Optimization of PCR-based methods for rapid detection of Campylobacter jejuni, Campylobacter coli and Yersinia enterocolitica serovar 0:3 in wastewater samples

PCR-based methods were evaluated for their adequacy to assess the removal of pathogens from wastewater samples. For the development and optimization of the methods, samples were taken at two different sites from two different constructed wetlands. Campylobacter jejuni/coli and Yersinia enterocolitica serogroup 0:3 were selected as model pathogens and Enterococcus faecalis as a standard microbiological indicator. The chosen PCR protocols were optimized for wastewater DNA extracts in order to obtain high sensitivity and reproducibility independently of the background flora. All PCR protocols were successfully performed and reproducible with a background of up to 10(10) nontarget cells per reaction. Five cells of Y. enterocolitica, 50 cells of C. jejuni/coli, and 500 cells of E. faecalis per 100ml treated water could be detected. The method detection limit in the settled wastewater was higher: 200 cells per 100ml for Y. enterocolitica, 2000 cells per 100ml for C. jejuni/coli, and 20,000 cells per 100ml for E. faecalis. C. jejuni/coli and Y. enterocolitica PCRs were adapted to municipal wastewater, with higher loads of potential PCR inhibitors. Sensitivity was lower for this type of wastewater: 200 cells of Y. enterocolitica and 2000 cells of C. jejuni/coli were detected per 100ml treated wastewater, 2500 cells of Y. enterocolitica and 25,000 cells of C. jejuni/coli per 100ml settled wastewater. The developed PCR methods enable the detection of C. jejuni/coli, Y. enterocolitica serogroup 0:3 and E. faecalis within 12h. They show specificity, reproducibility and low detection limits for the investigated pathogens.     

Surface modifications of silicon nitride for cellular biosensor applications

Thin films of silicon nitride (Si₃N₄) can be used in several kinds of micro-sized biosensors as a material to monitor fine environmental changes related to the process of bone formation in vitro. We found however that Si₃N₄ does not provide optimal conditions for osseointegration as osteoblast-like MG-63 cells tend to detach from the surface when cultured over confluence. Therefore Si₃N₄ was modified with self-assembled monolayers bearing functional end groups of primary amine (NH₂) and carboxyl (COOH) respectively. Both these modifications enhanced the interaction with confluent cell layers and thus improve osseointegration over Si₃N₄. Furthermore it was observed that the NH₂ functionality increased the adsorption of fibronectin (FN), promoted cell proliferation, but delayed the differentiation. We also studied the fate of pre-adsorbed and secreted FN from cells to learn more about the impact of above functionalities for the development of provisional extracellular matrix on materials interface. Taken together our data supports that Si₃N₄ has low tissue integration but good cellular biocompatibility and thus is appropriate in cellular biosensor applications such as the ion-sensitive field effect transistor (ISFET). COOH and NH₂ chemistries generally improve the interfacial tissue interaction with the sensor and they are therefore suitable substrates for monitoring cellular growth or matrix deposition using electrical impedance spectroscopy.     

Effects of coping-oriented couples therapy on depression: A randomized clinical trial.

The aim of this study was to evaluate the effectiveness of treating depression with coping-oriented couples therapy (COCT) as compared with cognitive-behavioral therapy (CBT; A. T. Beck, C. Ward, & M. Mendelson, 1961) and interpersonal psychotherapy (IPT; M. M. Weissman, J. C. Markowitz, & G. L. Klerman, 2000). Sixty couples, including 1 clinically depressed partner, completed pre- and posttest questionnaires as well as follow-up assessments at 6-month intervals over the subsequent 1.5 years. Effects of the 3 treatments on depressive symptomatology assessed by the Beck Depression Inventory (A. T. Beck, A. J. Rush, B. L. Shaw, & G. Emery, 1979) and Hamilton Rating Scale for Depression (M. Hamilton, 1960); recovery rates; and relapse rates were examined. Additionally, changes in relationship quality were evaluated. Results suggest that the COCT is as effective in improving depressive symptomatology as are the well-established, evidenced-based CBT and IPT approaches. The COCT did not demonstrate a significantly better outcome with regard to self-reported relationship satisfaction or dyadic coping; however, it did produce significant improvements in partners' expressed emotion, changes that were not seen in other treatment conditions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The Hallagerbakken housing project in Holmlia,Norway: when welfare became business

