Bradykinin B(1) receptor antagonist R954 inhibits eosinophil activation/proliferation/migration and increases TGF-beta and VEGF in a murine model of asthma

In the present study the effects of bradykinin receptor antagonists were investigated in a murine model of asthma using BALB/c mice immunized with ovalbumin/alum and challenged twice with aerosolized ovalbumin. Twenty four hours later eosinophil proliferation in the bone marrow, activation (lipid bodies formation), migration to lung parenchyma and airways and the contents of the pro-angiogenic and pro-fibrotic cytokines TGF-beta and VEGF were determined. The antagonists of the constitutive B(2) (HOE 140) and inducible B(1) (R954) receptors were administered intraperitoneally 30 min before each challenge. In sensitized mice, the antigen challenge induced eosinophil proliferation in the bone marrow, their migration into the lungs and increased the number of lipid bodies in these cells. These events were reduced by treatment of the mice with the B(1) receptor antagonist. The B(2) antagonist increased the number of eosinophils and lipid bodies in the airways without affecting eosinophil counts in the other compartments. After challenge the airway levels of VEGF and TGF-beta significantly increased and the B(1) receptor antagonist caused a further increase. By immunohistochemistry techniques TGF-beta was found to be expressed in the muscular layer of small blood vessels and VEGF in bronchial epithelial cells. The B(1) receptors were expressed in the endothelial cells. These results showed that in a murine model of asthma the B(1) receptor antagonist has an inhibitory effect on eosinophils in selected compartments and increases the production of cytokines involved in tissue repair. It remains to be determined whether this effects of the B(1) antagonist would modify the progression of the allergic inflammation towards resolution or rather towards fibrosis. (C) 2009 Elsevier Ltd. All rights reserved.

Evidences of the cooperative role of the chemokines CCL3, CCL4 and CCL5 and its receptors CCR1+and CCR5+in RANKL+ cell migration throughout experimental periodontitis in mice

Periodontal disease (PD) is characterized by the inflammatory bone resorption in response to the bacterial challenge, in a host response that involves a series of chemokines supposed to control cell influx into periodontal tissues and determine disease outcome. In this study, we investigated the role of chemokines and its receptors in the immunoregulation of experimental PD in mice. Aggregatibacter actinomycetemcomitans-infected C57BI/6 (WT) mice developed an intense inflammatory reaction and severe alveolar bone resorption, associated with a high expression of CCL3 and the migration of CCR5+, CCR1+ and RANKL+ cells to periodontal tissues. However, CCL3KO-infected mice developed a similar disease phenotype than WT strain, characterized by the similar expression of cytokines (TNF-alpha, IFN-gamma and IL-10), osteoclastogenic factors (RANKL and OPG) and MMPs (MMP-1, MMP-2, MMP-3, TIMP-1 and TIMP-3), and similar patterns of CCR1+, CCR5+ and RANKL+ cell migration. The apparent lack of function for CCL3 is possible due the relative redundancy of chemokine system, since chemokines such as CCL4 and CCL5, which share the receptors CCR1 and CCR5 with CCL3, present a similar kinetics of expression than CCL3. Accordingly, CCL4 and CCL5 kinetics of expression after experimental periodontal infection remain unaltered regardless the presence/absence of CCL3. Conversely, the individual absence of CCR1 and CCR5 resulted in a decrease of leukocyte infiltration and alveolar bone loss. When CCR1 and CCR5 were simultaneously inhibited by met-RANTES treatment a significantly more effective attenuation of periodontitis progression was verified, associated with lower values of bone loss and decreased counts of leukocytes in periodontal tissues. Our results suggest that the absence of CCL3 does not affect the development of experimental PD in mice, probably due to the presence of homologous chemokines CCL4 and CCL5 that overcome the absence of this chemokine. In addition, our data demonstrate that the absence of chemokine receptors CCR1+ and CCR5+ attenuate of inflammatory bone resorption. Finally, our data shows data the simultaneous blockade of CCR1 and CCR5 with MetRANTEs presents a more pronounced effect in the arrest of disease progression, demonstrating the cooperative role of such receptors in the inflammatory bone resorption process throughout experimental PD. (C) 2009 Elsevier Inc. All rights reserved.

