Methyltransferase Contingencies in the Pathway of Everninomicin D Antibiotics and Analogues

Everninomicins are orthoester oligosaccharide antibiotics with potent activity against multidrug-resistant bacterial pathogens. Everninomicins act by disrupting ribosomal assembly in a distinct region in comparison to clinically prescribed drugs. We employed microporous intergeneric conjugation with Escherichia coli to manipulate Micromonospora for targeted gene-replacement studies of multiple putative methyltransferases across the octasaccharide scaffold of everninomicin effecting the A₁, C, F, and H rings. Analyses of gene-replacement and genetic complementation mutants established the mutability of the everninomicin scaffold through the generation of 12 previously unreported analogues and, together with previous results, permitted assignment of the ten methyltransferases required for everninomicin biosynthesis. The in vitro activity of A₁- and H-ring-modifying methyltransferases demonstrated the ability to catalyze late-stage modification of the scaffold on an A₁-ring phenol and H-ring C-4’ hydroxy moiety. Together these results establish the potential of the everninomicin scaffold for modification through mutagenesis and in vitro modification of advanced biosynthetic intermediates.     

Adenosine Receptor Agonist HE-NECA Enhances Antithrombotic Activities of Cangrelor and Prasugrel in vivo by Decreasing of Fibrinogen Density in Thrombus

Blood platelets’ adenosine receptors (AR) are considered to be a new target for the anti-platelet therapy. This idea is based on in vitro studies which show that signaling mediated by these receptors leads to a decreased platelet response to activating stimuli. In vivo evidence for the antithrombotic activity of AR agonists published to date were limited, however, to the usage of relatively high doses given in bolus. The present study was aimed at verifying if these substances used in lower doses in combination with inhibitors of P2Y₁₂ could serve as components of dual anti-platelet therapy. We have found that a selective A_2A agonist 2-hexynyl-5’-N-ethylcarboxamidoadenosine (HE-NECA) improved the anti-thrombotic properties of either cangrelor or prasugrel in the model of ferric chloride-induced experimental thrombosis in mice. Importantly, HE-NECA was effective not only when applied in bolus as other AR agonists in the up-to-date published studies, but also when given chronically. In vitro thrombus formation under flow conditions revealed that HE-NECA enhanced the ability of P2Y₁₂ inhibitors to decrease fibrinogen content in thrombi, possibly resulting in their lower stability. Adenosine receptor agonists possess a certain hypotensive effect and an ability to increase the blood–brain barrier permeability. Therefore, the effects of anti-thrombotic doses of HE-NECA on blood pressure and the blood–brain barrier permeability in mice were tested. HE-NECA applied in bolus caused a significant hypotension in mice, but the effect was much lower when the substance was given in doses corresponding to that obtained by chronic administration. At the same time, no significant effect of HE-NECA was observed on the blood–brain barrier. We conclude that chronic administration of the A_2A agonist can be considered a potential component of a dual antithrombotic therapy. However, due to the hypotensive effect of the substances, dosage and administration must be elaborated to minimize the side-effects. The total number of animals used in the experiments was 146.     

Origin of Violet‐Blue Emission in Ti‐Doped Gahnite

ZnAl₂O₄ doped with Ti⁴⁺ (2%) was synthesized by the hydrothermal method at 220°C at pressure of 25 bars. An average grain size of the as‐prepared sample was 3 nm, the samples with biggest grain size were obtained after annealing at 300°C, 500°C, 600°C, 700°C, and 900°C, diameter of the latter was about 33 nm. IR spectroscopy indicated that ZnAl₂O₄ was partially inverted. The degree of the inversion decreases with increase in the annealing temperature but increases with increasing Ti⁴⁺ content. Absorption and emission spectra as well as emission decay profiles were recorded at 300 and 77 K. The observed spectra are due to charge‐transfer O^2??Ti⁴⁺ transitions. Color of the emission depends on the nanocrystal size and with increase in its diameter changes from violet to blue, accordingly the absorption bands exhibit redshift. The calculations based on Density Functional Theory confirmed the experimental results. 3d electrons of titanium ions form the bottom of the ZnAl₂O₄:Ti⁴⁺ conduction band, oxygen, aluminum or zinc vacancies create additional levels in the gahnite energy band gap. It was also found that in ZnAl₂O₄ aluminum or zinc vacancy induces magnetism with relatively high magnetic moment close to 1 μ_B per vacancy.     

Synthesis and Surface Active Properties of Novel Carbohydrate-based Cationic Surfactants

The synthesis of new cationic carbohydrate surfactants is presented in this paper. The obtained surfactants have structures that are typical for saponins, which contain fatty amide hydrophobic chains and hydrophilic heads with cationic carbohydrate units. Their surface active properties and biodegradability have been studied. For two types, the biodegradability was above 85% and comparable to standard carbohydrate surfactants.     

New Hormaphididae(Hemiptera,Aphidomorpha)from the Baltic Amber and its Palaeogeographic Significance

The present study describes a new genus and species Unicohormaphis sorini, which belongs to the subfamily Cerataphidinae. The contemporary distribution of aphids belonging to the Hormaphididae family is summarized and analyzed. According to the obtained data, the climate warming in Eocene was conducive to the expansion of the family over the areas of Europe. Subsequent climate changes, resulting from epeirogenic and orogenic movements towards the end of Eocene, led to a considerable cooling and drying of climate in the northern hemisphere. These processes contributed to the shrinking down of areas covered by subtropical forests, which constituted the habitat of Hormaphididae (Cerataphidinae). Retaining their way of life, the Cerataphidinae were forced to limit their distribution, and the representatives of the family in Europe became completely extinct.     

