergentes liés au rétrocharriage des nappes alpines. Une deuxième étape est identifiée par le développement d’une phase hydrothermale liée à un système de failles extensives et décrochantes dextres à direction principale E-W, NE-SW et NW-SE. Leur caractérisation minéralogique a permis la mise en évidence des phases cristallines de néoformation liées à cet événement constituées par : K-feldspath (microcline), chlorites (Fe+Mg), épidotes, prehnite, zéolites (laumontite), sphène, calcite. Dans ce contexte, pour obtenir une meilleure caractérisation de cet événement hydrothermal on a utilisé des géothermomètres sur chlorites, sensible aussi à la pression et a la a(H2O), qui ont donné des valeurs descendantes comprises entre 450-200°C. Les derniers mouvements sont mis en évidence par le développement d’une série de plans majeurs de failles à gouge, qui forment une structure en sigmoïdes d’épaisseur kilométrique reconnaissable à l’échelle de la vallée et caractérisée par des mouvements transpressifs avec une composante décrochante dextre toujours importante. Cette phase de déformation forme un système conjugué de failles avec direction moyenne E-W qui coupent la zone des racines des nappes alpines, la zone du Canavese et le corps ultramafique de Finero. Ce système se déroule de manière subparallèle à l'axe de la vallée le long de plusieurs dizaines de kilomètres. Une analyse complète et détaillée des gouges de faille par XRD a montré que la fraction argileuse (<2 µm) de ces gouges contient une partie de néoformation très importante constituée par, des illites, des chlorites et des interstratifiés de type illite/smectite ou chlorite/smectite. Des datations avec méthode K-Ar sur ces illites ont donné des valeurs comprises entre 12 et 4 Ma qui représentent l'âge de cette dernière déformation cassante. L'application de la méthode de la cristallinité de l'illite (C.I.) a permis d'évaluer les conditions thermiques qui caractérisent le déroulement de cette dernière phase tectonique qui se produit sous conditions de température caractéristiques de l'anchizone et de la diagenèse. L'ensemble des structures de déformation qu'on vient de décrire s'insère parfaitement dans le contexte de convergence oblique entre la plaque adriatique et celle européenne qui à produit l'orogène alpin. On peut considérer les structures tectoniques du Val Vigezzo-Centovalli comme l'expression d'une zone majeure de cisaillement "Simplo-Insubrienne". L'empilement structural et les structures tectoniques affleurantes dans la région sont le résultat de l'interaction entre un régime tectonique transpressif et un régime transtensif. Ces deux champs de tension sont antagonistes entre eux mais sont reliés, de toute façon, à une seule phase décrochante dextre principale, due à une convergence oblique entre deux plaques. À l'échelle de l'évolution géodynamique on peut distinguer différentes étapes au cours desquelles les structures de ces deux régimes tectoniques interagissent en manière différente. En accord avec les données géophysiques et les reconstructions paléodynamiques prises dans la littérature on considère que la ligne Rhône-Simplon-Centovalli représente l'évidence de surface de la suture majeure profonde entre la plaque Adriatique et celle Européenne. Les vitesses de soulèvement qui ont été calculées dans cette étude pour cette région des Alpes donnent une valeur moyenne de 0.8 mm/a qui est tout à fait comparable avec les données proposées par la littérature sur cette zone. La zone Val Vigezzo-Centovalli peut être donc considérée comme un carrefour géologique où se croisent différentes phases tectoniques qui représentent les évidences de surface d'une suture profonde majeure entre deux plaques dans un contexte de collision continentale. ABSTRACT: A wide and complex tectonic zone known as Centovalli line, crosses the Central Alps sector between Domodossola and Locarno. This area, formed by the Vigezzo Valley and Centovalli valley, constitutes the southernmost termination of the Lepontin dome and represents a portion of the alpine nappes root zone. It belongs to a large and complex shear-zone, partly associated with hydrothermal phenomena of alpine age (<20 My), which includes the Insubric Line and the Simplon fault zone. Vigezzo Valley and Centovalli constitute a real crossroads between the mains alpines tectonics lines as well as a zone of juxtaposition of the Southalpine basement with the Austroalpin and Pennique root zone. The deformation phases and the geological structures that can be studied between approximately 35 My and the present. The detailed field study showed the presence of many brittle and ductile deformation structures and fault rocks such as mylonites, cataclasites, pseudotachylites, kakirites, mineralized faults, fault gouges and folds. In the field we could distinguish at least four folds generations related to the various deformation phases. The number and the complexity of these structures indicate a very complicated history, comprising several different stages, that sometimes are related and even superimposed. Part of these deformation structures affect also the sedimentary deposits of quaternary age, in particular the silts and sands lake deposit. These sediments constitute the remainders of a lake basin ascribed to the interglacial Riss/Würm (Eemien, 67.000-120.000 years) and outcroping in the central part of the studied area, in the Eastern