Triggered by the urgent need to tackle the global crisis of multidrug-resistant bacterial infections, in this work, we present a way to overcome chloramphenicol resistance by introducing modifications based on the glycosylation of its hydroxyl groups. The synthesized derivatives demonstrate complete resistance to the action of recombinant chloramphenicol acetyltransferase (CAT) from Escherichia coli and efficacy against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli ESBL, and Pseudomonas aeruginosa ATCC 27853. Glycosylation gives chloramphenicol an additional advantage - the stable glycosidic form is less toxic to human dermal fibroblasts and has significantly better water solubility than non-glycosylated chloramphenicol. Using a specific glycosidase, chloramphenicol can be almost immediately released from the stable prodrug at the site of polybacterial infections.

The invasive pest Cameraria ohridella annually colonizes trees of the genus Aesculus across Europe, causing dark brown damage called mines that gradually cover the leaf surface. This study aimed to compare the VOC profiles emitted by chestnut tree leaves with varying susceptibility to the pest and assess whether these profiles change due to larval feeding. The research involved a susceptible tree (Ae. turbinata) and resistant trees (Ae. glabra and Ae. parviflora). Over three growing seasons (2014, 2015, 2016), leaf damage and the profile of VOCs were analyzed biweekly from May to September. Leaf damage increased progressively in Ae. turbinata during all seasons. The VOC profiles differed both qualitatively and quantitatively among the trees and across years. More VOCs were identified in 2014 and 2015 than in 2016. The susceptible Ae. turbinata exhibited the highest VOC diversity in 2014, aligning with severe leaf damage—profiles of VOCs in Ae. turbinata were different from resistant trees. Statistical analysis revealed that in 2014 and 2015, differences in the profile of VOCs between susceptible and resistant trees were apparent near pest-feeding dates. In 2016, VOCs significantly distinguished the profile of susceptible trees that were present before the appearance of the first generation of the pest.

Direct analysis of aromatic glycosidic precursors in plants has posed an analytical challenge for decades. Traditional techniques, such as SPE-GC/MS, primarily provided information on volatile aglycones released through hydrolysis. However, the application of high-resolution mass spectrometry combined with liquid chromatography has enabled the direct analysis of intact glycosides without the need for derivatization or hydrolysis. Advances in soft ionization methods, such as DART, offer a novel approach to exploring the hidden aromatic potential in grapes without chromatographic separation. In this work, we present a novel and rapid method for screening aromatic glycosidic precursors in white grapes using high-resolution mass spectrometry (Orbitrap) combined with the soft ionization DART method with nitrogen plasma. Optimization of N2-DART ionization parameters, including grid voltage, gas temperature, and Dip-it sampler speed, performed on selected synthetic glycosidic precursors, allowed the establishment of characteristic ionization patterns and evaluation of 15 groups of glycosidic precursors. The results from the profiling analysis using the N2-DART-Orbitrap-MS method are comparable to those obtained by HPLC/Orbitrap-MS method. This new analytical approach, N2-DART-Orbitrap-MS, reduces drastically analysis time by eliminating the need for chromatographic separation when screening glycoside precursors, uses a convenient Dip-it tips for sampling. It also allows for deeper exploration of ionization using nitrogen plasma, applied for the first time in the analysis of glycoside precursors, demonstrating the applicability of this method for the rapid characterization and screening of glycosidically bound aroma compounds in plants.

The chapter is devoted to the specificity of the analysis of volatile food flavor compounds. The specificity of food aroma compounds is described including the odor thresholds problem of flavor compound instability and influence of food matrix on the binding and release of aroma compounds. Distinction between volatiles and aroma compounds is discussed, together with strategies for the sensory guided analysis of key odorants, and food off-flavors and taints. The methods of isolation of aroma compounds are divided into methods used in the extraction of free aroma compounds and methods used for the extraction and analysis of bound aroma compounds. Free aroma compounds’ analysis comprises solvent extraction methods including supercritical fluid extraction, distillation methods with a special emphasis on simultaneous distillation extraction, headspace analysis focused on static headspace, and factors affecting methods’ sensitivity. The largest group of methods discussed is methods based on sorption mechanisms, where SPME is discussed in more detail, as the most frequently used technique in the analysis of volatile flavor compounds. The specificity of aroma research is reflected in the sub-chapter devoted to specific sample preparation for gas chromatography–olfactometry, and in a sub-chapter on sample preparation techniques for electronic noses. The last sub-chapter discusses the approaches in the sample preparation, fractionation, and analysis of glycosidally bound aroma compounds.

