Experimental inhibition of porcupine-mediated Wnt O-acylation attenuates kidney fibrosis.

Activated Wnt signaling is critical in the pathogenesis of renal fibrosis, a final common pathway for most forms of chronic kidney disease. Therapeutic intervention by inhibition of individual Wnts or downstream Wnt/β-catenin signaling has been proposed, but these approaches do not interrupt the functions of all Wnts nor block non-canonical Wnt signaling pathways. Alternatively, an orally bioavailable small molecule, Wnt-C59, blocks the catalytic activity of the Wnt-acyl transferase porcupine, and thereby prevents secretion of all Wnt isoforms. We found that inhibiting porcupine dramatically attenuates kidney fibrosis in the murine unilateral ureteral obstruction model. Wnt-C59 treatment similarly blunts collagen mRNA expression in the obstructed kidney. Consistent with its actions to broadly arrest Wnt signaling, porcupine inhibition reduces expression of Wnt target genes and bolsters nuclear exclusion of β-catenin in the kidney following ureteral obstruction. Importantly, prevention of Wnt secretion by Wnt-C59 blunts expression of inflammatory cytokines in the obstructed kidney that otherwise provoke a positive feedback loop of Wnt expression in collagen-producing fibroblasts and epithelial cells. Thus, therapeutic targeting of porcupine abrogates kidney fibrosis not only by overcoming the redundancy of individual Wnt isoforms but also by preventing upstream cytokine-induced Wnt generation. These findings reveal a novel therapeutic maneuver to protect the kidney from fibrosis by interrupting a pathogenic crosstalk loop between locally generated inflammatory cytokines and the Wnt/β-catenin signaling pathway.

Radiolabeled inhibitors as probes for imaging mutant IDH1 expression in gliomas: Synthesis and preliminary evaluation of labeled butyl-phenyl sulfonamide analogs.

INTRODUCTION: Malignant gliomas frequently harbor mutations in the isocitrate dehydrogenase 1 (IDH1) gene. Studies suggest that IDH mutation contributes to tumor pathogenesis through mechanisms that are mediated by the neomorphic metabolite of the mutant IDH1 enzyme, 2-hydroxyglutarate (2-HG). The aim of this work was to synthesize and evaluate radiolabeled compounds that bind to the mutant IDH1 enzyme with the goal of enabling noninvasive imaging of mutant IDH1 expression in gliomas by positron emission tomography (PET). METHODS: A small library of nonradioactive analogs were designed and synthesized based on the chemical structure of reported butyl-phenyl sulfonamide inhibitors of mutant IDH1. Enzyme inhibition assays were conducted using purified mutant IDH1 enzyme, IDH1-R132H, to determine the IC50 and the maximal inhibitory efficiency of the synthesized compounds. Selected compounds, 1 and 4, were labeled with radioiodine ((125)I) and/or (18)F using bromo- and phenol precursors, respectively. In vivo behavior of the labeled inhibitors was studied by conducting tissue distribution studies with [(125)I]1 in normal mice. Cell uptake studies were conducted using an isogenic astrocytoma cell line that carried a native IDH1-R132H mutation to evaluate the potential uptake of the labeled inhibitors in IDH1-mutated tumor cells. RESULTS: Enzyme inhibition assays showed good inhibitory potency for compounds that have iodine or a fluoroethoxy substituent at the ortho position of the phenyl ring in compounds 1 and 4 with IC50 values of 1.7 μM and 2.3 μM, respectively. Compounds 1 and 4 inhibited mutant IDH1 activity and decreased the production of 2-HG in an IDH1-mutated astrocytoma cell line. Radiolabeling of 1 and 4 was achieved with an average radiochemical yield of 56.6 ± 20.1% for [(125)I]1 (n = 4) and 67.5 ± 6.6% for [(18)F]4 (n = 3). [(125)I]1 exhibited favorable biodistribution characteristics in normal mice, with rapid clearance from the blood and elimination via the hepatobiliary system by 4 h after injection. The uptake of [(125)I]1 in tumor cells positive for IDH1-R132H was significantly higher compared to isogenic WT-IDH1 controls, with a maximal uptake ratio of 1.67 at 3 h post injection. Co-incubation of the labeled inhibitors with the corresponding nonradioactive analogs, and decreasing the normal concentrations of FBS (10%) in the incubation media substantially increased the uptake of the labeled inhibitors in both the IDH1-mutant and WT-IDH1 tumor cell lines, suggesting significant non-specific binding of the synthesized labeled butyl-phenyl sulfonamide inhibitors. CONCLUSIONS: These data demonstrate the feasibility of developing radiolabeled probes for the mutant IDH1 enzyme based on enzyme inhibitors. Further optimization of the labeled inhibitors by modifying the chemical structure to decrease the lipophilicity and to increase potency may yield compounds with improved characteristics as probes for imaging mutant IDH1 expression in tumors.

