p38 alpha MAP Kinase Controls IL-17 Synthesis in Vogt-Koyanagi-Harada Syndrome and Experimental Autoimmune Uveitis

PURPOSE. Interleukin (IL)-17, which is responsible for the initial influx of leukocytes into the target tissue, was recently described as the main cytokine involved in autoimmune diseases. Vogt-Koyanagi-Harada (VKH) syndrome is a significant cause of noninfectious blindness in the world. Herein the authors aimed at unraveling the involvement of IL-17 in VKH and in experimental autoimmune uveitis, focusing on the signaling pathways involved in IL-17 synthesis. METHODS. Mice were immunized with 161-180 peptide and pertussis toxin. Draining lymph node cells, harvested 21 days after immunization, were cultured in the presence or absence of p38 alpha mitogen-activated protein kinase (MAPK) inhibitor (SB203580) and assayed for cytokine production and quantification of CD4(+)IL-17(+) cells. Mice received intraocular injections of SB203580, and disease severity was evaluated by histologic examination of the enucleated eyes at day 21. CD4(+) lymphocytes from MSK-1/2-deficient mice, human CD4(+) cells silenced with MSK1 siRNA, or peripheral blood mononuclear cells (PBMCs) from VKH patients were cultured in the presence or absence of p38 alpha MAPK inhibitor and then assayed for IL-17, IFN-gamma, and IL-4 production. RESULTS. The inhibition of p38 alpha MAPK fully blocked the synthesis of IL-17 by PBMCs from VKH patients and lymphocytes from EAU mice. The absence of the msk1/2 gene resulted in failure to produce IL-17 by murine and human lymphocytes. Interestingly, intraocular injections of SB203580 in EAU mice did not suppress development of the disease. CONCLUSIONS. These data show that p38 alpha MAPK-MSK1/2 is involved in the control of IL-17 synthesis by CD4(+) T cells and that inhibition of p38 alpha MAPK in vitro suppresses IL-17 synthesis but that inhibition of this kinase in vivo did not protect from EAU. (Invest Ophthalmol Vis Sci. 2010;51:3567-3574) DOI: 10.1167/iovs.09-4393

Alboserpin, a Factor Xa Inhibitor from the Mosquito Vector of Yellow Fever, Binds Heparin and Membrane Phospholipids and Exhibits Antithrombotic Activity

The molecular mechanism of factor Xa (FXa) inhibition by Alboserpin, the major salivary gland anticoagulant from the mosquito and yellow fever vector Aedes albopictus, has been characterized. cDNA of Alboserpin predicts a 45-kDa protein that belongs to the serpin family of protease inhibitors. Recombinant Alboserpin displays stoichiometric, competitive, reversible and tight binding to FXa (picomolar range). Binding is highly specific and is not detectable for FX, catalytic site-blocked FXa, thrombin, and 12 other enzymes. Alboserpin displays high affinity binding to heparin (K(D) similar to 20 nM), but no change in FXa inhibition was observed in the presence of the cofactor, implying that bridging mechanisms did not take place. Notably, Alboserpin was also found to interact with phosphatidylcholine and phosphatidylethanolamine but not with phosphatidylserine. Further, annexin V (in the absence of Ca(2+)) or heparin outcompetes Alboserpin for binding to phospholipid vesicles, suggesting a common binding site. Consistent with its activity, Alboserpin blocks prothrombinase activity and increases both prothrombin time and activated partial thromboplastin time in vitro or ex vivo. Furthermore, Alboserpin prevents thrombus formation provoked by ferric chloride injury of the carotid artery and increases bleeding in a dose-dependent manner. Alboserpin emerges as an atypical serpin that targets FXa and displays unique phospholipid specificity. It conceivably uses heparin and phosphatidylcholine/phosphatidylethanolamine as anchors to increase protein localization and effective concentration at sites of injury, cell activation, or inflammation.

