Enzymatic Synthesis of Poly(alkylene succinate)s: Influence of Reaction Conditions

Application of lipases (preferentially Candida antarctica Lipase B, CALB) for melt polycondensation of aliphatic polyesters by transesterification of activated dicarboxylic acids with diols allows to displace toxic metal and metal oxide catalysts. Immobilization of the enzyme enhances the activity and the temperature range of use. The possibility to use enzyme-catalyzed polycondensation in melt is studied and compared to results of polycondensations in solution. The experiments show that CALB successfully catalyzes polycondensation of both, divinyladipate and dimethylsuccinate, respectively, with 1,4-butanediol. NMR spectroscopy, relative molar masses obtained by size exclusion chromatography, MALDI-TOF MS and wide-angle X-ray scattering are employed to compare the influence of synthesis conditions for poly(butylene adipate) (PBA) and poly(butylene succinate) (PBS). It is shown that the enzymatic activity of immobilized CALB deviates and influences the molar mass. CALB-catalyzed polycondensation of PBA in solution for 24 h at 70 °C achieves molar masses of up to Mw~60,000 g/mol, higher than reported previously and comparable to conventional PBA, while melt polycondensation resulted in a moderate decrease of molar mass to Mw~31,000. Enzymatically catalyzed melt polycondensation of PBS yields Mw~23,400 g/mol vs. Mw~40,000 g/mol with titanium(IV)n-butoxide. Melt polycondensation with enzyme catalysis allows to reduce the reaction time from days to 3–4 h

Automated analysis of acetaminophen toxicity on 3D HepaRG cell culture in microbioreactor

Real-time monitoring of bioanalytes in organotypic cell cultivation devices is a major research challenge in establishing stand-alone diagnostic systems. Presently, no general technical facility is available that offers a plug-in system for bioanalytics in diversely available organotypic culture models. Therefore, each analytical device has to be tuned according to the microfluidic and interface environment of the 3D in vitro system. Herein, we report the design and function of a 3D automated culture and analysis device (3D-ACAD) which actively perfuses a custom-made 3D microbioreactor, samples the culture medium and simultaneously performs capillary-based flow ELISA. A microstructured MatriGrid ® has been explored as a 3D scaffold for culturing HepaRG cells, with albumin investigated as a bioanalytical marker using flow ELISA. We investigated the effect of acetaminophen (APAP) on the albumin secretion of HepaRG cells over 96 h and compared this with the albumin secretion of 2D monolayer HepaRG cultures. Automated on-line monitoring of albumin secretion in the 3D in vitro mode revealed that the application of hepatotoxic drug-like APAP results in decreased albumin secretion. Furthermore, a higher sensitivity of the HepaRG cell culture in the automated 3D-ACAD system to APAP was observed compared to HepaRG cells cultivated as a monolayer. The results support the use of the 3D-ACAD model as a stand-alone device, working in real time and capable of analyzing the condition of the cell culture by measuring a functional analyte. Information obtained from our system is compared with conventional cell culture and plate ELISA, the results of which are presented herein

Mycobacterium microti: not just a coincidental pathogen for cats

Public interest in animal tuberculosis is mainly focused on prevention and eradication of bovine tuberculosis in cattle and wildlife. In cattle, immunodiagnostic tests such as the tuberculin skin test or the interferon gamma (IFN-γ) assay have been established and are commercially available. Feline tuberculosis is rather unknown, and the available diagnostic tools are limited. However, infections with Mycobacterium tuberculosis complex members need to be considered an aetiological differential diagnosis in cats with granulomatous lymphadenopathy or skin nodules and, due to the zoonotic potential, a time-efficient and accurate diagnostic approach is required. The present study describes 11 independent cases of Mycobacterium microti infection in domestic cats in Switzerland. For three cases, clinical presentation, diagnostic imaging, bacteriological results, immunodiagnostic testing, and pathological features are reported. An adapted feline IFN-γ release assay was successfully applied in two cases and appears to be a promising tool for the ante mortem diagnosis of tuberculosis in cats. Direct contact with M. microti reservoir hosts was suspected to be the origin of infection in all three cases. However, there was no evidence of M. microti infection in 346 trapped wild mice from a presumptive endemic region. Therefore, the source and modalities of infection in cats in Switzerland remain to be further elucidated

