Expanding the Toolbox of R-Selective Amine Transaminases by Identification and Characterization of New Members

Amine transaminases (ATAs) are used to synthesize enantiomerically pure amines, which are building blocks for pharmaceuticals and agrochemicals. R-selective ATAs belong to the fold type IV PLP-dependent enzymes, and different sequence-, structure- and substrate scope-based features have been identified in the past decade. However, our knowledge is still restricted due to the limited number of characterized (R)-ATAs, with additional bias towards fungal origin. We aimed to expand the toolbox of (R)-ATAs and contribute to the understanding of this enzyme subfamily. We identified and characterized four new (R)-ATAs. The ATA from Exophiala sideris contains a motif characteristic for d -ATAs, which was previously believed to be a disqualifying factor for (R)-ATA activity. The crystal structure of the ATA from Shinella is the first from a Gram-negative bacterium. The ATAs from Pseudonocardia acaciae and Tetrasphaera japonica are the first characterized (R)-ATAs with a shortened/missing N-terminal helix. The active-site charges vary significantly between the new and known ATAs, correlating with their diverging substrate scope.     

Untersuchung des Datenverkehrs aktueller Smart-TVs

Das Internet der Dinge (IoT) bezeichnet die Anbindung von Gegenständen des alltäglichen Gebrauchs an das Internet. Der Fernseher ist als Smart-TV bereits Teil des Internets. Einige Untersuchungen haben hier in jüngster Vergangenheit deutliche Missstände in Bezug auf Datenschutz und Datensicherheit aufgezeigt. Der Beitrag fasst die Ergebnisse einer aktuellen, umfangreichen Untersuchung von fünf Smart-TVs zusammen.     

Toward More Comprehensive Chinese Internet Users’ Studies: Translation and Validation of the Chinese–Mandarin Version of the 8-Item Information Retrieval on the Web Self-Efficacy Scale (Ch-IROWSE)

In this article, we fill a dearth in the literature of Internet self-efficacy (ISS) measurement. We introduce the translation and validation of the Chinese-Mandarin version of the 8-item Information Retrieval On the Web Self-Efficacy scale (IROWSE scale), which has been originally developed in the French language (Rodon & Meyer, 2017). After back-forward translation procedure and content clarity, we ran a test of this new version. We checked the intrinsic psychometric properties including reliability, factor structure (CFA), and criterion-related validity regarding skills for retrieving information on the Web, general attitude toward the Web, experience, gender, and age. Assessment of construct validity indicated, as expected, a one-factor model for the eight items (with coefficient alpha equal to 0.90). Its sum-scores were not confounded with skill levels or general attitudes, and it founds its sources in direct experiences. Furthermore, research would take into account the Internet World population more broadly beyond the West with this new version.     

Sheets of branched poly(lactic acid) obtained by one-step reactive extrusion–calendering process: physical aging and fracture behavior

The architectural modifications of a linear poly(D,L-Lactide) acid (PD,L-LA) commercial grade were induced by a one-step reactive extrusion–calendering process using a styrene-glycidyl acrylate copolymer as reactive agent. The melt degradation was counteracted by chain extension and branching reactions, leading to a stabilization of the melt properties and an increase in the molecular weight. For such modified samples [poly(lactic acid) (PLA)-reactive extrusion (REX)], the rate of physical aging at 30 °C was investigated during 1 week in order to simulate industrial storage conditions. Fracture behavior of “de-aged” and “controlled aged” (1 week) samples was investigated using the essential work of fracture (EWF) methodology and the critical tip opening displacement at the crack propagation onset, respectively. These analyses were complemented by digital image correlation analysis and inspection of the fractured surfaces by scanning electronic microscopy. As a result of the architectural modifications, the entanglement network density was increased. Those accounted for a slight decrease in the physical aging rate. Under uniaxial loading, aged reactive extrusion (REX) samples exhibited multiple crazing, leading to a slight increase in strain at break. Nevertheless, as a result of a similar dynamic environment of the entangled polymer coils, de-aged REX samples disclosed similar mechanical properties as compared to their neat counterparts. Regarding de-aged samples, the EWF analysis revealed no changes in the work required for the onset of crack propagation. However, the energy consumed up to the onset of crack propagation of aged PLA-REX samples decreased due to an apparently decreased network extensibility, promoting a premature craze–crack transition.     

PhosphoTyrosyl Phosphatase Activator of Plasmodium falciparum: Identification of Its Residues Involved in Binding to and Activation of PP2A

In Plasmodium falciparum (Pf), the causative agent of the deadliest form of malaria, a tight regulation of phosphatase activity is crucial for the development of the parasite. In this study, we have identified and characterized PfPTPA homologous to PhosphoTyrosyl Phosphatase Activator, an activator of protein phosphatase 2A which is a major phosphatase involved in many biological processes in eukaryotic cells. The PfPTPA sequence analysis revealed that five out of six amino acids involved in interaction with PP2A in human are conserved in P. falciparum. Localization studies showed that PfPTPA and PfPP2A are present in the same compartment of blood stage parasites, suggesting a possible interaction of both proteins. In vitro binding and functional studies revealed that PfPTPA binds to and activates PP2A. Mutation studies showed that three residues (V²⁸³, G²⁹² and M²⁹⁶) of PfPTPA are indispensable for the interaction and that the G²⁹² residue is essential for its activity. In P. falciparum, genetic studies suggested the essentiality of PfPTPA for the completion of intraerythrocytic parasite lifecycle. Using Xenopus oocytes, we showed that PfPTPA blocked the G2/M transition. Taken together, our data suggest that PfPTPA could play a role in the regulation of the P. falciparum cell cycle through its PfPP2A regulatory activity.     

