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The ability to distinguish among the subspecies of Pasteurella multocida isolates is important epidemiologically; however, classification at the subspecies level based on the results of conventional biochemical tests (fermentation of sorbitol and dulcitol) is reportedly not accurate in all cases. Therefore, we developed a rapid, multiplex PCR assay to differentiate among the 3 subspecies of P. multocida. The PCR assay includes the P. multocida species–specific primers KMT1SP6 and KMT1T7 as an internal amplification control, with a newly designed gatD (galactitol-1-phosphate-5-dehydrogenase)-specific primer pair (unique for subsp. gallicida), and primers targeting a 16S rRNA gene region specific for subsp. septica. The subspecies specificity of the PCR was demonstrated by applying the test to a collection of 70 P. multocida isolates, including the Heddleston serovar reference strains; all isolates and strains were assigned correctly. The PCR assay is a sensitive, specific, and highly effective method for the identification of P. multocida subspecies, and an alternative to biochemical test–based differentiation. A possible relationship was noticed between P. multocida subspecies and lipopolysaccharide (LPS) genotype; all but one of the subsp. gallicida strains were isolated only from avian hosts and represented L1 LPS genotype. Subsp. multocida and subsp. septica isolates were classified into 5 and 4 different LPS genotypes, respectively, of which L3 was the only LPS genotype shared between these 2 subspecies.

Megatrends are slow processes that are hardly noticeable initially, but later cause long-term global effects. The European Environmental Agency (EEA) has set 11 global megatrends. Some of them - globalization, risk of pandemic, technological development and climate change - have major impacts on microfungi in a vulnerable region of Europe, the Pannonian Biogeographical Region. Due to the globalization, high amount of inocula is imported with tropical fruits, soil and packaging materials. Due to technological development, these fungi gain new habitats in the immediate surroundings of humans. In most of the cases these are considered to be extreme environments for which fungi can increasingly adapt. As a result of climate change, non-native species are more likely to colonize the natural habitats in the region. Some of these fungal species have great effect on human health and agriculture. The effects of global megatrends on fungi raise new issues not only from the point of view of economy and health, but also from plant protection and environmental perspectives. Because of the increasing presence of these fungal species, it is important to take them into account during the development of adaptation strategies.

Consuming “sprouted seeds” is one of the most important factors of a healthy diet. An experiment was conducted in the University of Debrecen, Research Centre of Nyíregyháza (Hungary) in 2014 to analyse some morphological traits of four winter wheat genotypes (Triticum aestivum L.) and one spelt (T. spelta) variety. Our results showed that the spelt wheat variety “Franckenkorn” could maintain higher root length throughout the experimental period. On average, both “Perbetei” and “Franckenkorn” varieties could maintain higher root number compared to the other varieties. The extensive breeding line “1401 HK” had the highest shoot length throughout the whole experiment, being significantly higher than the landrace variety “Perbetei” and both of the varieties “KG Bendegúz” and “KG Kunhalom”. It could be concluded that “KG Bendegúz” cultivar and “Perbetei” landrace seem to be the most suitable for aquaculture techniques. In addition, “1401 HK” breeding line can be the most suitable for the production of juice since the minimal required shoot length (12 cm) was achieved within the shortest period of time after sowing (9 days). This breeding line and “Franckenkorn” can also be suitable for production of “wheatgrass”, because it is consumed without roots. Further research is needed to evaluate nutritional values of these genotypes.

This study presents the results of a Europe-wide training and Quality Control (QC) exercise carried out within the framework of the European Aerobiology Society’s QC Working Group. The main aim of this exercise was to examine the feasibility of carrying out a QC exercise for fungal spore monitoring in Europe, using a similar methodology to the one previously used for pollen. The QC survey was conducted in two parts: (1) Coordinators of national and regional aerobiological networks in Europe involved in the monitoring of atmospheric fungal spores were invited to complete a questionnaire survey related to their network and asked whether they were interested in taking part in an external inter-laboratory QC exercise; (2) Participating networks performed an inter-laboratory ring test with the same sample slide in order to determine the reproducibility of identifying and counting two fungal spore taxa (Alternaria and Epicoccum) in air samples collected by a Hirst-type volumetric spore trap. Participants were instructed to read five separate longitudinal transects in the “effective collecting area” of the slide. Reproducibility of analysis was determined following the method previously used in the European Aerobiology Society’s QC exercises for pollen. Thirty-two counters from 16 national or regional networks in Europe participated in the QC exercise. Coefficients of Variation (CV%) ranged from 23.0 to 22.5 when reading one transect and from 14.0 to 16.0 when reading five transects for Alternaria and Epicoccum, respectively. Considering a CV% of 30 as the limit for fungal spores, no significant differences were observed between the absolute errors from two, three, four and five transects. The only significant difference was between one and five transects. We recommend that fungal spore analysis should be carried out on about 5% of the slide (two transects in this study) because results were not significantly different to five transects.

Photosystem II (PSII) uses solar energy to oxidize water and delivers electrons for life on Earth. The photochemical reaction center of PSII is known to possess two stationary states. In the open state (PSIIO), the absorption of a single photon triggers electron-transfer steps, which convert PSII into the charge-separated closed state (PSIIC). Here, by using steady-state and time-resolved spectroscopic techniques on Spinacia oleracea and Thermosynechococcus vulcanus preparations, we show that additional illumination gradually transforms PSIIC into a light-adapted charge-separated state (PSIIL). The PSIIC-to-PSIIL transition, observed at all temperatures between 80 and 308 K, is responsible for a large part of the variable chlorophyll-a fluorescence (Fv) and is associated with subtle, dark-reversible reorganizations in the core complexes, protein conformational changes at noncryogenic temperatures, and marked variations in the rates of photochemical and photophysical reactions. The build-up of PSIIL requires a series of light-induced events generating rapidly recombining primary radical pairs, spaced by sufficient waiting times between these events--pointing to the roles of local electric-field transients and dielectric relaxation processes. We show that the maximum fluorescence level, Fm, is associated with PSIIL rather than with PSIIC, and thus the Fv/Fm parameter cannot be equated with the quantum efficiency of PSII photochemistry. Our findings resolve the controversies and explain the peculiar features of chlorophyll-a fluorescence kinetics, a tool to monitor the functional activity and the structural-functional plasticity of PSII in different wild-types and mutant organisms and under stress conditions. VC American Society of Plant Biologists 2021. All rights reserved

Background & Aims Liver fibrosis is a multifactorial trait that develops in response to chronic liver injury. Our aim was to characterize the genetic architecture of carbon tetrachloride (CCl4)-induced liver fibrosis using the Hybrid Mouse Diversity Panel, a panel of more than 100 genetically distinct mouse strains optimized for genome-wide association studies and systems genetics. Methods Chronic liver injury was induced by CCl4 injections twice weekly for 6 weeks. Four hundred thirty-seven mice received CCl4 and 256 received vehicle, after which animals were euthanized for liver histology and gene expression. Using automated digital image analysis, we quantified fibrosis as the collagen proportionate area of the whole section, excluding normal collagen. Results We discovered broad variation in fibrosis among the Hybrid Mouse Diversity Panel strains, demonstrating a significant genetic influence. Genome-wide association analyses revealed significant and suggestive loci underlying susceptibility to fibrosis, some of which overlapped with loci identified in mouse crosses and human population studies. Liver global gene expression was assessed by RNA sequencing across the strains, and candidate genes were identified using differential expression and expression quantitative trait locus analyses. Gene set enrichment analyses identified the underlying pathways, of which stellate cell involvement was prominent, and coexpression network modeling identified modules associated with fibrosis. Conclusions Our results provide a rich resource for the design of experiments to understand mechanisms underlying fibrosis and for rational strain selection when testing antifibrotic drugs., Graphical abstract

