A New Face of the Old Gene: Deletion of the PssA, Encoding Monotopic Inner Membrane Phosphoglycosyl Transferase in Rhizobium leguminosarum, Leads to Diverse Phenotypes That Could Be Attributable to Downstream Effects of the Lack of Exopolysaccharide.

The biosynthesis of subunits of rhizobial exopolysaccharides is dependent on glycosyltransferases, which are usually encoded by large gene clusters. PssA is a member of a large family of phosphoglycosyl transferases catalyzing the transfer of a phosphosugar moiety to polyprenol phosphate; thus, it can be considered as priming glycosyltransferase commencing synthesis of the EPS repeating units in Rhizobium leguminosarum. The comprehensive analysis of PssA protein features performed in this work confirmed its specificity for UDP-glucose and provided evidence that PssA is a monotopic inner membrane protein with a reentrant membrane helix rather than a transmembrane segment. The bacterial two-hybrid system screening revealed interactions of PssA with some GTs involved in the EPS octasaccharide synthesis. The distribution of differentially expressed genes in the transcriptome of the ΔpssA mutant into various functional categories indicated complexity of cell response to the deletion, which can mostly be attributed to the lack of exopolysaccharide and downstream effects caused by such deficiency. The block in the EPS biosynthesis at the pssA step, potentially leading to an increased pool of UDP-glucose, is likely to be filtered through to other pathways, and thus the absence of EPS may indirectly affect the expression of proteins involved in these pathways.

Genomic and Metabolic Characterization of Plant Growth-Promoting Rhizobacteria Isolated from Nodules of Clovers Grown in Non-Farmed Soil.

The rhizosphere microbiota, which includes plant growth-promoting rhizobacteria (PGPR), is essential for nutrient acquisition, protection against pathogens, and abiotic stress tolerance in plants. However, agricultural practices affect the composition and functions of microbiota, reducing their beneficial effects on plant growth and health. Among PGPR, rhizobia form mutually beneficial symbiosis with legumes. In this study, we characterized 16 clover nodule isolates from non-farmed soil to explore their plant growth-promoting (PGP) potential, hypothesizing that these bacteria may possess unique, unaltered PGP traits, compared to those affected by common agricultural practices. Biolog profiling revealed their versatile metabolic capabilities, enabling them to utilize a wide range of carbon and energy sources. All isolates were effective phosphate solubilizers, and individual strains exhibited 1-aminocyclopropane-1-carboxylate deaminase and metal ion chelation activities. Metabolically active strains showed improved performance in symbiotic interactions with plants. Comparative genomics revealed that the genomes of five nodule isolates contained a significantly enriched fraction of unique genes associated with quorum sensing and aromatic compound degradation. As the potential of PGPR in agriculture grows, we emphasize the importance of the molecular and metabolic characterization of PGP traits as a fundamental step towards their subsequent application in the field as an alternative to chemical fertilizers and supplements.

Exopolysaccharide Biosynthesis in Rhizobium leguminosarum bv. trifolii Requires a Complementary Function of Two Homologous Glycosyltransferases PssG and PssI.

The Pss-I region of Rhizobium leguminosarum bv. trifolii TA1 comprises more than 20 genes coding for glycosyltransferases, modifying enzymes, and polymerization/export proteins, altogether determining the biosynthesis of symbiotically relevant exopolysaccharides. In this study, the role of homologous PssG and PssI glycosyltransferases in exopolysaccharide subunit synthesis were analyzed. It was shown that the glycosyltransferase-encoding genes of the Pss-I region were part of a single large transcriptional unit with potential downstream promoters activated in specific conditions. The ΔpssG and ΔpssI mutants produced significantly lower amounts of the exopolysaccharide, while the double deletion mutant ΔpssIΔpssG produced no exopolysaccharide. Complementation of double mutation with individual genes restored exopolysaccharide synthesis, but only to the level similar to that observed for the single ΔpssI or ΔpssG mutants, indicating that PssG and PssI serve complementary functions in the process. PssG and PssI interacted with each other in vivo and in vitro. Moreover, PssI displayed an expanded in vivo interaction network comprising other GTs involved in subunit assembly and polymerization/export proteins. PssG and PssI proteins were shown to interact with the inner membrane through amphipathic helices at their C-termini, and PssG also required other proteins involved in exopolysaccharide synthesis to localize in the membrane protein fraction.

