Analysis of bacterial diversity in river biofilms using 16S rDNA PCR-DGGE: methodological settings and fingerprints interpretation

Reliability of bacterial diversity assessment using polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) analysis of 16S rDNA fragments was evaluated for a particular complex microbial assemblage: river epilithic biofilm. By comparing 3 routine protocols on replicates of one river biofilm sample, we found that common DNA extraction procedures gave comparable diversity (from 28.0 to 30.7 bands detected) and community composition (> 75% of homology) despite differences in the total amount of extracted DNA (from 0.9 to 4.2 microg). Therefore methodological improvements only concerned electrophoretic separation of DNA fragments (range of denaturing gradient from 35% to 70% and migration time=18h) and standardisation of DNA amounts used (PCR-template=50 ng, gel loading=700 ng). Using such a standardised methodology we found a good reproducibility of all steps of the procedure. When an Escherichia coli strain was introduced as a contaminant in a biofilm sample, we were able to recover ribotypes from the strain. As concerns fields sampling, a satisfactory repeatability of banding patterns from neighbouring pebbles (sampling point) allowed discriminating between the biofilm intrasite variability (various points from a cross-profile). These trials confirmed that PCR-DGGE is suitable to assess a reliable genetic fingerprint of epilithic biofilms in the river. Phylogenetic analysis of 40 partial sequences of 16S rDNA from DGGE gels of two sets of river biofilms samples proved evidences for the retrieval of DNA fragments related to phototroph Eukarya. However, in both cases plastidial 16S rDNA represented less than 25% of the analysed operational taxonomic units. Taking into account that Cyanobacteria, as members of the Bacteria, were also detected, sequence analysis of relevant bands from the pattern is required to target "bacteria", i.e. the functional group of prokaryotic microorganisms to which one commonly refers as a key component in sustaining the nutrient turnover.     

Evaluation of drainage system around a lined pilot cavern for underground cryogenic LNG storage

A lined pilot cavern for underground cryogenic LNG (liquefied natural gas) storage was constructed in granite in Daejeon, Korea in 2003 and commissioned in 2004. As the hydrostatic pressure of groundwater and thermal stress due to the formation of ice lenses may damage the containment system, rock drainage around the pilot cavern is needed to maintain the stability of the containment system. Once the drainage works were completed, the level of groundwater around the pilot cavern was controlled using drainage holes to form an ice ring as a second barrier to prevent the leakage of LNG from the cavern. In order to establish the drainage system for the pilot cavern, 15 boreholes were drilled into the rock. Fractures in the rock mass around the pilot cavern were characterized to determine the most appropriate orientation for the drainage holes. The major joints acting as conduits for inflow water were designed for efficient drainage. After the 15 drainage holes were drilled and their efficiency tested during the dry season in April 2003, it was found that there was a problem with the inflow of water through the main joint along the right-hand wall of the cavern, indicating the system was less efficient on the right-hand side. Hence, three more boreholes were drilled in the correct direction on the right-hand wall of the cavern. A second drainage test was conducted during the rainy season in August. This showed some seepage into the bottom of the pilot cavern. In order to reduce this seepage, two more additional drainage holes were drilled and grouted below the concrete invert. Although the drainage system was very efficient, weak points in the system were found by testing and changes were made to the system to improve its efficiency.     

Exposure levels to brominated compounds in seawater swimming pools treated with chlorine

Despite evidence of formation of brominated compounds in seawater swimming pools treated with chlorine, no data about exposure levels to these compounds have been reported. To address this issue, a survey has been carried out in four establishments (representing 8 pools) fed with seawater and devoted to relaxing and cure treatments (thalassotherapy centres located in Southeast of France). Carcinogenic and mutagenic brominated disinfection byproducts (trihalomethanes -THM- and halogenated acetic acids -HAA-) were quantified at varying levels, statistically related to organic loadings brought by bathers, and not from marine organic matter, and also linked to activities carried out in the pools (watergym vs swimming). Bromoform and dibromoacetic acid, the most abundant THM and HAA detected, were measured at levels up to 18-fold greater than the maximum contaminant levels of 60 and 80 μg/L fixed by US.EPA in drinking waters. The correlations between these disinfection byproducts and other environmental factors such as nitrogen, pH, temperature, free residual chlorine, UV₂₅₄, chloride and bromide concentrations, and daily frequentation were examined. Because thalassotherapy and seawater swimming pools (hotels, cruise ships,…) are increasing in use around the world and because carcinogenic and mutagenic brominated byproducts may be produced in chlorinated seawater swimming pools, specific care should be taken to assure cleanliness of users (swimmers and patients taking the waters) and to increase water circulation through media filters to reduce levels of brominated byproducts.     

