Coactivated platelet-derived growth factor receptor {alpha} and epidermal growth factor receptor are potential therapeutic targets in intimal sarcoma.

Intimal sarcoma (IS) is a rare, malignant, and aggressive tumor that shows a relentless course with a concomitant low survival rate and for which no effective treatment is available. In this study, 21 cases of large arterial blood vessel IS were analyzed by immunohistochemistry and fluorescence in situ hybridization and selectively by karyotyping, array comparative genomic hybridization, sequencing, phospho-kinase antibody arrays, and Western immunoblotting in search for novel diagnostic markers and potential molecular therapeutic targets. Ex vivo immunoassays were applied to test the sensitivity of IS primary tumor cells to the receptor tyrosine kinase (RTK) inhibitors imatinib and dasatinib. We showed that amplification of platelet-derived growth factor receptor α (PDGFRA) is a common finding in IS, which should be considered as a molecular hallmark of this entity. This amplification is consistently associated with PDGFRA activation. Furthermore, the tumors reveal persistent activation of the epidermal growth factor receptor (EGFR), concurrent to PDGFRA activation. Activated PDGFRA and EGFR frequently coexist with amplification and overexpression of the MDM2 oncogene. Ex vivo immunoassays on primary IS cells from one case showed the potency of dasatinib to inhibit PDGFRA and downstream signaling pathways. Our findings provide a rationale for investigating therapies that target PDGFRA, EGFR, or MDM2 in IS. Given the clonal heterogeneity of this tumor type and the potential cross-talk between the PDGFRA and EGFR signaling pathways, targeting multiple RTKs and aberrant downstream effectors might be required to improve the therapeutic outcome for patients with this disease.

Hepatitis C virus-linked mitochondrial dysfunction promotes hypoxia-inducible factor 1 alpha-mediated glycolytic adaptation.

Hepatitis C virus (HCV) infection induces a state of oxidative stress by affecting mitochondrial-respiratory-chain activity. By using cell lines inducibly expressing different HCV constructs, we showed previously that viral-protein expression leads to severe impairment of mitochondrial oxidative phosphorylation and to major reliance on nonoxidative glucose metabolism. However, the bioenergetic competence of the induced cells was not compromised, indicating an efficient prosurvival adaptive response. Here, we show that HCV protein expression activates hypoxia-inducible factor 1 (HIF-1) by normoxic stabilization of its alpha subunit. In consequence, expression of HIF-controlled genes, including those coding for glycolytic enzymes, was significantly upregulated. Similar expression of HIF-controlled genes was observed in cell lines inducibly expressing subgenomic HCV constructs encoding either structural or nonstructural viral proteins. Stabilization and transcriptional activation of HIF-1alpha was confirmed in Huh-7.5 cells harboring cell culture-derived infectious HCV and in liver biopsy specimens from patients with chronic hepatitis C. The HCV-related HIF-1alpha stabilization was insensitive to antioxidant treatment. Mimicking an impairment of mitochondrial oxidative phosphorylation by treatment of inducible cell lines with oligomycin resulted in stabilization of HIF-1alpha. Similar results were obtained by treatment with pyruvate, indicating that accumulation of intermediate metabolites is sufficient to stabilize HIF-1alpha. These observations provide new insights into the pathogenesis of chronic hepatitis C and, possibly, the HCV-related development of hepatocellular carcinoma.

Peroxisome proliferator-activated receptor-alpha-null mice have increased white adipose tissue glucose utilization, GLUT4, and fat mass: Role in liver and brain.