Engineering the specificity of Streptococcus pyogenes sortase A by loop grafting.

Sortases are a group of enzymes displayed on the cell-wall of Gram-positive bacteria. They are responsible for the attachment of virulence factors onto the peptidoglycan in a transpeptidation reaction through recognition of a pentapeptide substrate. Most housekeeping sortases recognize one specific pentapeptide motif; however, Streptococcus pyogenes sortase A (SpSrtA WT) recognizes LPETG, LPETA and LPKLG motifs. Here, we examined SpSrtA's flexible substrate specificity by investigating the role of the ß7/ß8 loop in determining substrate specificity. We exchanged the ß7/ß8 loop in SpSrtA with corresponding ß7/ß8 loops from Staphylococcus aureus (SaSrtA WT) and Bacillus anthracis (BaSrtA WT). While the BaSrtA-derived variant showed no enzymatic activity toward either LPETG or LPETA substrates, the activity of the SaSrtA-derived mutant toward the LPETA substrate was completely abolished. Instead, the mutant had an improved activity toward LPETG, the preferred substrate of SaSrtA WT.

PssJ Is a Terminal Galactosyltransferase Involved in the Assembly of the Exopolysaccharide Subunit in Rhizobium Leguminosarum bv. Trifolii.

Rhizobium leguminosarum bv. trifolii produces exopolysaccharide (EPS) composed of glucose, glucuronic acid, and galactose residues at a molar ratio 5:2:1. A majority of genes involved in the synthesis, modification, and export of exopolysaccharide are located in the chromosomal Pss-I region. In the present study, a ΔpssJ deletion mutant was constructed and shown to produce EPS lacking terminal galactose in the side chain of the octasaccharide subunit. The lack of galactose did not block EPS subunit translocation and polymerization. The in trans delivery of the pssJ gene restored the production of galactose-containing exopolysaccharide. The mutant was compromised in several physiological traits, e.g., motility and biofilm production. An impact of the pssJ mutation and changed EPS structure on the symbiotic performance was observed as improper signaling at the stage of molecular recognition, leading to formation of a significant number of non-infected empty nodules. Terminal galactosyltransferase PssJ was shown to display a structure typical for the GT-A class of glycosyltransferases and interact with other GTs and Wzx/Wzy system proteins. The latter, together with PssJ presence in soluble and membrane protein fractions indicated that the protein plays its role at the inner membrane interface and as a component of a larger complex.

Contamination of the urban environment with excrements of companion animals as an underestimated source of Staphylococcus species posing a threat to public health.

The aim of the study was to assess the incidence, resistance, virulence, and genotypic characteristics of Staphylococcus spp. residing in the gastrointestinal tract of dogs and cats, as a group of animals causing potential contamination of the urban space. A high percentage of strains resistant to penicillin (58%), oxacillin (9%) and tetracycline (60%) were found. All isolates resistant to penicillin, kanamycin, or chloramphenicol carried genes responsible for individual resistance (blaZ, aph(3')-IIIa, and cat (pC194)/cat (pC223), respectively. The mecA gene was detected in 45% of the oxacillin-resistant Staphylococcus pseudintermedius strains. The amplification of DNA fragments surrounding rare restriction sites analysis demonstrated high heterogeneity of genotypic profiles correlating with phenotypic resistance profiles. Multilocus sequence typing analysis classified the methicillin-resistant S. pseudintermedius strains as ST71, ST890, and the totally new ST1047. The presence of a high level of resistance among Staphylococcus strains may suggest a potential risk of transfer of these bacteria between companion animals and humans.

Mgl2 Is a Hypothetical Methyltransferase Involved in Exopolysaccharide Production, Biofilm Formation, and Motility in Rhizobium leguminosarum bv. trifolii.