This work considers dust deposition and the heavy metal (HM) content on leaves of urban trees (Acer platanoides L. ‘Globosum,’ Fraxinus excelsior L. ‘Westhof’s Glorie’ and Tilia tomentosa Moench.) in order to estimate the trees’ capacity to remove dust and HM from the air. Leaves were collected from the Buda Arboretum and from different streets of heavy traffic in Budapest, Hungary, during 2015 and 2016. At each site, five trees were sampled by collecting 6 leaves from each tree from the height of 2–3 m. Dust deposits on the leaves were removed by soaking the fresh foliage in distilled water for 20 h and then washed with ultrasound shaking. Afterward, the leaves were dried to constant weight and then they were digested in nitric acid–hydrogen peroxide treatment, and their Pb, Fe, Ni, Zn and Cu contents were measured using an inductively coupled plasma (ICP AS) spectrometer. The removed dust deposit was dried, and after a similar digestion treatment the Pb, Fe, Ni, Zn and Cu contents were measured using an AURORA AI 1200 AAS appliance. The HM deposit was calculated in mg m–2 leaf surface area. In 2015, the amount of foliar dust deposit from spring to autumn increased from 86.3 to 270.2 mg m–2. The most efficient tree species in trapping dust on their leaves was the silver linden (98.5–123.5 mg m−2), followed by the Norway maple (74.2–84.8 mg m−2) and the common ash (62.8–74.6 mg m−2). The deposit of HM elements showed seasonal differences: the quantity of Fe and Pb deposit on autumnal leaves increased five- to tenfold, while other heavy metals did not show accumulation. Silver linden with its pubescent (hairy) leaf surface proved to be most efficient in entrapping and retaining dust and heavy metals. The 60–100% higher Pb and Fe content of autumnal leaves indicate that over the season leaves may absorb Fe and Pb from the foliar dust. Our results confirmed that the foliar dust is a potential indicator for monitoring the HM content in the air. We also show that foliar dust deposits should be considered when estimating the capacity of urban trees to clean the air.

The occurrence of members of the Pasteurellaceae and Neisseriaceae families was studied in dogs and cats. A total of 110 nasal and pharyngeal swab samples from 47 dogs and 8 cats were collected. Most of the strains were identified by 16S rDNA sequencing, except Frederiksenia canicola and Pasteurella multocida where species-specific polymerase chain reactions were applied. The most frequently isolated species was F. canicola, which occurred only in dogs, mainly in the pharyngeal cavity. The second commonest bacterium, P. multocida was found in both types of samples and in both hosts. Other species from the family Pasteurellaceae, such as Haemophilus haemoglobinophilus, Pasteurella canis and P. dagmatis, were detected only in dogs. All isolated species belonging to the family Neisseriaceae, mainly representing Neisseria weaveri, were found only in the pharyngeal cavity. Neisseria weaveri and N. zoodegmatis could be detected in both hosts. Neisseria dumasiana and N. canis were isolated from dogs, while N. shayeganii only from a cat. For phylogenetic analysis, rpoB gene sequencing was performed, where the strains were on monophyletic branches and clearly separated from each other. In this study, recently described species such as F. canicola, N. shayeganii and N. dumasiana were detected that had never been isolated in Hungary before.

Allergic diseases are considered as important human health issues as they substantially restrict many allergic people. Trees such as the plane tree can pose a certain threat to allergy sufferers. Due to the frequent planting of these trees in large cities, their pollen can affect the increase in the frequency of local allergy symptoms. This study compares the plane tree pollen seasons in Poland, in Bialystok, Bydgoszcz, Warsaw, Zielona Gora, Piotrkow Trybunalski, Opole, Olsztyn, Szczecin and in Hungary, in Budapest, Debrecen, Gyor, Kaposvar, Kecskemet, Miskolc, Nyiregyhaza and Pecs in 2019. The investigations were carried out using the volumetric method (Hirst type pollen sampler). Seasonal Pollen Index (SPI) was estimated as the sum of daily average pollen concentrations in the given season. The season ranges from March to May, depending on the geographical latitude. Diagnosis of plane tree pollen allergy is made difficult due to some cross-reactivity with birch, but also alder, hazel, hornbeam, oak, beech, sweet chestnut, and to some extent with grass pollen. In 2019 the pollen season of Platanus started first in Hungary, in Pecs on the April 1st; in Poland the pollen season started in Szczecin and Zielona Gora on the April 4th. At the latest, a pollen season ended in Poland, in Warsaw and Bydgoszcz until May 11th–12th, similarly in Hungary – until May 11th in Budapest and Kaposvar. The differences of pollen seasons duration were very considerable, from 15 to 40 days. Also the differences of the highest airborne concentration especially between both countries were extremely considerable (2105 pollen grains/m3 in Nyíregyháza and 3 pollen grains/m3 in Białystok. The maximum values of seasonal pollen count in Polish cities occurred between April 24th and May 1st, and in Hungarian cities between April 9th and 16th. The highest plane tree pollen allergen hazard occurred in 2019 undoubtedly in Hungary: in Pecs, Kaposvar and Nyiregyhaza, and was ten times higher than in Poland. The highest variability in the analysed seasons was found in the peak value and annual total.

Starmerella bacillaris (synonym Candida zemplinina) is an important non-Saccharomyces yeast in winemaking with valuable oenological properties, accompanying Saccharomyces species in sweet wine fermentation, and has also been suggested for application as combined starter culture in dry or sweet wines. In this study, the major metabolites and nitrogen utilization of these yeasts are evaluated in the musts with high or extremely high sugar concentration. The change in the metabolic footprint of Saccharomyces cerevisiae, Saccharomyces uvarum and Starmerella bacillaris strains was compared when they were present as pure cultures in chemically defined grape juice medium with 220 and 320 g/L of sugar, to represent a fully matured and an overripe grape. Surprisingly, the extreme sugar concentration did not result in a considerable change in the rate of sugar consumption; only a shift of the sugar consumption curves could be noticed for all species, especially for Starmerella bacillaris. At the extreme sugar concentration, Starmerella bacillaris showed excellent glycerol production, moderate nitrogen demand together with a noticeable proline utilisation. The change in the overall metabolite pattern of Starmerella bacillaris allowed clear discrimination from the change of the Saccharomyces species. In this experiment, the adequacy of this non-Saccharomyces yeast for co-fermentation in juices with high sugar concentration is highlighted. Moreover, the results suggest that Starmerella bacillaris has a more active adaptation mechanism to extremely high sugar concentration.

Siliceous marine microfossils were unexpectedly discovered during the analysis of flower honey samples from Poland and Tunisia. The microfossils were represented by protist with siliceous skeletons: silicoflagellates, diatoms, and endoskeletal dinoflagellates. This is the first record of such microfossils in honeys. Based on the high percent of anemophilous pollen grains and spores in the sample, it was hypothesized that silicoflagellates were deposited from the air onto the nectariferous flowers, then bees harvested them with the nectar. Based on the comparison of pollen content of honeys and flowering calendar of Tunisia, the harvest time of honey was identified as a period between 1 April and 31 May 2011. Trajectory analysis of air masses in this period confirmed that siliceous microfossils could be aerosolized by wind from the rocks of the so-called Tripoli Formation of Messinian age (6–7 Ma). Similar to the Tunisian case, the Polish trajectory simulation also supports the hypothesis of atmospheric transport of silicoflagellates from outcrops of Oligocene age in the Polish Outer Carpathians. In the case of diatom content of honey, however, the source can be both natural (wind) and artificial (diatomaceous earth filters). For a correct determination, natural sources of siliceous bioparticles, such as wind transport from nearby outcrops should be also considered. Silicoflagellates could be used as complementary indicators of the geographical origin of honeys collected in areas characterized by diatomite outcrops, supporting the results obtained with other methods, thus, such indicators merit further studies within the area of honey authenticity.

To understand antibiotic resistance in pathogenic bacteria, we need to monitor environmental microbes as reservoirs of antimicrobial resistance genes (ARGs). These bacteria are present in the air and can be investigated with the whole metagenome shotgun sequencing approach. This study aimed to investigate the feasibility of a method for metagenomic analysis of microbial composition and ARGs in the outdoor air. Air samples were collected with a Harvard impactor in the PM10 range at 50 m from a hospital in Budapest. From the DNA yielded from samples of PM10 fraction single‐end reads were generated with an Ion Torrent sequencer. During the metagenomic analysis, reads were classified taxonomically. The core bacteriome was defined. Reads were assembled to contigs and the ARG content was analyzed. The dominant genera in the core bacteriome were Bacillus, Acinetobacter, Leclercia and Paenibacillus. Among the identified ARGs best hits were vanRA, Bla1, mphL, Escherichia coli EF‐Tu mutants conferring resistance to pulvomycin; BcI, FosB, and mphM. Despite the low DNA content of the samples of PM10 fraction, the number of detected airborne ARGs was surprisingly high., A standardized method for the metagenomic analysis of airborne samples would help to estimate the public health risks of airborne antimicrobial resistance genes (ARGs). We conducted a metagenomic analysis to reveal the core bacteriome and the ARG profile of three urban air samples collected with a Harvard impactor. The core bacteriome was dominated by Bacillus, Acinetobacter, Leclercia and Paenibacillus species. We detected several antimicrobial resistance genes. All of them were associated with non‐pathogenic bacteria.