The late 1970s and early 1980s witnessed one of the most influential shifts within the organisational system of production for Norwegian mass housing. Although the increased availability of resources following the Second World War and the discovery of North Sea oil put the Norwegian government in a favourable position for investing in state-sponsored housing programmes, such investments did not happen. Instead, the strong public responsibility for affordable housing as a right of all citizens was weakened, and housing evolved into a commodity to be traded primarily on the free market.

Hallagerbakken, a housing project within the Holmlia satellite town—the last large housing development in the south of Oslo—has been influenced by several of the changes that occurred in the housing sector during this period. The resulting hybrid housing typology represented an innovation in the Norwegian setting, and, as a result, the project provides a starting point for re-evaluating some of the shifts towards a more market-oriented reality within architecture and the built environment.     

Preparation and characterization of cross-linked phospholipid bilayer capillary coatings for protein separations

Analysis of protein and peptide mixtures via capillary electrophoresis is hindered by nonspecific adsorption of analytes to the capillary walls, resulting in poor separations and quantitative reproducibility. Phospholipid bilayer (PLB) coatings are very promising for improving protein and peptide separations due to the native resistance to nonspecific protein adsorption offered by PLBs; however, these coatings display limited chemical and temporal stability. Here, we show the preparation and characterization of a highly cross-linked, polymerized phospholipid capillary coating prepared using bis-SorbPC. Poly(bis-SorbPC) PLB coatings are prepared in situ within fully enclosed fused silica capillaries via self-assembly and radical polymerization. Polymerization of the PLB coating stabilizes the membrane against desorption from the surface and migration in an electric field, improves the temporal and chemical stability, and allows for the separation of both cationic and anionic proteins, while preserving the native resistance to nonspecific protein adsorption of natural PLBs.     

Supercritical antisolvent versus coevaporation: preparation and characterization of solid dispersions

The objective of this work was to improve the dissolution rate and aqueous solubility of oxeglitazar. Solid dispersions of oxeglitazar in PVP K17 (polyvinilpyrrolidone) and poloxamer 407 (polyoxyethylene-polyoxypropylene block copolymer) were prepared by supercritical antisolvent (SAS) and coevaporation (CoE) methods. Drug-carrier formulations were characterized by powder X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, gas chromatography, UV/VIS spectroscopy and in vitro dissolution tests. The highest dissolution rate (nearly 3-fold higher than raw drug) was achieved by preparation of drug/PVP K17 coevaporate. Oxeglitazar/PVP K17 solid dispersions were stabilized by hydrogen bonding but contained higher amount of residual dichloromethane (DCM) than poloxamer 407 formulations regardless of the method of preparation. SAS prepared oxeglitazar/poloxamer 407 dissolved more than two times faster than raw drug. However, unlike PVP K17, poloxamer 407 did not form a single phase amorphous solid solution with oxeglitazar which has been manifested in higher degrees of crystallinity, too. Among the two techniques, evaluated in this work, conventional coevaporation resulted in higher amorphous content but SAS reduced residual solvent content more efficiently.     

Biocompatibility of ultra-high molecular weight polyethylene (UHMW-PE) stabilized with α-tocopherol used for joint endoprostheses assessed in vitro