part of Santa Maria Maggiore plain. These sediments show a whole series of deformation structures such as inverse fault planes, combined shortening structures and true folds. These faults and folds would represent the surface evidence of a probably active tectonic deformation in quaternary time. Another rock formation attracted all our attention. It is a body of monogenic peridotite breccia which outcrops in discontinuity along the southernmost slope and the bottom of the Vigezzo valley on approximately 20 km. This breccia lies indifferently on the basement (Finero and Orselina units) or on the lake sediments. They are crossed by fault planes which developed slikenside and fault gouges whose orientation is the same of the faults gouges in the alpine basement. This breccia results from the weathering and the surface modelling of an original tectonic breccia which borders the external part of Finero peridotite body. This breccia deformation structures, like those of the lake sediments, were regarded as the surface interaction of active quaternary tectonics in the area. So the last brittle deformation phases which affects this area seems to be actives in quaternary time. Theoverall picture of the studied area on a regional scale enables us to point out a complex shear-zone directed E-W, parallel to the axis of the Centovalli and Vigezzo Valley. The field analysis indicates that this shear-zone began under ductile conditions and evolved in several stages to brittle faulting under surface conditions. The analysis of the geodynamic evolution of the area allows to define three different stages which mark the transition of this alpine basement root zone, from deep P-T conditions to P-T surface conditions. In this context on regional scale three principal deformation phases, which characterize these three stages can be distinguished. The oldest phase consisted of the amphibolitie facies mylonites, associated to dextral strikeslip movements. They are then replaced by green-schists facies mylonites and backfolds related to the backthrusting of the alpines nappes. A second episode is caracterized by the development of an hydrothermal phase bound to an extensive fault and dextral strike-slip fault system, with E-W, NW-SE and SE-NW principal directionsThe principal neoformed mineral phases related to this event are: K-feldspar (microcline), chlorites (Fe+Mg), epidotes prehnite, zéolites (laumontite), sphene and calcite. In this context, to obtain a better characterization of this hydrothermal event, we have used an chlorite geothermometer, sensitive also to the pressure and has the a(H2O), which gave downward values ranging between 450-200°C. The last movements are caracterized by the development of important gouge fault plans, which form a sigmoid structure of kilometric thickness which is recognizable at the valley scale, and is characterized by transpressive movements always with a significant dextral strike-slip component. This deformation phase forms a combined faults system with an average E-W direction, which cuts trough the alpine root zone, the Canavese zone and the Finero ultramafic body. This fault system takes place subparallel to the axis of the valley over several tens of kilometers. A complete and detailed XRD analysis of the gouges fault showed that the clay fraction (<2µm) contains a very significant neo-formation of illite, chlorites and mixed layered clays such as illite/smectite or chlorite/smectite. The K-Ar datings of the illite fraction <2µm gave values ranging between 12 and 4 My and the illite fraction <0.2µm gave more recents values until to 2,4-0 My.This values represent the age of this last brittle deformation. The application of the illite crystallinity method (C.I.) allowed evaluating the thermal conditions which characterize this tectonic phase that occured under temperature conditions of the anchizone and diagenesis. The whole set of deformation structures which we just described, perfectly fit the context of oblique convergence between the Adriatic and the European plate that produced the alpine orogen. We can regard the Vigezzo valley and Centovalli tectonic structures as the expression of a major "Simplo-Insubric" shear-zone. Structural stacking and tectonic structures that outcrop in the studied area, are the result of the interaction between a transpressive and a transtensve tectonic phases. These two tension fields are antagonistic but they are also connected, in any event, with only one principal dextral strike-slip movement, caused by an oblique convergence between two plates. On the geodynamic evolution scale we can distinguish various stages during which these two tectonic structures fields interact in various ways. In agreement with the geophysical data and the paleodynamic recostructions taken in the literature we considers that the Rhone-Simplon-Centovalli line are the surface feature of the major collision between the Adriatique and the European plate at depth. The uplift speeds we calculated in this study for this Alpine area give an average value of 0.8 mm/a, which is in good agreement with the data suggested by the literature on this zone. TheVigezzo Valley and Centovalli zone can therefore be regarded as a geological crossroad where various tectonic phases are superimposed. They represent the evidences of a major and deeper suture between two plates in a continental collision context.

