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The article refers to resources, problems and challenges of autism diagnosis and support system in Poland. The resources include: the increasing number of specialists, diagnostic and therapeutic centres, well-established course of education for people working with youths, standardised and normalised diagnostic tools. The diagnostic process is not without some areas in need of our focus: the tendency of some specialists to make unauthorised diagnosis, overshadowing; underestimation of comorbidity of ASD with other disorders. The challenges refer to introducing an effective system of monitoring the services provided in form of certification and control in order to prevent their abuse, initiating category of temporary diagnosis; paying more attention on individual’s resources, better cooperation among specialists, teachers and families, developing and unifying diagnostic standards.

The aim of the presented research was to assess the level of language development of early school age children in two aspects: language production and comprehension. For this purpose, 458 children from five Subcarpathian schools were examined. The Intelligence and Development Scales (IDS) test module and the Language Development Test (Test Rozwoju Jezykowego, TRJ) subtests were used in the study. The child’s gender, place of living and class were assumed to influence the level of development of language production and comprehension. Research results indicate a relatively large number of children with a low score in both language production and comprehension, as well as slightly better language skills of girls compared to boys. In addition, there is a need for checking the language development of children also at subsequent levels of education to identify those with difficulties in this area and implement proper support.

Hydrogels, based on natural polymers, such as hyaluronic acid, are gaining an increasing popularity because of their biological activity. The antibacterial effect of ozone is widely known and used, but the instability the gas causes, severely limits its application. Ozone entrapment in olive oil by its reaction with an unsaturated bond, allows for the formation of stable, therapeutically active ozone derivatives. In this study, we obtained an innovative hydrogel, based on hyaluronic acid containing micro/nanocapsules of ozonated olive oil. By combination of the biocompatible polymer with a high regenerative capacity and biologically active ingredients, we obtained a hydrogel with regenerative properties and a very weak inhibitory effect against both bacterial commensal skin microbiota and pathogenic Candida-like yeasts. We assessed the stability and rheological properties of the gel, determined the morphology of the composite, using scanning electron microscopy (SEM) and particle size by the dynamic light scattering (DLS) method. We also performed Attenuated total reflectance Fourier transform infrared (FTIR-ATR) spectroscopy. The functional properties, including the antimicrobial potential were assessed by the microbiological analysis and in vitro testing on the HaCat human keratinocyte cell line. The studies proved that the obtained emulsions were rheologically stable, exhibited an antimicrobial effect and did not show cytotoxicity in the HaCat keratinocyte model.

Cultural heritage objects are very frequently visited by tourists, the presence of which is considered to affect the bacterial and fungal aerosol levels in these facilities. Therefore, this study had two main aims with the first to determine the morning/evening changes in the concentration of bacterial and fungal aerosol as well as particulate matter in the selected historical objects of Krakow: churches, crypts, libraries, and museums. The second aim was to establish the most possible factors affecting the quality of air in the examined sites using data mining techniques: principal component analysis (PCA) and cluster analysis (CA). Bacterial and fungal aerosol levels were measured in ten historical objects by six-stage Andersen impactor. Particulate matter (PM10, PM4, PM2.5 and PM1), temperature, and relative humidity (RH) were also assessed. Unlike dusts, the microbial aerosol concentrations were not always significantly lower in the morning than in the evening. For bacteria, these relationships varied depending on the site, while for fungi, the morning concentrations were mostly higher than the evening ones. Both PCA and CA indicated that the bacterial and fungal aerosol concentrations were not associated with the particulate matter levels, which might suggest different sources of those two air pollution components. Temperature and RH appear to be more important in shaping the microbial aerosol levels and fungi are more dependent on RH than bacteria.

Mycobionts of many lichen genera appear to demonstrate strong selectivity in the choice of algal partner. The biological properties of a photobiont and its availability in an environment significantly determine the habitat requirements of lichens. Flexibility in photobiont choice extends the ecological amplitude of lichens; therefore, it may constitute an important adaptive strategy for colonization of extreme habitats. The photobiont inventory of the three epigeic lichens most resistant to soil pollution, i.e., Cladonia cariosa, C. rei, and the hyperaccumulator Diploschistes muscorum, was examined to verify whether and to what extent algal composition depends on the type of habitat and substrate enrichment with heavy metals. Photobionts Asterochloris and Trebouxia were identified in the studied lichen species; however, the presence of Trebouxia was directly related to anthropogenic sites with technogenic substrates, and the proportion of lichen specimens with these algae clearly depended on the level of heavy-metal soil pollution and the habitat type. The total number of algal haplotypes increased with increasing soil pollution, and the richness was associated more with soil pollution than with a given lichen species. Additionally, a large number of lichen individuals bearing multiple algal genotypes at polluted sites were recorded. Although Cladonia lichens were previously thought to be restricted to Asterochloris, they are able to start the relichenization process with Trebouxia under specific habitat conditions and to establish a stable association with these algae when colonization of disturbed sites takes place. Comparative analysis of the internal transcribed spacer (ITS) rDNA sequences revealed as many as 13 haplotypes of Trebouxia, and phylogenetic analysis grouped them into two different clades. Such a high level of genetic diversity indicates that Trebouxia is well adapted to metal pollution and could be an alternative photosynthetic partner for certain lichens, especially in polluted sites.

