STARTVerso1: A randomized trial of faldaprevir plus pegylated interferon/ribavirin for chronic HCV genotype-1 infection.

BACKGROUND & AIMS The efficacy and tolerability of faldaprevir, a potent hepatitis C virus (HCV) NS3/4A protease inhibitor, plus peginterferon and ribavirin was assessed in a double-blind, placebo-controlled phase 3 study of treatment-naïve patients with HCV genotype-1 infection. METHODS Patients were randomly assigned (1:2:2) to peginterferon/ribavirin plus: placebo (arm 1, n=132) for 24 weeks; faldaprevir (120 mg, once daily) for 12 or 24 weeks (arm 2, n=259); or faldaprevir (240 mg, once daily) for 12 weeks (arm 3, n=261). In arms 2 and 3, patients with early treatment success (HCV RNA <25 IU/mL at week 4 and undetectable at week 8) stopped all treatment at week 24. Other patients received peginterferon/ribavirin until week 48 unless they met futility criteria. The primary endpoint was sustained virologic response 12 weeks post-treatment (SVR12). RESULTS SVR12 was achieved by 52%, 79%, and 80% of patients in arms 1, 2, and 3, respectively (estimated difference for arms 2 and 3 versus arm 1: 27%, 95% confidence interval 17%-36%; and 29%, 95% confidence interval, 19%-38%, respectively; P<.0001 for both). Early treatment success was achieved by 87% (arm 2) and 89% (arm 3) of patients, of whom 86% and 89% achieved SVR12. Adverse event rates were similar among groups; few adverse events led to discontinuation of all regimen components. CONCLUSIONS Faldaprevir plus peginterferon/ribavirin significantly increased SVR12, compared with peginterferon/ribavirin, in treatment-naïve patients with HCV genotype-1 infection. There do not seem to be any differences in responses of patients given once-daily 120 or 240 mg faldaprevir.

Conditioned Medium of Demineralized Freeze-Dried Bone Activates Gene Expression in Periodontal Fibroblasts in Vitro.

BACKGROUND: Demineralized bone matrix (DBM) is used for the treatment of osseous defects. Conditioned medium from native bone chips can activate transforming growth factor (TGF)-β signaling in mesenchymal cells. The aim of the study was to determine whether processing of native bone into DBM affects the activity of the conditioned medium. METHODS: Porcine cortical bone blocks were subjected to defatting, different concentrations of hydrochloric acid and various temperatures. DBM was lyophilized, ground, and placed into culture medium. Human gingiva and periodontal fibroblasts were exposed to the respective conditioned medium (DBCM). Changes in the expression of TGF-β target genes were determined. RESULTS: DBCM altered the expression of TGF-β target genes, e.g., adrenomedullin, pentraxin 3, KN Motif And Ankyrin Repeat Domains 4, interleukin 11, NADPH oxidase 4, and BTB (POZ) Domain Containing 11, by at least five-fold. The response was observed in fibroblasts from both sources. Defatting lowered the activity of DBCM. The TGF-β receptor type I kinase inhibitor SB431542, but not the inhibitor of bone morphogenetic protein receptor dorsomorphin, blocked the effects of DBCM on gene expression. Moreover, conditioned medium obtained from commercial human DBM modulated the expression of TGF-β target genes. CONCLUSION: The findings suggest that the conditioned medium from demineralized bone matrix can activate TGF-β signaling in oral fibroblasts. KEYWORDS: TGF-beta superfamily proteins; bone; bone substitutes; bone transplantation; conditioned media; freeze drying

Tumour border configuration in colorectal cancer: proposal for an alternative scoring system based on the percentage of infiltrating margin.

AIMS Information on tumour border configuration (TBC) in colorectal cancer (CRC) is currently not included in most pathology reports, owing to lack of reproducibility and/or established evaluation systems. The aim of this study was to investigate whether an alternative scoring system based on the percentage of the infiltrating component may represent a reliable method for assessing TBC. METHODS AND RESULTS Two hundred and fifteen CRCs with complete clinicopathological data were evaluated by two independent observers, both 'traditionally' by assigning the tumours into pushing/infiltrating/mixed categories, and alternatively by scoring the percentage of infiltrating margin. With the pushing/infiltrating/mixed pattern method, interobserver agreement (IOA) was moderate (κ = 0.58), whereas with the percentage of infiltrating margins method, IOA was excellent (intraclass correlation coefficient of 0.86). A higher percentage of infiltrating margin correlated with adverse features such as higher grade (P = 0.0025), higher pT (P = 0.0007), pN (P = 0.0001) and pM classification (P = 0.0063), high-grade tumour budding (P < 0.0001), lymphatic invasion (P < 0.0001), vascular invasion (P = 0.0032), and shorter survival (P = 0.0008), and was significantly associated with an increased probability of lymph node metastasis (P < 0.001). CONCLUSIONS Information on TBC gives additional prognostic value to pathology reports on CRC. The novel proposed scoring system, by using the percentage of infiltrating margin, outperforms the 'traditional' way of reporting TBC. Additionally, it is reproducible and simple to apply, and can therefore be easily integrated into daily diagnostic practice.