In this study, functional characterization of the mgl2 gene located near the Pss-I exopolysaccharide biosynthesis region in Rhizobium leguminosarum bv. trifolii TA1 is described. The hypothetical protein encoded by the mgl2 gene was found to be similar to methyltransferases (MTases). Protein homology and template-based modeling facilitated prediction of the Mgl2 structure, which greatly resembled class I MTases with a S-adenosyl-L-methionine-binding cleft. The Mgl2 protein was engaged in exopolysaccharide, but not lipopolysaccharide, synthesis. The mgl2 deletion mutant produced exopolysaccharide comprised of only low molecular weight fractions, while overexpression of mgl2 caused overproduction of exopolysaccharide with a normal low to high molecular weight ratio. The deletion of the mgl2 gene resulted in disturbances in biofilm formation and a slight increase in motility in minimal medium. Red clover (Trifolium pratense) inoculated with the mgl2 mutant formed effective nodules, and the appearance of the plants indicated active nitrogen fixation. The mgl2 gene was preceded by an active and strong promoter. Mgl2 was defined as an integral membrane protein and formed homodimers in vivo; however, it did not interact with Pss proteins encoded within the Pss-I region. The results are discussed in the context of the possible involvement of the newly described potential MTase in various metabolic traits, such as the exopolysaccharide synthesis and motility that are important for rhizobial saprophytic and symbiotic relationships.

Diversity and plant growth promoting properties of rhizobia isolated from root nodules of Ononis arvensis.

This is the first report describing isolates from root nodules of Ononis arvensis (field restharrow). The aim of this investigation was to describe the diversity, phylogeny, and plant growth promoting features of microsymbionts of O. arvensis, i.e., a legume plant growing in different places of the southern part of Poland. Twenty-nine bacterial isolates were characterized in terms of their phenotypic properties, genome fingerprinting, and comparative analysis of their 16S rRNA, nodC and acdS gene sequences. Based on the nodC and 16S rRNA gene phylogenies, the O. arvensis symbionts were grouped close to bacteria of the genera Rhizobium and Mesorhizobium, which formed monophyletic clusters. The acdS gene sequences of all the isolates tested exhibited the highest similarities to the corresponding gene sequences of genus Mesorhizobium strains. The presence of the acdS genes in the genomes of rhizobia specific for O. arvensis implies that these bacteria may promote the growth and development of their host plant in stress conditions. The isolated bacteria showed a high genomic diversity and, in the BOX-PCR reaction, all of them (except three) exhibited DNA fingerprints specific only for them. Our studies showed that restharrow isolates formed effective symbiotic interactions with their native host (O. arvensis) and Ononis spinosa but not with Trifolium repens and Medicago sativa belonging to the same tribe Trifolieae as Ononis species and not with Lotus corniculatus, representing the tribe Loteae.

Multilocus sequence analysis supports the taxonomic position of Astragalus glycyphyllos symbionts based on DNA-DNA hybridization.

In this study, the phylogenetic relationship and taxonomic status of six strains, representing different phenons and genomic groups of Astragalus glycyphyllos symbionts, originating from Poland, were established by comparative analysis of five concatenated housekeeping gene sequences (atpD, dnaK, glnA, recA and rpoB), DNA-DNA hybridization and total DNA G+C content. Maximum-likelihood phylogenetic analysis of combined atpD, dnaK, glnA, recA and rpoB sequence data placed the studied bacteria into the clade comprising the genus Mesorhizobium. In the core gene phylograms, four A. glycyphyllos nodule isolates (AG1, AG7, AG15 and AG27) formed a cluster common with Mesorhizobium ciceri, whereas the two other A. glycyphyllos symbionts (AG17 and AG22) were grouped together with Mesorhizobium amorphae and M. septentrionale. The species position of the studied bacteria was clarified by DNA-DNA hybridization. The DNA-DNA relatedness between isolates AG1, AG7, AG15 and AG27 and reference strain M. ciceri USDA 3383T was 76.4-84.2%, and all these A. glycyphyllos nodulators were defined as members of the genomospecies M. ciceri. DNA-DNA relatedness for isolates AG17 and AG22 and the reference strain M. amorphae ICMP 15022T was 77.5 and 80.1%, respectively. We propose that the nodule isolates AG17 and AG22 belong to the genomic species M. amorphae. Additionally, it was found that the total DNA G+C content of the six test A. glycyphyllos symbionts was 59.4-62.1 mol%, within the range for species of the genus Mesorhizobium.