Immobilization of aloin encapsulated into liposomes in Layer-by-layer films for transdermal drug delivery

Layer-by-layer (LbL) films have been exploited in drug delivery systems that may be used in the form of patches, but the encapsulation of poor water soluble drugs and their release with a controlled rate are still major challenges to be faced. In this paper, we demonstrate the controlled release of aloin (barbaloin), an important component of the widely used Aloe vera, encapsulated into liposomes and immobilized in LbL films with a polyelectrolyte. With a systematic study using fluorescence spectroscopy of aloin release from solutions and from LbL films with different phospholipid liposomes, we inferred that optimized release was achieved with aloin incorporated into palmitoyl oleyl phosphatidyl glycerol (POPG) or dipalmitoyl phosphatidyl glycerol (DPPG) liposomes immobilized in LbL films. Significantly, with this optimized system aloin was almost completely released within 30 h, with a small release rate at the end, which followed a sharp release in the first 5 h. Upon comparing the rates of the distinct systems, we conclude that the main factors controlling the release are the electrostatic interactions involving the negatively charged phospholipids. Because these interactions can be tuned in LbL films, the approach used here opens the way for new drug delivery systems to be developed with fine control of the drug release.     

Determination of alcohols and volatile organic acids in anaerobic bioreactors for H2 production by near infrared spectroscopy

In recent years, near infrared (NIR) spectroscopy has been investigated as a tool for monitoring anaerobic digesters, but several adversities in its application have been reported. This study proposes the application of NIR for the determination of alcohols and volatile organic acids from H₂ production bioreactors and evaluates different approaches to optimize the prediction models. Partial least squad (PLS) models were developed using samples from anaerobic batch reactors fed with crude glycerol for wastewater treatment. The analytes predicted were: methanol, ethanol, 1-butanol, acetic, propanoic, butyric, isocaproic and total volatile organic acids (VFA). The optimization of the predictive capacity of the models was achieved through the orthogonal signal correction (OSC) preprocessing and the selection of variables performed by the genetic algorithm (GA). The application of the proposed models were based on the following figures of merit: accuracy, precision, linearity, limits of detection and quantitation, measurement interval, sensitivity, selectivity, signal-to-noise ratio and bias. Despite the low selectivity (maximum of 0.12%), the models presented high sensitivity [γ^?1 = 0.19 (mg L^?1)^?1], low LOQ (1 mg L^?1) and correlation between reference and predicted values (r) at least 0.93, except for propanoic acid (r_pred = 0.85). The F-test revealed that the selection of variables by GA significantly improved the accuracy and linearity of the prediction models for methanol, acetic acid, isocaproic acid and VFA. NIR spectroscopy has proved to be a powerful tool for monitoring H₂ production bioreactors since provides fast, low cost and multicomponent information.     

A new enzymatically produced 1-lactulose: A pilot study to test the bifidogenic effects

In a novel pilot plant scale process, lactulose was synthesized enzymatically, operating a maximum batch volume of 170 L. The test product was subjected to a randomized, double-blind, placebo-controlled pilot feeding study. Participants consumed a milk serum drink for 14 days, equalling a daily intake of about 3 g d⁻¹ galactosyl-fructoses. Additionally, the effect of industrially produced lactulose and a placebo drink without lactulose was determined. So far, clinical studies on the prebiotic action of lactulose only focussed on the effects of industrially generated lactulose. The intake of enzymatically synthesized lactulose significantly increased (p = 0.001) faecal bifidobacterial counts, while there was no difference with industrially produced lactulose. Analysis of the compounds showed that the drink with enzymatically synthesized lactulose contained an additional disaccharide, β-1,1-galactosyl-fructose. Presumably, this sugar has a bifidogenic effect and may represent a novel prebiotic.     

Maternal Flaxseed Oil During Lactation Enhances Bone Development in Male Rat Pups

Flaxseed oil is an alpha linolenic acid source important in the growth and body development stage; furthermore, this acid acts on adipose tissue and bone health. The aim of this study was to evaluate body composition, fatty acid composition, hormone profile, retroperitoneal adipocyte area and femur structure of pups at weaning, whose mothers were fed a diet containing flaxseed oil during lactation. After birth, pups were randomly assigned: control (C, n = 12) and flaxseed oil (FO, n = 12), rats whose mothers were treated with diet containing soybean or flaxseed oil. At 21 days, the pups were weaned and body mass, length, body composition, biochemical parameter, leptin, osteoprotegerin, osteocalcin, fatty acids composition, intra‐abdominal fat mass and femur structure were analyzed. FO showed (p < 0.05): higher body mass (+12 %) and length (+9 %); body fat mass (g, +45 %); bone mineral density (+8 %), bone mineral content (+55 %) and bone area (+35 %), osteocalcin (+173 %) and osteoprotegerin (+183 %). Arachidonic acid was lower (p < 0.0001), alpha‐linolenic and eicosapentaenoic were higher (p < 0.0001). Intra‐abdominal fat mass was higher (+25 %), however, the retroperitoneal adipocytes area was lower (?44 %). Femur mass (+10 %), distance between epiphyses (+4 %) and bone mineral density (+13 %) were higher. The study demonstrates that adequate flaxseed oil content during a lactation diet plays an important role in the development of pups.