Molecular Insights into an Antibiotic Enhancer Action of New Morpholine-Containing 5-Arylideneimidazolones in the Fight against MDR Bacteria

In the search for an effective strategy to overcome antimicrobial resistance, a series of new morpholine-containing 5-arylideneimidazolones differing within either the amine moiety or at position five of imidazolones was explored as potential antibiotic adjuvants against Gram-positive and Gram-negative bacteria. Compounds (7–23) were tested for oxacillin adjuvant properties in the Methicillin-susceptible S. aureus (MSSA) strain ATCC 25923 and Methicillin-resistant S. aureus MRSA 19449. Compounds 14–16 were tested additionally in combination with various antibiotics. Molecular modelling was performed to assess potential mechanism of action. Microdilution and real-time efflux (RTE) assays were carried out in strains of K. aerogenes to determine the potential of compounds 7–23 to block the multidrug efflux pump AcrAB-TolC. Drug-like properties were determined experimentally. Two compounds (10, 15) containing non-condensed aromatic rings, significantly reduced oxacillin MICs in MRSA 19449, while 15 additionally enhanced the effectiveness of ampicillin. Results of molecular modelling confirmed the interaction with the allosteric site of PBP2a as a probable MDR-reversing mechanism. In RTE, the compounds inhibited AcrAB-TolC even to 90% (19). The 4-phenylbenzylidene derivative (15) demonstrated significant MDR-reversal “dual action” for β-lactam antibiotics in MRSA and inhibited AcrAB-TolC in K. aerogenes. 15 displayed also satisfied solubility and safety towards CYP3A4 in vitro.     

Amorphous/crystalline Fe55Ni20Cu5P10Si5B5 composite produced by two-component melt–spinning

ABSTRACT

This work presents unique microstructure and thermal properties of the Fe₅₅Ni₂₀Cu₅P₁₀Si₅B₅ alloy produced by novel modification of the melt-spinning method, where two precursor alloys are simultaneously ejected i.e. Fe₄₀Ni₄₀P₁₀Si₅B₅ and Fe₇₀Cu₁₀P₁₀Si₅B₅, forming a composite ribbon. The investigation of melting the precursors by differential scanning calorimetry confirms the liquid miscibility of the Fe₇₀Cu₁₀P₁₀Si₅B₅ alloy. The two-component melt-spun composite obtained from the two alloys is compared to the alloys produced from a homogeneous liquid. The electron microscopy, X-ray diffraction and differential scanning calorimetry show different microstructure of the composite in comparison with the traditionally melt-spun alloys and our results reveal that the composite alloy inherits the thermal properties of the precursors.

This paper is part of a Thematic Issue on The Crystallographic Aspects of Metallic Alloys.     

Phytochemical Molecules from the Decarboxylation of Gomphrenins in Violet Gomphrena globosa L.—Floral Infusions from Functional Food

Herein, the generation of decarboxylated derivatives of gomphrenin pigments exhibiting potential health-promoting properties and the kinetics of their extraction during tea brewing from the purple flowers of Gomphrena globosa L. in aqueous and aqueous citric acid solutions were investigated. Time-dependent concentration monitoring of natural gomphrenins and their tentative identification was carried out by LC-DAD-ESI-MS/MS. The high content of acylated gomphrenins and their principal decarboxylation products, 2-, 15-, 17-decarboxy-gomphrenins, along with minor levels of their bidecarboxylated derivatives, were reported in the infusions. The identification was supported by the determination of molecular formulas of the extracted pigments by liquid chromatography coupled with high-resolution mass spectrometry (LCMS-IT-TOF). The influence of plant matrix on gomphrenins’ stability and generation of their derivatives, including the extraction kinetics, was determined by studying the concentration profiles in the primary and diluted infusions. Isolated and purified acylated gomphrenins from the same plant material were used for the preliminary determination of their decarboxylated derivatives. The acylated gomphrenins were found to be more stable than nonacylated ones. Citric acid addition had a degradative influence on natural gomphrenins mainly during the longer tea brewing process (above 15 min); however, the presence of plant matrix significantly increased the stability for betacyanins’ identification.     

