Fine characterization of immunological mechanisms mediated by the major allergens of Parietaria judaica and hypoallergenic hybrid, rPjEDcys

Purpose: Allergy is a hypersensitivity disease IgE-mediated, affecting more than 25% of the population. The symptoms of IgE-mediated allergies reactions can be transiently ameliorated pharmacologically, but the only curative treatment of allergies is Allergen-Specific Immunotherapy (SIT). Recombinant hypoallergenic allergen derivatives with reduced allergenic activity have been engineered to reduce side effects during SIT. Parietaria judaica (Pj) pollen contains two major allergens belonging to the family of Lipid Tranfer Proteins (Par j 1 and Par j 2). By means of DNA recombinant technology, a hybrid hypoallergenic (PjEDcys), expressing disulphide bond variants of Par j 1 and Par j 2, was generated. The aim of this research project is to study the immunological mechanisms activated by the major allergens of Parietaria judaica, Par j 1 and Par j 2, and hypoallergenic hybrid rPjEDcys. Moreover, the project I am involved is trying to address the question whether this engineered hypoallergenic derivative can be a potential products for safer Allergen Specific Immunotherapy (SIT). Methods: Par j 1, Par j 2 and PjEDcys were produced as recombinant proteins. Human Peripheral Blood Mononuclear Cell (PBMC) from P. judaica allergic patients were stimulated in vitro with wild-type recombinant allergens and hybrid. PBMC proliferation assay, cytokine secretion assay, magnetic cell sorting of different subset of regulatory T cells, multiparametric flow cytometric analysis and molecular characterization using Real Time-PCR on sorted cells allow to study the biological properties of wild-type recombinant allergens and hybrid hypoallergenic derivate. Results: In vitro analysis suggested that PjEDcys have a reduced allergenity and maintained T cells reactivity. PBMC of P. judaica allergic patients stimulated in vitro with the hybrid and the wild-type recombinant allergens scored a percentage of proliferating CD4+ and CD56+ cell higher than unstimulated sample. Consistent with these data, cytokine secretion assay on CD4+ cells demonstrated that PBMC stimulation with rPjEDcys showed a percentage of IL-5 and IL-13 secreting T CD4+ cells lower than the wild-type allergens. Both rPjEDcys and wild-type stimulation promote the secretion of IFN- \u3b3 and IL-10 by T CD4+ cells. Finally whit the aim to study which subset of regulatory cells respond to wild-tipe allergens and hypoallergenic hybrid new experiment are setting. Discussion: In this experimental setting, the use of the major allergens of Pj and the hybrid polypeptides, rPjEDcys allows me to study the immunological mechanisms activated by the two different antigen stimulation and to investigate differences between the wild-type allergen and the hypoallergenic mutant rPjEDcys. Our data showed that CD4+ cells are clearly the predominant cell population proliferating in response to mixture of Par j 1 and Par j 2 allergens. The hypoallergenic derivate rPjEDcys retain the ability to stimulate CD4+ cells proliferation like the mixture of allergens (rPar j 1 and rPar j 2). Moreover these results highlighted a particular interesting datum; the mixture of allergens and the rPjEDcys hybrid showed the ability to stimulate an innate immune response, inducing CD56+ cells proliferative response. Cytokine secretion assay demonstrate that rPjEDcys reduce the secretion of IL-5 and IL-13, Th2 cytokines with a critical role in the development of allergy, compared to wild-type allergens. This may reflect the different biological function exerted by rPjEDcys. Conclusion: Collectivelly, our findings demonstrate that PjEDcys show a reduced allergenicity but maintained its immunogenicity and maybe it is also capable to regulate and redirect the immune response. These results suggest that PjEDcys represent a useful approach for immunotherapy of allergic disease

Proportional Venn diagram and determinants of allergic respiratory diseases in Italian adolescents

