Nine orders of magnitude dynamic range: picomolar to millimolar concentration measurement in capillary electrophoresis with laser induced fluorescence detection employing cascaded avalanche photodiode photon counters

The dynamic range of capillary electrophoresis analysis is ultimately limited by molecular shot noise at low concentrations and by concentration-induced band broadening at high concentrations. We report a system that approaches these fundamental limits. A laser-induced fluorescence detector is reported that employs a cascade of four fiber-optic beam splitters connected in series to generate a primary signal and four attenuated signals, each monitored by a single-photon counting avalanche photodiode. Appropriate scaling of the signals from the five photodiodes produces a linear optical calibration curve for 5-carboxyl-tetramethylrhodamine from the concentration detection limit of 1 pM to the upper limit of 1 mM. Mass detection limits are 120 yoctomoles (70 molecules) injected into the instrument. The very-wide dynamic range instrument was used to study the metabolic products of the fluorescently labeled glycosphingolipid tetramethylrhodamine labeled GM1 (GM1-TMR) produced by single cells isolated from the rat cerebellum.     

Direct detection of point mutations in nonamplified human genomic DNA

Ultrasensitive detection protocols not requiring polymerase chain reaction (PCR)-mediated target DNA amplification are expected to significantly improve our possibilities in several research and diagnostic applications for which minute cell quantities are available. For this reason we have tested a nanoparticle-enhanced surface plasmon resonance imaging (SPRI) sensing strategy to detect point mutations in nonamplified genomic DNA. We have used genomic DNAs, not subject to costly, time-consuming, and prone to contamination PCR-based amplification procedures, obtained from both healthy individuals and homozygous or heterozygous patients affected by β-thalassemia, in order to demonstrate the specificity and the sensitivity of the described sensing strategy. The assay we describe is ultrasensitive and convenient. Attomolar concentrations of target genomic DNA are detected, DNAs from healthy individuals and homozygous or heterozygous patients affected by β-thalassemia are discriminated, and only simple manipulations of the genetic samples are required before the analysis. The proposed ultrasensitive detection of DNA point mutations involved in genomic disorders possibly represents an important advantage in several biomedical applications.     

High survival of frozen cells irradiated with gamma radiation

Cell storage in liquid nitrogen (LN) offers the most secure method of cell preservation even if cryopreserved cells are exposed to natural background of ionising radiation (IR). A lot of experiments have demonstrated that IR can induce damages in living cells, but only a little information regarding the response of cryopreserved cells is available. To investigate the effect of IR on frozen and unfrozen cells, peripheral blood mononuclear cells were directly irradiated at room temperature, then immediately frozen, or frozen and then irradiated in LN with different doses of gamma rays. After thawing, cells were incubated and death fraction was evaluated at different time points. Interestingly, the percentages of dead cells induced by IR gradually increased with both dose radiation and incubation time and were significantly lower for cells irradiated at -196°C than those irradiated at room temperature.     

Finite Element Modeling to Predict Cure-Induced Microcracking in Three-Dimensional Woven Composites

Realistic finite element models of 3D woven composites are constructed utilizing micro-scale numerical modeling to accurately represent the geometry of as-woven textile fabrics. The models are used to predict microcracking of carbon fiber / epoxy composites during resin curing. Numerical predictions of the stress concentration areas correlate well with the observations of microcracking obtained by micro-computed tomography.     

Evaluation of various extraction techniques for obtaining bioactive extracts from pine seeds

Evaluation was made of four extraction techniques (maceration with magnetic stirring, ultrasound-assisted extraction, microwave-assisted extraction, and extraction with pressurised liquids) to obtain polyphenolic extracts from pine seeds. The technique and temperature conditions that produce extracts containing the largest quantity of polyphenols were determined. Extraction assisted by microwave and extraction with pressurised liquids at temperatures >100 °C gave the best results. The correlation between the antioxidant activity of the extracts and the quantity of polyphenols that they contain has also been determined; a direct relationship between these two variables was found in the majority of extracts.The general biological activity of the extracts was also evaluated using the etiolated wheat coleoptile bioassay; extracts obtained with pressurised liquids, and with maceration under agitation, were the most bioactive. An inverse relationship between the degree of bioactivity and the extraction temperature was found.     