Comparison of the capacity of enamel matrix derivative gel and enamel matrix derivative in liquid formulation to adsorb to bone grafting materials.

BACKGROUND The use of an enamel matrix derivative (EMD) has been shown to enhance periodontal regeneration (e.g., formation of root cementum, periodontal ligament, and alveolar bone). However, in certain clinical situations, the use of EMD alone may not be sufficient to prevent flap collapse or provide sufficient stability of the blood clot. Data from clinical and preclinical studies have demonstrated controversial results after application of EMD combined with different types of bone grafting materials in periodontal regenerative procedures. The aim of the present study is to investigate the adsorption properties of enamel matrix proteins to bone grafts after surface coating with either EMD (as a liquid formulation) or EMD (as a gel formulation). METHODS Three different types of grafting materials, including a natural bone mineral (NBM), demineralized freeze-dried bone allograft (DFDBA), or a calcium phosphate (CaP), were coated with either EMD liquid or EMD gel. Samples were analyzed by scanning electron microscopy or transmission electron microscopy (TEM) using an immunostaining assay with gold-conjugated anti-EMD antibody. Total protein adsorption to bone grafting material was quantified using an enzyme-linked immunosorbent assay (ELISA) kit for amelogenin. RESULTS The adsorption of amelogenin to the surface of grafting material varied substantially based on the carrier system used. EMD gel adsorbed less protein to the surface of grafting particles, which easily dissociated from the graft surface after phosphate-buffered saline rinsing. Analyses by TEM revealed that adsorption of amelogenin proteins were significantly farther from the grafting material surface, likely a result of the thick polyglycolic acid gel carrier. ELISA protein quantification assay demonstrated that the combination of EMD liquid + NBM and EMD liquid + DFDBA adsorbed higher amounts of amelogenin than all other treatment modalities. Furthermore, amelogenin proteins delivered by EMD liquid were able to penetrate the porous surface structure of NBM and DFDBA and adsorb to the interior of bone grafting particles. Grafting materials coated with EMD gel adsorbed more frequently to the exterior of grafting particles with little interior penetration. CONCLUSIONS The present study demonstrates a large variability of adsorbed amelogenin to the surface of bone grafting materials when enamel matrix proteins were delivered in either a liquid formulation or gel carrier. Furthermore, differences in amelogenin adsorption were observed among NBM, DFDBA, and biphasic CaP particles. Thus, the potential for a liquid carrier system for EMD, used to coat EMD, may be advantageous for better surface coating.

The story of Community preference for food security

"Préférence communautaire" is an in-built notion of the CAP since its inception with the Treaty of Rome (1957). Its’ simple objective laid down at the Stresa Conference in 1958 is to prefer community produce over imports wherever possible, while at the same time promoting agricultural exports and FDI (“vocation exportatrice de l’Europe”). Does this contrast or correlate with the notion of “food sovereignty” which originated in 1996 as a notion of small farmer self-sufficiency (Via Campesina), and which now has found its way into the official EC discourse? Recent CAP reforms indeed seem to continue banking on border protection and on the occasional export subsidy. Nonetheless, coming together with claims to mitigate climate change, “food sovereignty” à la CAP fails to acknowledge efficiency losses at home and negative spillover effects on the right to food of food exporting developing countries. This chapter asks whether new non-tariff and domestic support measures are just new wine in the old cask of fortress Europe, together with the FDI promotion instruments of the FED and others. Might the increasing dynamics and new challenges of agricultural trade and investment lead to lower market and production shares for European farms? It concludes that in the medium term the WTO Green Box has the only legal and effective tools to promote EU agriculture and food.

