The adjustment of China endemic Heptacodium miconioides Rehd. to temperate zone of Poland.

Heptacodium miconioides is an increasingly popular ornamental plant, originally being endemic to China. The late and long flowering determines its ecological and ornamental value in cultivation. The aims of this research were to define and distinguish phenological phases of the development of Heptacodium miconioides in the temperate zone region and identification of anatomical changes within the stem during autumn in relation to phenological phases and climatic conditions. Phenological observations were carried out in Wroclaw during 2012-2013, as well as in Warsaw (Poland, 52.6°N, 20.5°E) during 2018-2021. During the last year of research an analysis of the anatomical structure was carried out for young stems that bore flowers that year, as well as older, 2- to 6-year-old ones. The material was collected H1 - 10.09., H2 - 28.09., H3 - 16.10., H4 - 3.11., H5 - 21.11. The width of annual increments in subsequent years was determined; length, width and vessel density in early and latewood for subsequent rings of annual growth was measured, as well as the width of the phloem in 1-6-year-old stems (2016-2021). In the vegetative stage three main stages of development were distinguished (leaf buds have the green tips; full autumn discoloration of leaves; leaves falling). In the generative phase, which lasted on average from August 22nd to January the 7th five main phases of development were distinguished (flowering, unripe fruits, ripe fruits, spreading of seeds). Increased average temperature during winter and spring had an effect on the growth pattern: early phenological stages occurred sooner and foliage development lasted 44 days longer. Flowering occurred at a similar date at both observed locations and climatic conditions. This year's shoots flowering on a radial section with axial symmetry, were slightly flattened and in clusters arranged regularly to match the shape. Heptacodium develops 2-6 years old shoots with radial symmetry. The growth ring boundaries are distinct, the wood semi-rings porous, with marked differences in the structure of the primary and secondary shoot. Lignification of tissues before winter ends during late leaf-fall phase. The research indicated the adaptive potential of Heptacodium in response to climatic conditions of temperate zone.

Overexpression of Bacterial Beta-Ketothiolase Improves Flax (Linum usitatissimum L.) Retting and Changes the Fibre Properties.

Beta-ketothiolases are involved in the beta-oxidation of fatty acids and the metabolism of hormones, benzenoids, and hydroxybutyrate. The expression of bacterial beta-ketothiolase in flax (Linum usitatissimum L.) results in an increase in endogenous beta-ketothiolase mRNA levels and beta-hydroxybutyrate content. In the present work, the effect of overexpression of beta-ketothiolase on retting and stem and fibre composition of flax plants is presented. The content of the components was evaluated by high-performance liquid chromatography, gas chromatography-mass spectrometry, Fourier-transform infrared spectroscopy, and biochemical methods. Changes in the stem cell walls, especially in the lower lignin and pectin content, resulted in more efficient retting. The overexpression of beta-ketothiolase reduced the fatty acid and carotenoid contents in flax and affected the distribution of phenolic compounds between free and cell wall-bound components. The obtained fibres were characterized by a slightly lower content of phenolic compounds and changes in the composition of the cell wall. Based on the IR analysis, we concluded that the production of hydroxybutyrate reduced the cellulose crystallinity and led to the formation of shorter but more flexible cellulose chains, while not changing the content of the cell wall components. We speculate that the changes in chemical composition of the stems and fibres are the result of the regulatory properties of hydroxybutyrate. This provides us with a novel way to influence metabolic composition in agriculturally important crops.

Frankfurter-Type Sausage Enriched with Buckwheat By-Product as a Source of Bioactive Compounds.