The kinetics and mechanism of the oxidation of hydrazine by [Ru-III(pic)(3)] (pic(-) = picolinate anion) were studied spectrophotometrically as a function of hydrazine concentrations at a constant pH of 5.5. The rate of the reaction was followed by monitoring the appearance of the MLCT band of [Ru-II(pic)(3)](-) at 466 nm. The reaction rate was found to be first-order for Ru(III)-complex concentration. The observed rate constant (k(obs)) values increased linearly with increase in the hydrazine concentration. The oxidation reaction was studied at varied temperature (20-40 degrees C). The Delta H-not equal (73 +/- 3 kJ mol(-1 )kJmol(-1)) and the negative Delta S-not equal (34 +/- 7 Jdeg(-1)mol(-1)) values are consistent with the proposed outer-sphere electron transfer process operative in the oxidation of hydrazine with [Ru-III(pic)(3)]. A working mechanism involving the formation of an active diazene (HN = NH) intermediate followed by its rapid two-electron oxidation by [Ru-III(pic)(3)] in a kinetically inconsequential step producing N-2 as sole product is proposed for an overall four-electron oxidation of hydrazine to dinitrogen.

One method of managing used car tires is decomposition by thermochemical conversion methods. By conducting the process at temperatures of 450–750 °C, three fractions are obtained from tires: oil, gas, and solid. The liquid product of the pyrolysis of used car tires is pyrolysis oil, which consists of aromatic, polyaromatic, and aliphatic hydrocarbons. Unconventional building materials were obtained from tire pyrolysis oil and the environmental impact was evaluated. Blocks made from pyrolysis oil showed mechanical strength of up to about 1700 N. No heavy metals or polycyclic aromatic hydrocarbons, which were found in the crude heavy-PO fraction, were detected in the filtrates after incubation of the block obtained from the heavy-PO fraction at 240 °C. The highest inhibition of Sorghum saccharatum shoot (74.4%) and root (57.5%) growth was observed for solid materials from the medium-PO fraction obtained at 240 °C. The most favorable values of the parameters for the process of obtaining blocks based on post-PO were an annealing temperature of 180 °C, time of 20 h, and mass ratio of catalyst to catalyzed oil of 0.045.

It was found that anionic Pd(II) complexes of type [CA](2)[PdCl4] and [CA](2)[Pd2Cl6] (CA = imidazolium or pyridinium cation) are effective catalysts for copper-free Sonogashira coupling in an aqueous medium without additional ligands or co-catalysts. Both types of substrates, containing electron donor or electron acceptor groups, formed the corresponding products in good to excellent yield. The most active complex, [bmpy](2)[Pd2Cl6] (bmpy = 1-butyl-4-methylpyridinium cation), was employed to the hydroarylation of the Sonogashira product in a one-pot process, using microwave radiation, NaBPh4 as a phenyl source and water as a proton donor. In these reactions the presence of electron donor substituents facilitated a higher conversion of diarylacetylenes to triarylethenes than electron acceptor groups. (C) 2022 Elsevier B.V. All rights reserved.