Lactoferrin: An adjuvant to enhance the efficacy of the BCG vaccine

Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB), is a disease with world wide consequences, affecting nearly a third of the world's population. The established vaccine for TB; an attenuated strain of Mycobacterium bovis Calmette Guerin (BCG), has existed virtually unchanged since 1921. Intensive research is focused on developing a TB vaccine that can surpass and improve the existing BCG vaccine. Lactoferrin, an iron binding protein found in mucosal secretions and granules of neutrophils was hypothesized to be an ideal adjuvant to enhance the efficacy of the BCG vaccine. Specifically, Lactoferrin enhanced the ratio of IL-12:IL-10 production from macrophages stimulated with LFS or infected with BCG, indicating the potential to affect T-cell development in vivo. Five different vaccination protocols were investigated for generation of host protective responses against MTB infection using Lactoferrin admixed to the BCG vaccine. Mice immunized and boosted at 2 weeks with BCG/Lactofefrin increased host protection against MTB infection by decreasing organ bacterial load and reducing lung histopathology. The observed postchallenge results paralleled with increasing production of IFN-γ, IL-2, TNF-α, and IL-12 from BCG stimulated splenocytes. In vitro studies examined possible mechanisms of Lactoferrin action on BCG infected macrophages and dendritic cells. Addition of Lactoferrin to BCG infected macrophages and dendritic cells increased stimulation of presensitized CD3+ and CD4+ T-cells. Analysis by fluorescent activated cell sorting (FACS) revealed an increase in surface expression of MHC I and decreased ratio of CD80/86 from BCG infected macrophages cultured with Lactoferrin. In contrast, Lactoferrin decreased surface expression of MHC I, MHC II, CD80, CD86, and CD40, but increased CD 11c, from BCG infected dendritic cells, indicating involvement of adhesion molecules. Overall, these studies indicate that Lactoferrin is a useful and effective adjuvant to improve efficacy of the BCG vaccine by enhancing generation of mycobacterial antigen specific T-cell responses through promotion of antigen presentation and T-cell stimulation.^

Major ion concentrations and chlorine isotope composition of water-soluble chloride and structurally bound chloride in DSDP/ODP serpentinized ultramafic rocks

Eight DSDP/ODP cores were analyzed for major ion concentrations and d37Cl values of water-soluble chloride (d37Clwsc) and structurally bound chloride (d37Clsbc) in serpentinized ultramafic rocks. This diverse set of cores spans a wide range in age, temperature of serpentinization, tectonic setting, and geographic location of drilled serpentinized oceanic crust. Three of the cores were sampled at closely spaced intervals to investigate downhole variation in Cl concentration and chlorine isotope composition. The average total Cl content of all 86 samples is 0.26±0.16 wt.% (0.19±0.10 wt.% as water-soluble Cl (Xwsc) and 0.09±0.09 wt.% as structurally bound Cl (Xsbc)). Structurally bound Cl concentration nearly doubles with depth in all cores; there is no consistent trend in water-soluble Cl content among the cores. Chlorine isotope fractionation between the structurally bound Cl**- site and the water-soluble Cl**- site varies from -1.08? to +1.16?, averaging to +0.21?. Samples with negative fractionations may be related to reequilibration of the water-soluble chloride with seawater post-serpentinite formation. Six of the cores have positive bulk d37Cl values (+0.05? to +0.36?); the other two cores (173-1068A (Leg-Hole) and 84-570) have negative bulk d37Cl values (-1.26? and -0.54?). The cores with negative d37Cl values also have variable Cl**-/SO4**2- ratios, in contrast to all other cores. The isotopically positive cores (153-920D and 147-895E) show no isotopic variation with depth; the isotopically negative core (173-1068A) decreases by ~1? with depth for both the water-soluble and structurally bound Cl fractions. Non-zero bulk d37Cl values indicate Cl in serpentinites was incorporated during original hydration and is not an artifact of seawater infiltration during drilling. Cores with positive d37Cl values are most likely explained by open system fractionation during hydrothermal alteration, with preferential incorporation of 37Cl from seawater into the serpentinite and loss of residual light Cl back to the ocean. Fluid / rock ratios were probably low as evidenced by the presence of water-soluble salts. The two isotopically negative cores are characterized by a thick overlying sedimentary package that was in place prior to serpentinization. We believe the low d37Cl values of these cores are a result of hydration of ultramafic rock by infiltrating aqueous pore fluids from the overlying sediments. The resulting serpentinites inherit the characteristic negative d37Cl values of the pore waters. Chlorine stable isotopes can be used to identify the source of the serpentinizing fluid and ultimately discern chemical and tectonic processes involved in serpentinization.