The geodiamolide H, derived from Brazilian sponge Geodia corticostylifera, regulates actin cytoskeleton, migration and invasion of breast cancer cells cultured in three-dimensional environment

We are investigating effects of the depsipeptide geodiamolide H, isolated from the Brazilian sponge Geodia corticostylifera, on cancer cell lines grown in 3D environment. As shown previously geodiamolide H disrupts actin cytoskeleton in both sea urchin eggs and breast cancer cell monolayers. We used a normal mammary epithelial cell line MCF 10A that in 3D assay results formation of polarized spheroids. We also used cell lines derived from breast tumors with different degrees of differentiation: MCF7 positive for estrogen receptor and the Hs578T, negative for hormone receptors. Cells were placed on top of Matrigel. Spheroids obtained from these cultures were treated with geodiamolide H. Control and treated samples were analyzed by light and confocal microscopy. Geodiamolide H dramatically affected the poorly differentiated and aggressive Hs578T cell line. The peptide reverted HsS78T malignant phenotype to polarized spheroid-like structures. MCF7 cells treated by geodiamolide H exhibited polarization compared to controls. Geodiamolide H induced striking phenotypic modifications in Hs578T cell line and disruption of actin cytoskeleton. We investigated effects of geodiamolide H on migration and invasion of Hs578T cells. Time-lapse microscopy showed that the peptide inhibited migration of these cells in a dose-dependent manner. Furthermore invasion assays revealed that geodiamolide H induced a 30% decrease on invasive behavior of Hs578T cells. Our results suggest that geodiamolide H inhibits migration and invasion of Hs578T cells probably through modifications in actin cytoskeleton. The fact that normal cell lines were not affected by treatment with geodiamolide H stimulates new studies towards therapeutic use for this peptide.

3D Cell-Migration Studies using Two-Photon Engineered Polymer Scaffolds

We use two-photon polymerization to fabricate 3D scaffolds with precise control over pore size and shape for studying cell migration in 3D. These scaffolds allow movement of cells in all directions. The fabrication, imaging, and quantitative analysis method developed here can be used to do systematic cell studies in 3D.

Synthesis of the Macrolactone of Migrastatin and Analogues with Potent Cell-Migration Inhibitory Activity

The synthesis of the macrolactone core of migrastatin 2, its potent anti-metastasis analogue 34, and ester derivatives 35 and 38 are reported. The approach involves the use of a dihydroxylation reaction to establish the desired C-8 stereocenter followed by a metathesis cyclization reaction. The effects of the compounds on the migration and invasion of human breast cancer cells were evaluated by using the wound-healing and the Boyden-chamber cell-migration and cell-invasion assays. The results revealed a high potency of the macrolactones 2 and 34 and the ester analogues 35 and 38, which suggests they have potential as antimetastatic agents.

The Sphene-Centered Ocellar Texture: An Effect of Grain-Supported Flow and Melt Migration in a Hyperdense Magma Mush

The sphene-centered ocellar texture consists of leucocratic ocelli with sphene (titanite) crystals at the center, enclosed in a biotite-rich matrix. This texture has been recognized worldwide in hybrid intermediate rocks. On the basis of structural, petrological, and geochronological data from selected outcrops of the Variscan Ribadelago pluton (NW Iberian Massif), we propose that the ocelli were formed by migration and accumulation of a residual melt through a plagioclase- and biotite-dominated crystalline framework. At the late stage of crystallization, the magma acted as a hyperdense suspension and reacted to the pressure gradient caused by the regional stress field, entering the domain of grain-supported flow. Microstructures reveal that aligned crystal domains arose in the crystal framework from the shearing and compaction of the crystal mush and behaved as magmatic microshears. Relative displacement of adjacent crystal clusters along these microshears corresponded to the onset of Reynolds dilatancy that generated an expansion of the crystal mush, involving melt migration and pore aperture. The mineralogy of the ocelli, dominated by andesine and sphene, represents the composition of the migrating melt. The chemistry of this late, Ti-rich melt stems from the incongruent melting of biotite. Magmatic sphene from the ocelli yields a U-Pb age of 317 +/- 1 Ma, which represents the final crystallization of the hybridized magmatic system. Moreover, this texture offers an opportunity to better understand the rheological behavior of highly crystallized magmas.