How cell size and number are determined during organ development remains a fundamental question in cell biology. Here, we identified a GRAS family transcription factor, called SCARECROW-LIKE28 (SCL28), with a critical role in determining cell size in Arabidopsis. SCL28 is part of a transcriptional regulatory network downstream of the central MYB3Rs that regulate G2 to M phase cell cycle transition. We show that SCL28 forms a dimer with the AP2-type transcription factor, AtSMOS1, which defines the specificity for promoter binding and directly activates transcription of a specific set of SIAMESE-RELATED (SMR) family genes, encoding plant-specific inhibitors of cyclin-dependent kinases and thus inhibiting cell cycle progression at G2 and promoting the onset of endoreplication. Through this dose-dependent regulation of SMR transcription, SCL28 quantitatively sets the balance between cell size and number without dramatically changing final organ size. We propose that this hierarchical transcriptional network constitutes a cell cycle regulatory mechanism that allows to adjust cell size and number to attain robust organ growth.

The DNA of all organisms is constantly damaged by physiological processes and environmental conditions. Upon persistent damage, plant growth and cell proliferation are reduced. Based on previous findings that RBR1, the only Arabidopsis homolog of the mammalian tumor suppressor gene retinoblastoma, plays a key role in the DNA damage response in plants, we unravel here the network of RBR1 interactors under DNA stress conditions. This led to the identification of homologs of every DREAM component in Arabidopsis, including previously not recognized homologs of LIN52. Interestingly, we also discovered NAC044, a mediator of DNA damage response in plants and close homolog of the major DNA damage regulator SOG1, to directly interact with RBR1 and the DREAM component LIN37B. Consistently, not only mutants in NAC044 but also the double mutant of the two LIN37 homologs and mutants for the DREAM component E2FB showed reduced sensitivities to DNA-damaging conditions. Our work indicates the existence of multiple DREAM complexes that work in conjunction with NAC044 to mediate growth arrest after DNA damage.

Funding Information: Funding: O.K. was supported by the ÚNKP-20-3—New National Excellence Program of the Ministry for Innovation and Technology from the source of the National Research, Development and Innovation Fund. M.A.A., H.S. and R.M. are grateful to the Academy of Finland (CleanSchool-project, grant no. 330150) for financial support. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. The genus Chaetomium is a frequently occurring fungal taxon world-wide. Chaetomium and Chaetomium-like species occur in indoor environments, where they can degrade cellulose-based building materials, thereby causing structural damage. Furthermore, several species of this genus may also cause adverse effects on human health. The aims of this research were to identify Chaetomium and Chaetomium-like strains isolated from indoor environments in Hungary and Finland, two geographi-cally distant regions of Europe with drier and wetter continental climates, respectively, and to study their morphological and physiological properties, as well as their extracellular enzyme activities, thereby comparing the Chaetomium and Chaetomium-like species isolated from these two different regions of Europe and their properties. Chaetomium and Chaetomium-like strains were isolated from flats and offices in Hungary, as well as from schools, flats, and offices in Finland. Fragments of the translation elongation factor 1α (tef1α), thesecond largest subunit of RNA polymerase II (rpb2) and β-tubulin (tub2) genes, as well as the internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster were sequenced, and phylogenetic analysis of the sequences performed. Morphological examinations were performed by stereomicroscopy and scanning electron microscopy. Thirty-one Chaetomium sp. strains (15 from Hungary and 16 from Finland) were examined during the study. The most abundant species was Ch. globosum in both countries. In Hungary, 13 strains were identified as Ch. globosum, 1 as Ch. cochliodes, and 1 as Ch. interruptum. In Finland, 10 strains were Ch. globosum, 2 strains were Ch. cochliodes, 2 were Ch. rectangulare, and 2 isolates (SZMC 26527, SZMC 26529) proved to be representatives of a yet undescribed phylogenetic species from the closely related genus Dichotomopilus, which we formally describe here as the new species Dichotomopilus finlandicus. Growth of the isolates was examined at different temperatures (4, 15, 20, 25, 30, 37, 35, 40, and 45◦ C), while their extracellular enzyme production was determined spectrophotometrically.

Since 2010, outbreaks of haemorrhagic septicaemia (HS) caused by Pasteurella (P.) multocida capsular type B (PmB) emerged in Germany. In 2017, we noticed a close spatiotemporal relationship between HS outbreak sites and wolf (Canis lupus) territories. Thus, the main objectives of our study were to investigate the molecular epidemiology of German PmB-HS-isolates and to assess the role of wolves as putative vectors of this pathogen. We collected 83 PmB isolates from HS outbreaks that occurred between 2010 and 2019 and sampled 150 wolves, which were found dead in the years 2017 to 2019, revealing another three PmB isolates. A maximum-likelihood-based phylogeny of the core genomes of 65 PmB-HS-isolates and the three PmB-wolf-isolates showed high relatedness. Furthermore, all belonged to capsular:LPS:MLST genotype B:L2:ST122RIRDC and showed highly similar virulence gene profiles, but clustered separately from 35 global ST122RIRDC strains. Our data revealed that German HS outbreaks were caused by a distinct genomic lineage of PmB-ST122 strains, hinting towards an independent, ongoing epidemiologic event. We demonstrated for the first time, that carnivores, i.e., wolves, might harbour PmB as a part of their oropharyngeal microbiota. Furthermore, the results of our study imply that wolves can carry the pathogen over long distances, indicating a major role of that animal species in the ongoing epidemiological event of HS in Germany.

Hydrothermal vents have been key to our understanding of the limits of life, and the metabolic and phylogenetic diversity of thermophilic organisms. Here we used environmental metagenomics combined with analysis of physico-chemical data and 16S rRNA amplicons to characterize the diversity, temperature optima, and biogeographic distribution of sediment-hosted microorganisms at the recently discovered Auka vents in the Gulf of California, the deepest known hydrothermal vent field in the Pacific Ocean. We recovered 325 metagenome assembled genomes (MAGs) representing 54 phyla, over 1/3 of the currently known phylum diversity, showing the microbial community in Auka hydrothermal sediments is highly diverse. Large scale 16S rRNA amplicon screening of 227 sediment samples across the vent field indicates that the MAGs are largely representative of the microbial community. Metabolic reconstruction of a vent-specific, deeply branching clade within the Desulfobacterota (Tharpobacteria) suggests these organisms metabolize sulfur using novel octaheme cytochrome-c proteins related to hydroxylamine oxidoreductase. Community-wide comparison of the average nucleotide identity of the Auka MAGs with MAGs from the Guaymas Basin vent field, found 400 km to the Northwest, revealed a remarkable 20% species-level overlap between vent sites, suggestive of long-distance species transfer and sediment colonization. An adapted version of a recently developed model for predicting optimal growth temperature to the Auka and Guaymas MAGs indicates several of these uncultured microorganisms could grow at temperatures exceeding the currently known upper limit of life. Extending this analysis to reference data shows that thermophily is a trait that has evolved frequently among Bacteria and Archaea. Combined, our results show that Auka vent field offers new perspectives on our understanding of hydrothermal vent microbiology.

The ErbB-3 BINDING PROTEIN 1 (EBP1) drives growth, but the mechanism of how it acts in plants is little understood. Here, we show that EBP1 expression and protein abundance in Arabidopsis (Arabidopsis thaliana) are predominantly confined to meristematic cells and are induced by sucrose and partially dependent on TARGET OF RAPAMYCIN (TOR) kinase activity. Consistent with being downstream of TOR, silencing of EBP1 restrains, while overexpression promotes, root growth, mostly under sucrose-limiting conditions. Inducible overexpression of RETINOBLASTOMA RELATED (RBR), a sugar-dependent transcriptional repressor of cell proliferation, depletes meristematic activity and causes precocious differentiation, which is attenuated by EBP1. To understand the molecular mechanism, we searched for EBP1- and RBR-interacting proteins by affinity purification and mass spectrometry. In line with the double-stranded RNA-binding activity of EBP1 in human (Homo sapiens) cells, the overwhelming majority of EBP1 interactors are part of ribonucleoprotein complexes regulating many aspects of protein synthesis, including ribosome biogenesis and mRNA translation. We confirmed that EBP1 associates with ribosomes and that EBP1 silencing hinders ribosomal RNA processing. We revealed that RBR also interacts with a set of EBP1-associated nucleolar proteins as well as factors that function in protein translation. This suggests EBP1 and RBR act antagonistically on common processes that determine the capacity for translation to tune meristematic activity in relation to available resources.