Adding the natural antioxidant α-tocopherol to ultra-high molecular weight polyethylene (UHMW-PE) can remarkably delay the oxidation of hip cups made thereof. However, α-tocopherol is likely to undergo different chemical transformations during manufacturing and sterilization of hip cups than in human metabolism. Therefore, the biocompatibility of the putative transformation products has to be investigated. In-vitro tests with L929 mice fibroblast-cells gave no evidence for cytotoxicity. To further ensure the biocompatibility, in-vitro tests with human cells were carried out in this study. Two different human cell lines, one adherent cell line, HF-SAR, and one suspension culture, GSJO, were tested on UHMW-PE-tablets (diameter: 15 mm; thickness: 2 mm; processed according to standard procedures for artificial hip-cups) with and without α-tocopherol with respect to cell viability, proliferation and morphology by means of cell counting, WSt-1 proliferation assay and scanning electron microscopy. Similar proliferation rates were found with both polyethylene samples. Further, we found intact morphology in light and electron microscopy on each substrate. The morphologic characteristics of skin fibroblasts were not changed by any material. Normal adherence and spreading of the fibroblasts was found on controls of glass, as well as on polystyrene and on stabilized and unstabilized polyethylene. The characteristic behaviour as suspension of the GSJO cells remained unchanged. The mitochondrial activity, as studied by WST-1 cell proliferation reagent, was identical on each substrate during the whole observation period of 7 days. Christian Wolf—in partial fulfillment of a Ph.D. (Dr.mont.) thesis at the University of Leoben     

Physicochemical characteristics and bronchial epithelial cell cytotoxicity of Folpan 80 WG® and Myco 500®, two commercial forms of folpet

Background

Pesticides, in particular folpet, have been found in rural and urban air in France in the past few years. Folpet is a contact fungicide and has been widely used for the past 50 years in vineyards in France. Slightly water-soluble and mostly present as particles in the environment, it has been measured at average concentration of 40.1 μg/m³ during its spraying, 0.16–1.2 μg/m³ in rural air and around 0.01 μg/m³ in urban air, potentially exposing both the workers and the general population. However, no study on its penetration by inhalation and on its respiratory toxicity has been published. The objective of this study was to determine the physicochemical characteristics of folpet particles (morphology, granulometry, stability) in its commercial forms under their typical application conditions. Moreover, the cytotoxic effect of these particles and the generation of reactive oxygen species were assessed in vitro on respiratory cells.

Results

Granulometry of two commercial forms of folpet (Folpan 80WG^® and Myco 500^®) under their typical application conditions showed that the majority of the particles (>75%) had a size under 5 μm, and therefore could be inhaled by humans. These particles were relatively stable over time: more than 75% of folpet remained in the particle suspension after 30 days under the typical application conditions. The inhibitory concentration (IC₅₀) on human bronchial epithelial cells (16HBE14o-) was found to be between 2.89 and 5.11 μg/cm² for folpet commercial products after 24 h of exposure. Folpet degradation products and vehicles of Folpan 80 WG^® did not show any cytotoxicity at tested concentrations. At non-cytotoxic and subtoxic concentrations, Folpan 80 WG^® was found to increase DCFH-DA fluorescence.

Conclusion

These results show that the particles of commercial forms of folpet are relatively stable over time. Particles could be easily inhaled by humans, could reach the conducting airways and are cytotoxic to respiratory cells in vitro. Folpet particles may mediate its toxicity directly or indirectly through ROS-mediated alterations. These data constitute the first step towards the risk assessment of folpet particles by inhalation for human health. This work confirms the need for further studies on the effect of environmental pesticides on the respiratory system.     

High friction on a bubble mattress

Reducing the friction of liquid flows on solid surfaces has become an important issue with the development of microfluidics systems, and more generally for the manipulation of fluids at small scales. To achieve high slippage of liquids at walls, the use of gas as a lubricant--such as microbubbles trapped in superhydrophobic surfaces--has been suggested. The effect of microbubbles on the effective boundary condition has been investigated in a number of theoretical studies, which basically show that on flat composite interfaces the magnitude of the slippage is proportional to the periodicity of the gaseous patterns. Recent experiments aiming to probe the effective boundary condition on superhydrophobic surfaces with trapped bubbles have indeed shown high slippage in agreement with these theoretical predictions. Here, we report nanorheology measurements of the boundary flow on a surface with calibrated microbubbles. We show that gas trapped at a solid surface can also act as an anti-lubricant and promote high friction. The liquid-gas menisci have a dramatic influence on the boundary condition, and can turn it from slippery to sticky. It is therefore essential to integrate the control of menisci in fluidic microsystems designed to reduce wall friction.