High-Throughput Screening in Protein Engineering: Recent Advances and Future Perspectives.

Over the last three decades, protein engineering has established itself as an important tool for the development of enzymes and (therapeutic) proteins with improved characteristics. New mutagenesis techniques and computational design tools have greatly aided in the advancement of protein engineering. Yet, one of the pivotal components to further advance protein engineering strategies is the high-throughput screening of variants. Compartmentalization is one of the key features allowing miniaturization and acceleration of screening. This review focuses on novel screening technologies applied in protein engineering, highlighting flow cytometry- and microfluidics-based platforms.

Yeast and human mitochondrial helicases.

Mitochondria are semiautonomous organelles which contain their own genome. Both maintenance and expression of mitochondrial DNA require activity of RNA and DNA helicases. In Saccharomyces cerevisiae the nuclear genome encodes four DExH/D superfamily members (MSS116, SUV3, MRH4, IRC3) that act as helicases and/or RNA chaperones. Their activity is necessary for mitochondrial RNA splicing, degradation, translation and genome maintenance. In humans the ortholog of SUV3 (hSUV3, SUPV3L1) so far is the best described mitochondrial RNA helicase. The enzyme, together with the matrix-localized pool of PNPase (PNPT1), forms an RNA-degrading complex called the mitochondrial degradosome, which localizes to distinct structures (D-foci). Global regulation of mitochondrially encoded genes can be achieved by changing mitochondrial DNA copy number. This way the proteins involved in its replication, like the Twinkle helicase (c10orf2), can indirectly regulate gene expression. Here, we describe yeast and human mitochondrial helicases that are directly involved in mitochondrial RNA metabolism, and present other helicases that participate in mitochondrial DNA replication and maintenance. This article is part of a Special Issue entitled: The Biology of RNA helicases - Modulation for life.

Phenotypic characterization of Astragalus glycyphyllos symbionts and their phylogeny based on the 16S rDNA sequences and RFLP of 16S rRNA gene.

In this study, the nitrogen fixing Astragalus glycyphyllos symbionts were characterized by phenotypic properties, restriction fragment length polymorphism (RFLP), and sequences of 16S rDNA. The generation time of A. glycyphyllos rhizobia in yeast extract mannitol medium was in the range 4-6 h. The studied isolates exhibited a low resistance to antibiotics, a moderate tolerance to NaCl, assimilated di- and trisaccharides, and produced acid in medium containing mannitol as a sole carbon source. In the cluster analysis, based on 86 phenotypic properties of A. glycyphyllos symbionts and the reference rhizobia, examined isolates and the genus Mesorhizobium strains were placed on a single branch, clearly distinct from other lineages of rhizobial genera. By the comparative analysis of 16S rRNA gene sequences and 16S rDNA-RFLP, A. glycyphyllos nodulators were also identified as the members of the genus Mesorhizobium. On the 16S rDNA sequence phylogram, the representatives of A. glycyphyllos nodule isolates formed a robust, monophyletic cluster together with the Mesorhizobium species at 16S rDNA sequence similarity of these bacteria between 95 and 99 %. Similarly, the cluster analysis of the combined RFLP-16S rDNA patterns, obtained with seven restriction endonucleases, showed that A. glycyphyllos rhizobia are closely related to the genus Mesorhizobium bacteria. The taxonomic approaches used in this paper allowed us to classify the studied bacteria into the genus Mesorhizobium.

Loss of Y in regulatory T lymphocytes in the tumor micro-environment of primary colorectal cancers and liver metastases.

