Evidence for a radical mechanism of the dechlorination of chlorinated propenes mediated by the tetrachloroethene reductive dehalogenase of Sulfurospirillum muftivorans

The reductive dehalogenation of chlorinated propenes was studied with the tetrachloroethene reductive dehalogenase purified from Sulfurospirillum multivorans to obtain indications for a radical mechanism of this reaction. When reduced methyl viologen (MV), which is a radical cation, was applied as electron donor for the reduction of different chloropropenes, a significant part of MV could not be rereduced with Ti(III) citrate, indicating that a part of the MV was consumed in a side reaction. Mass spectrometric analysis of assays with MV as electron donor revealed the formation of side products, the masses of which might account for the formation of adducts from a chloropropenyl radical and reduced methyl viologen. With Ti(III) citrate as sole electron donor, 2,3-dichloropropene was reduced and as a side product, 2,5-dichloro-1,5-hexadiene was formed demonstrating that the reductive dechlorination of 2,3-dichloropropene proceeds via a radical reaction mechanism. The results support different dehalogenation mechanisms forthe reductive dechlorination of chloropropenes and halogenated ethenes.     

Progress Toward an Integration of Process–Structure–Property–Performance Models for “Three-Dimensional (3-D) Printing” of Titanium Alloys

Electron beam direct manufacturing, synonymously known as electron beam additive manufacturing, along with other additive “3-D printing” manufacturing processes, are receiving widespread attention as a means of producing net-shape (or near-net-shape) components, owing to potential manufacturing benefits. Yet, materials scientists know that differences in manufacturing processes often significantly influence the microstructure of even widely accepted materials and, thus, impact the properties and performance of a material in service. It is important to accelerate the understanding of the processing–structure–property relationship of materials being produced via these novel approaches in a framework that considers the performance in a statistically rigorous way. This article describes the development of a process model, the assessment of key microstructural features to be incorporated into a microstructure simulation model, a novel approach to extract a constitutive equation to predict tensile properties in Ti-6Al-4V (Ti-64), and a probabilistic approach to measure the fidelity of the property model against real data. This integrated approach will provide designers a tool to vary process parameters and understand the influence on performance, enabling design and optimization for these highly visible manufacturing approaches.     

Optimal conditions for determination of zinc bacitracin,polymyxin B,oxytetracycline and sulfacetamide in animal feed by micellar electrokinetic capillary chromatography

A separation technique for zinc bacitracin, polymyxin B, oxytetracycline and sulfacetamide in animal feedstuffs by micellar electrokinetic capillary chromatography (MEKC) was developed. The running buffer was 20 mmol l^?1 borate, 20 mmol l^?1 phosphate, pH 8.4, containing 20 mmol l^?1 sodium dodecylsulphate and 10% (v/v) methanol. MEKC was performed at 25°C; the applied voltage was 25 kV with a running pressure of 10 mbar. Simultaneous UV detection for all analytes was at 215 nm. The method was validated for specificity, accuracy, linearity, precision and robustness. It was shown to be specific, accurate (recoveries were 99.7?±?0.3, 99.9?±?0.9, 99.8?±?1.0 and 99.5?±?0.4, respectively, for oxytetracycline-, sulfacetamide-, polymyxin B- and zinc bacitracin-spiked samples of feed for cow, pigs, chicken and cattle), linear over the tested range (correlation coefficients ≥0.9987) and precise (RSDs below 1.8% for each analyte). The method was applied to determine zinc bacitracin, polymyxin B, oxytetracycline and sulfacetamide as additives in animal feed.     

The Manufacture of Xeno- and Feeder-Free Clinical-Grade Human Embryonic Stem Cell Lines: First Step for Cell Therapy

Human embryonic stem cells (hESCs) are increasingly used in clinical trials as they can change the outcome of treatment for many human diseases. They are used as a starting material for further differentiation into specific cell types and to achieve the desirable result of the cell therapy; thus, the quality of hESCs has to be taken into account. Therefore, current good manufacturing practice (cGMP) has to be implemented in the transport of embryos, derivation of inner cell mass to xeno-free, feeder-free and defined hESC culture, and cell freezing. The in-depth characterization of hESC lines focused on safety, pluripotency, differentiation potential and genetic background has to complement this process. In this paper, we show the derivation of three clinical-grade hESC lines, MUCG01, MUCG02, and MUCG03, following these criteria. We developed and validated the system for the manufacture of xeno-free and feeder-free clinical-grade hESC lines that present high-quality starting material suitable for cell therapy according to cGMP.     

