Association of nuclear matrix proteins with granular and threaded nuclear bodies in cell lines undergoing apoptosis.

The granules which appear in the nucleolar area in apoptotic HL-60 cells after camptothecin administration (Zweyer et al., Exp. Cell Res. 221,27-40, 1995) were detected also in several other cell lines induced to undergo apoptosis by different stimuli, such as MOLT-4 treated with staurosporine, K-562 incubated with actinomycin D, P-815 exposed to temperature causing heat shock, Jurkat cells treated with EGTA, U-937 growing in the presence of cycloheximide and tumor necrosis factor-alpha, and HeLa cells treated with etoposide. Using immunoelectron microscopy techniques, we demonstrate that, besides the already described nuclear matrix proteins p125 and p160, these granules contain other nucleoskeletal polypeptides such as proliferating cell nuclear antigen, a component of ribonucleoprotein particles, a 105-kDa constituent of nuclear spliceosomes, and the 240-kDa nuclear mitotic apparatus-associated protein referred to as NuMA. Moreover, we also found in the granules SAF-A/hn-RNP-U and SATB1 proteins, two polypeptides that have been reported to bind scaffold-associated regions DNA sequences in vitro, thus mediating the formation of looped DNA structures in vivo. Fibrillarin and coilin are not present in these granules or the PML protein. Thus, the granules seen during the apoptotic process apparently are different from coiled bodies or other types of nuclear bodies. Furthermore, these granules do not contain chromatin components such as histones and DNA. Last, Western blotting analysis revealed that nuclear matrix proteins present in the granules are not proteolytically degraded except for the NuMA polypeptide. We propose that these granules might represent aggregates of nuclear matrix proteins forming during the apoptotic process. Moreover, since the granules are present in several cell lines undergoing apoptosis, they could be considered a previously unrecognized morphological hallmark of the apoptotic process.

Artificial Urinary Sphincter based on smart materials

Objective: There are only a few established artificial urinary sphincters for treatment of incontinence. We have developed a new device composed by three parts: the actuator, three contractile rings and a control unit. The actuator is made of Nitinol fibers, driven by microprocessor. The fibers are linked to the rings placed around the urethra. They function with alternance in their open and closed position. This concept is called piano concept. With this set-up, the constant compression on the urethra is strongly reduced. Methods: Six male sheep have been used for this study. The sphincter was open each hour for a period of 10 min., to guaranty urination. The bladder was filled with water while one cuff was closed and bladder pressure was monitored. The animals were sacrificed. Two biopsies around two cuffs of each explant and all three cuffs from each explant including urethra were analyzed. Urethra not surrounded by a cuff was taken as control. Results: The pressure exerted by the sphincter around the urethra provided continence. Simulated incontinence occurred at a pressure of 1bar measured on the bladder wall using a pressure probe. The closing force of the cuff was approx. 0·7N. No difference in tissue structure and organization of the urethra with and without artificial sphincter was observed. Conclusions: This device has several advantages compared to other urinary sphincters. It is easy to implant, has no hydraulic nature and reduces ischemic injury of the urethra by the alternance of urethral part compressed. Proof of concept in vivo has been demonstrated. Other studies are planned to determine long-term outcome.

Lateral condyle fracture of the humerus in children treated with bioabsorbable materials