Buckwheat by-products may be used as promising food ingredients due to their nutritional composition. Buckwheat husk (BH) may be used in meat products as a source of valuable compounds. In this study, the addition of BH to the quality of frankfurter-type sausages was investigated, aiming to reduce buckwheat waste and to develop nutritionally enriched sausages. For the purpose of this study, a range of measurements, as well as observations, have been carried out. This included the following: pH, weight losses, yield, the instrumental color and texture measurement, protein digestibility, polyphenols, amino acid, trace elements analysis, and the organoleptic evaluation. Compared with no BH sausages, the cooking losses of sausages with 3% BH were higher, while storage losses were lower. BH increased the hardness of sausages after two weeks of storage. The growing addition of BH resulted in a decrease in L* and b*. This change of color resulted in its lower level of consumer acceptability. BH addition did not reduce the protein digestibility. The total amino acid content increased with the increasing husk addition, from 161.8 mg/kg to 228.0 mg/kg. Moreover, BH increased the content of manganese, calcium, potassium and magnesium. This research suggested that incorporation of BH improved the nutritional value of sausages with minimal changes in technological and sensory properties.

The Antimicrobial Properties of Poplar and Aspen-Poplar Propolises and Their Active Components against Selected Microorganisms, including Helicobacter pylori.

There is a noticeable interest in alternative therapies where the outcome is the eradication of the Gram-negative bacterium, Helicobacter pylori (H. pylori), for the purpose of treating many stomach diseases (chronic gastritis and peptic ulcers) and preventing stomach cancer. It is especially urgent because the mentioned pathogen infects over 50% of the world's population. Recent studies have shown the potential of natural products, such as medicinal plant and bee products, on the inhibition of H. pylori growth. Propolis is such a bee product, with known antimicrobial activities. The main scope of the study is the determination of the antimicrobial activity of ethanolic extracts from 11 propolis samples (mostly from Poland, Ukraine, Kazakhstan, and Greece) against H. pylori, as well as selected bacterial and yeast species. The most effective against H. pylori was the propolis from Ukraine, with an MIC = 0.02 mg/mL while the rest of samples (except one) had an MIC = 0.03 mg/mL. Moreover, significant antimicrobial activity against Gram+ bacteria (with an MIC of 0.02-2.50 mg/mL) and three yeasts (with an MIC of 0.04-0.63 mg/mL) was also observed. A phytochemical analysis (polyphenolic profile) of the propolis samples, by ultra-high-performance liquid chromatography-diode array detector-mass spectrometry (UPLC-DAD-MS), was performed. An evaluation of the impact of the propolis components on antimicrobial activity, consisting of statistical analyses (principal component analysis (PCA) and hierarchical fuzzy clustering), was then performed. It was observed that the chemical composition characteristics of the poplar propolis correlated with higher antibacterial activity, while that of the poplar and aspen propolis correlated with weaker antibacterial activity. To summarize the activity in vitro, all tested propolis samples indicate that they can be regarded as useful and potent factors in antimicrobial therapies, especially against H. pylori.

Nanoapatites Doped and Co-Doped with Noble Metal Ions as Modern Antibiofilm Materials for Biomedical Applications against Drug-Resistant Clinical Strains of Enterococcus faecalis VRE and Staphylococcus aureus MRSA.

The main aim of our research was to investigate antiadhesive and antibiofilm properties of nanocrystalline apatites doped and co-doped with noble metal ions (Ag+, Au+, and Pd2+) against selected drug-resistant strains of Enterococcus faecalis and Staphylococcus aureus. The materials with the structure of apatite (hydroxyapatite, nHAp; hydroxy-chlor-apatites, OH-Cl-Ap) containing 1 mol% and 2 mol% of dopants and co-dopants were successfully obtained by the wet chemistry method. The majority of them contained an additional phase of metallic nanoparticles, in particular, AuNPs and PdNPs, which was confirmed by the XRPD, FTIR, UV-Vis, and SEM-EDS techniques. Extensive microbiological tests of the nanoapatites were carried out determining their MIC, MBC value, and FICI. The antiadhesive and antibiofilm properties of the tested nanoapatites were determined in detail with the use of fluorescence microscopy and computer image analysis. The results showed that almost all tested nanoapatites strongly inhibit adhesion and biofilm production of the tested bacterial strains. Biomaterials have not shown any significant cytotoxic effect on fibroblasts and even increased their survival when co-incubated with bacterial biofilms. Performed analyses confirmed that the nanoapatites doped and co-doped with noble metal ions are safe and excellent antiadhesive and antibiofilm biomaterials with potential use in the future in medical sectors.

