Development of a control banding tool for nanomaterials

Control banding (CB) can be a useful tool for managing the potential risks of nanomaterials. The here proposed CB, which should be part of an overall risk control strategy, groups materials by hazard and emission potential. The resulting decision matrix proposes control bands adapted to the risk potential levels and helps define an action plan. If this plan is not practical and financially feasible, a full risk assessment is launched. The hazard banding combines key concepts of nanomaterial toxicology: translocation across biological barriers, fibrous nature, solubility, and reactivity. Already existing classifications specific to the nanomaterial can be used "as is." Otherwise, the toxicity of bulk or analogous substances gives an initial hazard band, which is increased if the substance is not easily soluble or if it has a higher reactivity than the substance. The emission potential bands are defined by the nanomaterials' physical form and process characteristics. Quantities, frequencies, and existing control measures are taken into account during the definition of the action plan. Control strategies range from room ventilation to full containment with expert advice. This CB approach, once validated, can be easily embedded in risk management systems. It allows integrating new toxicity data and needs no exposure data. [Authors]

Serial protein labeling with infrared maleimide dyes to identify cysteine modifications

Cysteine thiol modifications are increasingly recognized to occur under both physiological and pathophysiological conditions, making their accurate detection, identification and quantification of growing importance. However, saturation labeling of thiols with fluorescent dyes results in poor protein recuperation and therefore requires the use of large quantities of starting material. This is especially important in sequential dye-labeling steps when applied for an identification of cysteine modifications. First, we studied the effects of different detergents during labeling procedure, i.e. Tween 20, Triton X-100 and CHAPS, on protein yield and composition. Tween 20 and Triton X-100 resulted in yields of around 50% labeled proteins compared to only 10% with PBS alone and a most diversified 2-DE protein pattern. Secondly, Tween 20 was used for serial protein labeling with maleimid fluorophores, first to conjugate to accessible thiols and after a reduction to label with another fluorophore previously masked di-sulphide and/or oxidized proteins in frontal cortex autopsy tissue of a subject with mild Alzheimer's disease. Two-DE DIGE revealed a complex protein pattern of readily labeled thiols and di-sulphide and/or oxidized proteins. Seventeen proteins were identified by MALDI-TOF and by peptide fingerprints. Several proteins were oxidized and involved in Alzheimer's disease. However methionine oxidation was prevalent. Infrared DIGE may provide an additional tool for an identification of oxidation susceptible proteins.

Two component Emden sphere

The theory of a self-gravitating gas sphere is given. The gravitational field is generated by two components, each of which is an independent isothermal gas. Various quantities of interest, such as density profiles, core radii of both components, masses, free-free luminosity, surface brightness, central surface density, and overestimate of central mass density, are given for different values of both parameters which arise naturally (ratio of central densities, ratio of rms velocities). Fundamental changes appear when comparison is made with a theory in which the second component is a 'test component'. Procedures are given for the complete analysis of real astrophysical configurations such as clusters of galaxies or globular clusters.

Effect of tibial tray inclination on the femoro-tibial contact pattern of a mobile total knee arthroplasty

Introduction: The posterior inclination of the tibial component is an important factor that can affect the success of total knee arthroplasty. It can reduce the posterior impingement and thus increase the range of flexion, but it may also induce instability in flexion, anterior impingement between the polyethylene of postero-stabilizing knee prosthesis, and anterior conflict with the cortical bone and the stem. Although the problem is identified, there is still a debate on the ideal inclination angle and the surgical technique to avoid an excessive posterior inclination. The aim of this study was to predict the effect of a posterior inclination of the tibial component on the contact pattern on the tibial insert, using a numerical musculoskeletal model of the knee joint. Methods: A 3D finite element model of the knee joint was developed to simulate an active and loaded squat movement after total knee arthroplasty. Flexion was actively controlled by the quadriceps muscle and muscle activations were estimated from EMG data and were synchronized by a feedback algorithm. Two inclinations of the tibial tray were considered: a posterior inclination of 0° or 10°. During the entire range of flexion, the following quantities were calculated: the tibiofemoral and patello-femoral contact force, and the contact pattern on polyethylene insert. The antero-posterior displacement of the contact pattern was also measured. Abaqus 6.7 was used for all analyses. Results: The tibio-femoral and patello-femoral contact forces increased during flexion and reached respectively 4 and 7 BW (bodyweight) at 90° of flexion. They were slightly affected by the inclination of the tibial tray. Without posterior inclination, the contact pattern on the tibial insert remained centered. The contact pressure was lower than 5 MPa below 60° of flexion, but exceeded 20 MPa at 90° of flexion. The posterior inclination displaced the contact point posteriorly by 2 to 4 mm. Conclusion: The inclination of the tibial tray displaced the contactpattern towards the posterior border of the tibial insert. However, even for 10° of inclination, the contact center remained far from the posterior border (12 mm). There was no instability predicted for this movement.