This study was aimed to determine the prevalence of antibiotic-resistant Escherichia coli in the Szreniawa river with detailed aims of: (i) assessment of differences in the number of microbiological indicators of water quality in a diurnal cycle in a vicinity of the sewage treatment plant (STP); (ii) determination of prevalence of antimicrobial resistant E. coli isolated from three sites located at varying locations toward the STP; (iii) evaluation of the presence of extended-spectrum beta lactamase (ESBL)-determining genes in waterborne E. coli isolated from three sites of Szreniawa and (iv) genetic similarity assessment among the E. coli populations. Bacteriological contamination (coliforms, E. coli, E. faecalis) was assessed using membrane filtration. Fifty E. coli strains, the species of which was confirmed by MALDI-TOF analysis, were subjected to antimicrobial resistance tests using standard disk-diffusion method. Double disk synergy test was used to assess the ESBL production and PCR tests were conducted to detect the ESBL-conferring genes and evaluate the genetic diversity. A significant variation in the number of bacteriological indicators was observed both within and between the sampling sites, suggesting the effect of effluent from the STP, point discharge of household sewage and agricultural runoff on the water contamination. The resistance to amoxicillin/clavulanate (90%) and ampicillin (36%) was most prevalent. Multidrug resistance was observed in 40% of strains but no ESBL-producing strains were observed phenotypically. However, the presence of three ESBL-determining genes (TEM, OXA and CTX-M) was detected in 24, 10 and 8% of strains, respectively. A number of factors caused considerable pollution of the river and numerous multidrug resistant E. coli strains were isolated.

Frequent occurrence of microbial resistance to biocides makes it necessary to find alternative antimicrobial substances for modern veterinary medicine. The aim of this study was to obtain biodegradable silver nanoparticle-containing (AgNPs) foils synthesized using non-toxic chemicals and evaluation of their activity against bacterial pathogens isolated from oral cavities of cats, dogs and horses. Silver nanoparticle foils were synthesized using sodium alginate, and glucose, maltose and xylose were used as reducing agents. The sizes of AgNPs differed depending on the reducing agent used (xylose < maltose < glucose). Foil without silver nanoparticles was used as control. Bacterial strains were isolated from cats, dogs and horses by swabbing their oral cavities. Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and extended-spectrum beta-lactamase (ESBL) producing E. coli were isolated on selective chromogenic microbiological media. The bactericidal effect of AgNPs foils obtained using non-toxic chemical compounds against E. coli, ESBL, S. aureus and MRSA isolated from oral cavities of selected animals was confirmed in this study. No statistically significant differences were observed between the foils obtained with different reducing agents. Therefore, all types of examined foils proved to be effective against the isolated bacteria.

Environmental aquatic pollution with antibiotics is a global challenge that affects even pristine mountain environments. Monitoring the concentration of antibiotics in water is critical to water resource management. In this review, we present the sources and degradation routes of antibiotics polluting surface waters, with particular focus on mountain environments and pristine areas. This pollution is strongly related to anthropopressure resulting from intensive tourism. An important aspect of the threat to the environment is water containing antibiotics at sub-inhibitory concentrations, which affects bacterial populations. Antibiotics are ecological factors driving microbial evolution by changing the bacterial community composition, inhibiting or promoting their ecological functions, and enriching and maintaining drug resistance. We paid attention to the stability of antibiotics and their half-lives in water related to biotic and abiotic degradation, which results from the structures of molecules and environmental conditions. Wastewater treatment combined with advanced treatment techniques significantly increase the efficiency of antibiotic removal from wastewater. Modern methods of wastewater treatment are crucial in reducing the supply of antibiotics to aquatic environments and enhancing the possibility of economic and safe reuse of wastewater for technical purposes. We provide a perspective on current research investigating antibiotic emergence in mountain areas and identify knowledge gaps in this field.