Composition and Antimicrobial Activity of Ilex Leaves Water Extracts.

Plants from the Ilex genus are known for properties such as antimicrobial and anti-inflammatory activity, can act as antiobesity agents and thus can be helpful in medicine. Some holly species, such as Ilex paraguariensis (widely known in the form of popular beverage: yerba mate), have been investigated, while others have been partially researched or remain unknown. Therefore, we performed qualitative and quantitative phytochemical analyses and screened antimicrobial properties of lesser-studied species (I. aquifolium L., I. aquifolium 'Argentea Marginata' and I. × meserveae 'Blue Angel'). I. paraguariensis was used as a standard species for comparison purposes. Investigations were performed on water extracts due to their expected activity and composition. Antimicrobial research included evaluating minimal inhibitory, bactericidal (Staphylococcus aureus and Escherichia coli) and fungicidal concentration (Candida albicans, Alternaria alternata, Fusarium oxysporum, and Aspergillus niger) of extracts. The influence of the extracts on the production, eradication, and viability of bacterial biofilms was also analysed. It was established that Ilex paraguariensis possesses the richest profile of hydroxycinnamic acids derivatives in terms of component concentration and diversity. Ilex spp., especially I. × meserveae, contain a slightly higher amount of flavonoids and more different flavonoid derivatives than I. paraguariensis. However, the strongest antibacterial activity was shown by I. aquifolium L. and its cultivar 'Argentea Marginata' in terms of minimal inhibitory, bactericidal and fungicidal concentration, and biofilm assays. Extracts from both species significantly reduced the biofilm viability of S. aureus as well, which may be of use in the production of multicomponent lavaseptics, antiseptics, diuretics (supporting urinary tract infection therapy) and, due to their action on fungi, additives to growth media for specific fungi. The significant content of saponins enables Ilex extracts to be used as natural emulsifiers, for example, in cosmetics. Moreover, relatively high chlorogenic acid and rutin content may suggest use of Ilex spp. to treat obesity, digestive problems, in chemoprevention, and as preservatives in the food industry.

Linseed Silesia, Diverse Crops for Diverse Diets. New Solutions to Increase Dietary Lipids in Crop Species.

The aim of the work was to compare the new variety of oil flax (Silesia) with already cultivated varieties in terms of plant productivity, oil content, fatty acid composition and significant secondary metabolites. The analyzed linseed varieties are characterized by low (Linola), medium (Silesia) and high (Szafir) content of omega-3 fatty acids. Special attention was paid to the quality of the oil and the characteristics that determine its stability (reduction of susceptibility to oxidation). A number of antioxidant compounds of secondary metabolism (simple phenols, phenolic acids, flavonoids, tannins) were identified in the linseed oils. All of these compounds can affect lipid oxidation by a mechanism that attenuates initiating radicals such as hydroxyl or forms an oxidizing primary product such as peroxides. Chelation of metal ions may also be involved in lipid oxidation. We propose a mechanism that encompasses all these processes and facilitates understanding of the complex relationships between them. The general thesis is that the ratio of polyunsaturated fatty acids is associated with a better metabolic state of flaxseed, and thus with a higher nutritional value. In addition, we find a number of specialized secondary metabolites characteristic of the flax studied, which could be useful for chemotaxonomy.

Biofilm eradication and antifungal mechanism of action against Candida albicans of cationic dicephalic surfactants with a labile linker.