During our experiments in 2017/2018 crop year at Látókép Experimental Farm of University of Debrecen, we examined the effect of different cultivars and increased dosages of artificial fertilizer (control, N90PK, N150PK) on quality of wheat gluten. The gluten quality parameters of winter wheat were significantly influenced by fertilizing and cultivar. N90PK dosage improved significantly the measured parameters, further fertilizer application had no statistically provable effect. Application of artificial fertilizers increased significantly the gluten spread of the samples, compared to the control ones. KG Kunhalom had significantly higher crude protein (CP), wet gluten (WGC), dry gluten, gluten ratio and Zeleny index (ZI) parameters, this means that KG Kunhalom had the best protein and gluten quality potential. Also only KG Kunhalom with N150PK fertilizing treatment complied with the Hungarian premium class. According to Hungarian standard, samples with control treatment and Hybiza with any fertilizing treatment were unfit to bakery use. Using Pearson’s correlation analysis results, CP and ZI were in tight positive; WGC was in medium positive, while gluten index was in loose negative correlation with fertilizing. In the case of growing wheat for baking use, there is a need to put great emphasis on selecting the right cultivar and agrotechnology practices.

Background and Objectives: West Nile virus (WNV) is one of the most important viral zoonotic infections in Hungary; however, no transfusion-transmitted WNV infections have been confirmed s...

Previously, a novel cell type, the multinucleated giant hemocyte (MGH) was identified in the ananassae subgroup of Drosophilidae. These cells share several features with mammalian multinucleated giant cells, a syncytium of macrophages formed during granulomatous inflammation. We were able to show that MGHs also differentiate in Zaprionus indianus, an invasive species belonging to the vittiger subgroup of the family, highly resistant to a large number of parasitoid wasp species. We have classified the MGHs of Z. indianusas giant hemocytes belonging to a class of cells which also include elongated blood cells carrying a single nucleus and anuclear structures. They are involved in encapsulating parasites, originate from the lymph gland, can develop by cell fusion, and generally carry many nuclei, while possessing an elaborated system of canals and sinuses, resulting in a spongiform appearance. Their nuclei are all transcriptionally active and show accretion of genetic material. Multinucleation and accumulation of the genetic material in the giant hemocytes represents a two-stage amplification of the genome, while their spongy ultrastructure substantially increases the contact surface with the extracellular space. These features may furnish the giant hemocytes with a considerable metabolic advantage, hence contributing to the mechanism of the effective immune response.

This study describes the prevalence of capsule biosynthesis genes, LPS genotypes, virulence associated genes and the analysis of the outer membrane protein (ompA) sequence of Pasteurella multocida isolates (n = 180) from different locations in Hungary, from various host species, including humans. When combining capsular types with LPS genotypes, eight capsule - LPS genotype combinations were detected. A: L3 was the most dominant in bovine and porcine isolates, A: L1 in feline and human isolates, while D: L3 was the most common among strains from small ruminants. The P. multocida toxin encoding gene toxA was highly prevalent among small ruminant and porcine strains, while in human, feline and bovine isolates it could not be detected. Combination of the tested virulence associated genes (hgbA, nanH, hgbB, tbpA, pfhA, hsf1, hsf2, tadD, ptfA) classified our P. multocida isolates into 13 different virulence gene profiles (VGPs). These VGPs showed an association with host species. Analysis of the ompA sequence data confirmed this distribution by host species, which may indicate that host adaptation is taking place. The typing scheme used in this study may be useful in epidemiological investigations.

During our experiments in 2017/2018 crop year at Látókép Experimental Farm of University of Debrecen we studied the effect of different forecrops (sweet corn, sunflower) and increased dosages of artificial fertilizers (control, N90PK, N150PK) on rheological properties of wheat. Both levels of artificial fertilizers significantly improved valorigraphic water absorption (WA), quality number (VQN) and dough-stability (DST), moreover alveographic L and W value. Applying artificial fertilizers valorigraphic mixing-tolerance (DMT) and dough softening (DS) values were decreased significantly comparing to the control ones. Sweet corn as a forecrop had significantly favourable effect on VQN, DDT, DST, DS and DMT; promylographic ductility; alveographic L values comparing to sunflower. Fertilizing x forecrop interaction affected in a significant way the DMT and P/L value. In addition, fertilizing x cultivar interaction had significant effect on alveographic L, promylographic ductility and ratio. Using Pearson’s correlation analysis results, fertilizer dosages were in strong positive correlation with VQN and DDT; alveographic L and W. The alveographic W was in strong positive correlation with promylographic energy (r=0,842**) and DST (r=0,863**), while the L was in medium positive correlation with promylographic ductility (r=0,744**). Our results proved that, the wheat flour’s rheological parameters are significantly affected by fertilizing dose, forecrop and cultivar

When we discuss the genetics of tumors, we cannot fail to remember that in the second decade of the twentieth century, more precisely in 1914, Theodore Boveri defined for the first time the chromosomal bases of cancer. In the last 30 years, progresses in genetics have only confirmed Boveri's remarkable predictions made more than 80 years ago. Before the cloning of the retinoblastoma 1 (RB1) gene, the existence of a genetic component in most, if not all, solid childhood tumors were well known. The existence of familial tumor aggregations has been found much more frequently than researchers expected to find at random. Sometimes, the demonstration of this family predisposition was very difficult, because the survival of children diagnosed as having a certain tumor, up to an age at which reproduction and procreation is possible, was very rare. In recent years, advances in the diagnosis and treatment of these diseases have made it possible for these children to survive until the age when they were able to start their own families, including the ability to procreate. Four distinct groups of so-called cancer genes have been identified: oncogenes, which promote tumor cell proliferation; tumor suppressor genes, which inhibit this growth/proliferation; anti-mutational genes, with a role in deoxyribonucleic acid (DNA) stability; and micro-ribonucleic acid (miRNA) genes, with a role in the posttranscriptional process.

Significance Rock varnish is a prominent feature of desert landscapes and the canvas for many prehistoric petroglyphs. How it forms—and, in particular, the basis for its extremely high manganese content—has been an enduring mystery. The work presented here establishes a biological mechanism for this manganese enrichment, underpinned by an apparent antioxidant strategy that enables microbes to survive in the harsh environments where varnish forms. The understanding that varnish is the residue of life using manganese to thrive in the desert illustrates that, even in extremely stark environments, the imprint of life is omnipresent on the landscape., Desert varnish is a dark rock coating that forms in arid environments worldwide. It is highly and selectively enriched in manganese, the mechanism for which has been a long-standing geological mystery. We collected varnish samples from diverse sites across the western United States, examined them in petrographic thin section using microscale chemical imaging techniques, and investigated the associated microbial communities using 16S amplicon and shotgun metagenomic DNA sequencing. Our analyses described a material governed by sunlight, water, and manganese redox cycling that hosts an unusually aerobic microbial ecosystem characterized by a remarkable abundance of photosynthetic Cyanobacteria in the genus Chroococcidiopsis as the major autotrophic constituent. We then showed that diverse Cyanobacteria, including the relevant Chroococcidiopsis taxon, accumulate extraordinary amounts of intracellular manganese—over two orders of magnitude higher manganese content than other cells. The speciation of this manganese determined by advanced paramagnetic resonance techniques suggested that the Cyanobacteria use it as a catalytic antioxidant—a valuable adaptation for coping with the substantial oxidative stress present in this environment. Taken together, these results indicated that the manganese enrichment in varnish is related to its specific uptake and use by likely founding members of varnish microbial communities.