When the Line is Crossed... : Paths to Control and Sanction Behaviour Necessitating a State Reaction

The article presents a special form of a European comparative synopsis. For this case examples have been chosen ranging from administrative or minor (criminal) offences to increasingly serious offences and offenders. In this way it can be comparatively demonstrated how the criminal justice systems studied handle specific cases and whether they do so in a similar or different way.

Y-90-Ibritumomab Tiuxetan (Zevalin (R)) Consolidation of First Remission In Advanced-Stage Follicular Non-Hodgkin's Lymphoma: Updated Results After a Median Follow-up of 66.2 Months From the International, Randomized, Phase III First-Line Indolent Trial (FIT) In 414 Patients

The FIT trial was conducted to evaluate the safety and efficacy of 90Y-ibritumomab tiuxetan (0.4 mCi/kg; maximum dose 32 mCi) when used as consolidation of first complete or partial remission in patients with previously untreated, advanced-stage follicular lymphoma (FL). Patients were randomly assigned to either 90Y-ibritumomab treatment (n = 207) or observation (n = 202) within 3 months (mo) of completing initial induction therapy (chemotherapy only: 86%; rituximab in combination with chemotherapy: 14%). Response status prior to randomization did not differ between the groups: 52% complete response (CR)/CR unconfirmed (CRu) to induction therapy and 48% partial response (PR) in the 90Y-ibritumomab arm vs 53% CR/CRu and 44% PR in the control arm. The primary endpoint was progression-free survival (PFS) of the intent-to-treat (ITT) population. Results from the first extended follow-up after a median of 3.5 years revealed a significant improvement in PFS from the time of randomization with 90Y-ibritumomab consolidation compared with control (36.5 vs 13.3 mo, respectively; P < 0.0001; Morschhauser et al. JCO. 2008; 26:5156-5164). Here we report a median follow-up of 66.2 mo (5.5 years). Five-year PFS was 47% in the 90Y-ibritumomab group and 29% in the control group (hazard ratio (HR) = 0.51, 95% CI 0.39-0.65; P < 0.0001). Median PFS in the 90Y-ibritumomab group was 49 mo vs 14 mo in the control group. In patients achieving a CR/CRu after induction, 5-year PFS was 57% in the 90Y-ibritumomab group, and the median had not yet been reached at 92 months, compared with a 43% 5-year PFS in the control group and a median of 31 mo (HR = 0.61, 95% CI 0.42-0.89). For patients in PR after induction, the 5-year PFS was 38% in the 90Y-ibritumomab group with a median PFS of 30 mo vs 14% in the control group with a median PFS of 6 mo (HR = 0.38, 95% CI 0.27-0.53). Patients who had received rituximab as part of induction treatment had a 5-year PFS of 64% in the 90Y-ibritumomab group and 48% in the control group (HR = 0.66, 95% CI 0.30-1.47). For all patients, time to next treatment (as calculated from the date of randomization) differed significantly between both groups; median not reached at 99 mo in the 90Y-ibritumomab group vs 35 mo in the control group (P < 0.0001). The majority of patients received rituximab-containing regimens when treated after progression (63/82 [77%] in the 90Y-ibritumomab group and 102/122 [84%] in the control group). Overall response rate to second-line treatment was 79% in the 90Y-ibritumomab group (57% CR/CRu and 22% PR) vs 78% in the control arm (59% CR/CRu, 19% PR). Five-year overall survival was not significantly different between the groups; 93% and 89% in the 90Y-ibritumomab and control groups, respectively (P = 0.561). To date, 40 patients have died; 18 in the 90Y-ibritumomab group and 22 in the control group. Secondary malignancies were diagnosed in 16 patients in the 90Y-ibritumomab arm vs 9 patients in the control arm (P = 0.19). There were 6 (3%) cases of myelodysplastic syndrome (MDS)/acute myelogenous leukemia (AML) in the 90Y-ibritumomab arm vs 1 MDS in the control arm (P = 0.063). In conclusion, this extended follow-up of the FIT trial confirms the benefit of 90Y-ibritumomab consolidation with a nearly 3 year advantage in median PFS. A significant 5-year PFS improvement was confirmed for patients with a CR/CRu or a PR after induction. Effective rescue treatment with rituximab-containing regimens may explain the observed no difference in overall survival between both patient groups who were - for the greater part - rituximab-naïve.