Detecting cracks in the tooth root of gears

A crack in the tooth root is the least desirable damage caused to gear units and often leads to failure of gear unit operation. A possible damage in gear units can be identified by monitoring vibrations. In relation to that, different methods of time signals analysis are presented. Signals have been obtained by experiments. Significant changes in tooth stiffness are the result of a fatigue crack in a tooth root. As a consequence, dynamic response differs from the one in concern to an undamaged tooth. Amplitudes of time signal are, by frequency analysis, presented as a function of frequencies in spectrum with time frequency analysis.     

Influence of Long-Distance Bicycle Riding on Serum/Urinary Biomarkers of Prostate Cancer

Herein, we present a study focused on the determination of the influence of long-distance (53 km) bicycle riding on levels of chosen biochemical urinary and serum prostate cancer (PCa) biomarkers total prostate-specific antigen (tPSA), free PSA (fPSA) and sarcosine. Fourteen healthy participants with no evidence of prostate diseases, in the age range from 49–57 years with a median of 52 years, underwent physical exercise (mean race time of 150 ± 20 min, elevation increase of 472 m) and pre- and post-ride blood/urine sampling. It was found that bicycle riding resulted in elevated serum uric acid (p = 0.001, median 271.76 vs. 308.44 µmol/L pre- and post-ride, respectively), lactate (p = 0.01, median 2.98 vs. 4.8 mmol/L) and C-reactive protein (p = 0.01, 0.0–0.01 mg/L). It is noteworthy that our work supports the studies demonstrating an increased PSA after mechanical manipulation of the prostate. The subjects exhibited either significantly higher post-ride tPSA (p = 0.002, median 0.69 vs. 1.1 ng/mL pre- and post-ride, respectively) and fPSA (p = 0.028, median 0.25 vs. 0.35 ng/mL). Contrary to that, sarcosine levels were not significantly affected by physical exercise (p = 0.20, median 1.64 vs. 1.92 µmol/mL for serum sarcosine, and p = 0.15, median 0.02 µmol/mmol of creatinine vs. 0.01 µmol/mmol of creatinine for urinary sarcosine). Taken together, our pilot study provides the first evidence that the potential biomarker of PCa—sarcosine does not have a drawback by means of a bicycle riding-induced false positivity, as was shown in the case of PSA.     

DNA detection using a triple readout optical/AFM/MALDI planar microwell plastic chip

A ready-to-spot disposable DNA chip for specific and sensitive detection of DNA was developed. Plastic copolymeric substrate chemistry was optimized to selectively couple the target DNA with the active chip surface. At the same time, the developed substrate limits the unspecific adsorption of probe DNA molecules or additional polar contaminants in the test samples to the chip surface. The combination of glycidyl and n-butyl methacrylates was found to best fit the requirements of the assay. The fabricated DNA microarrays have mechanical properties similar to those of the glass or silicon substrates and, at the same time, provide chemically reactive surfaces that do not require lengthy chemical modification. An additional advantage of the plastic microchip is its compatibility with different analytical readout techniques, such as mass spectrometry (MALDI-TOF/MS), optical detection (fluorescence and enzyme-induced metal deposition), and imaging techniques (atomic force microscopy). These multiple readout techniques have given us the ability to compare the sensitivity, selectivity, and robustness of current state-of-the-art bioanalytical methods on the same platform exemplified by successful DNA-based detection of human cytomegalovirus. The obtained sensitivity for enzymatically enhanced silver deposition (10(-15) M) surpasses that of conventional fluorescence readouts. In addition, the assay's dynamic range (10(-6)-10(-15) M), reproducibility, and reliability of the DNA probe detection speaks for the silver deposition method. At compromised sensitivity (10(-9) M), the length of the DNA probes could be checked and, alternatively, DNA single point polymorphisms could be analyzed.     