Funding Acknowledgements Type of funding sources: None. Background Short QT syndrome (SQTS) is a rare, autosomal dominant disease causing sudden cardiac death (SCD). Current guidelines recommend genetic testing. Associated variants in KCNQ1, KCNH2, KCNJ2 and SLC4A3 genes have been reported. Purpose We report a family with a variant in the SLC4A3 gene with several presentations of SCD and high clinical penetrance of SQTS. Methods We performed a post-mortem genetic testing in the index patient in whom prior ECG was available. Subsequently, clinical and electrophysiological work-up and cascade screening (CS) of the detected suspected variant was carried out in available relatives. Results The index patient had suffered a SCD at the age of 17 (figure, upper panel, arrow). A previously registered ECG showed a shortened QTc of 340ms (figure, lower panel). Autopsy revealed no structural heart disease. Post-mortem genetic testing revealed variants in the LDB3, MYH7 and a novel heterozygous missense variant, p.(Ser1039Arg) also in the SLC4A3 gene. Although predictive bioinformatic algorithms (AlignGVGD, SIFT, MutationTaster, Polyphen2) showed conflicting classifications, family history was notable for SCD without post-mortem genetic work-up in three second degree relatives (figure, upper panel, patients 207, 208 and 305, age of death 33, 25 and 33 years respectively). CS was performed in first and second degree relatives of the index patient and was highly suggestive for disease association of the variant in the SLC4A3 gene with co-segregation in all clinically affected family members. Only one patient with the variant had a normal QTc (figure, upper panel, patient 202) of 407ms, however this patient was on regular QT-prolonging medication (risperidone and loperamide). Conclusion Genetic testing revealed a novel in the SLC4A3 gene, which was recently implicated in the pathogenesis of the SQTS. Although predictive bioinformatic algorithms yielded conflicting results, CS of family members suggests a likely pathogenicity (class IV) of the variant. Further CS or functional tests are necessary to establish causality. Abstract Figure. ECG of index patient and family tree

Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): NEW NATIONAL EXCELLENCE PROGRAM OF THE MINISTRY FOR INNOVATION AND TECHNOLOGY FROM THE SOURCE OF THE NATIONAL RESEARCH, DEVELOPMENT AND INNOVATION FUND. Introduction and purpose Acute β-adrenergic receptor (β-AR) stimulation shortens the ventricular action potential (AP). This effect is mainly regulated by the β- adrenergic stimulation of the cardiac potassium currents. Our aim was to investigate the extent of calcium/calmodulin-dependent protein kinase II (CaMKII) involvement in mediating the effect of β-AR activation on the inward rectifier potassium current – I K1 . Methods We carried out our experiments on isolated cardiomyocytes originating from canine left ventricles. The inward rectifier potassium current – I K1 was measured under a "canonical" AP under action potential voltage clamp conditions. Data were collected in four study groups [1] Control conditions (CTRL) [2] Inhibition of CaMKII with 1 µM KN-93 (KN-93) [3] Inhibition of PKA with 3 µM H-89 (H-89) [4] Acute β-adrenergic stimulation with 10 nM isoproterenol (ISO) [5] β-adrenergic stimulation with CaMKII inhibition (KN-93 + ISO) [6] β-adrenergic stimulation with PKA inhibition (H-89 + ISO) [7] β-adrenergic stimulation with inhibited PKA and CaMKII (KN-93 + H-89 + ISO) Results I K1 current amplitude did not differ among the studied groups, the total carried charge however was significantly, about 30 % larger in the ISO group compared to CTRL, and about 20 % larger compared to KN-93 + ISO. Under beta- adrenergic stimulation, I K1 starts to activate earlier during the AP plateau. I K1 density was about 3 times greater both at +20 mV and at 0 mV membrane potential under the command "canonical" AP in ISO compared to CTRL. Similarly, I K1 density was about 60 % and 90 % larger at +20 mV and at 0 mV, respectively, in KN-93 + ISO compared to KN-93. Similar results have been obtained by conventional voltage-clamp technique. Conclusion Based on the results of our researches the CaMKII activation plays an important role in β -adrenergic stimulation the I K1 potassium current.

The study of stemflow fungi began over 50 years ago. Past work has been performed in different climatic regions of the world, with different sampling methods, by mycologists focusing on different taxonomical groups. Therefore, we aim to synthesize this work to delineate major conclusions and emerging hypothesis. Here, we present: (1) a systematic compilation of observations on stemflow conidial concentration, flux, and species composition; (2) an evaluation of the methods underlying these observations; (3) a testable theory to understand spatiotemporal dynamics in stemflow (including honeydews) conidial assemblages, with a focus on their relationship to bark structure and microhabitats; and (4) a discussion of major hypotheses based on past observations and new data. This represents a knowledge gap in our understanding of fungal dispersal mechanisms in forests, in a spatially-concentrated hydrologic flux that interacts with habitats throughout the forest microbiome. The literature synthesis and new data represent observations for 228 fungal species’ conidia in stemflow collected from 58 tree species, 6 palm species, and 1 bamboo species. Hypothetical relationships were identified regarding stemflow production and conidial concentration, flux, and species composition. These relationships appear to be driven by bark physico-chemical properties, tree canopy setting, the diversity of in-canopy microenvironments (e.g., tree holes, bark fissures, and epiphytes), and several possible conidia exchange processes (teleomorph aerosols, epi-faunal exchanges, fungal colonization of canopy microhabitats, and droplet impacts, etc.). The review reveals a more complex function of stemflow fungi, having a role in self-cleaning tree surfaces (which play air quality-related ecoservices themselves), and, on the other hand, these fungi may have a role in the protection of the host plant.

Bony cranial ornamentation is developed by many groups of vertebrates, including ankylosaur dinosaurs. To date, the morphology and ontogenetic origin of ankylosaurian cranial ornamentation has primarily focused on a limited number of species from only one of the two major lineages, Ankylosauridae. For members of the sister group Nodosauridae, less is known. Here, we provide new details of the cranial anatomy of the nodosauridHungarosaurusfrom the Santonian of Europe. Based on a number of previously described and newly identified fragmentary skulls and skull elements, we recognize three different size classes ofHungarosaurus. We interpret these size classes as representing different stages of ontogeny. Cranial ornamentation is already well-developed in the earliest ontogenetic stage represented herein, suggesting that the presence of outgrowths may have played a role in intra- and interspecific recognition. We find no evidence that cranial ornamentation inHungarosaurusinvolves the contribution of coossified osteoderms. Instead, available evidence indicates that cranial ornamentation forms as a result of the elaboration of individual elements. Although individual differences and sexual dimorphism cannot be excluded, the observed variation inHungarosauruscranial ornamentation appears to be associated with ontogeny.

There is a dynamic increase in the number of international sports competitions aimed for deaf athletes. More than 50 World and European championships are held every year in various sports disciplines. The participationand number of deaf athletes delegated to these sports competitions depend on the budget available to national sports federations (NSFs) of deaf athletes. Many times, NSFs have been forced to withdraw from participation in a competition owing to lack of financial resources. In numerous countries, NSFs have to conduct online fundraising dedicated to their representatives’ involvement in sports competitions. The authors reviewed the source materials in internet on financing the sport of the hearing impaired. No scientific articles covering the funding aspect of national sports federations in the world were found. The authors relied on financial documents provided by the NSF (Poland, Hungary, Slovakia, Spain) and on websites indicated by these federations.The aim of the article was to characterize selected national organizations gathering deaf athletes from Europe, Asia and Oceania, and North America in terms of the budget they had at their disposal in years 2013–2017, the number of athletes associated in the NSF, and the number of Olympic competitors and the country’s medal position during the 2013 and 2017 Deaflympics.An overview of NSF websites with reference to “annual reports”, “financial reports”, “financial statements”, “newsletter”, “national teams of”, “number of athletes of”, demonstrated numerous deficiencies in the transparent presentation of the financial situation of NSFs. The content of NSF activity reports also provides limited information on the number of athletes and clubs associated in these federations. The article constitutes an introduction to a discussion on the financing aspects and development of deaf people’s sport worldwide