Estrés térmico por alta temperatura en soja (Glycine max (L.) Merr.) : análisis de la dinámica de producción y fijación de vainas y su efecto sobre la determinación del rendimiento

En Argentina, y debido al amplio contexto geográfico donde crece, el cultivo de soja está expuesto frecuentemente a episodios de alta temperatura combinados con baja disponibilidad hídrica. Cuando ocurren durante etapas críticas, estas condiciones ambientales disminuyen el rendimiento del cultivo. Sin embargo, son escasos o inexistentes los estudios que evalúan los mecanismos ecofisiológicos que determinan la partición reproductiva y la generación del rendimiento en respuesta a la combinación de estos estreses abióticos. Los objetivos de esta tesis fueron estudiar la generación del rendimiento en soja, enfatizando el estudio de la dinámica de producción de estructuras reproductivas, la estabilidad de la partición reproductiva y de los mecanismos de compensación entre número y peso de granos en respuesta a la interacción de altas temperaturas y distintos niveles hídricos en suelo. Para ello, durante dos campañas, dos genotipos (DM 4200 y DM 4870) fueron evaluados ante la combinación de dos niveles de temperatura (impuestos como episodios estrés térmico de corta duración) y agua edáfica (riego y secano) durante la etapa crítica de generación de vainas. Se cuantificaron la evolución de la biomasa aérea, la eficiencia de uso de radiación, la dinámica de producción y fijación de vainas, el rendimiento y sus componentes (número y peso de granos) y las relaciones fuente/destino durante las etapas de determinación del número y del peso de los granos. La temperatura máxima promedio durante los episodios de estrés térmico superó en 5,6 ºC a la registrada en tratamientos control. La tasa de fijación de vainas fue 38 por ciento inferior en condiciones de alta temperatura combinada con secano con respecto a los controles regados. El estrés térmico en fructificación redujo el número de granos en ambas condiciones hídricas, aunque en distinta magnitud (17 por ciento riego; 42 por ciento secano). En contraste con los controles, el componente peso de granos no logró compensar la reducción del número de granos en la combinación de estrés térmico y secano. En consecuencia, la alta temperatura con secano redujo la partición reproductiva (15 por ciento p menor a 0,05) y el rendimiento fue consistentemente afectado (45 por ciento inferior a los controles regados). Los resultados de este estudio demuestran que la caída en partición reproductiva y, por ende, en rendimiento final no obedeció a fallas reproductivas ya que el número de vainas iniciadas fue similar en todos los tratamientos. La supervivencia de vainas se asoció con las condiciones ambientales durante las etapas tempranas de su desarrollo, alteradas por el episodio de estrés térmico. Estos resultados ponen de manifiesto la importancia de discriminar los efectos del estrés abiótico según el patrón temporal de fijación de vainas.

Colletotrichum graminicola (Ces.) Wilson sobre pasto ovillo (Dactylis Glomerata L.) en la Argentina

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The influence of alkali metal chlorides on the loss of water from glass-ionomer dental cements

The water loss behaviour of a clinical glass-ionomer dental cement has been studied with and without the addition of alkali metal chlorides. Dehydrating conditions were provided by placing specimens in a desiccator over concentrated sulphuric acid. Cements were prepared using either pure water or an aqueous solution of metal chloride (LiCl, NaCl, KCl) at 1.0 mol/dm(3). In addition, NaCl at 0.5 mol/dm(3) was also used to fabricate cements. Disc-shaped specimens of size 6 mm diameter x 2 mm thickness were made, six performulation, and cured at 37 degrees C for 1 hour They were then exposed to desiccating conditions, and the mass measured at regular intervals. All formulations were found to lose water in a diffusion process that equilibrated after approximately 3 weeks. Diffusion coefficients ranged from 2.27 (0.13) x 10(9) with no additive to 1.85 (0.07) x 10(9) m(2)/s with 1.0 mol/dm(3) KCl. For the salts, diffusion coefficients decreased in the order LiCl > NaCl > KCl. There was no statistically significant difference between the diffusion coefficients for 1.0 and 0.5 mol/dm(3) NaCl. For all salts at 1.0 mol/dm(3) and also additive-free cements, equilibrium losses were, with statistical limits, the same, ranging from 6.23 to 6.34%. On the other hand, 0.5 mol/dm(3) NaCl lost significantly more water 7.05%.