Male sex is a risk factor for colorectal cancer (CRC) with higher illness burden and earlier onset. Thus, we hypothesized that loss of chromosome Y (LOY) in the tumor micro-environment (TME) might be involved in oncogenesis. Previous studies show that LOY in circulating leukocytes of aging men was associated with shorter survival and non-hematological cancer, as well as higher LOY in CD4 + T-lymphocytes in men with prostate cancer vs. controls. However, nothing is known about LOY in leukocytes infiltrating TME and we address this aspect here. We studied frequency and functional effects of LOY in blood, TME and non-tumorous tissue. Regulatory T-lymphocytes (Tregs) in TME had the highest frequency of LOY (22%) in comparison to CD4 + T-lymphocytes and cytotoxic CD8 + T-lymphocytes. LOY score using scRNA-seq was also linked to higher expression of PDCD1, TIGIT and IKZF2 in Tregs. PDCD1 and TIGIT encode immune checkpoint receptors involved in the regulation of Tregs function. Our study sets the direction for further functional research regarding a probable role of LOY in intensifying features related to the suppressive phenotype of Tregs in TME and consequently a possible influence on immunotherapy response in CRC patients.

RNA degradation in yeast and human mitochondria.

Expression of mitochondrially encoded genes must be finely tuned according to the cell's requirements. Since yeast and human mitochondria have limited possibilities to regulate gene expression by altering the transcription initiation rate, posttranscriptional processes, including RNA degradation, are of great importance. In both organisms mitochondrial RNA degradation seems to be mostly depending on the RNA helicase Suv3. Yeast Suv3 functions in cooperation with Dss1 ribonuclease by forming a two-subunit complex called the mitochondrial degradosome. The human ortholog of Suv3 (hSuv3, hSuv3p, SUPV3L1) is also indispensable for mitochondrial RNA decay but its ribonucleolytic partner has so far escaped identification. In this review we summarize the current knowledge about RNA degradation in human and yeast mitochondria. This article is part of a Special Issue entitled: Mitochondrial Gene Expression.

Seasonality and intensity of airborne Boletus-type spores in relation to land use and weather pattern.

Forests are a natural source of airborne bolete spores. The timing of sporulation and its intensity as well as the dispersal of airborne spores and in consequence their concentrations depend in particular on the type of land use determining the availability of matter on which they develop and on meteorological factors. The aim of this study was to perform a spatial and temporal analysis of the occurrence of Boletus-type spores in the warm temperate climate of the Northern Hemisphere. An assumption was made that the spore concentrations depend on the type of land cover and weather conditions. The volumetric method was applied to investigate differences in spore concentrations and using spore traps installed at different heights and at locations with different land cover types. Boletus-type spores occurred in the air at high concentrations in late summer and in the autumn. The season start dates and maximum concentrations did not differ significantly between sites and seasons, but the season intensity varied. Higher spore concentrations were usually found in the region with a larger proportion of green areas, including forests. An analysis of the diurnal cycles showed that within 24 h spore concentration reached high levels twice, which was especially noticeable in ground level monitoring. Air temperature and air humidity were the main weather factors affecting the occurrence of airborne spores. This research indicates that when studying the effects of different factors on the concentration of airborne basidiospores, many environmental elements should be analyzed, including the characteristics of habitats in which basidiomycetes grow. Climate, weather, geobotany, and land use type should be taken into account in analysis and interpretation of aeromycological phenomena.

Factors affecting the quality of life in hemifacial spasm patients.