The dynamic development of nanotechnology has enabled the development of innovative and novel techniques for the production and use of nanomaterials. One of them is the use of nanocapsules based on biodegradable biopolymer composites. Closing compounds with antimicrobial activity inside the nanocapsule cause the gradual release of biologically active substances into the environment, and the effect on pathogens is regular, prolonged and targeted. Known and used in medicine for years, propolis, thanks to the synergistic effect of active ingredients, has antimicrobial, anti-inflammatory and antiseptic properties. Biodegradable and flexible biofilms were obtained, the morphology of the composite was determined using scanning electron microscopy (SEM) and particle size was measured by the dynamic light scattering (DLS) method. Antimicrobial properties of biofoils were examined on commensal skin bacteria and pathogenic Candida isolates based on the growth inhibition zones. The research confirmed the presence of spherical nanocapsules with sizes in the nano/micrometric scale. The properties of the composites were characterized by infrared (IR) and ultraviolet (UV) spectroscopy. It has been proven that hyaluronic acid is a suitable matrix for the preparation of nanocapsules, as no significant interactions between hyaluronan and the tested compounds have been demonstrated. Color analysis and thermal properties, as well as the thickness and mechanical properties of the obtained films, were determined. Antimicrobial properties of the obtained nanocomposites were strong in relation to all analyzed bacterial and yeast strains isolated from various regions of the human body. These results suggest high potential applicability of the tested biofilms as effective materials for dressings to be applied on infected wounds.

The exposure to airborne dusts and microorganisms is associated with respiratory diseases and other health effects. Currently, research concerning fungal aerosol and particulate matter levels in animal enclosures of zoological gardens is very scarce. Such premises are characterized by numerous sources of air contamination, and they are both working environment and popular tourist attractions. This study aimed to assess the levels of fungal aerosol in animal premises (tapirs, giraffes, pygmy hippos, monkeys, meerkats, pheasants, ostriches, exotarium) of the Kraków Zoo. The bioaerosol samples were collected using a six-stage Andersen impactor to assess the concentrations and size distribution of airborne fungi. Particulate matter (PM10, PM4, PM2.5 and PM1) concentration was assessed using DustTrak Aerosol Monitor. PM concentrations were relatively low (0.048–0.204 mg/m3) and did not exceed the limit values for occupational exposure. Also, the levels of fungal aerosol in almost all cases did not exceed the limit values. The smallest concentration of the total fungal aerosol was observed in the rooms for hippos (approx. 970 CFU/m3) while the highest—in exotarium (16,800 CFU/m3). The share of respirable fraction of bioaerosol was quite high—from 62.6% in monkey houses to 89.2% in exotarium. Having in mind that bioaerosol-forming fungi may contain allergenic species, attention should be paid to thorough cleaning of animals and their premises, and maintaining appropriate levels of microclimatic parameters in the rooms.

Historical facilities such as churches, crypts, libraries and museums are crucial for preserving cultural heritage but at the same time, they are frequently visited tourist objects and working environment. The aim of this study was to assess whether there are differences in the concentration and size distribution of bacterial and fungal aerosol in the mentioned groups of historical objects in Kraków, as well as to determine if there is tendency for seasonal fluctuations of bioaerosol concentrations, and what environmental parameters affect them most significantly. The concentrations of bacterial and fungal aerosol were measured in ten historical objects—churches, crypts, libraries and museums—using a six-stage Andersen impactor. Microbiological analyses were accompanied by measurements of particulate matter (PM10, PM4, PM2.5 and PM1) and microclimatic parameters. The results showed that the studied objects differ in terms of bioaerosol levels, but the differences were not drastic and in terms of bacterial aerosol concentrations the objects can be arranged as follows: churches > libraries > museums > crypts, while for fungal aerosol the order would be: churches > museums > crypts > libraries. The concentrations of bioaerosol did not exceed the safety levels for human exposure, but the levels suggesting possible threat of biodeterioration and the presence of internal source of contamination were exceeded in nearly all sites. The share of respirable fraction of bioaerosol was high (42.1–90.5% for bacteria and 80.7–94.7% for fungi). Bioaerosol concentration was most significantly associated with indoor temperature, and the impact of particulate matter was only moderate.

Objectives: This study was aimed to assess the concentration of microbial aerosol and species composition of airborne staphylococci in 10 healthcare facilities in southern Poland including primary healthcare units and hospital wards; and to assess whether the selected components of microbial aerosol pose a threat of severe infections to either patients or the personnel. Methods: The study was conducted at monthly intervals over a period of one year. Air samples were collected by MAS-100 sampler. The number of mesophilic bacteria, mould fungi, actinomycetes and staphylococci was determined on general and selective media. The species identification of staphylococci was conducted using API tests for strains that were pre-selected based on macroscopic and microscopic observations. Results: A total number of 1,584 samples were collected during the sampling period. The numbers of airborne microorganisms varied between the examined premises and between the seasons of the year. The observed differences were statistically significant with one exception for actinomycetes and their differences between the examined premises. The concentrations of mesophilic bacteria varied from 5 to 297 CFU/m3 of air, for Staphylococcus the values ranged from 1 to 96 CFU/m3, for fungi - from 1 to 100 CFU/m3, and the number of actinomycetes ranged from 7 to 321 CFU/m3. Ten species of coagulase-negative staphylococci (CoNS) were identified among 55 isolates with S. saprophyticus and S. warneri being the most frequently detected (n = 14 and 13, respectively). S. haemolyticus, which is one of the most common causal agents of nosocomial infections was observed in four facilities (n = 5). Conclusions: The microbial concentrations varied both between the seasons of the year and between the examined facilities. The highest bioaerosol concentrations were observed in most crowded premises. The identified species of staphylococci, although not typically associated with human infections, are common causal agents of nosocomial infections and infections in immunocompromised people.