Identification of a novel determinant for membrane association in hepatitis C virus nonstructural protein 4B.

Nonstructural protein 4B (NS4B) plays an essential role in the formation of the hepatitis C virus (HCV) replication complex. It is a relatively poorly characterized integral membrane protein predicted to comprise four transmembrane segments in its central portion. Here, we describe a novel determinant for membrane association represented by amino acids (aa) 40 to 69 in the N-terminal portion of NS4B. This segment was sufficient to target and tightly anchor the green fluorescent protein to cellular membranes, as assessed by fluorescence microscopy as well as membrane extraction and flotation analyses. Circular dichroism and nuclear magnetic resonance structural analyses showed that this segment comprises an amphipathic alpha-helix extending from aa 42 to 66. Attenuated total reflection infrared spectroscopy and glycosylation acceptor site tagging revealed that this amphipathic alpha-helix has the potential to traverse the phospholipid bilayer as a transmembrane segment, likely upon oligomerization. Alanine substitution of the fully conserved aromatic residues on the hydrophobic helix side abrogated membrane association of the segment comprising aa 40 to 69 and disrupted the formation of a functional replication complex. These results provide the first atomic resolution structure of an essential membrane-associated determinant of HCV NS4B.

First-line chemotherapy with local treatment can prevent external-beam irradiation and enucleation in low-stage intraocular retinoblastoma.

PURPOSE: To evaluate the efficacy of first-line chemotherapy (CT) in preventing external-beam radiotherapy (EBR) and/or enucleation in patients with retinoblastoma (Rbl). PATIENTS AND METHODS: Twenty-four patients with newly diagnosed unilateral or bilateral Rbl received CT associated with local treatment (LT). Two to five courses of etoposide and carboplatin were administered at 3- to 4-week intervals, depending on tumor response, and were completed each time by LT. RESULTS: Tumor response was observed in all eyes. Twenty-one of 24 patients showed a complete response (CR) that persisted at a median follow-up (FU) of 31 months (range, 4 to 41 months). Among the three patients who relapsed, two were lost to FU and one died of progressive disease. CR was achieved by CT and LT alone in 15 (71.4%) of 21 patients with less advanced disease (groups I to III). Six other patients with advanced disease (groups IV and V) experienced treatment failure and needed salvage treatment by EBR and/or enucleation. The difference between the two patient groups with regard to disease stage was statistically significant (P <.0001). EBR could be avoided in 13 (68.4%) of 19 patients, who presented with groups I to III (15 eyes) and group V (one eye) disease, whereas enucleation could be avoided in only two (40%) of five. CONCLUSION: CT combined with intensive LT is effective in patients with groups I to III Rbl, permitting the avoidance of EBR in the majority of these young children and, thus, reducing the risk of long-term sequelae. This is in contrast with the disappointing results for patients with groups IV and V Rbl, in whom EBR and/or enucleation was needed.

Low-frequency drug-resistant HIV-1 and risk of virological failure to first-line NNRTI-based ART: a multicohort European case-control study using centralized ultrasensitive 454 pyrosequencing.

OBJECTIVES: It is still debated if pre-existing minority drug-resistant HIV-1 variants (MVs) affect the virological outcomes of first-line NNRTI-containing ART. METHODS: This Europe-wide case-control study included ART-naive subjects infected with drug-susceptible HIV-1 as revealed by population sequencing, who achieved virological suppression on first-line ART including one NNRTI. Cases experienced virological failure and controls were subjects from the same cohort whose viraemia remained suppressed at a matched time since initiation of ART. Blinded, centralized 454 pyrosequencing with parallel bioinformatic analysis in two laboratories was used to identify MVs in the 1%-25% frequency range. ORs of virological failure according to MV detection were estimated by logistic regression. RESULTS: Two hundred and sixty samples (76 cases and 184 controls), mostly subtype B (73.5%), were used for the analysis. Identical MVs were detected in the two laboratories. 31.6% of cases and 16.8% of controls harboured pre-existing MVs. Detection of at least one MV versus no MVs was associated with an increased risk of virological failure (OR = 2.75, 95% CI = 1.35-5.60, P = 0.005); similar associations were observed for at least one MV versus no NRTI MVs (OR = 2.27, 95% CI = 0.76-6.77, P = 0.140) and at least one MV versus no NNRTI MVs (OR = 2.41, 95% CI = 1.12-5.18, P = 0.024). A dose-effect relationship between virological failure and mutational load was found. CONCLUSIONS: Pre-existing MVs more than double the risk of virological failure to first-line NNRTI-based ART.