Assessment of cytotoxicity and immune compatibility of phytochemicals‐mediated biosynthesised silver nanoparticles using Cynara scolymus

The study was focused on the phytochemicals‐mediated biosynthesis of silver nanoparticles using leaf extracts and infusions from Cynara scolymus. To identify the antioxidant activity and total phenolic content, the 1,1‐diphenyl‐1‐picrylhydrazyl and Folin–Ciocalteau methods were applied, respectively. The formation and stability of the reduced silver ions were monitored by UV–vis spectrophotometer. The particle sizes of the silver nanoparticles were characterised using the dynamic light scattering technique and scanning electron microscope. The phase composition of the obtained silver nanoparticles was characterised by X‐ray diffraction. The silver nanoparticles suspension, artichoke infusion, and silver ions were separately tested towards potential cytotoxicity and pro‐inflammatory effect using mouse fibroblasts and human monocytes cell line, respectively. The total phenolic content and antioxidant activity of ethanol extract and infusion were found significantly higher as compared to aqueous extract and infusion. The UV–visible spectrophotometric analysis revealed the presence of the characteristic absorption band of the Ag nanoparticles. Moreover, it was found that with the increasing volume of plant extract, the average size of particles was increased. Biocompatibility results evidently showed that silver nanoparticles do not induce monocyte activation, however in order to avoid their cytotoxicity suspension at a concentration <2 ppm should be applied.Inspec keywords: pharmaceuticals, health and safety, renewable materials, toxicology, organic compounds, antibacterial activity, X‐ray diffraction, nanomedicine, nanoparticles, nanofabrication, suspensions, ultraviolet spectra, visible spectra, scanning electron microscopy, silver, particle sizeOther keywords: phytochemicals‐mediated biosynthesis, antioxidant activity, total phenolic content, dynamic light scattering technique, silver nanoparticles suspension, scanning electron microscopy, Cynara scolymus, 1,1 diphenyl‐1‐picrylhydrazyl method, cytotoxicity, immune compatibility, leaf extracts, UV‐vis spectrophotometry, particle size, Folin‐Ciocalteau methods, phase composition, X‐ray diffraction, artichoke infusion, pro‐inflammatory effect, mouse fibroblasts, human monocytes cell line, Ag     

The Effect of Exogenous Application of Quercetin Derivative Solutions on the Course of Physiological and Biochemical Processes in Wheat Seedlings

Quercetin, classified as a flavonoid, is a strong antioxidant that plays a significant role in the regulation of physiological processes in plants, which is particularly important in the case of biotic and abiotic stresses. The study investigated the effect of the use of potassium quercetin solutions in various concentrations (0.5%, 1.0%, 3.0% and 5.0%) on the physiological and biochemical properties of wheat seedlings. A pot experiment was carried out in order to determine the most beneficial dose of this flavonoid acting as a bio-stimulant for wheat plants. Spraying with quercetin derivative solutions was performed twice, and physiological measurements (chlorophyll content and fluorescence as well as gas exchange) were carried out on the first and seventh days after each application. The total phenolic compounds content and the total antioxidant capacity were also determined. It was shown that the concentrations of potassium quercetin applied have a stimulating effect on the course of physiological processes. In the case of most of the tested physiological parameters (chlorophyll content and fluorescence and gas exchange) and the total antioxidant capacity, no significant differences were observed in their increase as a result of application with concentrations of 3.0 and 5.0%. Therefore, the beneficial effect of quercetin on the analysed parameters is already observed when spraying with a concentration of 3.0%.     

Curcumin and Its New Derivatives: Correlation between Cytotoxicity against Breast Cancer Cell Lines,Degradation of PTP1B Phosphatase and ROS Generation

Breast cancer is the most common cancer of women—it affects more than 2 million women worldwide. PTP1B phosphatase can be one of the possible targets for new drugs in breast cancer therapy. In this paper, we present new curcumin derivatives featuring a 4-piperidone ring as PTP1B inhibitors and ROS inducers. We performed cytotoxicity analysis for twelve curcumin derivatives against breast cancer MCF-7 and MDA-MB-231 cell lines and the human keratinocyte HaCaT cell line. Furthermore, because curcumin is a known antioxidant, we assessed antioxidant effects in its derivatives. For the most potent cytotoxic compounds, we determined intracellular ROS and PTP1B phosphatase levels. Moreover, for curcumin and its derivatives, we performed real-time microscopy to observe the photosensitizing effect. Finally, computational analysis was performed for the curcumin derivatives with an inhibitory effect against PTP1B phosphatase to assess the potential binding mode of new inhibitors within the allosteric site of the enzyme. We observed that two tested compounds are better anticancer agents than curcumin. Moreover, we suggest that blocking the -OH group in phenolic compounds causes an increase in the cytotoxicity effect, even at a low concentration. Furthermore, due to this modification, a higher level of ROS is induced, which correlates with a lower level of PTP1B.     

Enzymatic Synthesis of Lignin‐Based Concrete Dispersing Agents

Lignin is the most abundant aromatic biopolymer, functioning as an integral component of woody materials. In its unmodified form it shows limited water solubility and is relatively unreactive, so biotechnological lignin valorisation for high‐performance applications is greatly underexploited. Lignin can be obtained from the pulp and paper industry as a by‐product. To expand its application, a new synthesis route to new dispersing agents for use as concrete additives was developed. The route is based on lignin functionalisation by enzymatic transformation. Screening of lignin‐modifying systems resulted in functionalised lignin polymers with improved solubility in aqueous systems. Through grafting of sulfanilic acid or p‐aminobenzoic acid by fungal laccases, lignin became soluble in water at pH≤4 or pH≤7, respectively. Products were analysed and evaluated in miniaturised application tests in cement paste and mortar. Their dispersing properties match the performance criteria of commercially available lignosulfonates. The study provides examples of new perspectives for the use of lignin.