Potential of the enzyme laccase for the synthesis and derivatization of antimicrobial compounds

Laccases [E.C. 1.10.3.2, benzenediol:dioxygen oxidoreductase] can oxidize phenolic substances, e.g. di- and polyphenols, hydroxylated biaryls, aminophenols or aryldiamines. This large substrate spectrum is the basis for various reaction possibilities, which include depolymerization and polymerization reactions, but also the coupling of different substance classes. To catalyze these reactions, laccases demand only atmospheric oxygen and no depletive cofactors. The utilization of mild and environmentally friendly reaction conditions such as room temperature, atmospheric pressure, and the avoidance of organic solvents makes the laccase-mediated reaction a valuable tool in green chemistry for the synthesis of biologically active compounds such as antimicrobial substances. In particular, the production of novel antibiotics becomes vital due to the evolution of antibiotic resistances amongst bacteria and fungi. Therefore, laccase-mediated homo- and heteromolecular coupling reactions result in derivatized or newly synthesized antibiotics. The coupling or derivatization of biologically active compounds or its basic structures may allow the development of novel pharmaceuticals, as well as the improvement of efficacy or tolerability of an already applied drug. Furthermore, by the laccase-mediated coupling of two different active substances a synergistic effect may be possible. However, the coupling of compounds that have no described efficacy can lead to biologically active substances by means of laccase. The review summarizes laccase-mediated reactions for the synthesis of antimicrobial compounds valuable for medical purposes. In particular, reactions with two different reaction partners were shown in detail. In addition, studies with in vitro and in vivo experimental data for the confirmation of the antibacterial and/or antifungal efficacy of the products, synthesized with laccase, were of special interest. Analyses of the structure–activity relationship confirm the great potential of the novel compounds. These substances may represent not only a value for pharmaceutical and chemical industry, but also for other industries due to a possible functionalization of surfaces such as wood or textiles

Inactivation of airborne bacteria by plasma treatment and ionic wind for indoor air cleaning

Airborne bacteria are a general problem in medical or health care facilities with a high risk for nosocomial infections. Rooms with a continuous airflow, such as operation theaters, are of particular importance due to a possible dissemination and circulation of pathogens including multidrug-resistant microorganisms. In this regard, a cold atmospheric-pressure plasma (CAP) may be a possibility to support usual disinfection procedures due to its decontaminating properties. The aim of this study was to determine the antimicrobial efficacy of a plasma decontamination module that included a dielectric barrier discharge for plasma generation. Experimental parameters such as an airflow velocity of 4.5 m/s and microbial contaminations of approximately 6,000 colony-forming units (cfu)/m3 were used to simulate practical conditions of a ventilation system in an operating theater. The apathogenic microorganism Escherichia coli K12 DSM 11250/NCTC 10538 and the multidrug-resistant strains E. coli 21181 and 21182 (isolated from patients) were tested to determine the antimicrobial efficacy. In summary, the number of cfu was reduced by 31–89% for the tested E. coli strains, whereby E. coli K12 was the most susceptible strain toward inactivation by the designed plasma module. A possible correlation between the number or kind of resistances and susceptibility against plasma was discussed. The inactivation of microorganisms was affected by plasma intensity and size of the plasma treatment area. In addition, the differences of the antimicrobial efficacies caused through the nebulization of microorganisms in front (upstream) or behind (downstream) the plasma source were compared. The presence of ionic wind had no influence on the reduction of the number of cfu for E. coli K12, as the airflow velocity was too high for a successful precipitation, which would be a prerequisite for an increased antimicrobial efficacy. The inactivation of the tested microorganisms confirms the potential of CAP for the improvement of air quality. The scale-up of this model system may provide a novel tool for an effective air cleaning process

Enhanced laccase-mediated transformation of diclofenac and flufenamic acid in the presence of bisphenol A and testing of an enzymatic membrane reactor

The inadequate removal of pharmaceuticals and other micropollutants in municipal wastewater treatment plants, as evidenced by their detection of these substances in the aquatic environment has led to the need for sustainable remediation strategies. Laccases possess a number of advantages including a broad substrate spectrum. To identify promoting or inhibitory effects of reaction partners in the remediation processes we tested not only single compounds-as has been described in most studies-but also mixtures of pollutants. The reaction of diclofenac (DCF) and flufenamic acid (FA), mediated by Trametes versicolor laccase resulted in the formation of products, which were more hydrophilic than the respective reactant (reactant concentration of 0.1 mM; laccase activity 0.5 U/ml). Analyses (HPLC, LC/MS) showed that the product 1a and 1b for DCF and FA, respectively, to be a para-benzoquinone imine derivative. The formation of 1a was enhanced by the addition of bisphenol A (BPA). After 6 days 97% more product was formed in the mixture of DCF and BPA compared with DCF tested alone. Product 1a was also detected in experiments with micropollutant-supplemented secondary effluent. Within 24 h 67% and 100% of DCF and BPA were transformed, respectively (25 U/ml). Experiments with a membrane reactor (volume 10 l; phosphate buffer, pH 7) were in good agreement with the results of the laboratory scale experiments (50 ml). EC50-values were also determined. The data support the use of laccases for the removal or detoxification of recalcitrant pollutants. Thus, the enzyme laccase may be a component of an additional environmentally friendly process for the treatment stage of wastewater remediation