Natural polysaccharides, including hyaluronic acid, find a wide range of applications in biomedical sciences. There is a growing interest in nanocomposites containing hyaluronic acid and nanoparticles such as nanometals or graphene. In this study, we prepared foils of pure sodium hyaluronate and sodium hyaluronate containing nanosilver, graphene oxide, nanosilver/graphene oxide and characterized their properties. UV-vis spectroscopy and scanning electron microscopy (SEM) confirmed the formation of 10–20 nm silver nanoparticles. The structural changes were investigated using Fourier transforms infrared (FTIR) spectra and size exclusion chromatography. The obtained results suggest changes in molecular weights in the samples containing nanoparticles, which was highest in a sample containing nanosilver/graphene oxide. We also assessed the mechanical properties of the foils (thickness, tensile strength and elongation at break) and their wettability. The foils containing nanosilver and nanosilver/graphene oxide presented bacteriostatic activity against E. coli, Staphylococcus spp. and Bacillus spp., which was not observed in the control and sample containing graphene oxide. The composites containing graphene oxide and nanosilver/graphene oxide exhibited a cytotoxic effect on human melanoma WM266-4 cell lines (ATCC, Manassas, VA, USA).

Summary The bacterial nodulation (nod) genes are essential in the formation process of root nodules. This study was aimed to verify the occurrence of nodule-associated bacteria in two pea varieties (“Tarchalska” and “Klif ”) inoculated with Rhizobium inoculants – Nitragine™ and a noncommercial one produced by the Polish Institute of Soil Science and Plant Cultivation (IUNG). The number of colonies isolated on yeast extract mannitol (YEM) agar from the nodules of “Klif ” inoculated with IUNG inoculants was significantly higher than the number of colonies isolated from other variants. Species identification was based on sequencing of 16S rDNA, which revealed that despite careful sterilization of nodules, sequences of other bacterial species were detected. Among them, one sequence belonged to Rhizobium leguminosarum (isolated from IUNG inoculant). To assess the presence of nodulation-capable Rhizobium, amplification of the nodC gene was performed, which revealed that of 29 samples, 19 were positive. The remaining isolates, including reference strain and bacteria isolated from Nitragine™, lacked this gene. The results show that pea nodules harbor a very diverse community of bacteria. The lack of nodC gene in some strains isolated from plants inoculated with Nitragine™ and with IUNG inoculant proves that even if R. leguminosarum are abundant, they may not be efficient in nodulation.

Currently, there are almost no studies concerning bioaerosol and particulate matter levels in animal enclosures of zoos. Numerous air contamination sources can be found there, and zoos are both working environments and popular tourist objects. The aim of this study was to assess bacterial aerosol levels in premises for animals (giraffes, monkeys, pheasants and ostriches) of the Krakow Zoo. Bioaerosol samples were collected using six-stage Andersen impactor to assess the concentration and size distribution of airborne mesophilic bacteria, Gram-negative bacteria and staphylococci. Particulate matter levels (PM10, PM4, PM2.5 and PM1) were assessed using DustTrak Aerosol Monitor. The results showed that the concentration of airborne bacteria varied significantly between enclosures for the analyzed animal groups and various groups of bacteria reached their maximum concentrations in premises for different animals. The mean share of respirable fraction of bacteria was quite high—68%, with values ranging from 12.8% for Gram-negative bacteria to 98% for mannitol-positive staphylococci, indicating potential harmfulness to the health of exposed people. Dust concentrations remained at similar, relatively low levels (maximum of 0.105 mg/m3 for PM10 in rooms for giraffes). This study showed that neither bacterial bioaerosol levels, nor dust concentrations exceed the permissible values provided by Polish guidelines, but comparing the values with other countries’ limits suggests that the air could be considered as contaminated. The animals are a significant source of bioaerosol; therefore, attention should be paid to thorough cleaning of animals themselves and their premises and to maintaining appropriate levels of microclimatic parameters prevailing in the rooms.