Late Quaternary sedimentation in the Paraiba Basin, Northeastern Brazil: Landform, sea level and tectonics in Eastern South America passive margin

Late Quaternary deposits in the northeastern Brazil have been scarcely investigated, despite their relevance to the discussion of the post-rift evolution of the South American passive margin within the context of landform, sea level and tectonic deformation. Sedimentological, stratigraphic and morphological characterization of these deposits, referred as Post-Barreiras Sediments, led to their distinction from underlying Early/Middle Miocene strata. Based on optically stimulated luminescence (OSL) dating, two sedimentary units (PB1 and PB2) were recognized and related to the time intervals between 74.8 +/- 9.3 and 30.8 +/- 6.9 ka, and 8.8 +/- 0.9 and 1.8 +/- 0.2 ka, respectively. Unit PB1 consists of indurated sandstones and breccias either with massive bedding or complex types of soft sediment deformation structures generated by contemporaneous seismic activity. Unit PB2 is composed of massive sands or sands related to structures developed by dissipation of dunes. The present work, focusing on the Post-Barreiras Sediments, discusses landform, sea level and tectonics of the eastern South American passive margin during the latest Quaternary. Non-deposition and sub-aerial exposure related to the Tortonian worldwide low sea level combined with tectonic quiescence followed the Miocene transgression. Tectonic deformation in the latest Pleistocene created space to accommodate unit PB1 in downthrown faulted blocks and, perhaps, also synclines produced by strike-slip deformation. Although deposition of this unit was simultaneous with the progressive fall in sea level that followed the Last Interglacial Maximum, punctuated rises combined with land subsidence led to marine deposition close to the modern coastline. Renewed subsidence in the Holocene gave rise to accommodation of the Post-Barreiras Sediments. Most of unit PB2 was deposited during the Holocene Transgression, but it is not composed of marine sediments, which suggests either an insignificant rise in relative sea level or aeolian reworking of thin transgressive sands. The data presented here lead to a review of the evolution of the South American passive margin based on assumptions of uniform sedimentation and undeformed planation surfaces over a wide coastal area of the northeastern Brazil. (C) 2011 Elsevier B.V. All rights reserved.

Sediment deformation in Miocene and post-Miocene strata, Northeastern Brazil: Evidence for paleoseismicity in a passive margin

Stratigraphic intervals characterized by varied and complex styles of soft-sediment deformation structures are well preserved in Miocene and Late Pleistocene to Holocene deposits of a sedimentary basin located in Northeastern Brazil. The Miocene strata, represented by the Barreiras Formation, record only brittle structures, including numerous faults and fractures with straight and high angle-dipping planes that are often filled with sands derived from overlying beds. Folds consisting of broad anticlines and synclines are also present in this unit. The late Pleistocene to Holocene deposits, named Post-Barreiras Sediments, contain an indurated sandy package with a large variety of ductile and brittle deformation structures (i.e., massive sandstones with isolated sand fragments and breccias, undulatory strata, sand dykes and diapirs, sinks and bowls, pebbly pockets, plunged sediment mixtures, fitted sand masses, cone-shaped cracks, fault grading and sedimentary enclaves). These features, confined to sharp-based stratigraphic horizons that progressively grade downward into undisturbed deposits, are related to seismic shocks of high surface-wave magnitude (i.e., Ms>5 or 6). Amalgamated seismites suggest that previously formed seismites were affected by subsequent seismic-wave propagation. Seismic waves caused by activity along one, or most likely, several tectonic structures would have propagated throughout the depositional environment, producing laterally extensive seismites. The close proximity to earthquake epicenters would have promoted pervasive re-sedimentation due to pore overpressure, resulting high volumes of massive sandstones and breccia. The similarity between deposits with correlatable strata from many other areas along the Brazilian coast allows raise the hypothesis that the seismic episodes might have affected sedimentation patterns in a large (i.e., extension of several hundreds of kilometers) geographic area. Thus, the modern seismicity recorded along Northeastern Brazil was recurrent during the Quaternary and, perhaps, also in the Pliocene. The estimated high magnitude of the seismic events and the great regional extent of the affected area demonstrate that the Brazilian coast experienced tectonic stress through the last geological episodes of its evolution, which would have favored sediment accumulation and penecontemporaneous re-sedimentation. This geological context is unexpected in a passive margin, inducing to revisit the debate on how active is a passive margin. (C) 2010 Elsevier B.V. All rights reserved.

Denudation history of the Bocaina Plateau, Serra do Mar, southeastern Brazil: Relationships to Gondwana breakup and passive margin development

The Bocaina Plateau, which is situated on the eastern flank of the continental rift of southeastern Brazil, is the highest part of the Serra do Mar. Topographic relief in this area is suggested to be closely related to its complex tectono-magmatic evolution since the breakup of Western Gondwana and opening of the South Atlantic Ocean. Apatite fission track ages and track length distributions from 27 basement outcrops were determined to assess these hypotheses and reconstruct the denudation history of the Bocaina Plateau. The ages range between 303 +/- 32 and 46 +/- 5 Ma, and are significantly younger than the stratigraphic ages. Mean track lengths vary from 13.44 +/- 1.51 to 11.1 +/- 1.48 mu m, with standard deviations between 1.16 and 1.83 mu m. Contrasting ages within a single plateau and similar ages at different altitudes indicate a complex regional tectonothermal evolution. The thermal histories inferred from these data imply three periods of accelerated cooling related to the Early Cretaceous continental breakup, Early Cretaceous alkaline magmatism, and the Paleogene evolution of the continental rift of southeastern Brazil. The oldest fission track ages (>200 Ma) were obtained in the Serra do Mar region, suggesting that these areas were a long-lived source of sediments for the Parana, Bauru, and Santos basins. (C) 2010 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Quaternary tectonics in a passive margin: Marajo Island, northern Brazil