Activation of the peroxisome proliferator-activated receptor (PPAR)-alpha increases lipid catabolism and lowers the concentration of circulating lipid, but its role in the control of glucose metabolism is not as clearly established. Here we compared PPARalpha knockout mice with wild type and confirmed that the former developed hypoglycemia during fasting. This was associated with only a slight increase in insulin sensitivity but a dramatic increase in whole-body and adipose tissue glucose use rates in the fasting state. The white sc and visceral fat depots were larger due to an increase in the size and number of adipocytes, and their level of GLUT4 expression was higher and no longer regulated by the fed-to-fast transition. To evaluate whether these adipocyte deregulations were secondary to the absence of PPARalpha from liver, we reexpresssed this transcription factor in the liver of knockout mice using recombinant adenoviruses. Whereas more than 90% of the hepatocytes were infected and PPARalpha expression was restored to normal levels, the whole-body glucose use rate remained elevated. Next, to evaluate whether brain PPARalpha could affect glucose homeostasis, we activated brain PPARalpha in wild-type mice by infusing WY14643 into the lateral ventricle and showed that whole-body glucose use was reduced. Hence, our data show that PPARalpha is involved in the regulation of glucose homeostasis, insulin sensitivity, fat accumulation, and adipose tissue glucose use by a mechanism that does not require PPARalpha expression in the liver. By contrast, activation of PPARalpha in the brain stimulates peripheral glucose use. This suggests that the alteration in adipocyte glucose metabolism in the knockout mice may result from the absence of PPARalpha in the brain.

Plasmodium vivax antigen discovery based on alpha-helical coiled coil protein motif.

Protein α-helical coiled coil structures that elicit antibody responses, which block critical functions of medically important microorganisms, represent a means for vaccine development. By using bioinformatics algorithms, a total of 50 antigens with α-helical coiled coil motifs orthologous to Plasmodium falciparum were identified in the P. vivax genome. The peptides identified in silico were chemically synthesized; circular dichroism studies indicated partial or high α-helical content. Antigenicity was evaluated using human sera samples from malaria-endemic areas of Colombia and Papua New Guinea. Eight of these fragments were selected and used to assess immunogenicity in BALB/c mice. ELISA assays indicated strong reactivity of serum samples from individuals residing in malaria-endemic regions and sera of immunized mice, with the α-helical coiled coil structures. In addition, ex vivo production of IFN-γ by murine mononuclear cells confirmed the immunogenicity of these structures and the presence of T-cell epitopes in the peptide sequences. Moreover, sera of mice immunized with four of the eight antigens recognized native proteins on blood-stage P. vivax parasites, and antigenic cross-reactivity with three of the peptides was observed when reacted with both the P. falciparum orthologous fragments and whole parasites. Results here point to the α-helical coiled coil peptides as possible P. vivax malaria vaccine candidates as were observed for P. falciparum. Fragments selected here warrant further study in humans and non-human primate models to assess their protective efficacy as single components or assembled as hybrid linear epitopes.

Purification and characterization of two enantioselective alpha-ketoglutarate-dependent dioxygenases, RdpA and SdpA, from Sphingomonas herbicidovorans MH.

Alpha-ketoglutarate-dependent (R)-dichlorprop dioxygenase (RdpA) and alpha-ketoglutarate-dependent (S)-dichlorprop dioxygenase (SdpA), which are involved in the degradation of phenoxyalkanoic acid herbicides in Sphingomonas herbicidovorans MH, were expressed and purified as His6-tagged fusion proteins from Escherichia coli BL21(DE3)(pLysS). RdpA and SdpA belong to subgroup II of the alpha-ketoglutarate-dependent dioxygenases and share the specific motif HXDX(24)TX(131)HX(10)R. Amino acids His-111, Asp-113, and His-270 and amino acids His-102, Asp-104, and His 257 comprise the 2-His-1-carboxylate facial triads and were predicted to be involved in iron binding in RdpA and SdpA, respectively. RdpA exclusively transformed the (R) enantiomers of mecoprop [2-(4-chloro-2-methylphenoxy)propanoic acid] and dichlorprop [2-(2,4-dichlorophenoxy)propanoic acid], whereas SdpA was specific for the (S) enantiomers. The apparent Km values were 99 microM for (R)-mecoprop, 164 microM for (R)-dichlorprop, and 3 microM for alpha-ketoglutarate for RdpA and 132 microM for (S)-mecoprop, 495 microM for (S)-dichlorprop, and 20 microM for alpha-ketoglutarate for SdpA. Both enzymes had high apparent Km values for oxygen; these values were 159 microM for SdpA and >230 microM for RdpA, whose activity was linearly dependent on oxygen at the concentration range measured. Both enzymes had narrow cosubstrate specificity; only 2-oxoadipate was able to replace alpha-ketoglutarate, and the rates were substantially diminished. Ferrous iron was necessary for activity of the enzymes, and other divalent cations could not replace it. Although the results of growth experiments suggest that strain MH harbors a specific 2,4-dichlorophenoxyacetic acid-converting enzyme, tfdA-, tfdAalpha-, or cadAB-like genes were not discovered in a screening analysis in which heterologous hybridization and PCR were used.