The aim of the study was to determine the antimicrobial resistance profile and the occurrence of extended-spectrum beta-lactamase genes and to analyze the genetic diversity of Escherichia coli strains isolated from the environment of horse riding centers. The study was conducted using E. coli strains isolated from the air, manure, and horse nostril swabs in three horse riding centers differing in the system of horse keeping-stable (OJK Pegaz and KJK Szary) and free-range (SKH Nielepice). Resistance to antibiotics was determined using the disk-diffusion method, and the PCR technique was employed to detect the extended-spectrum β-lactamase (ESBL) genes, while the genetic diversity of strains was assessed by rep-PCR. A total of 200 strains were collected during the 2-year study, with the majority isolated from KJK Szary, while the smallest number was obtained from SKH Nielepice. The strains were mostly resistant to ampicillin, aztreonam, and ticarcillin. The tested strains were most frequently resistant to one or two antibiotics, with a maximum of ten antimicrobials at the same time. Two multidrug-resistant (MDR) strains were detected in OJK Pegaz while in KJK Szary there were two MDR and one extensively drug-resistant (XDR) strain. The ESBL mechanism was most frequently observed in OJK Pegaz (20.31% of strains) followed by KJK Szary (15.53% of strains) and SKH Nielepice (15.15% of strains). Among the ESBL-determining genes, only blaTEM and blaCTXM-9 were detected-blaTEM was mostly found in KJK Szary (53.40% of strains), while the second detected gene-blaCTXM-9-was most frequent in SKH Nielepice (6.06% of strains). The rep-PCR genotyping showed high variation among the analyzed strains, whereas its degree differed between the studied facilities, indicating that the type of horse keeping (stable vs. free-range) affects the genetic diversity of the E. coli strains. Having regard to the fact that the tested strains of E. coli were derived from non-hospitalized horses that were not treated pharmacologically, we can assume that the observed antimicrobial resistance may be of both-natural origin, i.e., not the result of the selection pressure, and acquired, the source of which could be people present in the horse riding facilities, the remaining horses which were not included in the study, and air, as well as water, fodder, and litter of the animals. Therefore, it can be concluded that the studied horses are the source of resistant E. coli and it is reasonable to continue monitoring the changes in antimicrobial resistance in those bacteria.

This study aimed to determine the incidence of antibiotic-resistant strains of Escherichia coli in waters of three Lesser Poland rivers: Raba, Prądnik and Bialka. The samples were collected from 30 sites—10 from each river along their course. Membrane filtration method was used to determine the water quality based on the prevalence of fecal coliforms in water. The contaminated samples were used to isolate Escherichia coli for further studies. After confirming the systematic position of randomly isolated strains, the antibiotic resistance tests were performed using the disk-diffusion method following the recommendations of the National Reference Centre for Susceptibility Testing. Escherichia coli were present in all rivers. Complete antibiograms were obtained for 83 isolates; 30 strains originating from the Prądnik river, 37 from Raba and 16 from Bialka. A total of 67.47% strains were resistant to at least 1 tested antimicrobial (76.7% isolates from Prądnik, 73% from Raba and 37.5% from Bialka), 33.7% of all strains were resistant to at least 5 antimicrobial agents, and one strain was resistant to even 13 antibiotics. The highest resistance was observed for ampicillin and the lowest for imipenem, fosfomycin and meropenem. The obtained results showed that the resistant strains of Escherichia coli are widely spread in the water environment, even in protected areas.

Mountain areas in Poland are among the most frequented tourist destinations and such intensive tourism negatively affects the natural environment. The COVID-19 pandemic and the resulting lockdown restricted travel for a few months in 2020, providing a unique opportunity to observe the studied mountain environment without the impact of typical tourist traffic. This study is based on the determination of antibiotic content, hydrochemical parameters, enumeration of culturable bacterial water quality indicators, antimicrobial susceptibility tests together with extended spectrum beta-lactamase (ESBL) gene detection in waterborne E. coli and NGS-based bacterial community composition at six sites along the Bialka river valley (one of the most popular touristic regions in Poland) in three periods: in summer and winter tourist seasons and during the COVID-19 lockdown. The results of individual measurements showed decreased numbers of bacterial indicators of water contamination (e.g. numbers of E. coli dropped from 99 × 104 CFU/100 ml to 12 CFU/100 ml at the most contaminated site) and the share of antimicrobial resistant E. coli (total resistance dropped from 21% in summer to 9% during lockdown, share of multidrug resistant strains from 100 to 44%, and ESBL from 20% in summer to none during lockdown). Antibiotic concentrations were the highest during lockdown. The use of multivariate analysis (principal component analysis - PCA and heatmaps) revealed a clear pattern of tourism-related anthropogenic pressure on the water environment and positive impact of COVID-19 lockdown on water quality. PCA distinguished three major factors determining water quality: F1 shows strong effect of anthropogenic pressure; F2 describes the lockdown-related quality restoration processes; F3 is semi-natural and describes the differences between the most pristine and most anthropogenically-impacted waters.