Implications of diagnosis related groups (DRGs) for the management of hospitals.

Although tremendous advances have been made in the diagnosis and treatment of patients, hospital administrative systems have progressed relatively slowly. The types of information available to managers in industrial sectors are not available in the health sector. For this reason, many phenomena, such as the variations of average costs and lengths of stay between different hospitals, have remained poorly explained.The DRG system defines groups of patients that consume relatively homogeneous quantities of hospital resources. On the basis, it is possible to standardize average lengths of stay and average hospital costs in terms of the differences in case mix treated. Thus DRGs can serve as an explanation of variations in these factors between different hospitals, and also (but not only) for prospective reimbursement schems. As in a number of other European countries, a project has been set up in Switzerland to examine the possibilities of using DRGs in hospital management, planning and financing.

Business Sphere, Vol. 19, no.2

The first commercial quantities of a soybean oil with about three percent linolenic acid oil were produced in Iowa in 1994 through a collaboration with Pioneer Hi-Bred International, Inc. Continued to breed new soybean varieties with a focus on a even lower linolenic acid content of one percent. I was interested in understanding whether the oil would be stable enough to eliminate the need for chemical hydrogenation. During the process of increasing one percent linolenic acid seed varieties to obtain oil for testing, the Food and Drug Administration announced it would require labeling for trans fat beginning in 2006. Instead of using the seed of the new varieties to obtain oil for testing, it was used for further seed production so that commercial quantities of the one percent linolenic acid oil could be available as quickly as possible.