Detecting Weak Spectral Lines in Interferometric Data through Matched Filtering

Modern radio interferometers enable observations of spectral lines with unprecedented spatial resolution and sensitivity. In spite of these technical advances, many lines of interest are still at best weakly detected and therefore necessitate detection and analysis techniques specialized for the low signal-to-noise ratio (S/N) regime. Matched filters can leverage knowledge of the source structure and kinematics to increase sensitivity of spectral line observations. Application of the filter in the native Fourier domain improves S/N while simultaneously avoiding the computational cost and ambiguities associated with imaging, making matched filtering a fast and robust method for weak spectral line detection. We demonstrate how an approximate matched filter can be constructed from a previously observed line or from a model of the source, and we show how this filter can be used to robustly infer a detection significance for weak spectral lines. When applied to ALMA Cycle 2 observations of CH3OH in the protoplanetary disk around TW Hya, the technique yields a ≈53% S/N boost over aperture-based spectral extraction methods, and we show that an even higher boost will be achieved for observations at higher spatial resolution. A Python-based open-source implementation of this technique is available under the MIT license at http://github.com/AstroChem/VISIBLE

Enzymatic Synthesis of Poly(alkylene succinate)s: Influence of Reaction Conditions

Application of lipases (preferentially Candida antarctica Lipase B, CALB) for melt polycondensation of aliphatic polyesters by transesterification of activated dicarboxylic acids with diols allows to displace toxic metal and metal oxide catalysts. Immobilization of the enzyme enhances the activity and the temperature range of use. The possibility to use enzyme-catalyzed polycondensation in melt is studied and compared to results of polycondensations in solution. The experiments show that CALB successfully catalyzes polycondensation of both, divinyladipate and dimethylsuccinate, respectively, with 1,4-butanediol. NMR spectroscopy, relative molar masses obtained by size exclusion chromatography, MALDI-TOF MS and wide-angle X-ray scattering are employed to compare the influence of synthesis conditions for poly(butylene adipate) (PBA) and poly(butylene succinate) (PBS). It is shown that the enzymatic activity of immobilized CALB deviates and influences the molar mass. CALB-catalyzed polycondensation of PBA in solution for 24 h at 70 °C achieves molar masses of up to Mw~60,000 g/mol, higher than reported previously and comparable to conventional PBA, while melt polycondensation resulted in a moderate decrease of molar mass to Mw~31,000. Enzymatically catalyzed melt polycondensation of PBS yields Mw~23,400 g/mol vs. Mw~40,000 g/mol with titanium(IV)n-butoxide. Melt polycondensation with enzyme catalysis allows to reduce the reaction time from days to 3–4 h

Formation of Oligomeric and Macrocyclic Ureas Based on 2,6-Diaminopyridine

The conversion of 1,3-bis-(6-amino-pyridin-2-yl)-urea (<b>1</b>) with <i>N</i>,<i>N</i>′-carbonyldiimidazole at high temperatures in DMSO yielded a mixture of defined cyclic trimers and tetramers. On the basis of model reactions, exchange reactions were evidenced, which convert the cyclic tetramer into a stable cyclic trimer. Linear even numbered oligomers were obtained in acetone under reflux where side reactions were suppressed. The pronounced tendency of cyclization is attributed to a preferred folded conformation of the urea bond between two pyridyl units

Spatiotemporal Photopatterning on Polycarbonate Surface through Visible Light Responsive Polymer Bound DASA Compounds

Besides interesting applications in drug delivery, photoresponsive molecules have great potential to serve as an efficient basis for postfunctionalization photopatterning of polymer surfaces. To the best of our knowledge, only UV light sources have been exploited as a photoinducer for creating patterned templates with or without hydrogels. In this work, we present a practically facile method for grafting visible light responsive donor–acceptor stenhouse adducts (DASAs) on amino-functionalized polycarbonate surfaces. DASA grafted surfaces have shown excellent lithographic performance using visible light. The functionalized surfaces exhibit significant changes of their physical properties after being illuminated with visible light. By using suitable masks, well-defined patterns can be replicated with high precision and resolution. Since the DASA ligand synthesis and surface functionalization is not cumbersome, this method may serve as a facile protocol for obtaining photopatterned polymer surfaces for various applications

Semifluorinated PMMA Block Copolymers: Synthesis, Nanostructure, and Thin Film Properties

Diblock copolymers (BCP) with poly(methyl methacrylate) and poly(1H,1H,2H,2H-perfluorodecyl methacrylate) (PsfMA) blocks prepared by anionic polymerization in tetrahydrofuran at −78 °C and atom transfer radical polymerization (ATRP) at 60 °C, respectively, with stepwise varied composition over a wide range in the phase diagram are compared with respect to synthesis limits, phase separation behavior in bulk, and properties of thin films. Both methods yield BCPs with low dispersity (1.1–1.2) at molar masses below 100 kg mol$^{−1}$. Higher semifluorinated contents can be achieved by ATRP in 1,3-bis(trifluoromethyl)benzene which ensured solubility of PsfMA. BCPs obtained by anionic polymerization show a more distinct phase separation, that is, more regular nanostructures. Additionally, self-organization of the semifluorinated side chains occurs generating smectic layers which alters in turn the BCP morphology especially in thin films as compared to non-semifluorinated BCP. All BCPs show amphiphilic behavior and form micelles in organic solvents which can be used to deposit nanoparticles