Acoustic properties of polypropylene composites reinforced with stone groundwood

Currently, acoustic isolation is one of the problems raised with building construction in Spain. The publication of the Basic Document for the protection against noise of the Technical Building Code has increased the demand of comfort for citizens. This has created the need to seek new composite materials that meet the new required acoustical building codes. In this paper we report the results of the newly developed composites that are able to improve the acoustic isolation of airborne noise. These composites were prepared from polypropylene (PP) reinforced with mechanical pulp fibers from softwood (Pinus radiata). Mechanical and acoustical properties of the composites from mechanical pulp (MP) and polypropylene (PP) have been investigated and compared to fiberglass (FG) composites. MP composites had lower tensile properties compared with FG composites, although these properties can be improved by incorporation of a coupling agent. The results of acoustical properties of MP composites were reported and compared with the conventional composites based on fiberglass and gypsum plasterboards. Finally, we suggest the application of MP composites as a light-weight building material to reduce acoustic transmitions

Time of injection determines the effect of alpha-MSH antiserum on DA neurons in psychological stress

Male rats were subjected to "psychological stress" which consisted in 10 sec footshock on the first day followed 24 hr later by a 10 sec stay in the experimental chamber without shock. Intravenous antiserum against alpha-MSH markedly changed the functional state of mesencephalic and hypothalamic DA neurons (assessed by histochemical microfluorimetry) when administered before the second session but not when given before the first session. These observations reveal an interesting parallelism in the temporal characteristics of the effects of alpha-MSH on avoidance behavior and central DA systems.

Management of corn stalk waste as reinforcement for polypropylene injection moulded composites

The main objective of this study was the management of corn stalk waste as reinforcement for polypropylene (PP) injection moulded composites as an alternative to wood flour and fibers. In the first step, corn stalk waste was subjected to various treatments, and four different corn stalk derivatives (flour and fibers) able to be used as reinforcement of composite materials were prepared and characterized. These derivatives are corn stalk flour, thermo-mechanical, semi-chemical, and chemical fibers. They were characterized in terms of their yield, lignin content, Kappa number, fiber length/diameter ratio, fines, coarseness, viscosity, and the length at the break of a standard sheet of paper. Results showed that the corn stalk derivatives have different physico-chemical properties. In the second step, the prepared flour and fibers were explored as a reinforcing element for PP composites. Coupled and non-coupled PP composites were prepared and tested for tensile properties. For overall trend, with the addition of a coupling agent, tensile properties of composites significantly improved, as compared with non-coupled samples. In addition, a morphological study revealed the positive effect of the coupling agent on the interfacial bonding. The composites prepared with semichemical fiber gave better results in comparison with the rest of the corn stalk derivatives due to its chemical characteristics

Effects of alpha-phosphoglucomutase deficiency on cell wall properties and fitness in Streptococcus gordonii.

Streptococcus gordonii alpha-phosphoglucomutase, which converts glucose 6-phosphate to glucose 1-phosphate, is encoded by pgm. The pgm transcript is monocistronic and is initiated from a sigma(A)-like promoter. Mutants with a gene disruption in pgm exhibited an altered cell wall muropeptide pattern and a lower teichoic acid content, and had reduced fitness both in vitro and in vivo. In vitro, the reduced fitness included reduced growth, reduced viability in the stationary phase and increased autolytic activity. In vivo, the pgm-deficient strain had a lower virulence in a rat model of experimental endocarditis.