Fallopia baldschuanica (Polygonaceae) is an Asian plant growing wild in parts of Europe and North and Central America as an introduced taxon, in many countries it is considered a potentially invasive species. This article presents the list of 18 volatile organic compounds (VOCs) emitted by the flowers of F. baldchuanica and identified by headspace gas chromatography/mass spectrometry (HS-GC/MS) analyzes, and a list of flower-visiting and pollinating insects that have been observed in the city center of Wrocław (SW Poland). β-ocimene, heptanal, nonanal, α-pinene, 3-thujene, and limonene, were detected as the floral scent’s most important aroma compounds. F. baldschuanica also produces the aphid alarm pheromones, i.e., β-farnesene and limonene, that repels aphids. Additionally, the pollinators of F. baldschuanica were indicated, based on two years of observations in five sites in the urban area. It was found, that the pollinators of this plant with the highest species stability are: Diptera from families Syrphidae (Chrysotoxum bicinctum, Eristalis pertinax, Eupeodes corollae, Episyrphus balteatus, Eristalis tenax, Syrphus ribesii, Eristalis intricaria), Muscidae (Musca domestica), Sarcophagidae (Sarcophaga spp.), Calliphoridae (Lucilia sericata, Lucilia caesar), Hymenoptera from families Vespidae (Vespula vulgaris), and Apidae (Apis sp., Bombus sp.). The key role of VOCs in adaptation to plant expansion is discussed.

Eugenyl-β-D-glucopyranoside, also referred to as Citrusin C, is a natural glucoside found among others in cloves, basil and cinnamon plants. Eugenol in a form of free aglycone is used in perfumeries, flavourings, essential oils and in medicinal products. Synthetic Citrusin C was incubated with human saliva in several in vitro models together with substrate-specific enzyme and antibiotics (clindamycin, ciprofloxacin, amoxicillin trihydrate and potassium clavulanate). Citrusin C was detected using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Citrusin C was completely degraded only when incubated with substrate-specific A. niger glucosidase E.C 3.2.1.21 (control sample) and when incubated with human saliva (tested sample). The addition of antibiotics to the above-described experimental setting, stopped Citrusin C degradation, indicating microbiologic origin of hydrolysis observed. Our results demonstrate that Citrusin C is subjected to complete degradation by salivary/oral cavity microorganisms. Extrapolation of our results allows to state that in the human oral cavity, virtually all β-D-glucosides would follow this type of hydrolysis. Additionally, a new method was developed for an in vivo rapid test of glucosidase activity in the human mouth on the tongue using fluorescein-di-β-D-glucoside as substrate. The results presented in this study serve as a proof of concept for the hypothesis that microbial hydrolysis path of β-D-glucosides begins immediately in the human mouth and releases the aglycone directly into the gastrointestinal tract.

In mammalian cells, 3-hydroxybutyrate (3-HB) is not only an intermediate metabolite during the oxidation of fatty acids, but also an important signaling molecule. On the other hand, the information about the metabolism or function of this compound in plants is scarce. In our study, we show for the first time that this compound naturally occurs in flax. The expression of bacterial β-ketothiolase in flax affects expression of endogenous genes of the 3-HB biosynthesis pathway and the compound content. The increase in 3-HB content in transgenic plants or after control plants treatment with 3-HB resulted in upregulation of genes involved in chromatin remodeling. The observation that 3-HB is an endogenous activator of methyltransferase 3 (CMT3), decreased DNA methylation I (DDM1), DEMETER DNA glycosylase (DME), and an inhibitor of sirtuin 1 (SRT1) provides an example of integration of different genes in chromatin remodeling. The changes in chromatin remodeling gene expression concomitant with those involved in phenolics and the lignin biosynthesis pathway suggest potential integration of secondary metabolic status with epigenetic changes.

Helicobacter pylori is a bacterium known mainly of its ability to cause persistent inflammations of the human stomach, resulting in peptic ulcer diseases and gastric cancers. Continuous exposure of this bacterium to antibiotics has resulted in high detection of multidrug-resistant strains and difficulties in obtaining a therapeutic effect. The purpose of the present study was to determine the usability of bacterial cellulose (BC) chemisorbed with 3-bromopyruvate (3-BP) or sertraline (SER) to act against lawn H. pylori biofilms. The characterization of BC carriers was made using a N2 adsorption/desorption analysis, tensile strength test, and scanning electron microscopy (SEM) observations. Determination of an antimicrobial activity was performed using a modified disk-diffusion method and a self-designed method of testing antibacterial activity against biofilm microbial forms. In addition, bacterial morphology was checked by SEM. It was found that BC disks were characterized by a high cross-linking and shear/stretch resistance. Growth inhibition zones for BC disks chemisorbed with 2 mg of SER or 3-BP were equal to 26.5–27.5 mm and 27–30 mm, respectively. The viability of lawn biofilm H. pylori cells after a 4-h incubation with 2 mg SER or 3-BP chemisorbed on BC disks was ≥4 log lower, suggesting their antibacterial effect. SEM observations showed a number of morphostructural changes in H. pylori cells exposed to these substances. Concluding, SER and 3-BP chemisorbed on BC carriers presented a promising antibacterial activity against biofilm H. pylori cells in in vitro conditions.