Les fractures du condyle externe de l'enfant sont un traumatisme relativement fréquent qui survient le plus souvent chez l'enfant entre 5 et 15 ans. Elles sont classées en 3 grades : le grade 1 sans déplacement qui peuvent être traitées conservativement et les grades 2 (déplacées) et 3 (disloquées) qui nécessitent une prise en charge chirurgicale par réduction ouverte et embrochage. Le traitement traditionnel de ces fractures consistent en la mise en place de broches/vis métalliques qui doivent être retirées quelques semaines plus tard lors d'une seconde intervention. Depuis les années 90, des matériaux bio-résorbables à base d'acide polyglycoliques sont apparus en chirurgie orthopédique et traumatologique. En raison de la survenue de complications telles que réactions inflammatoires, formation de séromes ou ostéolyse, l'utilisation du matériel conventionnel a été préférée. Cependant, depuis quelques années, de nouveaux matériaux en acide polylactique sont apparus sur le marché. La période de résorption étant beaucoup plus lente, ces matériaux induisent des réactions beaucoup plus faibles, sans conséquences cliniques relevées jusqu'à présent. Chez l'adulte, de récentes études ont prouvés l'efficacité et l'absence d'effets secondaires liés à ces matériaux. Par contre, chez l'enfant, peu d'études à moyen terme ont été réalisées et l'impact sur l'os en croissance est peu connu, même si pour l'heure aucune conséquence clinique n'a été relevée. Dans le cadre de notre étude, nous avons comparé 2 groupes d'enfants traités pour des fractures du condyle externe du coude. Le premier groupe traité par du matériel conventionnel et le second par matériel résorbable. Les enfants ont tous étés suivis de manière très étroite durant la première année et la récupération clinique est similaire pour les 2 groupes. A 4 ans du traumatisme, les résultats fonctionnels sont identiques pour les 2 groupes et aucune complication liée au matériel résorbable n'a été mise en évidence. Par ailleurs, à 1 et 4 ans, les clichés radiologiques montrent l'absence de lésions liées au matériel résorbable. En conclusion, dans le cadre de notre travail nous avons pu montrer l'efficacité des matériaux résorbables dans la prise en charge des fractures du condyle externe du coude chez l'enfant. Ces matériaux permettent d'optimiser la prise en charge de ce type de fracture en prévenant un risque opératoire et anesthésique liés à une seconde intervention nécessaire au retrait du matériel d'ostéosynthèse conventionnel. Par ailleurs le coût lié à la prise en charge globale de ce type de fracture est moindre lors de l'utilisation de matériel résorbable.

Biofilm formation on bone grafts and bone graft substitutes: comparison of different materials by a standard in vitro test and microcalorimetry.

We analyzed the initial adhesion and biofilm formation of Staphylococcus aureus (ATCC 29213) and S. epidermidis RP62A (ATCC 35984) on various bone grafts and bone graft substitutes under standardized in vitro conditions. In parallel, microcalorimetry was evaluated as a real-time microbiological assay in the investigation of biofilm formation and material science research. The materials beta-tricalcium phosphate (beta-TCP), processed human spongiosa (Tutoplast) and poly(methyl methacrylate) (PMMA) were investigated and compared with polyethylene (PE). Bacterial counts (log(10) cfu per sample) were highest on beta-TCP (S. aureus 7.67 +/- 0.17; S. epidermidis 8.14 +/- 0.05) while bacterial density (log(10) cfu per surface) was highest on PMMA (S. aureus 6.12 +/- 0.2, S. epidermidis 7.65 +/- 0.13). Detection time for S. aureus biofilms was shorter for the porous materials (beta-TCP and processed human spongiosa, p < 0.001) compared to the smooth materials (PMMA and PE), with no differences between beta-TCP and processed human spongiosa (p > 0.05) or PMMA and PE (p > 0.05). In contrast, for S. epidermidis biofilms the detection time was different (p < 0.001) between all materials except between processed human spongiosa and PE (p > 0.05). The quantitative analysis by quantitative culture after washing and sonication of the material demonstrated the importance of monitoring factors like specific surface or porosity of the test materials. Isothermal microcalorimetry proved to be a suitable tool for an accurate, non-invasive and real-time microbiological assay, allowing the detection of bacterial biomass without removing the biofilm from the surface.

Properties of debris flow deposits and source materials compared: implications for debris flow characterization

In the Alps, debris flow deposits generally contain < 5% clay-size particles, and the role of the surface-charged < 2 mu m particles is often neglected, although these particles may have a significant impact on the rheological properties of the interstitial fluid. The objective of this study was to compare debris flow deposits and parent materials from two neighbouring catchments of the Swiss Alps, with special emphasis on the colloidal constituents. The catchments are small in area (4 km(2)), 2.5 km long, similar in morphology, but different in geology. The average slopes are 35-40%. The catchments were monitored for debris flow events and mapped for surface aspect and erosion activity. Debris flow deposits and parent materials were sampled, the clay and silt fractions extracted and the bulk density, < 2 mm fraction bulk density, particle size distribution, chemical composition, cation exchange capacity (CEC) and mineralogy analysed. The results show that the deposits are similar to the parent screes in terms of chemical composition, but differ in terms of: (i) particle size distribution; and (ii) mineralogy, reactivity and density of the < 2 mm fraction. In this fraction, compared with the parent materials the deposits show dense materials enriched in coarse monocrystalline particles, of which the smallest and more reactive particles were leached. The results suggest that deposit samples should not be considered as representative of source or flow materials, particularly with respect to their physical properties.