This study was aimed to determine the anthropogenic and natural factors affecting spatial and temporal changes in the physicochemical parameters and bacterial indicators of water quality in the river Białka. The impact of intensive development of the tourist infrastructure on the quality of river water and the potential health threats to tourists was also assessed. Water samples were collected over a period of 2.5 years, once per each month in four sites along the river. Temperature, electrolytic conductivity, pH, and water level were measured onsite; flow rate data were acquired from the Institute of Meteorology and Water Management; chemical analyses allowed to determine the amount of fourteen ions, while microbiological indicators included total and thermotolerant coliforms, total and thermotolerant Escherichia coli, and mesophilic and psychrophilic bacteria. The combination of hydrological, hydrochemical, and microbiological methods generated large amount of data, which were processed by multivariate statistical analysis. A downstream cumulative effect was observed in the contamination of the river water. Fecal coliforms and E. coli were detected in all sites, suggesting the source of fecal contamination even in the protected areas. Intensive development of a ski resort and the related infrastructure, together with the need to accommodate numerous tourists in the examined region, has an evident environmental impact. The resulting deterioration of water quality poses health risks to tourists, as water from the Białka river is used for a variety of purposes, including as a raw drinking water or for artificial snowing of ski slopes. The seasonal changes in the physicochemical parameters mainly result from varying natural factors that shape the water quality in the studied region. The differences in the number of analyzed microorganisms result from seasonal variation in touristic activity and are affected mostly by point sources of sewage inflow.

Despite intensive research no therapies targeted against the oncogenic EGFRvIII are present in the clinic. One of the reasons is the elusive nature of the molecular structure and activity of the truncated receptor. The recent publications indicate the EGF-bound wild-type EGFR to trans-phosphorylate the EGFRvIII initiating aberrant signaling cascade. The elevated stability of the mutant receptor contributes towards oncogenic potential, preventing termination of signaling by receptor degradation. Here, we show that inhibition of phosphatases leads to a marked increase in phosphorylation of wild-type EGFR and EGFRvIII, indicating that both undergo cyclic rounds of phosphorylation and dephosphorylation on all investigated tyrosine residues, including Tyr1045. Still, we observe elevated stability of the mutant receptor, suggesting phosphorylation as insufficient to cause degradation. Hyperphosphorylation of EGFRvIII was hindered only by EGFR tyrosine kinase inhibitors. Co-immunoprecipitation as well as semi-native Western blotting structural analyses together with functional investigation of EGFRvIII's phosphorylation following depletion of wild-type EGFR by shRNA or EGF-mediated degradation indicated homodimerization as the predominant quaternary structure of the mutant receptor. Dimers were observed only under non-reducing conditions, suggesting that homodimerization is mediated by covalent bonds. Previous reports indicated cysteine at position 16 to mediate covalent homodimerization. Upon its substitution to serine, we have observed impaired formation of dimers and lower phosphorylation levels of the mutated oncogene. Based on the obtained results we propose that EGFRvIII is predominantly regulated dynamically by phosphatases that counteract the process of trans-phosphorylation occurring within the homodimers.

Relict charcoal hearths (RCHs) increases soil fertility in forest ecosystems. However, the effects of RCHs on the activity and abundance of soil microorganisms remain unknown. In this paper, we analysed the impact of relict charcoal production on the soil enzymatic activity and composition of soil bacterial and fungal communities in Scots pine forests of the Manowo Forest District in northern Poland. Moreover, we determined the effect of relict charcoal production on the soil properties. Our research was conducted by comparing the physical, chemical, enzymatic and microbiological properties of charcoal-enriched and charcoal-free soils. Significant differences in physical properties were found between these two soil types in terms of their structure and water holding capacity. As expected, horizons enriched with charcoal were characterised by a significantly higher organic carbon content (4.7% on average compared to 2.2% in control horizons), and also by a considerably higher content of available phosphorus (an average of 64.07 mg·kg−1 compared to 36.21 mg·kg−1 in the control). Similarly, RCH horizons displayed a higher pH and higher contents of Ca and Na cations. These results indicated that RCH soils provided more favourable conditions for the soil microbiome, as reflected by the higher enzymatic activity and diversity of the microorganisms. Moreover, bacterial and fungal communities in RCH soils were more diverse and had greater species/genera richness, especially in the case of fungi. Members of the genus Rhodoplanes dominated the bacterial community at both RCH and non-RCH sites, followed by Streptomyces, Burkholderia, Skermanella, Tsukamurella and Candidatus Solibacter. Both culture- and next generation sequencing (NGS)-based analyses showed that soil fungal communities were dominated by Ascomycota, with Penicillium as the most abundant genus. Our results showed that hearth soils may represent a significant C pool in the forest ecosystem. This study supports the strategy of safeguarding such charcoal-enriched soils as precious C reservoirs and ecologically important biodiversity hotspots. Moreover, the application of charcoal may effectively increase the microbial diversity of forest soils, especially during the reforestation or re-cultivation of disturbed habitats.