The 2008 DGFI Realization of the ITRS: DTRF2008 (data)

The DTRF2008 is a realization of the International Terrestrial Reference System ITRS. The DTRF2008 consists of station positions and velocities of global distributed observing stations of the space geodetic observation techniques VLBI, SLR, GPS and DORIS. The DTRF2008 was released in May 2010 and includes the observation data of the techniques up to and including 2008. The observation data are processed and submitted by the corresponding international services: IGS (International GNSS Service, http://igscb.jpl.nasa.gov) IVS (International VLBI Service, http://ivscc.gsfc.nasa.gov) ILRS (International Laser Ranging Service, http://ilrs.gsfc.nasa.gov) IDS (International DORIS Service, http://ids-doris.org). The DTRF2008 is an independent ITRS realization, which is computed on the basis of the same input data as the ITRF2008 (IGN, Paris). Both realizations differ with respect to their computation strategies: while the ITRF2008 is based on the combination of solutions, the DTRF2008 is computed by the combination of normal equations. The DTRF2008 comprises the coordinates of 559 GPS-, 106 VLBI-, 122 SLR- and 132 DORIS-stations. The reference epoch is 1.1.2005, 0h UTC. The Earth Orientation Parameters (EOP) - that means the coordinates of the terrestrial and the celestial pole, UT1-UTC and the Length of Day (LOD) - were simultaneously estimated with the station coordinates. The EOP time series cover the period of 1983 to 2008. The station names are the official IERS indications: cdp numbers or 4-character IDs and DOMES numbers (http://itrf.ensg.ign.fr/doc_ITRF/iers_sta_list.txt). The solution is available in different file formats (SINEX and SSC), see below. A detailed description of the solution is given by Seitz M. et al. (2012). The results of a comparison of DTRF2008 and ITRF2008 is given by Seitz M. et al. (2013). More information as well as residual time series of the station positions can be made available by request.

Abundance of benthic foraminifera in late Paleocene to eraly Pliocene sediments of DSDP Hole 74-525A on the Walvis Ridge, South Atlantic (Table 1)

Benthic forammifers in the size-fraction greater than 0.073 mm were studied in 88 Paleocene to Pleistocene samples from Deep Sea Drilling Project Site 525 (Hole 525A, Walvis Ridge, eastern south Atlantic). Clustering of the samples on the basis of the 86 most abundant foramimfers (in total, 331 taxa were identified) allowed separating two major assemblage zones: the Paleocene to Eocene interval, and the Oligocene to Pleistocene interval. Each of these, in turn, were subdivided into three minor subzones as follows: lower upper Paleocene (approx. 62.4 to 57 8 Ma); upper upper Paleocene (56.6 to 56 2 Ma), lower and middle Eocene (55.3 to 46 8 Ma); upper Oligocene to middle Miocene (25.3 to 16 Ma), middle Miocene to Pliocene (15.7 to 4.2 Ma), and lower Pleistocene (0.4 to 0.02 Ma), with only minor differences with the previous zone. Some very abundant taxa span most of the column studies (Bolivina huneri, Cassidulina subglobosa, Eponides bradyi, E. weddellensis, Gavelinella micra, Oridorsalis umbonatus, etc.). Several of the faunal breaks recorded coincide with conspicuous minima in the specific diversity curve, thus suggesting that the corresponding turnovers signal the final stages of periods of faunal impoverishment. At least one major bottomwater temperature drop (as derived from delta18O data) is synchronous with a decrease in the forammiferal specific diversity. On the other hand, a specific diversity maximum in the middle Miocene might be associated with a delta13C increase at approx 16 to 12 Ma. Highest foraminiferal abundances (up to 600-800 individuals per gram of dry sediment) occurred in the late Paleocene and in the early Pleistocene, in coincidence with the lowest diversity figures calculated. The magnitude of the most important faunal turnover recorded, between the middle Eocene and the late Oligocene, is magnified in our data set by the large hiatus which separates the middle Eocene from the upper Oligocene sediments. Considerably smaller overturns occurred within the late Paleocene (in coincidence with changes in the specific diversity, absolute abundance of forammiferal tests, and delta13C), and in the middle Miocene (in coincidence with a specific diversity maximum and a delta13C excursion). New reformation on the morphology and the stratigraphic ranges of several species is furnished. For all the taxa recorded the number of occurrences, total number of individuals identified and first and last appearances are listed.