Concept for improved handling ensures effective contactless plasma treatment of patients with kINPen® MED

The nursing of patients with wounds is an essential part of medical healthcare. In this context, cold atmospheric-pressure plasma sources can be applied for skin decontamination and stimulation of wound healing. One of these plasma devices is the commercially available kINPen® MED (neoplas tools GmbH), a cold atmospheric-pressure plasma jet which is approved as a medical device, class-IIa. For the plasma treatment, a sterile disposable spacer is recommended to ensure a constant and effective distance between plasma and skin. The disadvantage of this spacer is its form and size which means that the effective axis/area is not visible for the attending doctor or qualified personnel and consequently it is a more or less intuitive treatment. In addition, the suggested perpendicular treatment is not applicable for the attending specialist due to lack of space or patient/wound positioning. A concept of a sensory unit was developed to measure the treatment distance and to visualize the effective treatment area for different angles. To determine the effective area for the plasma treatment, some exemplary methods were performed. Thus, the antimicrobial (Staphylococcus aureus DSM799/ATCC6538) efficacy, reactive oxygen species (ROS) distribution and (vacuum) ultraviolet ((V)UV) irradiation were determined depending on the treatment angle. Finally, a simplified first approach to visualize the effective treatment area at an optimal distance was designed and constructed to train attending specialists for optimal wound area coverage. © 2020 by the authors

Ring-Closure Mechanisms Mediated by Laccase to Synthesize Phenothiazines, Phenoxazines, and Phenazines

The green and environmentally friendly synthesis of highly valuable organic substances is one possibility for the utilization of laccases (EC 1.10.3.2). As reactants for the herein described syntheses, different o-substituted arylamines or arylthiols and 2,5-dihydroxybenzoic acid and its derivatives were used. In this way, the formation of phenothiazines, phenoxazines, and phenazines was achieved in aqueous solution mediated by the laccase of Pycnoporus cinnabarinus in the presence of oxygen. Two types of phenothiazines (3-hydroxy- and 3-oxo-phenothiazines) formed in one reaction assay were described for the first time. The cyclization reactions yielded C–N, C–S, or C–O bonds. The syntheses were investigated with regard to the substitution pattern of the reaction partners. Differences in C–S and C–N bond formations without cyclization are discussed

Plasma-activation of larger liquid volumes by an inductively-limited discharge for antimicrobial purposes

A new configuration of a discharge chamber and power source for the treatment of up to 1 L of liquid is presented. A leakage transformer, energizing two metal electrodes positioned above the liquid, limits the discharge current inductively by utilizing the weak magnetic coupling between the primary and secondary coils. No additional means to avoid arcing (electric short-circuiting), e.g., dielectric barriers or resistors, are needed. By using this technique, exceeding the breakdown voltage leads to the formation of transient spark discharges, producing non-thermal plasma (NTP). These discharges effected significant changes in the properties of the treated liquids (distilled water, physiological saline solution, and tap water). Considerable concentrations of nitrite and nitrate were detected after the plasma treatment. Furthermore, all tested liquids gained strong antibacterial efficacy which was shown by inactivating suspended Escherichia coli and Staphylococcus aureus. Plasma-treated tap water had the strongest effect, which is shown for the first time. Additionally, the pH-value of tap water did not decrease during the plasma treatment, and its conductivity increased less than for the other tested liquids. © 2019 by the authors

Kräuter für Nutz- und Heimtiere: Ratgeber für die Anwendung ausgewählter Heil- und Gewürzpflanzen