Marajo Island is located in a passive continental margin that evolved from rifting associated with the opening of the Equatorial South Atlantic Ocean in the Late Jurassic/Early Cretaceous period. This study, based on remote sensing integrated with sedimentology, as well as subsurface and seismographic data available from the literature, allows discussion of the significance of tectonics during the Quaternary history of marginal basins. Results show that eastern Marajo Island contains channels with evidence of tectonic control. Mapping of straight channels defined four main groups of lineaments (i.e. NNE-SSW, NE-SW, NW-SE and E-W) that parallel main normal and strike-slip fault zones recorded for the Amazon region. Additionally, sedimentological studies of late Quaternary and Holocene deposits indicate numerous ductile and brittle structures within stratigraphic horizons bounded by undeformed strata, related to seismogenic deformation during or shortly after sediment deposition. This conclusion is consistent with subsurface Bouguer mapping suggestive of eastern Marajo Island being still part of the Marajo graben system, where important fault reactivation is recorded up to the Quaternary. Together with the recognition of several phases of fault reactivation, these data suggest that faults developed in association with rift basins might remain active in passive margins, imposing important control on development of depositional systems. Copyright (C) 2007 John Wiley & Sons, Ltd.

Glypican-3 regulates migration, adhesion and actin cytoskeleton organization in mammary tumor cells through Wnt signaling modulation

Glypican-3 (GPC3) is a proteoglycan involved in migration, proliferation and cell survival modulation in several tissues. There are many reports demonstrating a downregulation of GPC3 expression in some human tumors, including mesothelioma, ovarian and breast cancer. Previously, we determined that GPC3 reexpression in the murine mammary adenocarcinoma LM3 cells induced an impairment of their in vivo invasive and metastatic capacities together with a higher susceptibility to in vitro apoptosis. Currently, the signaling mechanism of GPC3 is not clear. First, it was speculated that GPC3 regulates the insulin-like growth factor (IGF) signaling system. This hypothesis, however, has been strongly challenged. Recently, several reports indicated that at least in some cell types GPC3 serves as a selective regulator of Wnt signaling. Here we provide new data demonstrating that GPC3 regulates Wnt pathway in the metastatic adenocarcinoma mammary LM3 cell line. We found that GPC3 is able to inhibit canonical Wnt signals involved in cell proliferation and survival, as well as it is able to activate non canonical pathway, which directs cell morphology and migration. This is the first report indicating that breast tumor cell malignant properties can be reverted, at least in part, by GPC3 modulation of Wnt signaling. Our results are consistent with the potential role of GPC3 as a metastasis suppressor.

Intrachain Energy Migration to Weak Charge-Transfer State in Polyfluorene End-Capped with Naphthalimide Derivative

Polyfluorene end-capped with N-(2-benzothiazole)-1 8-naphthalimide (PF-BNI) is a highly fluorescent material with fluorescence emission modulated by solvent polarity Its low energy excited state is assigned as a mixed configuration state between the singlet S(1) of the fluorene backbone (F) with the charge transfer (CI) of the end group BNI The triexponential fluorescence decays of PF-BNI were associated with fast energy migration to form an intrachain charge-transfer (ICCT) state polyfluorene backbone decay and ICCT deactivation Time-resolved fluorescence anisotropy exhibited biexponential relaxation with a fast component of 12-16 ps in addition to a slow one in the range 0 8-1 4 ns depending on the solvent showing that depolarization occurs from two different processes energy migration to form the ICCT state and slow rotational diffusion motion of end segments at a longer time Results from femtosecond transient absorption measurements agreed with anisotropy decay and showed a decay component of about 16 ps at 605 nm in PF BNI ascribed to the conversion of S(1) to the ICCT excited state From the ratio of asymptotic and initial amplitudes of the transient absorption measurement the efficiency of intrachain ICCT formation is estimated in 0 5 which means that on average, half of the excited state formed in a BNI-(F)(n)-BNI chain with n = 32 is converted to its low energy intrachain charge-transfer (ICCT) state