Sour beer production: impact of pitching sequence of yeast and lactic acid bacteria

Lactic acid bacteria have long been used in brewing to acidify mash or wort and in the production of traditional sour beer styles (Lambic, Gueuze) using spontaneous fermentation. This approach is time consuming (some sour beers are matured for three years to obtain the appropriate flavour), so many brewers choose a faster approach using mixed cultures (yeast and bacteria). In this study, the influence of the pitching sequence of bacteria and yeast on the fermentation process was evaluated (by rate of fermentation and acidification, attenuation degree and ethanol concentration). The trials were performed using three procedures: (i) the addition of bacteria followed by the yeast; (ii) the addition of the yeast followed by the bacteria and (iii) the simultaneous addition of yeast and bacteria. In each trial the following time intervals were applied: 24, 48 and 72 h. Weight loss of samples and drop in pH were measured daily, and ethanol content, real extract and lactic acid concentration measured at the end of the process. It was shown that the order of addition of yeast and bacteria determines the success of the lactic acid fermentation. The appropriate yield of lactic acid (ca. 6 g/L) and the drop in pH (<3.6) can be achieved by inoculating the bacteria prior to the inoculation of yeast. When yeast was pitched before the bacteria, the lactic acid content was ca. 2 g/L and pH was 3.9–4.2. © 2019 The Institute of Brewing & Distilling     

Effects of various clarification treatments on phenolic compounds and color of apple juice

The purpose of this study was to investigate the changes that occur during conventional clarification using gelatin, bentonite, silica sol, and water-soluble chitosan on the phenolic compounds, antioxidant activity, and color of apple juice. The apple material used in this study was of two varieties: Sampion and Idared. The changes in the polyphenols composition (procyanidins, hydroxycinnamic derivatives, and dihydrochalcones) were monitored through the clarification process. Sampion apple control juices contained more total polyphenols than do Idared apple juices. In Sampion variety apple juice, the dominant polyphenols are the flavan-3-ols (86% of total polyphenols), followed by hydroxycinnamic acids (9.7%), dihydrochalcones (3.0%), and flavonols (1.3%). In Idared apple juice the hydroxycinnamic acids (especially chlorogenic acid) are dominant (about 48% of total polyphenols), followed by flavan-3-ols (40%). However, the concentration of polymeric procyanidins in Sampion apple juices was 62.8 and 46.3% less when the Profloc (chitosan) and gelatin treatments were used, respectively. Aktivbentonit and Puranit (bentonite) supplementary added in juices clarification have some protective effect on polymeric procyanidins only with Profloc treatment. That kind of effect was not observed in Idared apple juices with almost eight times smaller polymeric procyanidins concentration than in Sampion apple juices. The antioxidant activity, measured by the DPPH (1,1-diphenyl-2-picrylhydrazyl radical) method, ranged from 0.20 mg TEAC/mL in Idared apple juice to 0.30 mg TEAC/mL in Sampion apple juice, measured by the ABTS method, from 0.17 to 0.48 mg TEAC/mL, respectively. Clarification of apple juices with chosen clarifying agents has statistically no significant (p>0.05) influence on antioxidant capacity. This study suggests that chitosan can be used as a conventional clarifying aid of apple juices and that treatment has no impact on their biochemical parameters.     

Effect of lactic acid fermentation on antioxidant properties and phenolic acid contents of oyster (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis>) and chanterelle (<Emphasis Type="Italic">Cantharellus cibarius</Emphasis>) mushrooms

Fruiting bodies of Pleurotus ostreatus (oyster) and Cantharellus cibarius (chanterelle) mushrooms underwent acid fermentation using 3 strains of lactic acid bacteria (LAB) as starter cultures. Polyphenol contents, antioxidant activities, and phenolic acid contents in fresh, blanched, and fermented mushrooms were investigated. Fruiting bodies of oyster mushrooms exhibited higher total phenolic contents than chanterelle mushrooms. Blanching caused a decrease in polyphenol contents and antioxidant activities in both mushroom types. No important differences were observed in total phenolic compound contents (measured using Folin-Ciocalteau reagent) in mushrooms using different LAB strains. Lactobacillus plantarum was the most useful microorganism for lactic acid fermentation of fruiting bodies for reduction of the pH value. The highest concentrations of single phenolic acids: gallic, homogentisic, and ferulic acids were present in mushrooms fermented using L. plantarum.     