Large variations in prevalence of atopy and allergic diseases are reported worldwide in children, but in epidemiological studies the use of skin prick tests (SPT) and spirometry along with questionnaires is not common in the Mediterranean Area. The present work was aimed at evaluating the prevalence of current asthma (CA), rhinoconjunctivitis (RC), and eczema (E), with atopy and respiratory function, and the role of risk factors for allergic respiratory diseases. A total of 2150 Italian schoolchildren were cross-sectionally investigated through respiratory questionnaire, SPT, and spirometry. A proportional Venn diagram quantified the distribution of CA, RC, and E, stratifying for allergic sensitization to show differences in prevalence of allergic diseases among subjects with and without positive SPT. CA prevalence was 4.2%, RC 17.9%, and E 5.3%. CA and RC increased, while E decreased, with respect to previous local studies. Allergic sensitization prevalence (evaluated as positive response to at least one SPT) was 39.2%. A double Venn diagram identified 15 categories. Atopic CA was threefold more frequent than non-atopic CA. Atopic vs non-atopic RC and E were 9.6% vs 10.3% and 2.0% vs 3.3%, respectively. Atopic vs non-atopic RC associated with CA were 1.6% vs 0.5%; the same figures for RC associated with E were 0.8% vs 1.3%. Asymptomatic atopic subjects were 27.0%. Atopy, RC, parental asthma, and environmental risk factors were associated with CA. Atopy and environmental factors were risk factors also for RC. Asthma and traffic exposure were linked to reduced lung function. Respiratory allergic diseases are still increasing and largely concomitant in Italian adolescents. Atopy is more important for CA than RC. Avoiding exposures to measured environmental risk factors would prevent 41% of current asthma and 34% of rhinoconjunctivitis

Desarrollo de un Framework 3D para GameMaker

En este ensayo se describen los elementos del desarrollo de un Framework 3D para el motor gráfico GameMaker con el objetivo de construir un juego de estilo acción en tercera persona. Este trabajo requiere la integración en dicho motor pensado para las dos dimensiones, componentes tales como la importación de modelos 3D, las animaciones por esqueleto, la iluminación o las colisiones en un espacio tridimensional. Finalmente buscando la recreación de un viejo proyecto de videojuego con el fin de ejercer una comparación entre las dos versiones con más de seis años de diferencia entre ellas.In this essay, it describes the elements of the development of a 3D Framework for the GameMaker game engine with the objective of building a third person action game. This project requires the integration in the said engine thought for twodimension components such as 3D model importer, skeletal animations, illumination or collisions on a three-dimensional space. Looking for remaking an old proyect of a video game with the purpose of establishing a comparison between the two versions with six years of difference between them.En aquest assaig es descriuen els elements de el desenvolupament d'un Framework 3D per al motor gràfic GameMaker amb l'objectiu de construir un joc d'estil acció en tercera persona. Aquest treball requereix la integració en l'esmentat motor pensat per a les dues dimensions, components com ara la importació de models 3D, les animacions per esquelet, la il·luminació o les col·lisions en un espai tridimensional. Finalment buscant la recreació d'un vell projecte de videojoc per tal d'exercir una comparació entre les dues versions amb més de sis anys de diferència entre elles

Multiple in vitro and in vivo regulatory effects of budesonide in CD4+ T lymphocyte subpopulations of allergic asthmatics.