Bacterial cellulose is one of the most promising polymers of recent years. Herein, we present a possibility of BC application as a carrier of gentamycin antibiotic for the treatment and prevention of bone infections. We have shown that BC saturated with gentamycin significantly reduces the level of biofilm-forming bone pathogens, namely Staphylococcus aureus and Pseudomonas aeruginosa, and displays very low cytotoxicity in vitro against osteoblast cell cultures. Another beneficial feature of our prototype dressing is prolonged release of gentamycin, which provides efficient protection from microbial contamination and subsequent infection. Moreover, it seems that bacterial cellulose (BC) alone without any antimicrobial added, may serve as a barrier by significantly hampering the ability of the pathogen to penetrate to the bone structure. Therefore, a gentamycin-saturated BC dressing may be considered as a possible alternative for gentamycin collagen sponge broadly used in clinical setting. (c) 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 108B:30-37, 2020.

In this research, bacterial cellulose (BC), one of the most promising biopolymers of the recent years, was saturated with thyme, eucalyptus and clove essential oils (EOs) and applied against staphylococcal and pseudomonal biofilms formed on hydroxyapatite (HA). BC dressings were thoroughly analyzed with regard to their physical properties. Moreover, the exact composition and ability of particular EO molecules to adhere to HA was assessed. Additionally, cytotoxicity of oil-containing, cellulose-based dressings towards osteoblasts and fibroblasts as well as their impact on reactive oxygen species (ROS) production by macrophages was assessed. The results revealed the high ability of BC dressings to absorb and subsequently release EOs from within their microstructure; the highest number of compounds able to adhere to HA was found in the thyme EO. The eucalyptus EO displayed low, while thyme and clove EOs displayed high cytotoxicity towards fibroblast and osteoblast cell lines. The clove EO displayed the highest eradication ability toward staphylococcal, while the thyme EO against pseudomonal biofilm. Taken together, the results obtained indicate the suitability of EO-saturated BC dressings to eradicate pseudomonal and staphylococcal biofilm on HA surface and moreover, to not trigger reactive oxygen species production by immune system effector cells. However, due to cytotoxic effects of thyme and clove EOs towards cell lines in vitro, the eucalyptus EO-saturated BC dressing is of highest potential to be further applied.

Chronic leg ulceration is a disease usually associated with other comorbidities, and significantly reduces patient quality of life. Infected leg ulcers can lead to limb-threatening sequelae or mortality. Leg ulcerations are colonized by a number of microbes that are able to cause life-threating infections in susceptible patients. Wound exudate is a body fluid that collects metabolites from patient eukaryotic cells and from prokaryotic bacterial communities inhabiting the wound. This study aimed at identification of metabolites in exudates collected from chronic leg ulcers, and correlation of this metabolome with patient comorbidities and microbiological status of the wound. By means of NMR spectroscopy we detected 42 metabolites of microbial or patient origin. The metabolites that were in abundance in exudates analyzed were lactate, lysine, and leucine. Metabolites were associated with the presence of neutrophils in wounds and destruction of high quantities of microbes, but also with hypoxia typical for venous insufficiency. The combination of nuclear magnetic resonance spectroscopy technique and partial least squares discriminant analysis allowed us to further discriminate groups of metabolites with regards to potential clinical meaning. For example, to discriminate between S.aureus versus all other isolated microbial species, or between patients suffering from type I or II diabetes versus patients without diabetes. Therefore, wound exudate seems to be highly applicable material for discriminant analysis performed with the use of NMR technique to provide for rapid metabolomics of chronic wound status.