Prosodic Morphology : Gender in Arabic Perfect Active and Passive 3rd Person Singular Verbs

Prosodic /template Morphology, that "draws heavily on the theoretical apparatus and formalisms of the generative phonology model known as autosegmental phonology" (Katamba, F. 1993: 154), is the best analysis that can handle Arabic morphology. Verbs in Arabic are represented on three independent tiers: root tier, the skeletal tier and the vocalic melody tier (Katamba, F. 1993). Vowel morphemes, which are represented by diacritics, are inserted within the consonant morphemes, which are represented by primary symbols, to form words. The morpheme tier hypothesis paves the way to understand the nonconcatenative Arabic morphology. This paper analyzes gender in perfect active and passive 3rd person singular verbs on the basis of PM. The focus of the analysis shall be drawn heavily on the most common Arabic verbs; triconsonantal verbs, with brief introduction of the less common verbs; quadriconsonantal perfect active and passive masculine and feminine 3rd person singular verbs. I shall, too, cast the light on some vowel changes that some verbs undergo when voice changes.

Passive solar options for reducing heating demand and maintain indoor climate in a multifamily house in Sweden

This research was carried out by studying possible renovation of a two-storey detached multifamily building by using passive solar design options in a cold climate in Borlänge, Sweden where the heating Degree Days are 4451 (base 20°C). Borlänge`s housing company, Tunabyggen, plans to renovate the project house located inthe multicultural district, Jakobsgårdarna. The goal of the thesis was to suggest a redesign of the current building, decrease the heating energy use, by applying passive solar design and control strategies, in a most reasonable way. In addition ensure a better thermal comfort for the tenants in the dwellings. Literatures have been studied, from which can be inferred that passive design should be abasic design consideration for all housing constructions, because it has advantages to ensure thermal comfort, and reduce the energy use. In addition further savings can be achieved applying different types of control strategies, from which the house will be more personalized, and better adapted to the user’s needs.The proposed method is based on simulations by using TRNSYS software. First a proper building model was set up, which represents the current state of the project building. Then the thermal insulation and the windows were upgraded, based on today's building regulations. The developments of the passive solar options were accomplished in two steps. First of all the relevant basic passive design elements were considered, then those advantages were compared to the advantages of applying new conventional thermostat, and shading control strategies.The results show that there is significant potential with the different types of passive solar design; their usage depends primarily on the location of the site as well as the orientation of the project building. Applying the control strategies, such as thermostat, and shading control, along the thermal insulation upgrade, may lead to significant energy savings (around 40 %), by comparison to the reference building without any upgrade.

Passive crosslaminated timber buildings : Final report Cerbof-project no. 76

In this project, Stora Enso’s newly developed building system has been further developed to allow building to the Swedish passive house standard for the Swedish climate. The building system is based on a building framework of CLT (Cross laminated timber) boards. The concept has been tested on a small test building. The experience gained from this test building has also been used for planning a larger building (two storeys with the option of a third storey) with passive house standard with this building system. The main conclusions from the project are:  It is possible to build airtight buildings with this technique without using traditional vapour barriers. Initial measurements show that this can be done without reaching critical humidity levels in the walls and roof, at least where wood fibre insulation is used, as this has a greater capacity for storing and evening out the moisture than mineral wool. However, the test building has so far not been exposed to internal generation of moisture (added moisture from showers, food preparation etc.). This needs to be investigated and this will be done during the winter 2013-14.  A new fixing method for doors and windows has been tested without traditional fibre filling between them and the CLT panel. The door or window is pressed directly on to the CLT panel instead, with an expandable sealing strip between them. This has been proved to be successful.  The air tightness between the CLT panels is achieved with expandable sealing strips between the panels. The position of the sealing strips is important, both for the air tightness itself and to allow rational assembly.  Recurrent air tightness measurements show that the air tightness decreased somewhat during the first six months, but not to such an extent that the passive house criteria were not fulfilled. The reason for the decreased air tightness is not clear, but can be due to small movements in the CLT construction and also to the sealing strips being affected by changing outdoor temperatures.  Long term measurements (at least two years) have to be carried out before more reliable conclusions can be drawn regarding the long term effect of the construction on air tightness and humidity in the walls.  An economic analysis comparing using a concrete frame or the studied CLT frame for a three storey building shows that it is probably more expensive to build with CLT. For buildings higher than three floors, the CLT frame has economic advantages, mainly because of the shorter building time compared to using concrete for the frame. In this analysis, no considerations have been taken to differences in the influence on the environment or the global climate between the two construction methods.