Abstract BACKGROUND: Increased activation and increased survival of T lymphocytes characterise bronchial asthma. OBJECTIVES: In this study the effect of budesonide on T cell survival, on inducible co-stimulator T cells (ICOS), on Foxp3 and on IL-10 molecules in T lymphocyte sub-populations was assessed. METHODS: Cell survival (by annexin V binding) and ICOS in total lymphocytes, in CD4+/CD25+ and in CD4+/CD25- and Foxp3 and IL-10 in CD4+/CD25+ and in CD4+/CD25-cells was evaluated, by cytofluorimetric analysis, in mild intermittent asthmatics (n = 19) and in controls (n = 15). Allergen induced T lymphocyte proliferation and the in vivo effects of budesonide in mild persistent asthmatics (n = 6) were also explored. RESULTS: Foxp3 was reduced in CD4+/CD25- and in CD4+/CD25+ cells and ICOS was reduced in CD4+/CD25+ cells but it was increased in CD4+CD25-in asthmatics when compared to controls. In asthmatics, in vitro, budesonide was able to: 1) increase annexin V binding and to reduce ICOS in total lymphocytes; 2) increase annexin V binding and Foxp3 and to reduce ICOS in CD4+/CD25- cells; 3) reduce annexin V binding and to increase IL-10 and ICOS in CD4+/CD25+ cells; 4) reduce cell allergen induced proliferation. In vivo, budesonide increased ICOS in CD4+/CD25+ while it increased Foxp3 and IL-10 in CD4+/CD25+ and in CD4+/CD25- cells. CONCLUSIONS: Budesonide modulates T cell survival, ICOS, Foxp3 and IL-10 molecules differently in T lymphocyte sub-populations. The findings provided shed light on new mechanisms by which corticosteroids, drugs widely used for the clinical management of bronchial asthma, control T lymphocyte activation

Use of a comprehensive diagnostic algorithm for Anisakis allergy in a high seroprevalence Mediterranean setting

Background. Diagnosis of anisakis allergy (AA) is based on the skin prick test (SPT) and specific IgE (sIgE) determination. Anyway, false positivity cases are due to cross reactivity with numerous allergens. The aim of the study was to evaluate the reliability of a comprehensive diagnostic algorithm for the AA. Methods. An observational study was conducted on a sample of consecutive subjects accessing the allergology outpatient ambulatories of two hospitals located in Western Sicily. All the recruited outpatients were tested by Skin Prick Test performed using anisakis extracts by ALK-Abello (Madrid, Spain). Specific IgE dosage for anisakis extracts was then performed by using ImmunoCAP250 (Immunodiagnostics Uppsala, Sweden). Consequently, outpatients who tested positive to first line tests underwent sIgE testing for ascaris and tropomyosin. Lastly, outpatients positive to the first line were invited to be further tested by basophil activation test (BAT) by using Flow CAST kit and anisakis commercial extract (Buhlmann Laboratories AG, Schonenbuch, Switzerland), as confirmatory analysis. Results. One hundred and eleven outpatients with an anamnesis suggestive of sensitization to anisakis (AS) and 466 subjects with chronic urticaria (CU) were recruited in the study. Of these, 22 with AS and 41 with CU showed a sensitization to anisakis allergens. The diagnostic algorithm revealed that 8.8% of outpatients who tested positive to sIgE determination were affected by CU, while 82.5% of all the sIgE positivity was related to cross-reactivity. Overall, a genuine anisakis seroprevalence of 2.3% was documented. Within a sub-sample of 15 subjects with clinical symptoms related to AA, n. 8 showed a real positivity after BAT. A greater response to A. pegreffii allergens as compared to A. simplex was reported. Conclusions. Our preliminary findings support the high clinical specificity of BAT for AA diagnosis, suggesting implementing this method in a comprehensive diagnostic algorithm

Cogeneration: An option to facilitate load following in Small Modular Reactors

Nuclear Power Plants (NPPs) have been historically deployed to cover the base-load of the electricity demand. Nowadays some NPPs might perform daily load cycling operation (i.e. load following) between 50% and 100% of their rated power. With respect to the insertion of control rods or comparable action to reduce the nuclear power generation, a more efficient alternative might be the “Load Following by Cogeneration”, i.e. diverting the excess of power, respect to the electricity demand, to an auxiliary system. A suitable cogeneration system needs: 1. To have a demand of electricity and/or heat in the region of 500 MWe–1.5 GWt; 2. To meet a significant market demand; 3. To have access to adequate input to process; 4. To be flexible: cogeneration might operate at full load during the night when the request of electricity is low, and be turned off during the daytime. From the economic standpoint, it is essential that the investment in the auxiliary system is profitable. This paper provides a techno-economic assessment of systems potentially suitable for coupling with a NPP for load following. The results show that district heating, desalination and hydrogen might be technically and economically feasible