Autoantibody response to chromatographic fractions from oxidized LDL in unstable angina patients and healthy controls

Levels of autoantibodies to oxidized low-density lipoprotein (oxLDL) have been correlated to atherosclerosis; however, contradictory results have been shown. To better understand the role of autoantibodies to oxLDL in atherogenesis, and their potential to predict risk of developing coronary artery disease we investigated the antibody response of unstable angina (UA) patients and healthy controls against chromatographic separated fractions of oxLDL. Five major peaks were detected after chromatographic separation of oxLDL and 10 fractions were collected. Surprisingly, when the response to high molecular weight fractions was analysed, we observed a significant increase in the levels of autoantibodies in controls compared to UA. In contrast, when the autoantibody response to intermediate and low molecular weight fractions was analysed, we observed that the UA group showed consistently higher levels compared with controls. Our data demonstrates that within oxLDL there are major fractions that can be recognized by autoantibodies from either UA patients or healthy individuals, and that the use of total oxLDL as an antigen pool may mask the presence of some antigenic molecules and their corresponding antibodies. Further studies are needed, but the analysis of antibody profiles may indeed open up a novel approach for evaluation and prevention against atherosclerosis.

Development of an adenoviral vector with robust expression driven by p53

Here we introduce a new adenoviral vector where transgene expression is driven by p53. We first developed a synthetic promoter, referred to as PGTx beta containing a p53-responsive element, a minimal promoter and the first intron of the rabbit P-globin gene. Initial assays using plasmid-based vectors indicated that expression was tightly controlled by p53 and was 5-fold stronger than the constitutive CMV immediate early promoter/enhancer. The adenoviral vector, AdPG, was also shown to offer p53-responsive expression in prostate carcinoma cells LNCaP (wt p53), DU-145 (temperature sensitive mutant of p53) and PC3 (p53-null, but engineered to express temperature-sensitive p53 mutants). AdPG served as a sensor of p53 activity in LNCaP cells treated with chemotherapeutic agents. Since p53 can be induced by radiotherapy and chemotherapy, this new vector could be further developed for use in combination with conventional therapies to bring about cooperation between the genetic and pharmacologic treatment modalities. (c) 2007 Elsevier Inc. All rights reserved.

Fur controls iron homeostasis and oxidative stress defense in the oligotrophic alpha-proteobacterium Caulobacter crescentus

In most bacteria, the ferric uptake regulator (Fur) is a global regulator that controls iron homeostasis and other cellular processes, such as oxidative stress defense. In this work, we apply a combination of bioinformatics, in vitro and in vivo assays to identify the Caulobacter crescentus Fur regulon. A C. crescentus fur deletion mutant showed a slow growth phenotype, and was hypersensitive to H(2)O(2) and organic peroxide. Using a position weight matrix approach, several predicted Fur-binding sites were detected in the genome of C. crescentus, located in regulatory regions of genes not only involved in iron uptake and usage but also in other functions. Selected Fur-binding sites were validated using electrophoretic mobility shift assay and DNAse I footprinting analysis. Gene expression assays revealed that genes involved in iron uptake were repressed by iron-Fur and induced under conditions of iron limitation, whereas genes encoding iron-using proteins were activated by Fur under conditions of iron sufficiency. Furthermore, several genes that are regulated via small RNAs in other bacteria were found to be directly regulated by Fur in C. crescentus. In conclusion, Fur functions as an activator and as a repressor, integrating iron metabolism and oxidative stress response in C. crescentus.