PROTEOMICS OF OXIDATIVE LESIONS OCCURRING DURING TRANSFUSION-PURPOSED STORAGE OF ERYTHROCYTE CONCENTRATES

Erythrocyte concentrates (ECs) are the major labile blood product being transfused worldwide, aiming at curing anemia of diverse origins. In Switzerland, ECs are stored at 4 °C up to 42 days in saline-adenine-glucose-mannitol (SAGM). Such storage induces cellular lesions, altering red blood cells (RBCs) metabolism, protein content and rheological properties. A hot debate exists regarding the impact of the storage lesions, thus the age of ECs on transfusion-related clinical adverse outcomes. Several studies tend to show that poorer outcomes occur in patients receiving older blood products. However, no clear association was demonstrated up to date. While metabolism and early rheological changes are reversible through transfusion of the blood units, oxidized proteins cannot be repaired, and it is likely such irreversible damages would affect the quality of the blood product and the efficiency of the transfusion. In vivo, RBCs are constantly exposed to oxygen fluxes, and are thus well equipped to deal with oxidative challenges. Moreover, functional 20S proteasome complexes allow for recognition and proteolysis of fairly oxidized protein, and some proteins can be eliminated from RBCs by the release of microvesicles. The present PhD thesis is involved in a global research project which goal is to characterize the effect of processing and storage on the quality of ECs. Assessing protein oxidative damages during RBC storage is of major importance to understand the mechanisms of aging of stored RBCs. To this purpose, redox proteomic-based investigations were conducted here. In a first part, cysteine oxidation and protein carbonylation were addressed via 2D-DIGE and derivatization-driven immunodetection approaches, respectively. Then, the oxidized sub- proteomes were characterized through LC-MS/MS identification of proteins in spots of interest (cysteine oxidation) or affinity-purified carbonylated proteins. Gene ontology annotation allowed classifying targets of oxidation according to their molecular functions. In a third part, the P20S activity was evaluated throughout the storage period of ECs, and its susceptibility to highly oxidized environment was investigated. The potential defensive role of microvesiculation was also addressed through the quantification of eliminated carbonylated proteins. We highlighted distinct protein groups differentially affected by cysteine oxidation, either reversibly or irreversibly. In addition, soluble extracts showed a decrease in carbonylation at the beginning of the storage and membrane extracts revealed increasing carbonylation after 4 weeks of storage. Engaged molecular functions revealed that antioxidant (AO) are rather reversibly oxidized at their cysteine residue(s), but are irreversibly oxidized through carbonylation. In the meantime, the 20S proteasome activity is decreased by around 40 % at the end of the storage period. Incubation of fresh RBCs extracts with exogenous oxidized proteins showed a dose-dependent and protein-dependent inhibitory effect. Finally, we proved that the release of microvesicles allows the elimination of increasing quantities of carbonylated proteins. Taken together, these results revealed an oxidative pathway model of RBCs storage, on which further investigation towards improved storage conditions will be based. -- Les concentrés érythrocytaires (CE) sont le produit sanguin le plus délivré au monde, permettant de traiter différentes formes d'anémies. En Suisse, les CE sont stocké à 4 °C pendant 42 jours dans une solution saline d'adénine, glucose et mannitol (SAGM). Une telle conservation induit des lésions de stockage qui altèrent le métabolisme, les protéines et les propriétés rhéologique du globule rouge (GR). Un débat important concerne l'impact du temps de stockage des CE sur les risques de réaction transfusionnelles, certaines études tentant de démontrer que des transfusions de sang vieux réduiraient l'espérance de vie des patients. Cependant, aucune association concrète n'a été prouvée à ce jour. Alors que les modifications du métabolisme et changement précoces des propriétés rhéologiques sont réversibles suite à la transfusion du CE, les protéines oxydées ne peuvent être réparées, et il est probable que de telles lésions affectent la qualité et l'efficacité des produits sanguins. In vivo, les GR sont constamment exposés à l'oxygène, et sont donc bien équipés pour résister aux lésions oxydatives. De plus, les complexes fonctionnels de proteasome 20S reconnaissent et dégradent les protéines modérément oxydées, et certaines protéines peuvent être éliminées par les microparticules. Cette thèse de doctorat est imbriquée dans un projet de recherche global ayant pour objectif la caractérisation des effets de la préparation et du stockage sur la qualité des GR. Evaluer les dommages oxydatifs du GR pendant le stockage est primordial pour comprendre les mécanismes de vieillissement des produits sanguin. Dans ce but, des recherches orientées redoxomique ont été conduites. Dans une première partie, l'oxydation des cystéines et la carbonylation des protéines sont évaluées par électrophorèse bidimensionnelle différentielle et par immunodétection de protéines dérivatisées. Ensuite, les protéines d'intérêt ainsi que les protéines carbonylées, purifiées par affinité, sont identifiées par spectrométrie de masse en tandem. Les protéines cibles de l'oxydation sont classées selon leur fonction moléculaire. Dans une troisième partie, l'activité protéolytique du protéasome 20S est suivie durant la période de stockage. L'impact du stress oxydant sur cette activité a été évalué en utilisant des protéines exogènes oxydées in vitro. Le potentiel rôle défensif de la microvesiculation a également été étudié par la quantification des protéines carbonylées éliminées. Dans ce travail, nous avons observé que différents groupes de protéines sont affectés par l'oxydation réversible ou irréversible de leurs cystéines. De plus, une diminution de la carbonylation en début de stockage dans les extraits solubles et une augmentation de la carbonylation après 4 semaines dans les extraits membranaires ont été montrées. Les fonctions moléculaires engagées par les protéines altérées montrent que les défenses antioxydantes sont oxydées de façon réversible sur leurs résidus cystéines, mais sont également irréversiblement carbonylées. Pendant ce temps, l'activité protéolytique du protéasome 20S décroit de 40 % en fin de stockage. L'incubation d'extraits de GR en début de stockage avec des protéines oxydées exogènes montre un effet inhibiteur « dose-dépendant » et « protéine-dépendant ». Enfin, les microvésicules s'avèrent éliminer des quantités croissantes de protéines carbonylées. La synthèse de ces résultats permet de modéliser une voie oxydative du stockage des GRs, à partir de laquelle de futures recherches seront menées avec pour but l'amélioration des conditions de stockage.