Improved Synthesis of Buprenorphine from Thebaine and/or Oripavine via Palladium‐Catalyzed N‐Demethylation/Acylation and/or Concomitant O‐Demethylation

An improved preparation of buprenorphine via palladium‐catalyzed N‐demethylation/acylation is reported. Three routes were investigated and compared in overall yield. The first involved N‐demethylation/acylation of an advanced intermediate obtained from thebaine followed by hydrolysis of the N‐acetamide and alkylation with cyclopropylmethyl bromide and/or reduction of the N‐acetyl group with the Schwartz reagent followed by N‐alkylation. The second route employed cyclopropylcarboxylic acid anhydride in the N‐demethylation/acylation protocol and subsequent reduction of the cyclopropylcarboxamide by either lithium aluminum hydride or under hydrosilylation conditions. Both of these routes originated in thebaine and therefore required O‐demethylation as a final step. The third route employed an N‐demethylation/acylation sequence starting from oripavine rather than thebaine, thus avoiding the O‐demethylation. The routes are compared for overall efficiency and experimental and spectral data are provided for all new compounds.     

Combining reconstructive and discriminative subspace methods for robust classification and regression by subsampling

Linear subspace methods that provide sufficient reconstruction of the data, such as PCA, offer an efficient way of dealing with missing pixels, outliers, and occlusions that often appear in the visual data. Discriminative methods, such as LDA, which, on the other hand, are better suited for classification tasks, are highly sensitive to corrupted data. We present a theoretical framework for achieving the best of both types of methods: an approach that combines the discrimination power of discriminative methods with the reconstruction property of reconstructive methods which enables one to work on subsets of pixels in images to efficiently detect and reject the outliers. The proposed approach is therefore capable of robust classification with a high-breakdown point. We also show that subspace methods, such as CCA, which are used for solving regression tasks, can be treated in a similar manner. The theoretical results are demonstrated on several computer vision tasks showing that the proposed approach significantly outperforms the standard discriminative methods in the case of missing pixels and images containing occlusions and outliers.     

Synthesis of silver nanoparticles by Bacillus subtilis T‐1 growing on agro‐industrial wastes and producing biosurfactant

In this study, culture supernatnats of Bacillus subtilis T‐1 growing on brewery effluents and molasses was used for silver nanoparticles (Ag‐NPs) synthesis. The biosurfactant production of B. subtilis T‐1 was confirmed by the detection of genes in the genome and by the identification of the product in the supernatants. The genes for synthesis of surfactin (sfp, srfAA) and iturin (ituC) were noted by PCR reactions. Also, in examined culture supernatants the presence of C₁₃, C₁₄ and C₁₅ surfactin homologues with the sodiated molecules [M + Na]⁺ at m /z 1030, 1044 and 1058 was confirmed using LC/MS/MS analysis. The formation of NPs in the culture supernatants was confirmed by UV–vis spectroscopy. The dynamic light scattering measurements and transmission electron microscopy images showed the nanometric sizes of the biosynthesised Ag‐NPs which ranged from several nm to several tens of nm depending on the used culture supernatant. Biological properties of Ag‐NPs were evaluated by binding of Ag‐NPs with DNA isolated from the Escherichia coli ATCC 25922 and B. subtilis ATCC 6633. Biogenic Ag‐NPs were actively bound to DNA in increased concentration which could be the one important mode of antibacterial action of the Ag‐NPs.Inspec keywords: silver, nanoparticles, nanofabrication, materials preparation, microorganisms, antibacterial activity, industrial waste, agrochemicals, surfactants, breweries, genomics, genetics, chromatography, mass spectroscopic chemical analysis, ultraviolet spectroscopy, visible spectroscopy, spectrochemical analysis, light scattering, transmission electron microscopy, DNA, bonds (chemical), biochemistry, molecular biophysics, nanobiotechnology, biological techniques, particle size, enzymesOther keywords: silver nanoparticle synthesis, Bacillus subtilis T‐1 growth, agro‐industrial waste, biosurfactant production, brewery effluent, molasses, Ag‐NP synthesis, B. subtilis T‐1, gene detection, genome, supernatant product identification, surfactin synthesis, sfp, srfAA, iturin synthesis, ituC, PCR reaction, C₁₃ surfactin homologue, C₁₄ surfactin homologue, C₁₅ surfactin homologue, sodiated molecules, LC‐MS‐MS analysis, UV‐vis spectroscopy, dynamic light scattering measurement, transmission electron microscopy image, Ag‐NP nanometric size range, Ag‐NP biosynthesis, used culture supernatant dependence, biological properties, DNA isolation, Escherichia coli ATCC 25922, B. subtilis ATCC 6633, biogenic Ag‐NP‐DNA binding, Ag‐NP antibacterial action, Ag