Round robin test: wear of nine dental restorative materials in six different wear simulators - supplement to the round robin test of 2005.

OBJECTIVE: The purpose of the present study was to submit the same materials that were tested in the round robin wear test of 2002/2003 to the Alabama wear method. METHODS: Nine restorative materials, seven composites (belleGlass, Chromasit, Estenia, Heliomolar, SureFil, Targis, Tetric Ceram) an amalgam (Amalcap) and a ceramic (IPS Empress) have been submitted to the Alabama wear method for localized and generalized wear. The test centre did not know which brand they were testing. Both volumetric and vertical loss had been determined with an optical sensor. After completion of the wear test, the raw data were sent to IVOCLAR for further analysis. The statistical analysis of the data included logarithmic transformation of the data, the calculation of relative ranks of each material within each test centre, measures of agreement between methods, the discrimination power and coefficient of variation of each method as well as measures of the consistency and global performance for each material. RESULTS: Relative ranks of the materials varied tremendously between the test centres. When all materials were taken into account and the test methods compared with each other, only ACTA agreed reasonably well with two other methods, i.e. OHSU and ZURICH. On the other hand, MUNICH did not agree with the other methods at all. The ZURICH method showed the lowest discrimination power, ACTA, IVOCLAR and ALABAMA localized the highest. Material-wise, the best global performance was achieved by the leucite reinforced ceramic material Empress, which was clearly ahead of belleGlass, SureFil and Estenia. In contrast, Heliomolar, Tetric Ceram and especially Chromasit demonstrated a poor global performance. The best consistency was achieved by SureFil, Tetric Ceram and Chromasit, whereas the consistency of Amalcap and Heliomolar was poor. When comparing the laboratory data with clinical data, a significant agreement was found for the IVOCLAR and ALABAMA generalized wear method. SIGNIFICANCE: As the different wear simulator settings measure different wear mechanisms, it seems reasonable to combine at least two different wear settings to assess the wear resistance of a new material.

Cerebellar cortical layers: in vivo visualization with structural high-field-strength MR imaging.

Purpose: To perform in vivo imaging of the cerebellum with an in-plane resolution of 120 mm to observe its cortical granular and molecular layers by taking advantage of the high signal-to-noise ratio and the increased magnetic susceptibility-related contrast available at high magnetic field strength such as 7 T. Materials and Methods: The study was approved by the institutional review board, and all patients provided written consent. Three healthy persons (two men, one woman; mean age, 30 years; age range, 28-31 years) underwent MR imaging with a 7-T system. Gradient-echo images (repetition time msec/echo time msec, 1000/25) of the human cerebellum were acquired with a nominal in-plane resolution of approximately 120 mum and a section thickness of 1 mm. Results: Structures with dimensions as small as 240 mum, such as the granular and molecular layers in the cerebellar cortex, were detected in vivo. The detection of these structures was confirmed by comparing the contrast obtained on T2*-weighted and phase images with that obtained on images of rat cerebellum acquired at 14 T with 30 mum in-plane resolution. Conclusion: In vivo cerebellar imaging at near-microscopic resolution is feasible at 7 T. Such detailed observation of an anatomic area that can be affected by a number of neurologic and psychiatric diseases, such as stroke, tumors, autism, and schizophrenia, could potentially provide newer markers for diagnosis and follow-up in patients with such pathologic conditions. (c) RSNA, 2010.

Wear of two denture teeth materials in vivo-2-year results.