Hole-vibronic coupling in oligothiophenes: impact of backbone torsional flexibility on relaxation energies

Density functional theory calculations together with highly resolved gas-phase ultraviolet photoelectron spectroscopy have been applied to oligothiophene chains with up to eight thiophene rings. One of the important parameters governing the charge transport properties in the condensed phase is the amount of energy relaxation upon ionization. Here, we investigate the impact on this parameter of the backbone flexibility present in oligothiophenes as a result of inter-ring torsional motions. With respect to oligoacenes that are characterized by a coplanar and rigid backbone, the torsional flexibility in oligothiophenes adds to the relaxation energy and leads to the broadening of the first ionization peak, making its analysis more complex.     

Particle sizing with a fast polar nephelometer

We reported previously the design of a polar nephelometer that uses a rotational confocal imaging setup to enable fast scanning of the scattering phase function within a field of view of 55 degrees . The full dynamic range of the detection system can be used by increasing the signal-to-noise ratio by means of averaging successive scans. The calibration of the angular response of the instrument is achieved by obtaining the transfer function of the optical detection system using Rayleigh scatterers. Accurate particle sizing of individual polystyrene spheres (ranging from 1.5 to 9 micro m in diameter) in aqueous suspension is achieved by maximizing a correlation coefficient between precalculated tables of Mie phase functions and data obtained from the polar nephelometer. Good correlation is achieved between experimental and theoretical data, proving the functioning of the instrument as a fast and convenient particle sizer.     

Estimation of polyvinyl alcohol cryogel mechanical properties with four ultrasound elastography methods and comparison with gold standard testings

Tissue-mimicking phantoms are very useful in the field of tissue characterization and essential in elastography for the purpose of validating motion estimators. This study is dedicated to the characterization of polyvinyl alcohol cryogel (PVA-C) for these types of applications. A strict fabrication procedure was defined to optimize the reproducibility of phantoms having a similar elasticity. Following mechanical stretching tests, the phantoms were used to compare the accuracy of four different elastography methods. The four methods were based on a one-dimensional (1-D) scaling factor estimation, on two different implementations of a 2-D Lagrangian speckle model estimator (quasistatic elastography methods), and on a 1-D shear wave transient elastography technique (dynamic method). Young's modulus was investigated as a function of the number of freeze-thaw cycles of PVA-C, and of the concentration of acoustic scatterers. Other mechanical and acoustic parameters-such as the speed of sound, shear wave velocity, mass density, and Poisson's ratio-also were assessed. The Poisson's ratio was estimated with good precision at 0.499 for all samples, and the Young's moduli varied in a range of 20 kPa for one freeze-thaw cycle to 600 kPa for 10 cycles. Nevertheless, above six freeze-thaw cycles, the results were less reliable because of sample geometry artifacts. However, for the samples that underwent less than seven freeze-thaw cycles, the Young's moduli estimated with the four elastography methods showed good matching with the mechanical tensile tests with a regression coefficient varying from 0.97 to 1.07, and correlations R2 varying from 0.93 to 0.99, depending on the method.     

Parameter identification and computational simulation of polyurethane foaming process by finite pointset method

Numerical simulation of the polyurethane foaming process is a valuable method to analyze the molding process at an early stage of product development to shorten time-to-market cycles and cut costs by using fewer prototypes. However, this process involves highly coupled thermo-chemo-rheological modeling and needs adequate model parameters’ identification. A theoretical model including chemical reactions and thermo-rheological coupling of conservation equations was developed. Based on the theoretical model, three-dimensional numerical simulation for mold filling of the polyurethane foam was carried out by using Finite Pointset Method (FPM) to predict flow field, flow front advancement, temperature and density distributions during mold filling. A FOAMAT system was used to monitor foam height rise and reaction temperature on a cylindrical test tube and foam viscosity was measured by using a dynamic rotational rheometer with parallel-plate system. The parameters of the model were identified by an inverse analysis method which consists in determining the parameters by comparing the computed quantities to those measured experimentally. The overall modeling was validated by using short shot foams obtained with a panel mold cavity. Mold filling of an automotive underlay carpet cavity was investigated numerically. Flow front results were successfully compared to short shot foams obtained with the industrial mold cavity.     