Our research aims to expand the knowledge on relationships between the structure of cationic dicephalic surfactants-N,N-bis[3,3_-(dimethylamine)propyl]alkylamide dihydrochlorides and N,N-bis[3,3_-(trimethylammonio)propyl]alkylamide dibromides (alkyl: n-C9H19, n-C11H23, n-C13H27, n-C15H31)-and their antifungal mechanism of action on Candida albicans. The mentioned groups of amphiphilic substances are characterized by the presence of a weak, hydrochloride cationic center readily undergoing deprotonation, as well as a stable, strong quaternary ammonium group and alkyl chains capable of strong interactions with fungal cells. Strong fungicidal properties and the role in creation and eradication of biofilm of those compounds were discussed in our earlier works, yet their mechanism of action remained unclear. It was shown that investigated surfactants induce strong oxidative stress and cause increase in cell membrane permeability without compromising its continuity, as indicated by increased potassium ion (K+) leakage. Thus experiments carried out on the investigated opportunistic pathogen indicate that the mechanism of action of the researched surfactants is different than in the case of the majority of known surfactants. Results presented in this paper significantly broaden the understanding on multifunctional cationic surfactants and their mechanism of action, as well as suggest their possible future applications as surface coating antiadhesives, fungicides and antibiofilm agents in medicine or industry.

Impact of Plant Origin on Eurasian Propolis on Phenolic Profile and Classical Antioxidant Activity.

Propolis is a bee product with known medical properties, including antioxidant activity. The scope of the study is profiling 19 different Eurasian propolis samples (mostly from Russia and Kazakhstan, Kyrgyzstan, Poland, Ukraine, and Slovakia). Profiles of propolises were investigated by ultra-high-performance liquid chromatography-diode array detector-mass spectrometry (UPLC-DAD-MS). Classical antioxidant properties, which are based on electron donation mechanism, were assessed by DPPH, ferric reducing antioxidant power (FRAP), and oxygen radical absorbance capacity (ORAC) assays. Total phenolic and flavonoid contents were also evaluated by colorimetric tests. Most of the samples exhibited significant content of polyphenols (from 30.28 to 145.24 mg GAE/g of propolis) and flavonoids (from 10.45 to 82.71 mg GAE/g of propolis). Most of the propolis samples exhibited potent antiradical (DPPH test-from 8.83 to 64.47 mg GAE/g of propolis) and reducing activity (FRAP test-from 0.08 to 1.17 mmol Fe2+/g of propolis). Based on the occurrence of marker compounds, propolis samples were classified as poplar, aspen-birch, aspen-poplar, and aspen-birch-poplar type. Main markers present in propolis of poplar (e.g., chrysin, pinocembrin, galangin, and 3-O-acetyl-pinobanksin), birch (ermanin and acacetin) and aspen (2-acetyl-1,3-di-p-coumaroylglycerol) origin were used. DPPH, FRAP, and ORAC tests results were correlated with flavonoids, total polyphenols, or the polyphenols other than flavonoids content. In term of activity, poplar propolis type was variable, while aspen-birch-poplar type usually exhibited high DPPH and FRAP activity.

Metabolism of the Cyanogenic Glucosides in Developing Flax: Metabolic Analysis, and Expression Pattern of Genes.

Cyanogenic glucosides (CG), the monoglycosides linamarin and lotaustralin, as well as the diglucosides linustatin and neolinustatin, have been identified in flax. The roles of CG and hydrogen cyanide (HCN), specifically the product of their breakdown, differ and are understood only to a certain extent. HCN is toxic to aerobic organisms as a respiratory inhibitor and to enzymes containing heavy metals. On the other hand, CG and HCN are important factors in the plant defense system against herbivores, insects and pathogens. In this study, fluctuations in CG levels during flax growth and development (using UPLC) and the expression of genes encoding key enzymes for their metabolism (valine N-monooxygenase, linamarase, cyanoalanine nitrilase and cyanoalanine synthase) using RT-PCR were analyzed. Linola cultivar and transgenic plants characterized by increased levels of sulfur amino acids were analyzed. This enabled the demonstration of a significant relationship between the cyanide detoxification process and general metabolism. Cyanogenic glucosides are used as nitrogen-containing precursors for the synthesis of amino acids, proteins and amines. Therefore, they not only perform protective functions against herbivores but are general plant growth regulators, especially since changes in their level have been shown to be strongly correlated with significant stages of plant development.

Chemical profiling and cytotoxic activity of 150-year old original sample of Jerusalem Balsam.