Antimicrobial and Cytotoxic Properties of Bisquaternary Ammonium Bromides of Different Spacer Length

A group of cationic gemini surfactants (bisquaternary ammonium bromides) with different spacer chain lengths (8–6–8, 8–7–8, 8–8–8, 8–9–8) was investigated, paying special attention to antimicrobial and the cytotoxic properties as well as their antimicrobial activity during long‐term storage. It was shown that the compounds investigated exhibit excellent antimicrobial activity against Gram‐positive bacteria (Staphylococcus aureus) and Gram‐negative bacteria (Pseudomonas aeruginosa) as well as antifungal properties (Candida albicans). The gemini surfactants tested had the differential level of cytotoxicity against normal lymphocytes. It was shown that the spacer chain length plays an important role in antibacterial activity and influences the cytotoxicity. The gemini surfactants with shorter spacer chain length, that had higher critical micelle concentration, showed generally weaker antibacterial properties, but on the other hand, these exhibited lower level of cytotoxicity. Furthermore, the aqueous solution of gemini surfactants exhibited the same antimicrobial activity even after 3 months.     

The Effects of Vitamin D on the Expression of IL-33 and Its Receptor ST2 in Skin Cells; Potential Implication for Psoriasis

Interleukin 33 (IL-33) belongs to the IL-1 family and is produced constitutively by epithelial and endothelial cells of various organs, such as the skin. It takes part in the maintenance of tissue homeostasis, repair, and immune response, including activation of Th2 lymphocytes. Its involvement in pathogenesis of several inflammatory diseases including psoriasis was also suggested, but this is not fully understood. The aim of the study was to investigate expression of IL-33 and its receptor, ST2, in psoriasis, and the effects of the active form of vitamin D (1,25(OH)₂D₃) on their expression in skin cells. Here we examined mRNA and protein profiles of IL-33 and ST2 in 18 psoriatic patients and healthy volunteers by qPCR and immunostaining techniques. Potential effects of 1,25(OH)₂D₃ and its receptor (VDR) on the expression of IL-33 and ST2 were tested in cultured keratinocytes, melanocytes, fibroblasts, and basal cell carcinoma cells. It was shown that 1,25(OH)₂D₃ effectively stimulated expression of IL-33 and its receptor ST2’s mRNAs in a time-dependent manner, in keratinocytes and to the lesser extends in melanocytes, but not in fibroblasts. Furthermore, the effect of vitamin D on expression of IL-33 and ST2 was VDR-dependent. Finally, we demonstrated that the expression of mRNA for IL-33 was mainly elevated in the psoriatic skin but not in its margin. Interestingly, ST2 mRNA was downregulated in psoriatic lesion compared to both marginal tissue as well as healthy skin. Our data indicated that vitamin D can modulate IL-33 signaling, opening up new perspectives for our understanding of the mechanism of vitamin D action in psoriasis therapy.     

Diagnostic Utility of Genetic and Immunohistochemical H3-3A Mutation Analysis in Giant Cell Tumour of Bone

To validate the reliability and implementation of an objective diagnostic method for giant cell tumour of bone (GCTB). H3-3A gene mutation testing was performed using two different methods, Sanger sequencing and immunohistochemical (IHC) assays. A total of 214 patients, including 120 with GCTB and 94 with other giant cell-rich bone lesions, participated in the study. Sanger sequencing and IHC with anti-histone H3.3 G34W and G34V antibodies were performed on formalin-fixed, paraffin-embedded tissues, which were previously decalcified in EDTA if needed. The sensitivity and specificity of the molecular method was 100% (95% CI: 96.97–100%) and 100% (95% CI: 96.15–100%), respectively. The sensitivity and specificity of IHC was 94.32% (95% CI: 87.24–98.13%) and 100% (95% CI: 93.94–100.0%), respectively. P.G35 mutations were discovered in 2/9 (22.2%) secondary malignant GCTBs and 9/13 (69.2%) GCTB after denosumab treatment. We confirmed in a large series of patients that evaluation of H3-3A mutational status using direct sequencing is a reliable tool for diagnosing GCTB, and it should be incorporated into the diagnostic algorithm. Additionally, we discovered IHC can be used as a screening tool. Proper tissue processing and decalcification are necessary. The presence of the H3-3A mutation did not exclude malignant GCTB. Denosumab did not eradicate the neoplastic cell population of GCTB.