Notch signaling controls the balance of ciliated and secretory cell fates in developing airways

Although there is accumulated evidence of a role for Notch in the developing lung, it is still unclear how disruption of Notch signaling affects lung progenitor cell fate and differentiation events in the airway epithelium. To address this issue, we inactivated Notch signaling conditionally in the endoderm using a Shh-Cre deleter mouse line and mice carrying floxed alleles of the Pofut1 gene, which encodes an O-fucosyltransferase essential for Notch-ligand binding. We also took the same conditional approach to inactivate expression of Rbpjk, which encodes the transcriptional effector of canonical Notch signaling. Strikingly, these mutants showed an almost identical lung phenotype characterized by an absence of secretory Clara cells without evidence of cell death, and showed airways populated essentially by ciliated cells, with an increase in neuroendocrine cells. This phenotype could be further replicated in cultured wild-type lungs by disrupting Notch signaling with a gamma-secretase inhibitor. Our data suggest that Notch acts when commitment to a ciliated or non-ciliated cell fate occurs in proximal progenitors, silencing the ciliated program in the cells that will continue to expand and differentiate into secretory cells. This mechanism may be crucial to define the balance of differentiated cell profiles in different generations of the developing airways. It might also be relevant to mediate the metaplastic changes in the respiratory epithelium that occur in pathological conditions, such as asthma and chronic obstructive pulmonary disease.

C(k)-solvability near the characteristic set for a class of planar complex vector fields of infinite type

This paper deals with semi-global C(k)-solvability of complex vector fields of the form L = partial derivative/partial derivative t + x(r) (a(x) + ib(x))partial derivative/partial derivative x, r >= 1, defined on Omega(epsilon) = (-epsilon, epsilon) x S(1), epsilon > 0, where a and b are C(infinity) real-valued functions in (-epsilon, epsilon). It is shown that the interplay between the order of vanishing of the functions a and b at x = 0 influences the C(k)-solvability at Sigma = {0} x S(1). When r = 1, it is permitted that the functions a and b of L depend on the x and t variables, that is, L = partial derivative/partial derivative t + x(a(x, t) + ib(x, t))partial derivative/partial derivative x, where (x, t) is an element of Omega(epsilon).

THE POINCARE-BENDIXSON THEOREM ON THE KLEIN BOTTLE FOR CONTINUOUS VECTOR FIELDS

We present a version of the Poincare-Bendixson Theorem on the Klein bottle K(2) for continuous vector fields. As a consequence, we obtain the fact that K(2) does not admit continuous vector fields having a omega-recurrent injective trajectory.

Phase portraits of quadratic polynomial vector fields having a rational first integral of degree 3

In this paper, we classify all the global phase portraits of the quadratic polynomial vector fields having a rational first integral of degree 3. (C) 2008 Elsevier Ltd. All rights reserved.

Invariant theory and reversible-equivariant vector fields

In this paper we present results for the systematic study of reversible-equivariant vector fields - namely, in the simultaneous presence of symmetries and reversing symmetries - by employing algebraic techniques from invariant theory for compact Lie groups. The Hilbert-Poincare series and their associated Molien formulae are introduced,and we prove the character formulae for the computation of dimensions of spaces of homogeneous anti-invariant polynomial functions and reversible-equivariant polynomial mappings. A symbolic algorithm is obtained for the computation of generators for the module of reversible-equivariant polynomial mappings over the ring of invariant polynomials. We show that this computation can be obtained directly from a well-known situation, namely from the generators of the ring of invariants and the module of the equivariants. (C) 2008 Elsevier B.V, All rights reserved.

Nonexistence of global solutions for a class of complex vector fields on two-torus

The goal of this paper is study the global solvability of a class of complex vector fields of the special form L = partial derivative/partial derivative t + (a + ib)(x)partial derivative/partial derivative x, a, b epsilon C(infinity) (S(1) ; R), defined on two-torus T(2) congruent to R(2)/2 pi Z(2). The kernel of transpose operator L is described and the solvability near the characteristic set is also studied. (c) 2008 Elsevier Inc. All rights reserved.

Gevrey solvability near the characteristic set for a class of planar complex vector fields of infinite type

We study the Gevrey solvability of a class of complex vector fields, defined on Omega(epsilon) = (-epsilon, epsilon) x S(1), given by L = partial derivative/partial derivative t + (a(x) + ib(x))partial derivative/partial derivative x, b not equivalent to 0, near the characteristic set Sigma = {0} x S(1). We show that the interplay between the order of vanishing of the functions a and b at x = 0 plays a role in the Gevrey solvability. (C) 2008 Elsevier Inc. All rights reserved.