Vacuum Plasma Spray Deposition of Titanium Particle/Glass-Ceramic Matrix Biocomposites

The vacuum plasma spray technique (VPS) has been successfully employed to coat Ti-6A1-4V substrates with bioactive glasses and Ti-particle/glass-ceramic matrix biocomposites. The composites were prepared by sintering, under an Ar flow, green bars of bioactive glass powders and 30% volume Ti particles. The bioactive glasses have the two following compositions: SCB (48.8SiO₂−48.8CaO−2.4B₂O₃) and TSCB (46.6SiO₂−48.7CaO−2.5B₂O₃−2.2TiO₂) (mol%). The VPS bioactive coatings were characterized by means of scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD), and mechanical tests (Vickers indentations and tensile and shear tests). Their bioactivity was tested by soaking the samples in a simulated body fluid (SBF) and by analyzing the growth of hydroxylapatite (HA) by SEM, EDS, and XRD. Leaching tests of Ca, Si, and P in SBF were made by inductively coupled plasma-atomic emission spectroscopy (ICP-AES, Perkin-Elmer 5000) to study the in vitro bioactivity of the samples versus time. Each coating was found to be bioactive and well bonded to the substrate; the composites showed better mechanical properties than the pure glass matrices and the hydroxylapatite coatings deposited by the same VPS technique.     

Chemical analysis,antioxidant, antiinflammatory and anticholinesterase activities of Origanum ehrenbergii Boiss and Origanum syriacum L. essential oils

The essential oils of Origanum ehrenbergii and O. syriacum collected in Lebanon were analysed by GC and GC–MS and evaluated for their anticholinesterase, NO production inhibitory activities, and antioxidant properties. O.ehrenbergi essential oil was characterised by the presence of 37 components, representing 94.9% of the total oil of which thymol (19%) and p-cymene (16.1%) were the main abundant compounds. Thirty-six compounds characterised the O.syriacum essential oil, representing 90.6% of the total oil. The most abundant components were thymol (24.7%) and carvacrol (17.6%). O. ehrenbergii demonstrated interesting scavenging effects on DPPH with an IC₅₀ value of 0.99 μg/ml. In addition, both O. ehrenbergii and O. syriacum oils inhibited oxidation of linoleic acid after 30 min of incubation, as well as after 60 min of incubation with IC₅₀ values of 42.1 and 33.6 μg/ml, and 46.9 and 58.9 μg/ml, respectively. Interestingly, O. ehrenbergii oil inhibited NO production in the murine monocytic macrophage cell line RAW 264.7 with an IC₅₀ value of 66.4 μg/ml. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibition was assessed by modifications of the Ellman’s method. O. ehrenbergii exhibited a strong activity against both cholinesterases with IC₅₀ values of 0.3 μg/ml. The data suggest that O. ehrenbergii and O. syriacum oils could be used as a valuable new flavour with functional properties for food or nutriceutical products with particular relevance to supplements for the elderly.     

Factorial validity and consistency of the MBI-GS across occupational groups in Norway.

The present study tested the factorial validity of the Maslach Burnout Inventory-General Survey (MBI-GS) among 694 participants from 4 different occupational groups. Confirmatory factor analyses of the total sample, as well as multigroup analyses and analyses of each of the 4 occupational groups separately, indicated that the original 3-factor model of the MBI-GS provided a good fit to the data. Internal consistencies of the subscales of the MBI-GS were acceptable, and test-retest reliability indicated relative stability of scores over a 6-month interval. These results indicate that the proposed 3-factor structure of the MBI-GS, based on 16 items, can be replicated in the total sample as well as across different occupational groups in Norway. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Removal of viable and inactivated Cryptosporidium by dual- and tri-media filtration

The limited efficacy of disinfectants, other than ultraviolet irradiation and ozonation, as a barrier against Cryptosporidium parvum in drinking water treatment has underscored the increased importance of oocyst removal by filtration. Currently, no reliable surrogates have been identified for C. parvum removal by filtration. As a result, evaluations of the Cryptosporidium removal by treatment operations have been performed using oocysts. It has typically been assumed that chemically inactivated oocysts are suitable surrogates for viable oocysts. Measurements of electrophoretic mobility, however, have shown that chemical inactivation changes the surface charge of Cryptosporidium oocysts. The present bench-scale research indicated that formalin-inactivated oocysts are reliable surrogates for viable oocysts during both stable filter operation and periods where filtration processes are challenged, such as coagulation failure. This finding is important because of the practical difficulties associated with using viable oocysts in filtration investigations. Poor coagulation conditions severely compromised removal of viable and inactivated oocysts by dual- and tri-media filters compared to stable operating conditions and filter ripening, emphasizing the importance of optimized chemical pre-treatment (coagulation) for the successful removal of oocysts during filtration. The treatment optimization experiments also indicated that tri-media filters offered only marginally higher oocyst removals than dual-media filters.