BACKGROUND AND PURPOSE: Hemifacial spasm (HFS), a movement disorder manifested by unilateral spasms of the muscles innervated by the facial nerve, interferes with social life in about 90% of patients, causing social isolation and depression and having a significant impact on the quality of life. The aim of the study was to assess factors affecting the quality of life in patients with HFS in respect of influence of the severity of depression symptoms and botulinum toxin type A (BTX-A) therapy. MATERIAL AND METHODS: Eighty-five out of 129 patients included in the HFS database of the Movement Disorders Outpatient Clinic, Department of Neurology, University Hospital, Cracow who fulfilled the inclusion criteria and had no exclusion criteria (suffering from concomitant movement disorders, other severe chronic diseases or cognitive impairment) were studied. Demographic and clinical data (age at onset, disease duration and accompanying symptoms) were collected. Severity of HFS was assessed by the five-point clinical scale and seven-point Clinical Global Impression scale. Quality of life was assessed with the HFS-36 questionnaire and severity of depressive symptoms was evaluated with the Beck Depression Inventory. HFS-36 was performed twice, before BTX-A injection and two weeks later. RESULTS: The mean global score of HFS-36 was 47 ± 31 (maximum: 140 pts). Decreased HFS-36 score resulted from divergent deterioration in all subscales included in the questionnaire. Independent risk factors of deterioration in HFS-36 were increased severity of HFS and depressive symptoms as well as accompanying trismus. The HFS-36 score depended on the number and type of accompanying symptoms as well. Botulinum toxin type A therapy led to a significant improvement of HFS-36, particularly high in patients with multiple (> 4) HFS-related symptoms. CONCLUSIONS: The HFS-36 score depends mostly on severity of HFS, depressive symptoms and occurrence of accompanying trismus. It improves after BTX-A treatment.

The influence of botulinum toxin on auditory disturbances in hemifacial spasm.

BACKGROUND AND PURPOSE: Hemifacial spasm (HFS) is frequently accompanied by other symptoms, such as visual and auditory disturbances or pain. The aim of the study was to assess the occurrence of auditory symptoms accompanying HFS using subjective and objective methods, their relation with other HFS symptoms, and their resolution after botulinum toxin (BTX-A) treatment. MATERIAL AND METHODS: The occurrence of hypoacusis, ear clicks and tinnitus was assessed by questionnaire in 126 HFS patients from an electronic database which included medical data such as severity of HFS rated by clinical scale and magnetic resonance imaging focused on the presence of vascular nerve VII and VIII conflict. Forty consecutive patients treated with BTX-A and 24 controls matched by sex and age underwent laryngological examination including audiometry, tympanometry and acoustic middle ear reflex before injection and two weeks later. RESULTS: About 45.2% of patients complained of auditory disturbances (31.7% hypoacusis, 30.2% ear clicks and 7.1% tinnitus) on the side of HFS. Auditory disturbances correlated with severity of HFS symptoms but not with age, disease duration, or neurovascular conflict with nerves VII and VIII. We did not find abnormalities in audiometric and tympanometric assessment in patients in comparison with controls. No abnormalities were detected in brainstem evoked potentials comparing the sides with and without HFS symptoms. Tinnitus and absence of ipsilateral acoustic middle ear reflex occurred more often in patients with auditory symptoms than those without them. BTX-A treatment caused resolution of subjective acoustic symptoms without any improvement in audiometric assessment. CONCLUSIONS: Auditory disturbances accompanying HFS are probably caused by dysfunction of the Eustachian tube, which improves after BTX-A treatment.

Characteristics of the Surface Topography and Tribological Properties of Reinforced Aluminum Matrix Composite.

Due to their excellent synergistic properties, Aluminum Matrix Composites (AMC) have achieved a high degree of prominence in different industries. In addition to strength, the wear resistance of materials is also an important criterion for numerous applications. The wear resistance depends on the surface topography as well as the working conditions of the interacting parts. Therefore, extensive experiments are being conducted to improve the suitability of engineering materials (including AMC) for different applications. This paper presents research on manufactured aluminum metal matrix composites reinforced with 10 wt.% of Al2SiO5 (aluminum sillimanite). The manufactured and prepared samples were subjected to surface topography measurements and to tribological studies both with and without lubricant using a block-on-ring tester. Based on the results, analyses of the surface topography (i.e., surface roughness parameters, Abbott-Firestone curve, and surface defects) as well as of the tribological characteristics (i.a. friction coefficient, linear wear, and wear intensity) were performed. Differences in the surface topography of the manufactured elements were shown. The surface topography had a significant impact on tribological characteristics of the sliding joints in the tests where lubrication was and was not used. Better tribological characteristics were obtained for the surfaces characterized by greater roughness (determined on the basis of both the profile and surface texture parameters). In the case of tribological tests with lubrication, the friction coefficient as well as the wear intensity was significantly lower compared to tribological tests without lubrication. However, lower values of the friction coefficient and wear intensity were still recorded for the surfaces that were characterized by greater roughness. The obtained results showed that it is important to analyze the surface topography because surface characteristics influence tribological properties.