Exhaled carbon monoxide in asthmatics: a meta-analysis

<p>Abstract</p> <p>Background</p> <p>The non-invasive assessment of airway inflammation is potentially advantageous in asthma management. Exhaled carbon monoxide (eCO) measurement is cheap and has been proposed to reflect airway inflammation and oxidative stress but current data are conflicting. The purpose of this meta-analysis is to determine whether eCO is elevated in asthmatics, is regulated by steroid treatment and reflects disease severity and control.</p> <p>Methods</p> <p>A systematic search for English language articles published between 1997 and 2009 was performed using Medline, Embase and Cochrane databases. Observational studies comparing eCO in non-smoking asthmatics and healthy subjects or asthmatics before and after steroid treatment were included. Data were independently extracted by two investigators and analyzed to generate weighted mean differences using either a fixed or random effects meta-analysis depending upon the degree of heterogeneity.</p> <p>Results</p> <p>18 studies were included in the meta-analysis. The eCO level was significantly higher in asthmatics as compared to healthy subjects and in intermittent asthma as compared to persistent asthma. However, eCO could not distinguish between steroid-treated asthmatics and steroid-free patients nor separate controlled and partly-controlled asthma from uncontrolled asthma in cross-sectional studies. In contrast, eCO was significantly reduced following a course of corticosteroid treatment.</p> <p>Conclusions</p> <p>eCO is elevated in asthmatics but levels only partially reflect disease severity and control. eCO might be a potentially useful non-invasive biomarker of airway inflammation and oxidative stress in nonsmoking asthmatics.</p

How certain are greenhouse gas reductions from bioenergy? Life cycle assessment and uncertainty analysis of wood pellet-to-electricity supply chains from forest residues

Climate change and energy policies often encourage bioenergy as a sustainable greenhouse gas (GHG) reduction option. Recent research has raised concerns about the climate change impacts of bioenergy as heterogeneous pathways of producing and converting biomass, indirect impacts, uncertainties within the bioenergy supply chains and evaluation methods generate large variation in emission profiles. This research examines the combustion of wood pellets from forest residues to generate electricity and considers uncertainties related to GHG emissions arising at different points within the supply chain. Different supply chain pathways were investigated by using life cycle assessment (LCA) to analyse the emissions and sensitivity analysis was used to identify the most significant factors influencing the overall GHG balance. The calculations showed in the best case results in GHG reductions of 83% compared to coal-fired electricity generation. When parameters such as different drying fuels, storage emission, dry matter losses and feedstock market changes were included the bioenergy emission profiles showed strong variation with up to 73% higher GHG emissions compared to coal. The impact of methane emissions during storage has shown to be particularly significant regarding uncertainty and increases in emissions. Investigation and management of losses and emissions during storage is therefore key to ensuring significant GHG reductions from biomass

Heme oxygenase-1 and carbon monoxide in pulmonary medicine

Heme oxygenase-1 (HO-1), an inducible stress protein, confers cytoprotection against oxidative stress in vitro and in vivo. In addition to its physiological role in heme degradation, HO-1 may influence a number of cellular processes, including growth, inflammation, and apoptosis. By virtue of anti-inflammatory effects, HO-1 limits tissue damage in response to proinflammatory stimuli and prevents allograft rejection after transplantation. The transcriptional upregulation of HO-1 responds to many agents, such as hypoxia, bacterial lipopolysaccharide, and reactive oxygen/nitrogen species. HO-1 and its constitutively expressed isozyme, heme oxygenase-2, catalyze the rate-limiting step in the conversion of heme to its metabolites, bilirubin IXα, ferrous iron, and carbon monoxide (CO). The mechanisms by which HO-1 provides protection most likely involve its enzymatic reaction products. Remarkably, administration of CO at low concentrations can substitute for HO-1 with respect to anti-inflammatory and anti-apoptotic effects, suggesting a role for CO as a key mediator of HO-1 function. Chronic, low-level, exogenous exposure to CO from cigarette smoking contributes to the importance of CO in pulmonary medicine. The implications of the HO-1/CO system in pulmonary diseases will be discussed in this review, with an emphasis on inflammatory states