Interactive Vector Field Feature Identification

We introduce a flexible technique for interactive exploration of vector field data through classification derived from user-specified feature templates. Our method is founded on the observation that, while similar features within the vector field may be spatially disparate, they share similar neighborhood characteristics. Users generate feature-based visualizations by interactively highlighting well-accepted and domain specific representative feature points. Feature exploration begins with the computation of attributes that describe the neighborhood of each sample within the input vector field. Compilation of these attributes forms a representation of the vector field samples in the attribute space. We project the attribute points onto the canonical 2D plane to enable interactive exploration of the vector field using a painting interface. The projection encodes the similarities between vector field points within the distances computed between their associated attribute points. The proposed method is performed at interactive rates for enhanced user experience and is completely flexible as showcased by the simultaneous identification of diverse feature types.

Atmosphere and hydrological controls of the evapotranspiration over a floodplain forest in the Bananal Island region, Amazonia

This article discusses seasonal and interannual variations of the evapotranspiration (ET) rates in Bananal Island floodplain, Brazil. Measurements included ET and sensible heat flux using the eddy covariance method, atmospheric forcings (net radiation, Rn, vapor pressure deficit, VPD, wind speed and air temperature), soil moisture profiles, groundwater level and flood height, taken from November 2003 to December 2006. For the hydrological years (October-September) of 2003/2004, 2004/2005 and 2005/2006, the accumulated precipitation was 1692, 1471, 1914 mm and the accumulated ET was 1361, 1318 and 1317 mm, respectively. Seasonal analyses indicated that ET decreased in the dry season (average 3.7 mm day(-1)), despite the simultaneous increase in Rn, air temperature and VPD. The increase of ET in the wet season and particularly in the flood period (average 4.1 mm day(-1)) showed that the free water surface evaporation strongly influenced the energy exchange. Soil moisture, which was substantially depleted during the dry season, and adaptative vegetation mechanisms such as leaf senescence contributed to limit the dry season ET. Strong drainage within permeable sandy soils helped to explain the soil moisture depletion. These results suggest that the Bananal flooding area shows a different pattern in relation to the upland Amazon forests, being more similar to the savanna strictu senso areas in central Brazil. For example, seasonal ET variation was not in phase with Rn; the wet season ET was higher than the dry season ET; and the system stored only a tiny memory of the flooding period, being sensitive to extended drought periods.

Determination of controls on Holocene barrier progradation through application of OSL dating: The Ilha Comprida Barrier example, Southeastern Brazil

Barrier development during the Holocene is studied using the example of the Ilha Comprida, Southeastern Brazil. Aerial photos, facies analysis, and optically stimulated luminescence dating are used to define the barrier emergence and evolution. Optically stimulated luminescence ages and facies successions indicate that the Ilha Comprida probably began as a Holocene transgressive barrier island 6000 years ago, just before the last relative sea-level maximum. Since then the barrier has progradated through the addition of curved beach ridges. Based on beach ridge alignments, six units of growth are identified with two growth directions, transverse and longitudinal. Rates of progradation with transverse growth vary from 0.13 to 4.6 m/year. Rates of longitudinal growth to NE range from 5.2 to 30 m/year. Variation in coastal progradation rates and sediment retention during the last 6000 years is compared with climate, physiography and relative sea-level changes. The physiography, represented by pre-Cenozoic hills, is the major control on sediment retention and alternation between longitudinal and transverse growth. Climate variations, such as the Little Ice Age event, apparently control the formation of ridges types: beach ridges, foredunes, and blowouts. These results allow the use of the Ilha Comprida Barrier as an example to analyze the major controls on barriers progradation. (C) 2011 Elsevier B.V. All rights reserved.