Stone-ground wood pulp-reinforced polypropylene composites: water uptake and thermal properties

Two of the drawbacks of using natural-based composites in industrial applications are thermal instability and water uptake capacity. In this work, mechanical wood pulp was used to reinforce polypropylene at a level of 20 to 50 wt. %. Composites were mixed by means of a Brabender internal mixer for both non-coupled and coupled formulations. Differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) were used to determine the thermal properties of the composites. The water uptake behavior was evaluated by immersion of the composites in water until an equilibrium state was reached. Results of water absorption tests revealed that the amount of water absorption was clearly dependent upon the fiber content. The coupled composites showed lower water absorption compared to the uncoupled composites. The incorporation of mechanical wood pulp into the polypropylene matrix produced a clear nucleating effect by increasing the crystallinity degree of the polymer and also increasing the temperature of polymer degradation. The maximum degradation temperature for stone ground wood pulp–reinforced composites was in the range of 330 to 345 ºC

Tensile strength characteristics of polypropylene composites reinforced with stone groundwood fibers from softwood

The behavior of stone groundwood / polypropylene injection-molded composites was evaluated with and without coupling agent. Stone groundwood (SGW) is a fibrous material commonly prepared in a high yield process and mainly used for papermaking applications. In this work, the use of SGW fibers was explored as a reinforcing element of polypropylene (PP) composites. The surface charge density of the composite components was evaluated, as well as the fiber’s length and diameter inside the composite material. Two mixing extrusion processes were evaluated, and the use of a kinetic mixer, instead of an internal mixer, resulted in longer mean fiber lengths of the reinforcing fibers. On the other hand, the accessibility of surface hydroxyl groups of stone groundwood fibers was improved by treating the fibers with 5% of sodium hydroxide, resulting in a noticeable increase of the tensile strength of the composites, for a similar percentage of coupling agent. A new parameter called Fiber Tensile Strength Factor is defined and used as a baseline for the comparison of the properties of the different composite materials. Finally the competitiveness of stone groundwood / polypropylene / polypropylene-co-maleic anhydride system, which compared favorably to sized glass-fiber / polypropylene GF/PP and glass-fiber / polypropylene / polypropylene-co-maleic anhydride composite formulations, was quantified by means of the fiber tensile strength factor

Relations entre la pharmacocinétique de population de l'imatinib et l'alpha-1-glycoproteine acide chez des patients hémato-oncologiques

Introduction: La disposition de l'imatinib (Glivec®) implique des systèmes connus pour de grandes différences inter-individuelles, et l'on peut s'attendre à ce que l'exposition à ce médicament varie largement d'un patient à l'autre. L'alpha-1-glycoprotéine acide (AAG), une protéine circulante liant fortement l'imatinib, représente l'un de ces systèmes. Objectif: Cette étude observationnelle visait à explorer l'influence de l'AAG plasmatique sur la pharmacocinétique de l'imatinib. Méthode: Une analyse de population a été effectuée avec le programme NONMEM sur 278 échantillons plasmatiques issus de 51 patients oncologiques. L'influence des taux d'AAG sur la clairance (CL) et le volume de distribution (Vd) a ainsi été étudiée. Résultats: Un modèle à un compartiment avec absorption de premier ordre a permis de décrire les données. Une relation hyperbolique entre taux d'AAG et CL ou Vd a été observée. Une approche mécanistique a donc été élaborée, postulant que seule la concentration libre subissait une élimination du premier ordre, et intégrant la constante de dissociation comme paramètre du modèle. Cette approche a permis de déterminer une CLlibre moyenne de 1310 l/h et un Vd de 301 l. Par comparaison, la CLtotale déterminée initialement était de 14 l/h. La CLlibre est affectée par le poids corporel et le type de pathologie. Qui plus est, ce modèle a permis d'estimer in vivo la constante d'association entre imatinib et AAG (5.5?106 l/mol), ainsi que la fraction libre moyenne de l'imatinib (1.1%). La variabilité inter-individuelle estimée pour la disposition de l'imatinib (17% sur CLlibre et 66% sur Vd) diminuait globalement de moitié avec le modèle incorporant l'impact de l'AAG. Discussion-conclusion: De tels résultats clarifient l'impact de la liaison protéinique sur le devenir de l'imatinib. Des taux élevés d'AAG ont été présumés représenter un facteur de résistance à l'imatinib. Toutefois, cela est peu probable, notre modèle prédisant que la concentration libre reste inchangée. D'un autre côté, s'il est un jour démontré que l'imatinib requiert un programme de suivi thérapeutique (TDM), la mesure des concentrations libres, ou la correction des concentrations totales en fonction des taux d'AAG, devraient être envisagées pour une interprétation précise des résultats.