Assessment of elastic parameters of human skin using dynamic elastography

Sonoelastography and transient elastography are two ultrasound-based techniques that facilitate noninvasive characterization of the viscoelastic properties of soft tissues by investigating their response to shear mechanical excitation. Young's modulus is the principle assessment parameter. Because it defines local tissue stiffness, it is of major interest for the medical imaging and cosmetic industries as it could replace subjective palpation by yielding local, quantitative information. In this paper, we describe a new high-resolution device capable of measuring local Young's modulus in very thin layers (1-5 mm) and devoted to the in vivo evaluation of the elastic properties of human skin. It uses an ultrasonic probe (50 MHz) for tracking the displacements induced by a 300 Hz shear wave generated by a ring surrounding the transducer. The displacements are measured using a conventional cross-correlation technique between successive ultrasonic back-scattered echoes. First, this noninvasive technique has been experimentally proven to be accurate for investigating elasticity in different skin-mimicking phantoms. Second, data were acquired in vivo on human forearms. As expected, Young's modulus was found to be higher in the dermis than in the hypodermis and other soft tissues.     

Renal Clearable Theranostic Nanoplatforms for Gastrointestinal Stromal Tumors

Advances in molecular imaging modalities have accelerated the diagnosis and treatment of human diseases. However, tumors less than 1 cm in size still remain difficult to localize by conventional means because of the difficulty in specific targeting/delivery to the tumor site. Furthermore, high nonspecific uptake in the major organs and persistent background retention results in low tumor-to-background ratio. The targeting and therapy of gastrointestinal stromal tumors (GIST) using nonsticky and renal clearable theranostic nanoparticles (a.k.a. H-Dots) are demonstrated. H-Dots not only target GIST for image-guided surgery, but also tailor the fate of anticancer drugs such as imatinib (IM) to the tumor site resulting in efficient treatment of unresectable GIST. In addition, H-Dots can monitor targetability, pharmacokinetics, and drug delivery, while also showing therapeutic efficacy in GIST-bearing xenograft mice following surgical resection. More importantly, IM loaded H-Dots exhibit lower uptake into the immune system, improved tumor selectivity, and increased tumor suppression compared to free IM, which accumulates in the spleen/liver. Precisely designed H-Dots can be used as a promising theranostic nanoplatform that can potentially reduce the side effects of conventional chemotherapies.     

Cytocompatibility of titanium metal injection molding with various anodic oxidation post-treatments

Metal injection molding (MIM) is a near net shape manufacturing method that allows for the production of components of small to moderate size and complex shape. MIM is a cost-effective and flexible manufacturing technique that provides a large innovative potential over existing methods for the industry of implantable devices. Commercially pure titanium (CP-Ti) samples were machined to the same shape as a composite feedstock with titanium and polyoxymethylene, and these metals were injected, debinded and sintered to assess comparative biological properties. Moreover, we treated MIM-Ti parts with BIOCOAT®, BIODIZE® and BIOCER®, three different anodic oxidation techniques that treat titanium using acid, alkaline and anion enriched electrolytes, respectively. Cytocompatibility as well as morphological and chemical features of surfaces was comparatively assessed on each sample, and the results revealed that MIM-Ti compared to CP-Ti demonstrated a specific surface topography with a higher roughness. MIM-Ti and BIOCER® samples significantly enhanced cell proliferation, cell adhesion and cell differentiation compared to CP-Ti. Interestingly, in the anodization post-treatment established in this study, we demonstrated the ability to improve osseointegration through anionic modification treatment. The excellent biological response we observed with MIM parts using the injection molding process represents a promising manufacturing method for the future implantable devices in direct contact with bones.