In the present article, we propose a simple Antibiofilm Dressing's Activity Measurement (A.D.A.M.) test that allows to check in vitro a dressing's suitability against biofilm-related wound infections. To perform the test, three agar discs are covered with biofilm formed by the tested pathogen after which they are assembled one over another in the form of an agar plug and placed in the well of a 24-well plate. The top disc is covered with the analyzed dressing and the entire set is incubated for 24h. During this time, the investigated antimicrobial substance is released from the dressing and penetrates to subsequent biofilm-covered agar discs. Biofilm reduction is measured using 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) spectrometric assay and the results are compared to untreated control samples (agar plug covered with biofilm and without the dressing/or with a passive dressing placed on the top disc). Furthermore, in order to standardize the differences in penetrability of the drugs released from active dressings the results can be expressed as a dimensionless value referred to as the Penetrability Index. In summary, A.D.A.M. test is simple, cheap, can be performed practically in every clinical laboratory and takes no more time than routine microbiological diagnostics. Apart from measuring the released drug's activity, the A.D.A.M. test allows to assess drug penetrability (across three agar discs), reflecting real wound conditions, where microbes are frequently hidden under the necrotic tissue or cloth. In conclusion, the A.D.A.M. test produces a high volume of data that, when analyzed, can provide a researcher with a valuable hint concerning the applicability of active dressings against specific biofilm pathogens in a particular setting.

Flax (Linum usitatissimum) is a crop plant valued for its oil and fiber. Unfortunately, large losses in cultivation of this plant are caused by fungal infections, with Fusarium oxysporum being one of its most dangerous pathogens. Among the plant's defense strategies, changes in the expression of genes of the shikimate/phenylpropanoid/benzoate pathway and thus in phenolic contents occur. Among the benzoates, salicylic acid, and its methylated form methyl salicylate play an important role in regulating plants' response to stress conditions. Upon treatment of flax plants with the fungus we found that methyl salicylate content increased (4.8-fold of the control) and the expression profiles of the analyzed genes suggest that it is produced most likely from cinnamic acid, through the β-oxidative route. At the same time activation of some genes involved in lignin and flavonoid biosynthesis was observed. We suggest that increased methyl salicylate biosynthesis during flax response to F. oxysporum infection may be associated with phenylpropanoid pathway activation.

Four synthesized terpenyl-beta-D-glycopyranosides (geranyl, neryl, citronellyl, myrtenyl) were subjected to enzymatic (AR 2000, pH 5.5) and acid (citric buffer, pH 2.5) hydrolysis. Decrease of glycosides was measured by HPLC and the volatiles released - by comprehensive gas chromatography-mass spectrometry (GC x GC-ToF-MS). Enzymatic hydrolysis performed for 21 h yielded 100% degree of hydrolysis for all glycosides but citronellyl (97%). Degree of acid hydrolysis was highly dependent on type of aglycone and the conditions. The highest degree was achieved for geraniol, followed by citronellol and nerol. Myrtenylo-beta-D-glycopyranoside was the most resistant glycoside to hydrolysis. Acid hydrolysis degree was also related to temperature/time combination, the highest being for 100 degrees C and 2 h. In a result of enzymatic hydrolysis 85-91% of total peak areas was terpene aglycone, whereas for acid hydrolysis the area of released terpene aglycone did not exceed 1.3% of total peak area indicating almost complete decomposition/transformation of terpenyl aglycone. (C) 2015 Elsevier Ltd. All rights reserved.

The study aimed at evaluation of various types of alkali rinsing with regard to their efficacy in terms of removal, not only of bacteria but also bacterial metabolites, from cellulose matrices formed by three Komagataeibacter xylinus strains. Moreover, we tested the type of alkali rinsing on membrane cytotoxicity in vitro in fibroblast and osteoblast cells and we compared matrices' ability to induce oxidative stress in macrophages. We identified 11 metabolites of bacterial origin that remained in cellulose after rinsing. Moreover, our results indicated that the type of alkali rinsing should be adjusted to specific K. xylinus strains that are used as cellulose producers to obtain safe biomaterials in the context of low cytotoxicity and macrophage induction. The findings have translational importance and may be of direct significance to cellulose dressing manufacturers.