Meadow Saffron Used as a Spice

Introduction: Intoxications with colchicine usually occur by ingestion of meadow saffron leaves (Colchicum autumnale) which are mistakenly collected for alimentary purposes instead of the leaves of crow garlic (Allium ursinum). Colchicine, the main alkaloid of Colchicum autumnale, is present in all parts of the plant. We report a rarer source of mistake, i.e. between the flowers of Colchicum autumnale and Crocus sativus. The similarity in this case is limited to the appearance of the flowers, but Colchicum autumnale, which is also flowering in autumn, lacks the crimson stigma from which the saffron spice is derived from Crocus sativus. Case report: A 47-year-old woman collected the stamens of a flower resembling Crocus sativus for use as saffron. Her knowledge about Crocus sativus was limited to having seen this plant previously at a museum of saffron (Mund, Switzerland). She prepared a meal with rice using three pinches of ''saffron'' for ten tablespoons of rice. She and her 8-year-old child, both ate the usual amount of rice (6 and 2 tablespoons, respectively). The 2 brothers (4- and 9-years-old) only ate 3 teaspoons of rice each. A slightly bitter taste and the absence of a yellow colouration were peculiar. Three to four hours after the meal, the mother developed nausea and contacted the Swiss Toxicological Information Centre, suspecting a plant misidentification. All family members were referred to the regional university hospital for administration of oral activated charcoal. No other symptoms were reported, notably no symptoms in the 8-year-old boy and his brothers. Colchicine serum concentration (blood sample obtained 15 hours after ingestion) measured by HPLC-mass spectrometry was 0.36 mg/L for the mother, and 0.13 mg/L for the 8-year-old child, respectively (therapeutic levels: 0.30-2.5 mg/L). Conclusion: This report demonstrates that a significant amount of colchicine may be absorbed even after ingestion of very small quantities of Colchicum autumnale, which in this case was confused with Crocus sativus. Serum colchicine concentrations in the sub-/therapeutic range can be quantified by HPLC-mass spectrometry, which allows a very sensitive and specific detection of this alkaloid in blood and urine.

Antigen binding properties of purified immunoglobulin A and reconstituted secretory immunoglobulin A antibodies.

The hybridoma cell line ZAC3 expresses Vibrio cholerae lipopolysaccharide (LPS)-specific mouse IgA molecules as a heterogeneous population of monomeric (IgAm), dimeric (IgAd), and polymeric (IgAp) forms. We describe a gentle method combining ultrafiltration, ion-exchange chromatography, and size exclusion chromatography for the simultaneous and qualitative separation of the three molecular forms. Milligram quantities of purified IgA molecules were recovered allowing for direct comparison of the biological properties of the three forms. LPS binding specificity was tested after purification; IgAd and IgAp were found to bind strongly to LPS whereas IgAm did not. Secretory IgA (sIgA) could be reconstituted in vitro by combining recombinant secretory component (rSC) and purified IgAd or IgAp, but not IgAm. Surface plasmon resonance-based binding experiments using LPS monolayers indicated that purified reconstituted sIgA and IgA molecules recognize LPS with identical affinity (KA 1.0 x 10(8)M-1). Thus, this very sensitive assay provides the first evidence that the function of SC in sIgA complex is not to modify the affinity for the antigen. KA falls to 6.6 x 10(5) M-1 when measured by calorimetry using detergent-solubilized LPS and IgA, suggesting that the LPS environment is critical for recognition by the antibody.

Fuel Adjustment, April 2011

Article 2120 of the Standard Specifications for Highway and Bridge Construction Series 2009 provides for a fuel adjustment factor to be applied to payments and partial payments for quantities of certain items of excavation work as the work is done, in accordance with the specification, when indicated in the contract documents. A Current Price Index (CPI), in dollars per gallon (liter), will be established by the DOT for each month. The CPI will be the price of No. 2 High Sulfur Diesel, as reported by Oil Price Information Service using the first weekday for the month and the average of all prices reported for Des Moines. The Base Price Index (BPI) for each contract will be the CPI in effect during the month previous to the month of the letting of that contract. If the contract has metric units, divide the Price Index ($/gal) by 3.785412 to obtain $/liter.