Herbal formulations have been used in ethnomedicine and pharmacy around the world for thousands of years. One of them is Jerusalem Balsam (JB), which came into use in the seventeenth century. Today, people still produce and use it regularly as prophylactic supplement. JB has been widely used in Europe since the nineteenth century, as a remedy possessing antibacterial, antifungal and anti-inflammatory activities. The composition of the product was not known, although possible formulations were reported. In this study the original sample, which dated back to 1870, was investigated for chemical composition and cytotoxic activity. The obtained results were compared with results from more recently produced samples. Several tests were carried out, namely GC-MS, UPLC-PDA-Q-TOF-MS and MTT. Only the 150-year old sample showed a significant cytotoxic activity on cancer cell lines. At a concentration of 125 µg/mL after 72 h of incubation, the original sample inhibited almost 90% of cell metabolic activity, while contemporary samples showed none or little activity. None of the tested samples showed a significant impact on normal cells. These results may be attributed to the activities of benzoic acid and its derivatives, cinnamic acid derivatives, vanillin, group of sesquiterpenes and cembrene.

Temporal biosynthesis of flavone constituents in flax growth stages.

Previous studies showed that chalcone synthase (chs) silencing in flax (Linum usitatisimum) induces a signal transduction cascade that leads to extensive modification of plant metabolism. Result presented in the current study, performed on field grown flax plants - (across the whole vegetation period) demonstrates that, in addition to its role in tannin and lignin biosynthesis, the chs gene also participates in the regulation of flavone biosynthesis during plant growth. Apigenin and luteolin glycosides constitute the flavones, the major group of flavonoids in flax. Alterations in their levels correlate with plant growth, peaking at the flower initiation stage. Suppression of chs gene expression causes significant changes in the ratio of flavone constituents at the early stage of flax growth. A significant correlation between flavonoid 3'-hydroxylase (F3'H) gene expression and accumulation of luteolin glycosides has been found, indicating that flavone biosynthesis during flax growth and development is regulated by temporal expression of this gene. The lack of such a correlation between the flavone synthase (FNS) gene and flavone accumulation in the course of plant growth suggests that the main route of flavone biosynthesis is mediated by eriodictyol. This is the first report indicating the ratio of flavone constituents as a potent marker of flax growth stages and temporal expression of F3'H, the key gene of their biosynthesis.

Profile of Polyphenolic and Essential Oil Composition of Polish Propolis, Black Poplar and Aspens Buds.

In this work, we studied similarities and differences between 70% ethanol in water extract (70EE) and essential oils (EOs) obtained from propolis, black poplars (Populus nigra L.) and aspens (P. tremula L.) to ascertain which of these is a better indicator of the plant species used by bees to collect propolis precursors. Composition of 70EE was analyzed by UPLC-PDA-MS, while GC-MS was used to research the EOs. Principal component analyses (PCA) and calculations of Spearman's coefficient rank were used for statistical analysis. Statistical analysis exhibited correlation between chemical compositions of propolis and Populus buds' 70EE. In the case of EOs, results were less clear. Compositions of black poplars, aspens EOs and propolises have shown more variability than 70EE. Different factors such as higher instability of EOs compared to 70EE, different degradation pattern of benzyl esters to benzoic acid, differences in plant metabolism and bees' preferences may be responsible for these phenomena. Our research has therefore shown that 70EE of propolis reflected the composition of P. nigra or complex aspen⁻black poplar origin.

Impact of CAD-deficiency in flax on biogas production.

Global warming and the reduction in our fossil fuel reservoir have forced humanity to look for new means of energy production. Agricultural waste remains a large source for biofuel and bioenergy production. Flax shives are a waste product obtained during the processing of flax fibers. We investigated the possibility of using low-lignin flax shives for biogas production, specifically by assessing the impact of CAD deficiency on the biochemical and structural properties of shives. The study used genetically modified flax plants with a silenced CAD gene, which encodes the key enzyme for lignin synthesis. Reducing the lignin content modified cellulose crystallinity, improved flax shive fermentation and optimized biogas production. Chemical pretreatment of the shive biomass further increased biogas production efficiency.