Autotrophic nitrogen removal by anaerobic ammonium oxidizing (anammox) bacteria is an energy-efficient nitrogen removal process in wastewater treatment. However, full-scale deployment under mainstream conditions remains challenging for practitioners due to the high stress susceptibility of anammox bacteria towards fluctuations in dissolved oxygen (DO) and temperature. Here, we investigated the response of microbial biofilms with verified anammox activity to DO shocks under 20 °C and 14 °C. While pulse disturbances of 0.3 mg L−1 DO prompted only moderate declines in the NH4+ removal rates, 1.0 mg L−1 DO led to complete but reversible inhibition of the NH4+ removal activity in all reactors. Genome-centric metagenomics and metatranscriptomics were used to investigate the stress response on various biological levels. We show that temperature regime and strength of DO perturbations induced divergent responses from the process level down to the transcriptional profile of individual taxa. Community-wide gene expression differed significantly depending on the temperature regime in all reactors, and we found a noticeable impact of DO disturbances on genes involved in transcription, translation, replication and posttranslational modification at 20 °C but not 14 °C. Genome-centric analysis revealed that different anammox species and other key biofilm taxa differed in their transcriptional responses to distinct temperature regimes and DO disturbances., Communications Biology, 4, ISSN:2399-3642

Patients with chronic kidney disease (CKD) have elevated circulating levels of trimethylamine N-oxide (TMAO), a metabolite derived from gut microbes and associated with cardiovascular diseases. High circulating levels of TMAO and its dietary precursor, choline, predict increased risk for development of CKD in apparently healthy subjects, and studies in mice fed TMAO or choline suggest that TMAO can contribute to kidney impairment and renal fibrosis. Here we examined the interactions between TMAO, kidney disease, and cardiovascular disease in mouse models. We observed that while female hyperlipidemic apoE KO mice fed a 0.2% adenine diet for 14 weeks developed CKD with elevated plasma levels of TMAO, provision of a non-lethal inhibitor of gut microbial trimethylamine (TMA) production, iodomethylcholine (IMC), significantly reduced multiple markers of renal injury (plasma creatinine, cystatin C, FGF23, and TMAO), reduced histopathologic evidence of fibrosis, and markedly attenuated development of microalbuminuria. In addition, while the adenine-induced CKD model significantly increased heart weight, a surrogate marker for myocardial hypertrophy, this was largely prevented by IMC supplementation. Surprisingly, adenine feeding did not increase atherosclerosis and significantly decreased the expression of inflammatory genes in the aorta compared to the control groups, effects unrelated to TMAO levels. Our data demonstrate that inhibition of TMAO production attenuated CKD development and cardiac hypertrophy in mice, suggesting that TMAO reduction may be a novel strategy in treating CKD and its cardiovascular disease complications.

New palynological data from the deep-water Gușterița section in the Transylvanian Basin depict the late Miocene evolution of Lake Pannon between 11.0 and 10.5 Ma. Spores and pollen were used to analyze vegetation and climate while dinoflagellate cysts were used to reconstruct lake ecology. The section hosts primarily endemic brackish-water dinoflagellate cysts related to those in the Ponto-Caspian realm. The lake was fringed by swamps of taxodioid gymnosperms and riparian forests. The proximity of the Carpathians and Apuseni Mts. allowed the presence of montane vegetation with Picea and Abies. The azonal vegetation narrowed down the extent of the zonal mesophytic forests. Plants were growing under warm-temperate, humid climate with mean annual temperature of 13.3–21.3 °C and mean annual precipitation of 705–1613 mm. The decline of the thermophilous Taxodioideae is linked to the uplift of the mountains and volcanism, but the expansion of Lake Pannon contributed to the swamp-retreat too. The lake expansion was stepwise with the alternation of minor floodings and stabilization of the lake-level. This pattern is inferred from the oscillation of “Virgodinium”-dominated dinoflagellate cyst assemblages characteristic of transgressive periods and assemblages with Spiniferites indicating the slowing-down of the flooding and higher nutrient input from rivers. Active sediment transport into the deep basin via turbidity currents is inferred by the large amount of plant debris in the sedimentary organic matter. The introduction of new dinoflagellate cyst morphotypes with more pronounced ornamentation and thicker wall is possibly a phenotypic response of the dinoflagellates to increasing freshwater influence.

Background Short QT syndrome (SQTS) is a rare, autosomal dominant disease causing sudden cardiac death (SCD). Genetic testing is recommended according to current guidelines. Variants in KCNQ1, KCNH2, KCNJ2 and SLC4A3 genes have been reported in SQTS. Purpose We report implications of genetic testing and cascade screening (CS) in two families with phenotypical presentation of SQTS and novel genetic variants of unknown significance. Methods We performed a thorough clinical and electrophysiological work-up of the index patients of both families. In addition, genetic screening was conducted. Subsequently, segregation analysis of potentially pathogenic variants was carried out in available relatives. Results Index patient 1 presented with a history of recurrent syncope. His ECG showed a shortened QTc of 340ms. Family history was unremarkable. Structural heart disease was excluded by cardiac MRI and coronary angiography. Genetic testing detected a rare heterozygous missense variant in the KCNH2 gene (p.(Arg328Cys), frequency 0.053%), predicted to be pathogenic according to various prediction algorithms (Polyphen, SIFT, Align GVGD, mutation taster). CS of relatives did not confirm this variant as the causative mutation. Reanalysis of whole-exome sequencing data revealed a novel heterozygous missense variant, p.(Arg370Cys) in the recently identified SLC4A3 gene. A variant at the same position has previously been associated with SQTS. CS suggested disease association. The second index patient had a SCD at the age of 17. A previously registered ECG showed a shortened QTc of 340ms. Autopsy revealed no structural heart disease. Post-mortem genetic testing revealed variants in the LDB3, MYH7 and a novel heterozygous missense variant, p.(Ser1039Arg) also in the SLC4A3 gene. Family history was positive for SCD in three 2° relatives. The index patient's father had a positive phenotype with a QTc of 365ms. CS again suggested disease association of the variant in the SLC4A3 gene only. Conclusion Genetic testing revealed two novel variants in the SLC4A3 gene, which was recently implicated in the pathogenesis of the SQTS. Predictive bioinformatic algorithms to assess the pathogenicity of missense variants are of limited relevance, but genetic analysis of additional unaffected and affected family members may be instrumental to identify pathogenic DNA sequence variations. Family tree index patients 1 and 2 Funding Acknowledgement Type of funding source: None

Organic anion-transporting polypeptide 3A1 (OATP3A1) is a membrane transporter mediating the cellular uptake of various hormones such as estrone-3-sulfate, prostaglandins E1 and E2 and thyroxine. OATP3A1 is widely expressed in the human body and its presence in tissue-blood barriers, neurons and muscle cells marks it as a potential pharmacological target. Herein we demonstrate that an otherwise membrane impermeant, zwitterionic fluorescent coumarin probe, bearing a sulfonate function is a potent substrate of human OATP3A1, thus readily transported into HEK-293-OATP3A1 cells allowing functional investigation and the screen of drug interactions of the OATP3A1 transporter. At the same time, dyes lacking either the sulfonate motif or the coumarin scaffold showed a dramatic decrease in affinity or even a complete loss of transport. Furthermore, we observed a distinct inhibition/activation pattern in the OATP3A1-mediated uptake of closely related fluorescent coumarin derivatives differing only in the presence of the sulfonate moiety. Additionally, we detected a synergistic effect between one of the probes tested and the endogenous OATP substrate estrone-3-sulfate. These data, together with docking results indicate the presence of at least two cooperative substrate binding sites in OATP3A1. Besides providing the first sensitive probe for testing OATP3A1 substrate/inhibitor interactions, our results also help to understand substrate recognition and transport mechanism of the poorly characterized OATP3A1. Moreover, coumarins are good candidates for OATP3A1-targeted drug delivery and as pharmacological modulators of OATP3A1.

