Effect of Natural Graphite Fineness on the Performance and Electrical Conductivity of Cement Paste Mixes for Self-Sensing Structures.

Cementitious composites are the most widely used construction materials; however, their poor durability necessitates frequent monitoring and repairs. The emergence of self-sensing composites could reduce the need for costly and time-consuming structural inspections. Natural graphite, due to its low cost and wide availability, is a promising additive to generate an electrically conductive network which could ultimately lead to a self-sensing mechanism. Despite several studies using natural graphite as a conductive additive, the effect of its fineness on the cementitious composite's performance has not been explored. This study experimentally investigated the effect of three graphite products of varying fineness on the early age, mechanical, and electrical conductivity performance of cement pastes. The fluidity of the graphite-cement paste reduced significantly with increasing graphite fineness, and graphite did not affect the cement hydration. The finer the graphite, the lower the effect on the mechanical performance, as confirmed by compressive strength testing and micro-indentation. Electrical conductivity testing showed that the percolation threshold depended on the graphite fineness and was found at ~20 wt % for the fine and medium graphite, while it increased to 30-40 wt % for the coarse graphite. This is the first study that has investigated holistically the effect of graphite fineness on the performance of cement pastes and will pave the way for using this material as an additive for self-sensing structures.

Standardization of an ash (Fraxinus excelsior) pollen allergen extract.

BACKGROUND: Ash tree (Fraxinus excelsior) is the main representative of the Oleaceae family in temperate zones. Diagnosis of ash pollen allergy is made difficult due to (1) an overlapping pollinization period with Betulaceae, (2) non-inclusion in current diagnostic assays, and (3) some cross- reactivity with minor allergens from Betulaceae. The aim of this study was to calibrate an ash pollen in-house reference preparation (IHRP) in allergic patients in order to produce standardized products for diagnosis and immunotherapy purposes. METHODS: Ash pollen IHRP was extracted, ultrafiltered and freeze dried. Allergens in the extract were detected after 2-dimensional PAGE using specific sera and a monoclonal antibody. The Fra e 1 content of IHRP was evaluated by quantitative immunoprint. Forty-eight subjects from the North-East of France exhibiting clinical symptoms, a positive skin test and specific IgE levels > or =class 2 to ash pollen were recruited. IgE immunoprints were performed to select patients sensitized to the ash Fra e 1 allergen as opposed to cross-reacting allergens. Serial 10-fold dilutions of the IHRP were tested by skin prick tests in order to determine the concentration inducing a geometrical mean wheal diameter of 7 mm, said to correspond to an index of reactivity (IR) of 100 per millilitre. RESULTS: IgE-reactive molecules in IHRP comprise Fra e 1, Fra e 2, a 9-kDa molecule (presumably Fra e 3), as well as a doublet at 15 kDa and high molecular weight allergens. The 100 IR concentration of IHRP inducing a geometrical mean wheal diameter of 7 mm in 22 patients sensitized to Fra e 1 corresponds to the 1/126 (w/v) extraction ratio (i.e. 259 microg/ml of protein by Bradford) and contains 17 microg/ml of Fra e 1. The variability in total activity of 5 batches of standardized extracts was found to be significantly reduced when compared with 7 non-standardized extracts. CONCLUSION: An ash pollen IHRP was defined and molecularly characterized. Its successful standardization at 100 IR/ml in patients specifically sensitized to Fra e 1 allowed a skin reactivity-based calibration in properly diagnosed patients. Such a standardized ash pollen extract is a reliable tool to support immunotherapy of ash pollen allergy.

Cloning, expression, and clinical significance of the major allergen from ash pollen, Fra e 1.

BACKGROUND: Ash tree, an Oleaceae member, is considered an important source of pollen allergy in Central Europe. Fra e 1 is a protein of the Ole e 1-like family, which regulates pollen tube growth. It has been suggested to be a relevant allergen from ash pollen. OBJECTIVE: Cloning Fra e 1-cDNA and overproducing a properly folded recombinant allergen to analyze its clinical significance. METHODS: Fra e 1-encoding cDNA was amplified by PCR, cloned in Escherichia coli , and sequenced. The recombinant allergen was produced in Pichia pastoris and used in immunoblotting, ELISA, histamine release, and skin prick tests. Sera and blood cells from patients sensitized to ash pollen as well as anti-Ole e 1 monoclonal and polyclonal antisera were used. RESULTS: Recombinant Fra e 1 (rFra e 1) is a glycoprotein of 145 amino acids exhibiting 82%, 88%, and 91% identity with Syr v 1, Ole e 1, and Lig v 1, allergens of the Oleaceae family. It was secreted to the extracellular medium of the yeast cultures and purified by means of 3 chromatographic steps. IgG from Ole e 1-specific antibodies recognized rFra e 1. IgE antibodies from ash-sensitized patients bound to rFra e 1 with a prevalence of 75%. The recombinant allergen induced histamine release. Twenty-nine of 30 ash-sensitized patients were positive to rFra e 1 by skin prick tests. CONCLUSION: Fra e 1 is a relevant allergen in ash pollen sensitization. It has been efficiently produced in P pastoris and could be used in diagnosis.