Phenotypic traits of lichens can be greatly modified by environmental factors. Granulose thalli on soil and podetia, densely covered with granules, referring to common and widespread lichen Cladonia cervicornis subsp. verticillata were found near zinc smelter. The granules are stratified, filled with fungal medulla and heavily encrusted with calcium oxalate weddellite crystals, not observed on regularly developed thalli of the species. Phylogenetic analysis revealed that deformed granulose forms belong to this taxon, showing that the phenotypic plasticity of the lichens of Cladonia can lead to the emergence of features that do not coincide with the taxonomic definition of the species. The heavy-metal accumulation capacity of both granulose and regular form of primary and secondary lichen thallus, in relation to the element content in corresponding substrate, was determined. Granulose-modified thalli accumulate greater amounts of heavy metals than regular ones, meaning that the bioaccumulation property of a given species may be greatly affected by morphological modifications. The granulose forms are also characterised by considerably higher ratios of Cd, Pb and As concentrations in lichen samples in relation to the corresponding substrates than regular ones. This means that collection of variously formed thalli should be avoided in biomonitoring sampling procedures. The results indicate that a substantial part of the element load, in particular zinc, in the examined lichen thalli collected near the smelter originates from atmospheric fallout. Electronic supplementary material The online version of this article (10.1007/s11356-017-0639-1) contains supplementary material, which is available to authorized users.

Microorganisms may cause the almost complete degradation of a material structure, even though symptoms of drastic microbial damage may not be apparent macroscopically. Due to their enzymatic activities and outstanding adaptation abilities to various environmental conditions, actinomycetes constitute a particular threat to a variety of materials. They are able to use, and subsequently degrade any substrate, whether this happens in a matter of days, weeks or years. Given the frequency of occurrence and a number of features that enable Streptomyces to growth on materials, more studies should be undertaken on methods allowing for their proper and rapid identification. Taxonomic studies gain particular importance in the case of damaged valuable historic materials. This paper focuses on identification of actinomycetes isolated from various historic and contemporary materials, including textiles, parchment and ceramics. Strengths and weaknesses of approaches that have been chosen for actinomycetes taxonomic studies are described.

Bacteria of the genus Staphylococcus were isolated from air sampled from living spaces in Krakow (Poland). In total, 55 strains belonging to the genus Staphylococcus were isolated from 45 sites, and 13 species of coagulase-negative staphylococci were identified. The species composition of studied airborne microbiota contains Staphylococcus species that are rarely infectious to humans. Most commonly isolated species comprised S. hominis and S. warneri. The disk-diffusion tests showed that the collected isolates were most frequently resistant to erythromycin. The PCR technique was employed to search for genes conferring the resistance in staphylococci to antibiotics from the group of macrolides, lincosamides and streptogramins. The analyzed Staphylococcus isolates possessed simultaneously 4 different resistance genes. The molecular analysis with the use of specific primers allowed to determine the most prevalent gene which is mphC, responsible for the resistance to macrolides and for the enzymatic inactivation of the drug by phosphotransferase. The second most often detected gene was msrA1, which confers the resistance of staphylococci to macrolides and is responsible for active pumping of antimicrobial particles out of bacterial cells.

This study aimed to gain insight into the presence of antibiotics, occurrence of antimicrobial resistance and prevalence of extended-spectrum β-lactamase (ESBL) genes in Escherichia coli in surface water, based on the example of the Bialka river, located in one of the most attractive tourist destinations in Poland. Water samples were collected in three sites: in the Tatra National Park (TNP), by the sewage discharge from the local treatment plant (STP) and c.a. 3 km downstream (DSTP). The analyses included determination of antibiotic content, enumeration of bacterial indicators of poor water quality, isolation and identification of Escherichia coli, which was subjected to antimicrobial susceptibility tests and assessment of ESBL-determining genes. Fourteen antimicrobials out of 24 tested were detected in river waters in varying concentrations. Trimethoprim and ofloxacin were most frequently detected. Most antibiotics were absent in the TNP, the highest numbers and the highest concentrations of antibiotics were observed by the STP discharge to decrease their content downstream. Culture-based tests of microbiological contamination showed similar results. Resistance to ampicillin was most frequent (64.5% strains), followed by cefazolin (50%). 20.6% of strains were ESBL-positive, while ESBL-determining gene, blaTEM was detected in 23.8% of E. coli strains. The largest percentage of antibiotic resistant and MDR E. coli strains was detected nearby the STP, indicating that malfunctioning STP may contribute largely to river water contamination downstream, also having significant environmental and economic impact.

Parenthood is one of the most important roles people have in their lives. This role is unique when parents’ child is disabled, ill or suffers from developmental disorders. The empirical study presented in this article refers to the description of parenthood in the case of parents of children with autistic spectrum disorders. The author attempted to determine how mothers and fathers performed their parental roles as well as their meaning of life level and relationships between these variables. The study covered 80 parents of children with autistic spectrum disorders. The following research tools were used: the Parental Role Performance Questionnaire by Żaneta Stelter and the Purpose in Life test (PIL) by J.C Crumbach and L.T. Maholik. Analyses showed significant connections between the variables measured.