Dry Sliding Wear Studies on Sillimanite and B4C Reinforced Aluminium Hybrid Composites Fabricated by Vacuum Assisted Stir Casting Process.

This paper presents the results of studies to understand the influence of hybridisation on mechanical and tribological behaviour as well as dry sliding wear of aluminium metal matrix composites. Sillimanite and boron carbide (B4C) were used as primary and secondary reinforcements and pure aluminium was used as the matrix material. The composite was fabricated by using a vacuum assisted stir casting process. Different research instruments were used, including a scanning electron microscope with EDX spectrometer, a surface measurement device, a thermal image analyser, as well as a tribotester. The results show that tensile, impact strength and hardness of the hybridised composites are superior (a step ahead) than unreinforced and primary composites. The wear behaviour of the fabricated specimens was tested for the dry sliding wear behaviour under the load range of 10-50 N with the steps of 20 N for the sliding velocities 0.75, 1.5 and 2.25 m/s over a distance of 1000 m. The wear rate increased with load and decreased as the wt.% of reinforcement increased. The wear rate of the composite with 10 wt.% Al2SiO5 was approximately 44% lower than that of the composite with 5 wt.% Al2SiO5. The same dependence was noted for hybrid composite (5 wt.% Al2SiO5 + 5 wt.% B4C)-the wear rate was approximately 50.8% lower than that of the composite with 5 wt.% Al2SiO5 under the same test condition. The friction coefficient decreased as the weight percentage of the reinforcement (Al2SiO5 and B4C) increased due to the uniform distribution of the reinforcement on the surface of the composites. The main wear mechanism of the studied materials was abrasion wear. The wear mechanism of the composite had tribochemical type. It involved the oxidation and transfer of the material, which formed protective tribolayers ensuring an additional sliding process. The mechanism that played the main role in the wear process of the composites was a combination of abrasive, adhesive and oxidative wear.

Bioaerosols on the atmospheric super highway: An example of long distance transport of Alternaria spores from the Pannonian Plain to Poland.

Alternaria spores are pathogenic to agricultural crops, and the longest and the most severe sporulation seasons are predominantly recorded in rural areas, e.g. the Pannonian Plain (PP) in South-Central Europe. In Poland (Central Europe), airborne Alternaria spore concentrations peak between July and August. In this study, we test the hypothesis that the PP is the source of Alternaria spores recorded in Poland after the main sporulation season (September-October). Airborne Alternaria spores (2005-2019) were collected using volumetric Hirst spore traps located in 38 locations along the potential pathways of air masses, i.e. from Serbia, Romania and Hungary, through the Czech Republic, Slovakia and Ukraine, to Northern Poland. Three potential episodes of Long Distance Transport (LDT) were selected and characterized in detail, including the analysis of Alternaria spore data, back trajectory analysis, dispersal modelling, and description of local weather and mesoscale synoptic conditions. During selected episodes, increases in Alternaria spore concentrations in Poznan were recorded at unusual times that deviated from the typical diurnal pattern, i.e. at night or during morning hours. Alternaria spore concentrations on the PP were very high (>1000 spores/m3) at that time. The presence of non-local Ambrosia pollen, common to the PP, were also observed in the air. Air mass trajectory analysis and dispersal modelling showed that the northwest part of the PP, north of the Transdanubian Mountains, was the potential source area of Alternaria spores. Our results show that Alternaria spores are transported over long distances from the PP to Poland. These spores may markedly increase local exposure to Alternaria spores in the receptor area and pose a risk to both human and plant health. Alternaria spores followed the same atmospheric route as previously described LDT ragweed pollen, revealing the existence of an atmospheric super highway that transports bioaerosols from the south to the north of Europe.