The age of movements along the insubric line West of Locarno (Northern Italy and southern Switzerland)

Permo-Mesozoic Canavese sediments are pinched in between the pre-Alpine high-grade metamorphic Ivrea Zone and the Alpine metamorphosed Sesia Zone along the Insubric Line W of Locarno. According to the ``illite crystallinity'' these sediments were deformed under anchi- and epizonal conditions. Synkinematically formed white mica in the mylonitized Canavese sediments yields the following K-Ar age ranges: 60-76 Ma at the southwestern end, 28-43 Ma in the central part and 19-26 Ma in the northeastern part of the Insubric Line W of Locarno. The youngest age group dates the main uplift and dextral strike-slip movements of the Insubric Line, comprising mylonites in the NE and cataclasites in the SW. This activity correlates with Late Oligocene to Early Miocene rapid cooling and uplift of the Central Alps.

Validation and long-term evaluation of a modified on-line chiral analytical method for therapeutic drug monitoring of (R,S)-methadone in clinical samples.

Matrix effects, which represent an important issue in liquid chromatography coupled to mass spectrometry or tandem mass spectrometry detection, should be closely assessed during method development. In the case of quantitative analysis, the use of stable isotope-labelled internal standard with physico-chemical properties and ionization behaviour similar to the analyte is recommended. In this paper, an example of the choice of a co-eluting deuterated internal standard to compensate for short-term and long-term matrix effect in the case of chiral (R,S)-methadone plasma quantification is reported. The method was fully validated over a concentration range of 5-800 ng/mL for each methadone enantiomer with satisfactory relative bias (-1.0 to 1.0%), repeatability (0.9-4.9%) and intermediate precision (1.4-12.0%). From the results obtained during validation, a control chart process during 52 series of routine analysis was established using both intermediate precision standard deviation and FDA acceptance criteria. The results of routine quality control samples were generally included in the +/-15% variability around the target value and mainly in the two standard deviation interval illustrating the long-term stability of the method. The intermediate precision variability estimated in method validation was found to be coherent with the routine use of the method. During this period, 257 trough concentration and 54 peak concentration plasma samples of patients undergoing (R,S)-methadone treatment were successfully analysed for routine therapeutic drug monitoring.

Colloidal systems for the delivery of cyclosporin A to the anterior segment of the eye.

Due to the eye's specific anatomical and physiological conformation, the treatment of eye diseases is a real challenge for pharmaceutical therapy. The presence of efficient protective barriers (i.e., the conjunctival and corneal membranes) and protective mechanisms (i.e., blinking and nasolachrymal drainage) makes this organ particularly impervious to local drug therapy. To overcome these issues, numerous strategies have been envisioned using pharmaceutical technology. Many formulations currently on the market or still under development are emulsions or colloidal systems intended to enhance precorneal residence time and corneal penetration, causing a consequent increase in drug bioavailability after instillation. After a review of some recent developments in the field of cyclosporin A formulations for the eye, a novel micellar formulation of cyclosporine A based on a diblock methoxy-poly(ethylene glycol)-hexysubstituted poly(lactides) (MPEG-hexPLA) is described.

Pharmacokinetics and posterior segment biodistribution of ESBA105, an anti-TNF-alpha single-chain antibody, upon topical administration to the rabbit eye.