Introduction. Streptococcus agalactiae infections are among the most significant causes of neonatal invasive diseases. Proper screening and detection of pregnant women carrying GBS allows intrapartum administration of antibiotic prophylaxis and is an effective measure in preventing transmission of bacteria from mother to newborns. Material and methods. Sixty three bacterial strains were isolated from vaginal swabs from pregnant and nonpregnant women of reproductive age. Species were identified by colony morphology, haemolysis type, Gram staining and SLIDEX® Strepto Plus latex test. Antimicrobial resistance of 56 strains was determined using disk-diffusion method. The presence of molecular resistance determinants was assessed using PCR with specific primers, and capsular types were identified using multiplex PCR. Results. None of the strains were resistant to the first drug of choice, penicillin. A large percentage of isolates (78.6%) were resistant to doxycycline. The prevalence of resistance to macrolides and lincosamides, antibiotics used in women allergic to penicillin, was high. Those results corresponded with PCR tests, as tetM and ermA1 were most frequently detected genes (98.4 and 87.3%, respectively). 7.94% of strains possessed 7 different out of 13 tested genes determining resistance to different groups of antimicrobials. Among the capsular types, Ia, which proved to be associated with the most severe and invasive infections in mothers and neonates, was the most prevalent (65.08%). Conclusions. Even though they are susceptible to penicillin, multidrug resistance is common among S. agalactiae strains isolated from women of reproductive age and this resistance can be caused by more than one gene per single isolate

Certain lichens are effective colonisers of polluted sites. However, little is known about the tolerance of photobionts and the degree of mycobiont selectivity to photobionts relative to metal pollution. The present study recognises the genetic and anatomical diversity of Asterochloris photobionts in epigeic lichens, i.e. Cladonia cariosa, C. rei, and Diploschistes muscorum, in relation to a wide spectrum of soil pollution. In accordance with phylogenetic analysis, photobionts were clustered in 7 moderately- to well-supported clades, including 19 haplotypes. The mycobionts of all studied lichens demonstrated a low level of selectivity and were capable of associating with various Asterochloris lineages. This tendency was also expressed by the frequent (~25%) occurrence of multiple algal genotypes in a single thallus. This indicates that identified Asterochloris lineages are generally tolerant to heavy-metal pollution, and the low level of selectivity of mycobionts enables them to select the most suitable and/or available partner. The trend of increasing incidence of certain Asterochloris lineages and decreasing frequency of others along with increasing soil pollution was observed. This proves the superior adaptation of some photobionts to polluted sites. Such symbiotic plasticity constitute an adaptive feature necessary for the successful colonisation. High number of haplotypes at polluted sites could be the result of multiple introduction events from different areas during the initial stages of spontaneous succession. Regardless of the genetic pattern, Asterochloris cells were considerably smaller, and the density and compaction of cells in the algal layer were higher, in lichen specimens from polluted sites, indicating that photobiont characteristics may be closely dependent on heavy-metal pollution.

Introduction Atherosclerotic plaque formation is an inflammatory process that involves the recruitment of neutrophil granulocytes and the generation of reactive oxygen species (ROS). ROS formation by myeloperoxidase, a key enzyme in H2O2 degradation, can be modulated by addition of sodium thiocyanate (NaSCN). However, the therapeutic use of NaSCN to counteract atherogenesis has been controversial, because MPO oxidizes NaSCN to hypothiocyanous acid, which is a reactive oxygen species itself. Therefore, this study aimed to investigate the effect of NaSCN treatment on atherogenesis in vivo. Methods Apolipoprotein E knockout (ApoE−/−) mice on western-diet were treated with NaSCN for 8 weeks. Blood levels of total cholesterol, IL-10, and IL-6 were measured. Aortic roots from these mice were analyzed histologically to quantify plaque formation, monocyte, and neutrophil granulocyte infiltration. Oxidative damage was evaluated via an L-012 chemiluminescence assay and staining for chlorotyrosine in the aortic walls. Endothelial function was assessed by use of endothelium-dependent vasodilation in isolated aortic rings. Neointima formation was evaluated in wild-type mice following wire injury of the carotid artery. Results NaSCN treatment of ApoE-/- mice lead to a reduction of atherosclerotic plaque size in the aortic roots but had no effect on monocyte or granulocyte infiltration. Serum levels of the pro-inflammatory cytokine IL-6 decreased whereas anti-inflammatory IL-10 increased upon NaSCN treatment. In our experiments, we found oxidative damage to be reduced and the endothelial function to be improved in the NaSCN-treated group. Additionally, NaSCN inhibited neointima formation. Conclusion NaSCN has beneficial effects on various stages of atherosclerotic plaque development in mice.