Synthesis and properties of polyelectrolyte microgel particles

A series of cationic poly(N-isopropylacrylamide/4-vinylpyridine) [poly(NIPAM/4-VP)] polyelectrolyte co-polymer microgels have been prepared by surfactant free emulsion polymerization (SFEP) with varying compositions of 4-VP and NIPAM. The compositions of 4-VP were 15, 25, 35, 45, 55 wt.% relative to NIPAM. The temperature and pH responsive swelling–deswelling properties of these microgels have been investigated using dynamic light scattering (DLS) and electrophoretic mobility measurements. DLS results have shown that the particle diameter of the poly(NIPAM/4-VP) microgels decreases with increasing concentration (wt.%) of 4-VP over the 20–60 °C temperature range due to the increased amount of hydrophobic group. The particle size of all poly(NIPAM/4-VP) microgel series increases with decreasing pH, as the 4-VP units become more protonated at low pH below the pKa (5.39) of the monomer 4-VP. Electrophoretic mobility results have shown that electrophoretic mobility increases as the temperature/pH increases at a constant background ionic strength (1 × 10− 4 mol dm− 3 NaCl). These results are in good agreement with DLS results. The temperature/pH sensitivity of these microgels depends on the ratio of NIPAM/4-VP concentration in the co-polymer microgel systems. The combined temperature/pH responsiveness of these polyelectrolyte microgels can be used in applications where changes in particle size with small change in pH or temperature is of great consequence.

Cyclic voltammetry and spectroelectrochemical study of nickel and cobalt diphenyltetraazaannulene complexes

The absorption spectra. cyclic voltammetry and spectroelectrochemistry of [Ni(II)DPTAA] and [Co(II)DPTAA] (DPTAA = 6,13-diphenyldibenzo[b,i][1,4,8,11] tetraaza[14]annulene) complexes in DMF are reported in detail. The ligand oxidation is observed for [Ni(II)DPTAA] at +0.70 V vs. SCE whereas Ni2(+/+) occurs at - 1.60 V. For [Co(II)DPTAA], a ligand oxidation redox couple is seen at +0.56 V while the Co2+/+ and Co2+/3+ redox couples appear at -1.21 and +0.24 V, respectively. All observed redox couples are assigned to reversible one-electron processes on account of peak separations and scan-rate dependency. These processes were further investigated by spectroelectrochemistry for [Co(II)DPTAA]. For [Co(II)DPTAA], axial ligation of pyridine was found to shift the Co2+/3+ redox couple more negative. while the ligand oxidation was shifted to more positive potentials. From a spectrophotometric titration of [Co(II)DPTAA] with pyridine an equilibrium constant, K-f, was determined for the binding of pyridine to [Co(II)DPTAA]. This was found to be 10.2 dm(3) mol(-1), slightly lower than that of [Co(II)TAA], indicating the influence of the phenyl groups. From this value and shifts in the Co2+/3+ redox couple upon ligation, an equilibrium constant for the binding of pyridine to [Co(III)DPTAA], K'(f), was found to be 5.06 x 10(6) dm(3) mol(-1). (c) 2007 Elsevier B.V. All rights reserved.

Preparation of o/w emulsions stabilized by solid particles and their characterization by oscillatory rheology

The purpose of this investigation was to examine the preparation and characterisation of hexane-in-water emulsions stabilised by clay particles. These emulsions, called Pickering emulsions, are characterised by the adsorption of solid particles at the oil/water (o/w) interface. The development of an elastic film at the o/w interface following the adsorption of colloidal particles helps to promote emulsion stability. Three different solid materials were used: silica sand, kaolin, and bentonite. Particles were added to the liquid mixtures in the range of 0.5–10 g dm−3. Emulsions were prepared using o/w ratios of 0.1, 0.2, 0.3, and 0.4. The effect of sodium chloride, on the stability of the prepared emulsions, was assessed in the range of 0–0.5 mol dm−3. In addition the use of a cationic surfactant hexadecyl-trimethylammonium bromide (CTAB) as an aid to improving emulsion stability was assessed in the concentration range of 0–0.05% (w/v). Characterisation of emulsion stability was realised through measurements of rheological properties including non-Newtonian viscosity, the elastic modulus, G', the loss modulus, G", and complex modulus, G*. The stability of the emulsions was evaluated immediately after preparation and 4 weeks later. Using the stability criteria, that for highly stable emulsions: G' > G" and both G' and G" are independent of frequency (varpi) it was concluded that highly stable emulsions could be prepared using a bentonite concentration of 2% (or more); an o/w ratio greater than 0.2; a CTAB concentration of 0.01%; and a salt concentration of 0.05 M or less—though salt was required.