OBJECTIVE: (1) To quantify wear of two different denture tooth materials in vivo with two study designs, (2) to relate tooth variables to vertical loss. METHODS: Two different denture tooth materials had been used (experimental material=test; DCL=control). In study 1 (split-mouth, 6 test centers) 60 subjects received complete dentures, in study 2 (two-arm, 1 test center) 29 subjects. In study 1 the mandibular dentures were supported by implants in 33% of the subjects, in study 2 only in 3% of the subjects. Impressions of the dentures were taken and poured with improved stone at baseline and after 6, 12, 18 and 24 months. Each operator evaluated the wear subjectively. Wear analysis was carried out with a laser scanning device. Maximal vertical loss of the attrition zones was calculated for each tooth cusp and tooth. A mixed linear model was used to statistically analyse the logarithmically transformed wear data. RESULTS: Due to drop-outs and unmatchable casts, only 47 subjects of study 1 and 14 of study 2 completed the 2-year recall. Overall, 75% of all teeth present could be analysed. There was no statistically difference in the overall wear between the test and control material for either study 1 or study 2. The relative increase in wear over time was similar in both study designs. However, a strong subject effect and center effect were observed. The fixed factors included in the model (time, tooth, center, etc.) accounted for 43% of the variability, whereas the random subject effect accounted for another 30% of the variability, leaving about 28% of unexplained variability. More wear was consistently recorded in the maxillary teeth compared to the mandibular teeth and in the first molar teeth compared to the premolar teeth and the second molars. Likewise, the supporting cusps showed more wear than the non-supporting cusps. The amount of wear did not depend on whether or not the lower dentures were supported by implants. The subjective wear was correct in about 67% of the cases if it is postulated that a wear difference of 100μm should be subjectively detectable. SIGNIFICANCE: The clinical wear of denture teeth is highly variable with a strong patient effect. More wear can be expected in maxillary denture teeth compared to mandibular teeth, first molars compared to premolars and supported cusps compared to non-supported cusps. Laboratory data on the wear of denture tooth materials may not be confirmed in well-structured clinical trials probably due to the large inter-individual variability.