Funding Acknowledgements none Introduction Short QT syndrome (SQTS) is a rare, autosomal dominant disease causing ventricular fibrillation and sudden cardiac death. Genetic testing is recommended according to current guidelines. Mutations in KCNQ1, KCNH2, KCNJ2 and more recently SLC4A3 genes have been implicated in SQTS. These genes encode potassium channel subunits and a bicarbonate transporter regulating intracellular pH. A dominant mutation in this transporter can lead to increased intracellular pH and shortened action potential. Purpose We present a family with a short QT phenotype and recurrent syncope in whom a novel genetic variant was detected by whole-exome sequencing (WES), confirmed by cascade screening. Methods We performed a thorough work-up of the index patient including medical history, physical examination, 12-lead ECG, echocardiography, stress testing, coronary angiography, flecainide challenge, and genetic testing with NGS. QTc was determined using Bazett’s formula. CS of all 1° and two 2° relatives was performed. Results The ECG of the index patient showed a QTc of 340ms and characteristics compatible with a SQTS (figure). Clinical work-up was unremarkable. A first genetic search with next generation sequencing focusing on genes that have been previously involved in the pathogenesis of channelopathies detected a rare known heterozygous missense variant in the KCNH2 gene (Arg328Cys, frequency 0.053%), which was predicted to be pathogenic according to various prediction algorithms (Polyphen, SIFT, Align GVGD, mutation taster). ECG screening of all asymptomatic first-degree family members identified a SQT phenotype in the mother (QTc 355ms), but not in the father (QTc 380ms) and sister (410ms). The KCNH2 variant was found in the father and sister but not the affected mother, which excludes this variant as the causative mutation in this family. Therefore, reanalysis of WES data was performed and revealed a novel heterozygous missense variant p.(Arg370Cys) in the SLC4A3 gene, recently associated with SQTS. A mutation in this gene at the same position has been previously reported in SQTS. The p.(Arg370Cys) mutation was found in the mother but not in the unaffected father or sister. Furthermore the mutation was present in two affected maternal uncles (QTc 319ms and 342ms) supporting the assumption that this was the causative mutation in this family. Conclusions A novel genetic variant in the SLC4A3 gene leading to sQT phenotype could be detected using WES and cascade screening. Predictive bioinformatic algorithms to assess the pathogenicity of missense variants are of limited relevance, but genetic analysis of additional unaffected and affected family members may be instrumental to identify pathogenic DNA sequence variations. Abstract Figure. Pedigree and ECGs of the family

Background: Evidence suggests the renin-angiotensin system (RAS) plays key immunomodulatory roles. In particular, angiotensin-converting enzyme (ACE) has been shown to play a role in antimicrobial host defense. ACE inhibitors (ACEi) and angiotensin receptor blockers (ARB) are some of the most commonly prescribed medications, especially in patients undergoing invasive surgery. Thus, the current study assessed the immunomodulatory effect of RAS-modulation in a preclinical model of implant infection.Methods:In vitro antimicrobial effects of ACEi and ARBs were first assessed. C57BL/6J mice subsequently received either an ACEi (lisinopril; 16 mg/kg/day), an ARB (losartan; 30 mg/kg/day), or no treatment. Conditioned mice blood was then utilized to quantify respiratory burst function as well as Staphylococcus aureus Xen36 burden ex vivo in each treatment group. S. aureus infectious burden for each treatment group was then assessed in vivo using a validated mouse model of implant infection. Real-time quantitation of infectious burden via bioluminescent imaging over the course of 28 days post-procedure was assessed. Host response via monocyte and neutrophil infiltration within paraspinal and spleen tissue was quantified by immunohistochemistry for F4/80 and myeloperoxidase, respectively.Results: Blood from mice treated with an ACEi demonstrated a decreased ability to eradicate bacteria when mixed with Xen36 as significantly higher levels of colony forming units (CFU) and biofilm formation was appreciated ex vivo (p < 0.05). Mice treated with an ACEi showed a higher infection burden in vivo at all times (p < 0.05) and significantly higher CFUs of bacteria on both implant and paraspinal tissue at the time of sacrifice (p < 0.05 for each comparison). There was also significantly decreased infiltration and respiratory burst function of immune effector cells in the ACEi group (p < 0.05).Conclusion: ACEi, but not ARB, treatment resulted in increased S. aureus burden and impaired immune response in a preclinical model of implant infection. These results suggest that perioperative ACEi use may represent a previously unappreciated risk factor for surgical site infection. Given the relative interchangeability of ACEi and ARB from a cardiovascular standpoint, this risk factor may be modifiable.

African swine fever virus (ASFV) is an important animal pathogen causing substantial economic losses in the swine industry globally. At present, little is known about the molecular biology of ASFV, including its transcriptome organization. In this study, we applied cutting-edge sequencing approaches, namely the Illumina short-read sequencing (SRS) and the Oxford Nanopore Technologies long-read sequencing (LRS) techniques, together with several library preparation chemistries to analyze the ASFV dynamic transcriptome. SRS can generate a large amount of high-precision sequencing reads, but it is inefficient for identifying long RNA molecules, transcript isoforms and overlapping transcripts. LRS can overcome these limitations, but this approach also has shortcomings, such as its high error rate and the low coverage. Amplification-based LRS techniques produce relatively high read counts but also high levels of spurious transcripts, whereas the non-amplified cDNA and direct RNA sequencing techniques are more precise but achieve lower throughput. The drawbacks of the various technologies can be circumvented by the combined use of these approaches.

Autotrophic nitrogen removal by anaerobic ammonium oxidizing (anammox) bacteria is an energy-efficient nitrogen removal process in wastewater treatment. However, full-scale deployment under mainstream conditions remains challenging for practitioners due to the high stress susceptibility of anammox bacteria towards fluctuations in dissolved oxygen and temperature. Here, we investigated the response of microbial biofilms with verified anammox activity to oxygen shocks under favorable and cold temperature regimes. Genome-centric metagenomics and metatranscriptomics were used to investigate the stress response on various biological levels. We show that temperature regime and strength of oxygen perturbations induced divergent responses from the process level down to the transcriptional profile of individual taxa. Temperature induced distinct transcriptional states in compositionally identical communities and transient pulses of dissolved oxygen resulted in the upregulation of stress-response only under favorable temperatures. Anammox species and other key biofilm taxa display different transcriptional responses to the induced stress regimes.

In this paper we report the phenotypic and partial genetic characterisation of a novel bacterium strain isolated from a cat with severe nephritis. Multilocus sequence analysis was performed on the 16S rRNA and three housekeeping (recN, rpoB, infB) gene sequences obtained by PCR. In accordance with the results of phenotypic tests, the phylogenetic analyses confirmed the relatedness of the new strain (6036) to the family Pasteurellaceae. On the phylogenetic trees, strain 6036 appeared in a separate branch, closest to that of the type species (Frederiksenia canicola) of the genus Frederiksenia. These two bacteria shared 95.14 and 76.88% identity in their partial 16S rRNA and recN gene sequences, respectively. The rpoB- and infB-based phylogenetic analyses indicated that strain 6036 is most closely related to Bibersteinia trehalosi (with 90.58% identity) and [Haemophilus] felis ATCC 49733 (89.50% identity), respectively. The predicted genome identity values, based on the recN gene sequences, suggested that strain 6036 can be classified into the genus Frederiksenia as a novel species. A PCR method, specific to strain 6036, was developed to allow its rapid and accurate identification and differentiation from F. canicola and other species of Pasteurellaceae. The minimal inhibitory concentrations of 18 antimicrobial agents for strain 6036 were also determined.

International audience; Cell cycle entry and quiescence are regulated by the E2F transcription factors in association with RETINOBLASTOMA-RELATED (RBR). E2FB is considered to be a transcriptional activator of cell cycle genes, but its function during development remains poorly understood. Here, by studying E2FB-RBR interaction, E2F target gene expression, and epidermal cell number and shape in e2fb mutant and overexpression lines during leaf development in Arabidopsis (Arabidopsis thaliana), we show that E2FB in association with RBR plays a role in the inhibition of cell proliferation to establish quiescence. In young leaves, both RBR and E2FB are abundant and form a repressor complex that is reinforced by an autoregulatory loop. Increased E2FB levels, either by expression driven by its own promoter or ectopically together with DIMERIZATION PARTNER A, further elevate the amount of this repressor complex, leading to reduced leaf cell number. Cell overproliferation in e2fb mutants and in plants overexpressing a truncated form of E2FB lacking the RBR binding domain strongly suggested that RBR repression specifically acts through E2FB. The increased number of small cells below the guard cells and of fully developed stomata indicated that meristemoids preferentially hyperproliferate. As leaf development progresses and cells differentiate, the amount of RBR and E2FB gradually declined. At this stage, elevation of E2FB level can overcome RBR repression, leading to reactivation of cell division in pavement cells. In summary, E2FB in association with RBR is central to regulating cell proliferation during organ development to determine final leaf cell number.