Fuel Adjustment, May 2011

Article 2120 of the Standard Specifications for Highway and Bridge Construction Series 2009 provides for a fuel adjustment factor to be applied to payments and partial payments for quantities of certain items of excavation work as the work is done, in accordance with the specification, when indicated in the contract documents. A Current Price Index (CPI), in dollars per gallon (liter), will be established by the DOT for each month. The CPI will be the price of No. 2 High Sulfur Diesel, as reported by Oil Price Information Service using the first weekday for the month and the average of all prices reported for Des Moines. The Base Price Index (BPI) for each contract will be the CPI in effect during the month previous to the month of the letting of that contract. If the contract has metric units, divide the Price Index ($/gal) by 3.785412 to obtain $/liter.

Regional reproducibility of calibrated BOLD functional MRI: implications for the study of cognition and plasticity.

Calibrated BOLD fMRI is a promising alternative to the classic BOLD contrast due to its reduced venous sensitivity and greater physiological specificity. The delayed adoption of this technique for cognitive studies may stem partly from a lack of information on the reproducibility of these measures in the context of cognitive tasks. In this study we have explored the applicability and reproducibility of a state-of-the-art calibrated BOLD technique using a complex functional task at 7 tesla. Reproducibility measures of BOLD, CBF, CMRO2 flow-metabolism coupling n and the calibration parameter M were compared and interpreted for three ROIs. We found an averaged intra-subject variation of CMRO2 of 8% across runs and 33% across days. BOLD (46% across runs, 36% across days), CBF (33% across runs, 46% across days) and M (41% across days) showed significantly higher intra-subject variability. Inter-subject variability was found to be high for all quantities, though CMRO2 was the most consistent across brain regions. The results of this study provide evidence that calibrated BOLD may be a viable alternative for longitudinal and cognitive MRI studies.

Can we identify Walrasian allocations?

[eng] We consider a discrete time, pure exchange infinite horizon economy with two or more consumers and at least one concumption good per period. Within the framework of decentralized mechanisms, we show that for a given consumption trade at any period of time, say at time one, the consumers will need, in general, an infinite dimensional (informational) space to identigy such a trade as an intemporal Walrasian one. However, we show and characterize a set of enviroments where the Walrasian trades at each period of time can be achieved as the equilibrium trades of a sequence of decentralized competitive mechanisms, using only both current prices and quantities to coordinate decisions.

One hundred base pairs of 5' flanking sequence of a vaccinia virus late gene are sufficient to temporally regulate late transcription.

A vaccinia virus late gene coding for a major structural polypeptide of 11 kDa was sequenced. Although the 5' flanking gene region is very A+T rich, it shows little homology either to the corresponding region of vaccinia early genes or to consensus sequences characteristic of most eukaryotic genes. Three DNA fragments (100, 200, and 500 base pairs, respectively), derived from the flanking region and including the late gene mRNA start site, were inserted into the coding sequence of the vaccinia virus thymidine kinase (TK) early gene by homologous in vivo recombination. Recombinants were selected on the basis of their TK- phenotype. Cells were infected with the recombinant viruses and RNA was isolated at 1-hr intervals. Transcripts initiating either from the TK early promoter, or from the late gene promoter at its authentic position, or from the translocated late gene promoters within the early gene were detected by nuclease S1 mapping. Early after infection, only transcripts from the TK early promoter were detected. Later in infection, however, transcripts were also initiated from the translocated late promoters. This RNA appeared at the same time and in similar quantities as the RNA from the late promoter at its authentic position. No quantitative differences in promoter efficiency between the 100-, 200-, and 500-base-pair insertions were observed. We conclude that all necessary signals for correct regulation of late-gene expression reside within only 100 base pairs of 5' flanking sequence.

Objetivo: cuantificar la reproducción

Se estudian en este trabajo algunas magnitudes relacionadas con el enfoque de la reproducción social. Ante todo se hace hincapié en tres ideas fundamentales, las nociones de: a) Salidas menos entradas; b) Salidas dividido por entradas; c) Subsistemas. A continuación se subrayan los obstáculos para la cuantificación directa de estos conceptos, y se repasan las vías sugeridas para sortear las dificultades (por medio de constructos teóricos propuestos por Leontief, von Neumann y Sraffa). Luego se examinan dos nuevos indicadores: la ¿tasaespecífica de excedente¿, que se refiere a los bienes autorreproducibles, y el ¿coeficiente neto de reproducción¿, que se predica de todos los bienes básicos. De pasada se apuntan algunas pistas para establecer indicadores del mismogénero en campos como la economía ecológica y la economía feminista. Por último, se anotan algunas conjeturas relacionadas con la dirección adecuada del cambio técnico