Textural controls on induced polarization and permeability in granular media

The spatial resolution visualized with hydrological models and the conceptualized images of subsurface hydrological processes often exceed resolution of the data collected with classical instrumentation at the field scale. In recent years it was possible to increasingly diminish the inherent gap to information from point like field data through the application of hydrogeophysical methods at field-scale. With regards to all common geophysical exploration techniques, electric and electromagnetic methods have arguably to greatest sensitivity to hydrologically relevant parameters. Of particular interest in this context are induced polarisation (IP) measurements, which essentially constrain the capacity of a probed subsurface region to store an electrical charge. In the absence of metallic conductors the IP- response is largely driven by current conduction along the grain surfaces. This offers the perspective to link such measurements to the characteristics of the solid-fluid-interface and thus, at least in unconsolidated sediments, should allow for first-order estimates of the permeability structure.¦While the IP-effect is well explored through laboratory experiments and in part verified through field data for clay-rich environments, the applicability of IP-based characterizations to clay-poor aquifers is not clear. For example, polarization mechanisms like membrane polarization are not applicable in the rather wide pore-systems of clay free sands, and the direct transposition of Schwarz' theory relating polarization of spheres to the relaxation mechanism of polarized cells to complex natural sediments yields ambiguous results.¦In order to improve our understanding of the structural origins of IP-signals in such environments as well as their correlation with pertinent hydrological parameters, various laboratory measurements have been conducted. We consider saturated quartz samples with a grain size spectrum varying from fine sand to fine gravel, that is grain diameters between 0,09 and 5,6 mm, as well as corresponding pertinent mixtures which can be regarded as proxies for widespread alluvial deposits. The pore space characteristics are altered by changing (i) the grain size spectra, (ii) the degree of compaction, and (iii) the level of sorting. We then examined how these changes affect the SIP response, the hydraulic conductivity, and the specific surface area of the considered samples, while keeping any electrochemical variability during the measurements as small as possible. The results do not follow simple assumptions on relationships to single parameters such as grain size. It was found that the complexity of natural occurring media is not yet sufficiently represented when modelling IP. At the same time simple correlation to permeability was found to be strong and consistent. Hence, adaptations with the aim of better representing the geo-structure of natural porous media were applied to the simplified model space used in Schwarz' IP-effect-theory. The resulting semi- empiric relationship was found to more accurately predict the IP-effect and its relation to the parameters grain size and permeability. If combined with recent findings about the effect of pore fluid electrochemistry together with advanced complex resistivity tomography, these results will allow us to picture diverse aspects of the subsurface with relative certainty. Within the framework of single measurement campaigns, hydrologiste can than collect data with information about the geo-structure and geo-chemistry of the subsurface. However, additional research efforts will be necessary to further improve the understanding of the physical origins of IP-effect and minimize the potential for false interpretations.¦-¦Dans l'étude des processus et caractéristiques hydrologiques des subsurfaces, la résolution spatiale donnée par les modèles hydrologiques dépasse souvent la résolution des données du terrain récoltées avec des méthodes classiques d'hydrologie. Récemment il est possible de réduire de plus en plus cet divergence spatiale entre modèles numériques et données du terrain par l'utilisation de méthodes géophysiques, notamment celles géoélectriques. Parmi les méthodes électriques, la polarisation provoquée (PP) permet de représenter la capacité des roches poreuses et des sols à stocker une charge électrique. En l'absence des métaux dans le sous-sol, cet effet est largement influencé par des caractéristiques de surface des matériaux. En conséquence les mesures PP offrent une information des interfaces entre solides et fluides dans les matériaux poreux que nous pouvons lier à la perméabilité également dirigée par ces mêmes paramètres. L'effet de la polarisation provoquée à été étudié dans différentes études de laboratoire, ainsi que sur le terrain. A cause d'une faible capacité de polarisation des matériaux sableux, comparé aux argiles, leur caractérisation par l'effet-PP reste difficile a interpréter d'une manière cohérente pour les environnements hétérogènes.¦Pour améliorer les connaissances sur l'importance de la structure du sous-sol sableux envers l'effet PP et des paramètres hydrologiques, nous avons fait des mesures de laboratoire variées. En détail, nous avons considéré des échantillons sableux de quartz avec des distributions de taille de grain entre sables fins et graviers fins, en diamètre cela fait entre 0,09 et 5,6 mm. Les caractéristiques de l'espace poreux sont changées en modifiant (i) la distribution de taille des grains, (ii) le degré de compaction, et (iii) le niveau d'hétérogénéité dans la distribution de taille de grains. En suite nous étudions comment ces changements influencent l'effet-PP, la perméabilité et la surface spécifique des échantillons. Les paramètres électrochimiques sont gardés à un minimum pendant les mesures. Les résultats ne montrent pas de relation simple entre les paramètres pétro-physiques comme par exemples la taille des grains. La complexité des media naturels n'est pas encore suffisamment représenté par les modèles des processus PP. Néanmoins, la simple corrélation entre effet PP et perméabilité est fort et consistant. En conséquence la théorie de Schwarz sur l'effet-PP a été adapté de manière semi-empirique pour mieux pouvoir estimer la relation entre les résultats de l'effet-PP et les paramètres taille de graines et perméabilité. Nos résultats concernant l'influence de la texture des matériaux et celles de l'effet de l'électrochimie des fluides dans les pores, permettront de visualiser des divers aspects du sous-sol. Avec des telles mesures géo-électriques, les hydrologues peuvent collectionner des données contenant des informations sur la structure et la chimie des fluides des sous-sols. Néanmoins, plus de recherches sur les origines physiques de l'effet-PP sont nécessaires afin de minimiser le risque potentiel d'une mauvaise interprétation des données.

LC-MS method development and comparison of sampling materials for the analysis of organic gunshot residues

This study aimed at comparing the efficiency of various sampling materials for the collection and subsequent analysis of organic gunshot residues (OGSR). To the best of our knowledge, it is the first time that sampling devices were investigated in detail for further quantitation of OGSR by LC-MS. Seven sampling materials, namely two "swab"-type and five "stub"-type collection materials, were tested. The investigation started with the development of a simple and robust LC-MS method able to separate and quantify molecules typically found in gunpowders, such as diphenylamine or ethylcentralite. The evaluation of sampling materials was then systematically carried out by first analysing blank extracts of the materials to check for potential interferences and determining matrix effects. Based on these results, the best four materials, namely cotton buds, polyester swabs, a tape from 3M and PTFE were compared in terms of collection efficiency during shooting experiments using a set of 9 mm Luger ammunition. It was found that the tape was capable of recovering the highest amounts of OGSR. As tape-lifting is the technique currently used in routine for inorganic GSR, OGSR analysis might be implemented without modifying IGSR sampling and analysis procedure.