Technical Challenges in the Construction of Gothic Vaults: The Gothic Theory of Structural Design

The construction of a Gothic vault implied the solution of several technical challenges. The literature on Gothic vault construction is quite large and its growth continues steadily. The main challenge of any structure is that, during and after construction, it must be "safe", that is, it must not collapse. Indeed, it must be amply safe, able to support different loads for long periods of time. Masonry architecture has shown its structural safety for centuries or millennia. The Pantheon of Rome stands today after almost 2,000 years without having needed any structural reinforcement (of course, the survival of any building implies continuous maintenance) . Hagia Sophia in Istanbul, finished in the 6th century AD, has withstood not only the dead loads but also many severe earthquakes . Finally, the Gothic cathedrals, with their appearance of weakness, are• more than a half millennium old. The question arises of what the source of this amazing strength is and how the illiterate master masons were able to design such daring and safe structures . This question is usually evaded in manuals of Gothic architecture. This is quite surprising, the structure being a fundamental part of Gothic buildings. The present article aims to give such an explanation, which has been studied in detail elsewhere. In the first part, the Gothic design methods "V ill be discussed. In the second part, the validity of these methods wi11 be verified within the frame of the modern theory of masonry structures . References have been reduced to a minimum to make the text simpler and more direct.

Protonatable residues at the cytoplasmic end of transmembrane helix-2 in the signal transducer HtrI control photochemistry and function of sensory rhodopsin I.

Neutral residue replacements were made of 21 acidic and basic residues within the N-terminal half of the Halobacterium salinarium signal transducer HtrI [the halobacterial transducer for sensory rhodopsin I (SRI)] by site-specific mutagenesis. The replacements are all within the region of HtrI that we previously concluded from deletion analysis to contain sites of interaction with the phototaxis receptor SRI. Immunoblotting shows plasmid expression of the htrI-sopI operon containing the mutations produces SRI and mutant HtrI in cells at near wild-type levels. Six of the HtrI mutations perturb photochemical kinetics of SRI and one reverses the phototaxis response. Substitution with neutral amino acids of Asp-86, Glu-87, and Glu-108 accelerate, and of Arg-70, Arg-84, and Arg-99 retard, the SRI photocycle. Opposite effects on photocycle rate cancel in double mutants containing one replaced acidic and one replaced basic residue. Laser flash spectroscopy shows the kinetic perturbations are due to alteration of the rate of reprotonation of the retinylidene Schiff base. All of these mutations permit normal attractant and repellent signaling. On the other hand, the substitution of Glu-56 with the isosteric glutamine converts the normally attractant effect of orange light to a repellent signal in vivo at neutral pH (inverted signaling). Low pH corrects the inversion due to Glu-56 -> Gln and the apparent pK of the inversion is increased when arginine is substituted at position 56. The results indicate that the cytoplasmic end of transmembrane helix-2 and the initial part of the cytoplasmic domain contain interaction sites with SRI. To explain these and previous results, we propose a model in which (i) the HtrI region identified here forms part of an electrostatic bonding network that extends through the SRI protein and includes its photoactive site; (ii) alteration of this network by photoisomerization-induced Schiff base deprotonation and reprotonation shifts HtrI between attractant and repellent conformations; and (iii) HtrI mutations and extracellular pH alter the equilibrium ratios of these conformations.

Protein kinase cross-talk: membrane targeting of the beta-adrenergic receptor kinase by protein kinase C.

The beta-adrenergic receptor kinase (betaARK) is the prototypical member of the family of cytosolic kinases that phosphorylate guanine nucleotide binding-protein-coupled receptors and thereby trigger uncoupling between receptors and guanine nucleotide binding proteins. Herein we show that this kinase is subject to phosphorylation and regulation by protein kinase C (PKC). In cell lines stably expressing alpha1B- adrenergic receptors, activation of these receptors by epinephrine resulted in an activation of cytosolic betaARK. Similar data were obtained in 293 cells transiently coexpressing alpha1B- adrenergic receptors and betaARK-1. Direct activation of PKC with phorbol esters in these cells caused not only an activation of cytosolic betaARK-1 but also a translocation of betaARK immunoreactivity from the cytosol to the membrane fraction. A PKC preparation purified from rat brain phospborylated purified recombinant betaARK-1 to a stoichiometry of 0.86 phosphate per betaARK-1. This phosphorylation resulted in an increased activity of betaARK-1 when membrane-bound rhodopsin served as its substrate but in no increase of its activity toward a soluble peptide substrate. The site of phosphorylation was mapped to the C terminus of betaARK-1. We conclude that PKC activates betaARK by enhancing its translocation to the plasma membrane.