Dieser Ratgeber führt wissenschaftliche Erkenntnisse und traditionelles Hausmittelwissen zusammen, beleuchtet alles Wissenswerte zu über 50 Heilpflanzen und gibt konkrete Anwendungsbeispiele. Ziel ist es, altbewährte Pflanzenanwendungen wieder mehr in die moderne Tierhaltung einzubinden. Der anwenderorientierte Aufbau des Buches ermöglicht es dem Leser, Kenntnisse über die verschiedenen Zubereitungen und Anwendungen von Heilpflanzen zu erwerben und diese in der Praxis einzusetzen. Zubereitung, Aufbewahrung und Anwendung von Kräutern, sowie deren Wirkung und Einsatz bei einzelnen Tierarten werden ausführlich dargestellt. „Die Aufgabe heutiger Wissenschaft ist weniger die Suche nach neuen wirksamen Pflanzen, vielmehr die Überprüfung und Absicherung dieses althergebrachten Wissensschatzes im Lichte moderner Erkenntnisse. Das Autorenteam setzt sich aus jungen engagierten Wissenschaftlern und Tierärzten zusammen. Ihnen ist es ein großes Anliegen, dass die Erkenntnisse der Kräuterheilkunde möglichst vielen Tierhaltern – insbesondere ihren Tieren – von Nutzen sein werden. Es bleibt der Wunsch: die vielen praktischen Anleitungen mögen einen starken Impuls zur Wiederbelebung der Kräuterheilkunde bei Tieren geben.“ Dr. Gerhard Plakol

Effects of orally administered fumonisin B1 (FB1), partially hydrolysed FB1, hydrolysed FB1 and N-(1-deoxy-D-fructos-1-yl) FB1 on the sphingolipid metabolism in rats

Fumonisin B1 (FB1) is a Fusarium mycotoxin frequently occurring in maize-based food and feed. Alkaline processing like nixtamalisation of maize generates partially and fully hydrolysed FB1 (pHFB1 and HFB1) and thermal treatment in the presence of reducing sugars leads to formation of N-(1-deoxy-D-fructos- 1-yl) fumonisin B1 (NDF). The toxicity of these metabolites, in particular their effect on the sphingolipid metabolism, is either unknown or discussed controversially.We produced high purity FB1, pHFB1a+b, HFB1 and NDF and fed them to male Sprague Dawley rats for three weeks. Once a week, urine and faeces samples were collected over 24 h and analysed for fumonisin metabolites as well as for the sphinganine (Sa) to sphingosine (So) ratio by validated LC–MS/MS based methods. While the latter was significantly increased in the FB1 positive control group, the Sa/So ratios of the partially and fully hydrolysed fumonisins were indifferent from the negative control group. Although NDF was partly cleaved during digestion, the liberated amounts of FB1 did not raise the Sa/So ratio. These results show that the investigated alkaline and thermal processing products of FB1 were, at the tested concentrations, non-toxic for rats, and suggest that according food processing can reduce fumonisin toxicity for humans

The opposing homeobox genes Goosecoid and Vent1/2 self-regulate Xenopus patterning

We present a loss-of-function study using antisense morpholino (MO) reagents for the organizer-specific gene Goosecoid (Gsc) and the ventral genes Vent1 and Vent2. Unlike in the mouse Gsc is required in Xenopus for mesodermal patterning during gastrulation, causing phenotypes ranging from reduction of head structures—including cyclopia and holoprosencephaly—to expansion of ventral tissues in MO-injected embryos. The overexpression effects of Gsc mRNA require the expression of the BMP antagonist Chordin, a downstream target of Gsc. Combined Vent1 and Vent2 MOs strongly dorsalized the embryo. Unexpectedly, simultaneous depletion of all three genes led to a rescue of almost normal development in a variety of embryological assays. Thus, the phenotypic effects of depleting Gsc or Vent1/2 are caused by the transcriptional upregulation of their opposing counterparts. A principal function of Gsc and Vent1/2 homeobox genes might be to mediate a self-adjusting mechanism that restores the basic body plan when deviations from the norm occur, rather than generating individual cell types. The results may shed light on the molecular mechanisms of genetic redundancy

A whole-genome shotgun approach for assembling and anchoring the hexaploid bread wheat genome

Citation: Chapman, J. A., Mascher, M., Buluç, A., Barry, K., Georganas, E., Session, A., . . . Rokhsar, D. S. (2015). A whole-genome shotgun approach for assembling and anchoring the hexaploid bread wheat genome. Genome Biology, 16(1). doi:10.1186/s13059-015-0582-8Polyploid species have long been thought to be recalcitrant to whole-genome assembly. By combining high-throughput sequencing, recent developments in parallel computing, and genetic mapping, we derive, de novo, a sequence assembly representing 9.1 Gbp of the highly repetitive 16 Gbp genome of hexaploid wheat, Triticum aestivum, and assign 7.1 Gb of this assembly to chromosomal locations. The genome representation and accuracy of our assembly is comparable or even exceeds that of a chromosome-by-chromosome shotgun assembly. Our assembly and mapping strategy uses only short read sequencing technology and is applicable to any species where it is possible to construct a mapping population. © 2015 Chapman et al. licensee BioMed Central.Additional Authors: Muehlbauer, G. J.;Stein, N.;Rokhsar, D. S