PURPOSE: The purpose of this study was to characterize local distribution and systemic absorption of the tumor necrosis factor (TNF)-alpha inhibitory single-chain antibody fragment (scFv) ESBA105 following topical administration to the eye in vivo. METHODS: Rabbits received ESBA105 as topical eye drops in two dosing regimens. First, pharmacokinetics after the topical route of administration was compared to the intravenous (i.v.) route by means of applying the identical cumulative daily dose of ESBA105. In a second study rabbits received five eye drops daily for six consecutive days in a lower frequency topical dosing regimen. Kinetics and biodistribution of ESBA105 in ocular tissues and fluids as well as in sera were determined in all animals. RESULTS: After topical administration to the eye, ESBA105 quickly reaches therapeutic concentrations in all ocular compartments. Systemic exposure after topical administration is 25,000-fold lower than exposure after i.v. injection of the identical cumulative daily dose. ESBA105 levels in vitreous humor and neuroretina are significantly higher on topical administration than after i.v. injection. Absolute and relative intraocular biodistribution of ESBA105 is different with topical and systemic delivery routes. Compared to its terminal half-life in circulation (7 hours), the vitreal half-life of ESBA105 is significantly enhanced (16-24 hours). CONCLUSIONS: On topical administration, ESBA105 is efficiently absorbed and distributed to all compartments of the eye, whereby systemic drug exposure is very low. Based on its unique intraocular biodistribution and pharmacokinetics and the absolute intraocular levels reached, topical ESBA105 appears highly attractive for treatment of various ophthalmological disorders.

The fourth transmembrane segment of the Na,K-ATPase alpha subunit: a systematic mutagenesis study.

The Na,K-ATPase is a major ion-motive ATPase of the P-type family responsible for many aspects of cellular homeostasis. To determine the structure of the pathway for cations across the transmembrane portion of the Na,K-ATPase, we mutated 24 residues of the fourth transmembrane segment into cysteine and studied their function and accessibility by exposure to the sulfhydryl reagent 2-aminoethyl-methanethiosulfonate. Accessibility was also examined after treatment with palytoxin, which transforms the Na,K-pump into a cation channel. Of the 24 tested cysteine mutants, seven had no or a much reduced transport function. In particular cysteine mutants of the highly conserved "PEG" motif had a strongly reduced activity. However, most of the non-functional mutants could still be transformed by palytoxin as well as all of the functional mutants. Accessibility, determined as a 2-aminoethyl-methanethiosulfonate-induced reduction of the transport activity or as inhibition of the membrane conductance after palytoxin treatment, was observed for the following positions: Phe(323), Ile(322), Gly(326), Ala(330), Pro(333), Glu(334), and Gly(335). In accordance with a structural model of the Na,K-ATPase obtained by homology modeling with the two published structures of sarcoplasmic and endoplasmic reticulum calcium ATPase (Protein Data Bank codes 1EUL and 1IWO), the results suggest the presence of a cation pathway along the side of the fourth transmembrane segment that faces the space between transmembrane segments 5 and 6. The phenylalanine residue in position 323 has a critical position at the outer mouth of the cation pathway. The residues thought to form the cation binding site II ((333)PEGL) are also part of the accessible wall of the cation pathway opened by palytoxin through the Na,K-pump.

Disease-causing mutations improving the branch site and polypyrimidine tract: pseudoexon activation of LINE-2 and antisense Alu lacking the poly(T)-tail.

Cryptic exons or pseudoexons are typically activated by point mutations that create GT or AG dinucleotides of new 5' or 3' splice sites in introns, often in repetitive elements. Here we describe two cases of tetrahydrobiopterin deficiency caused by mutations improving the branch point sequence and polypyrimidine tracts of repeat-containing pseudoexons in the PTS gene. In the first case, we demonstrate a novel pathway of antisense Alu exonization, resulting from an intronic deletion that removed the poly(T)-tail of antisense AluSq. The deletion brought a favorable branch point sequence within proximity of the pseudoexon 3' splice site and removed an upstream AG dinucleotide required for the 3' splice site repression on normal alleles. New Alu exons can thus arise in the absence of poly(T)-tails that facilitated inclusion of most transposed elements in mRNAs by serving as polypyrimidine tracts, highlighting extraordinary flexibility of Alu repeats in shaping intron-exon structure. In the other case, a PTS pseudoexon was activated by an A>T substitution 9 nt upstream of its 3' splice site in a LINE-2 sequence, providing the first example of a disease-causing exonization of the most ancient interspersed repeat. These observations expand the spectrum of mutational mechanisms that introduce repetitive sequences in mature transcripts and illustrate the importance of intronic mutations in alternative splicing and phenotypic variability of hereditary disorders.