Almost all the species of the Chenopodiaceae family present in our flora flower from July–August to the autumn. Unfortunately, allergies do not take a vacation. Warm, dry July and August weather should limit pollen emissions. However, similarly to most plants in dry habitats, goosefoot are well adapted to such conditions and does not provide even a short reprieve to pollen allergic patients. However, goosefoot pollen does not have a very large allergenic significance; despite the long pollen season lasting about 3 months, pollen concentrations in the air are low and very rarely exceed the concentration of 30 grains/m3. This study compares Chenopodiaceae pollen seasons in Poland, Hungary and Slovakia in 2019. The investigations were carried out using the volumetric method (Hirst type pollen sampler). Seasonal pollen index was estimated as the sum of daily average pollen concentrations in the given season. The pollen season ranges from June to September, depending on the geographical latitude. In Hungary and Slovakia there are much longer pollen seasons than in Poland. Pollen of goosefoot family contains the panallergen profilins, which are responsible for cross-reactivity among pollen-sensitized patients. In 2019 the pollen season of goosefoot started first in Hungary, in Kaposvar on June 7th and in Slovakia, in Žilina, on June 8th; in Poland pollen season started much later, on June 14th in Szczecin and Opole. At the latest, a pollen season ended in Nitria (Slovakia) on October 16th; in Kecskemet (Hungary) on October 3rd. In Poland the season ended much earlier than in Hungary and Slovakia already on August 25th. The differences of pollen season durations are considerable, the number of days ranged from 72 to 128. The dynamics of the pollen seasons of goosefoot family show similarities within a given country and considerable differences between these countries. However, the differences of the highest airborne concentration between the countries are not considerable (25 pollen grains/m3 in Poland, 49 pollen grains/m3 in Hungary, and 30 pollen grains/m3 in Slovakia. The maximum values of seasonal pollen count in Polish cities occurred between July 26th and August 29th, in Hungarian cities between August 27th and 30th, and in Slovakian cities between August 7th and 28th. Pollen season was characterized by extremely different total annual pollen SPI, in Poland from 116 to 360; in Hungary and Slovakia within the limits 290 to 980. Droughts that occur more frequently during the summer facilitate the spread of species of the goosefoot family due to the possibility of these plants gaining new habitats.

Congenital cataracts are the prime cause for irreversible blindness in children. The global incidence of congenital cataract is 2.2–13.6 per 10,000 births, with the highest prevalence in Asia. Nearly half of the congenital cataracts are of familial nature, with a predominant autosomal dominant pattern of inheritance. Over 38 of the 45 mapped loci for isolated congenital or infantile cataracts have been associated with a mutation in a specific gene. The clinical and genetic heterogeneity of congenital cataracts makes the molecular diagnosis a bit of a complicated task. Hence, whole exome sequencing (WES) was utilized to concurrently screen all known cataract genes and to examine novel candidate factors for a disease-causing mutation in probands from 11 pedigrees affected with familial congenital cataracts. Analysis of the WES data for known cataract genes identified causative mutations in six pedigrees (55%) in PAX6, FYCO1 (two variants), EPHA2, P3H2,TDRD7 and an additional likely causative mutation in a novel gene NCOA6, which represents the first dominant mutation in this gene. This study identifies a novel cataract gene not yet linked to human disease. NCOA6 is a transcriptional coactivator that interacts with nuclear hormone receptors to enhance their transcriptional activator function., International Journal of Molecular Sciences, 21 (24), ISSN:1422-0067

Optic nerve hypoplasia (ONH) is a congenital optic nerve abnormality caused by underdevelopment of retinal ganglion cells (RGCs). Despite being a rare disease, ONH is the most common optic disc anomaly in ophthalmological practice. So far, mutations in several genes have been identified as causative, however many cases of ONH remain without a molecular explanation. The early transcription factor atonal basic-helix-loop-helix (bHLH) transcription factor 7 (ATOH7) is expressed in retinal progenitor cells and has a crucial role in RGC development. Previous studies have identified several mutations in the ATOH7 locus in cases of eye developmental diseases such as nonsyndromic congenital retinal nonattachment and persistent hyperplasia of the primary vitreous. Here we present two siblings with a phenotype predominated by bilateral ONH, with additional features of foveal hypoplasia and distinct vascular abnormalities, where whole-exome sequencing identified two compound heterozygous missense mutations affecting a conserved amino acid residue within the bHLH domain of ATOH7 (NM_145178.3:c.175G>A; p.(Ala59Thr) and c.176C>T; p.(Ala59Val)). ATOH7 expression constructs with patient single nucleotide variants were cloned for functional characterization. Protein analyses revealed decreased protein amounts and significantly enhanced degradation in the presence of E47, a putative bHLH dimerization partner. Protein interaction assays revealed decreased heterodimerization and DNA-binding of ATOH7 variants, resulting in total loss of transcriptional activation of luciferase reporter gene expression. These findings strongly support pathogenicity of the two ATOH7 mutations, one of which is novel. Additionally, this report highlights the possible impact of altered ATOH7 dimerization on protein stability and function.

Quantification of Abs toward a single epitope is critical to understanding immunobiological processes. In autoimmunity, the prognostic value of the serological profiles of patients draws much attention, but the detection of Abs toward a single epitope is not well controlled. Particularly, the rheumatoid arthritis (RA)–specific anti–citrullinated protein/peptide Abs (ACPA) are specific to a two-atom change on arginyl residues and are considered a heterogeneous family of Abs. As a model, we studied ACPA to decipher how peptide features used as immunosorbent impact Ab detection. We synthesized 30 peptides encompassing immunodominant epitopes of citrullinated fibrin differing by their length and biotin location and tested them using ELISA with 120 sera from RA and non-RA rheumatic disease controls, generating over 3000 experimental measurements. We showed that minor molecular changes in peptide chemical structure had dramatic consequences. Even when peptides exhibited the same epitope, measured Ab titers were extremely variable, and patients’ seropositivity was discordant in up to 50% of cases. The distance between epitope and biotin was the most critical parameter for efficient Ab detection irrespective of biotin position or peptide length. Finally, we identified a 15-mer peptide bearing a single citrullinated epitope detecting almost all ACPA-positive sera, thus revealing a high degree of homogeneity in RA autoimmune response. This integrative analysis deciphers the dramatic impact of the molecular design of peptide-based technologies for epitope-specific Ab quantification. It provides a model for assay development and highlights that the studies using such technologies can give a wrong perception of biological processes and therefore that medical use of data must be cautious.

Hyperandrogenic state in females is accompanied with metabolic syndrome, insulin resistance and vascular pathologies. A total of 67%–85% of hyperandrogenic women suffer also from vitamin D deficiency. We aimed to check a potential interplay between hyperandrogenism and vitamin D deficiency in producing insulin resistance and effects on coronary resistance arteries. Adolescent female rats were divided into four groups, 11–12 animals in each. Transdermal testosterone-treated and vehicle-treated animals were kept either on vitamin D-deficient or on vitamin D-supplemented diet for 8 weeks. Plasma sexual steroid, insulin, leptin and vitamin D plasma levels were measured, and oral glucose tolerance test was performed. In coronary arterioles, insulin receptor and vitamin D receptor expressions were tested by immunohistochemistry, and insulin-induced relaxation was measured in vitro on isolated coronary resistance artery segments. Testosterone impaired glucose tolerance, and it diminished insulin relaxation but did not affect the expression of insulin and vitamin D receptors in vascular tissue. Vitamin D deficiency elevated postprandial insulin levels and homeostatic model assessment insulin resistance. It also diminished insulin-induced coronary arteriole relaxation, while it raised the expression of vitamin D and insulin receptors in the endothelial and medial layers. Our conclusion is that both hyperandrogenism and vitamin D deficiency reduce sensitivity of coronary vascular tissue to insulin, but they do it with different mechanisms.

Model experiments under laboratory conditions were carried out to assess the accumulation tendency of Zr from a silica-zirconia core–shell nanoparticles, synthesised for the assay. Acute exposition tests were conducted with Daphnia magna and Danio rerio and the accumulation tendency of the nano and the molecular form was compared. Significant elevation of Zr was found in the tissue of the test organisms treated by the NP compared to the control, however, the retention of zebrafish was lower than that of the daphnids. Increased level of bioconcentration factor and the strong correlation of the redundancy analysis data suggest accumulation tendency in D. magna, yet long-term experiments are required to prove and further assess the environmental risk of food-chain bioconcentration. The BCF results were under the REACH limit for the D. rerio groups indicating the low short-term accumulation tendency of Zr from the NP. However, Zr level was significantly higher in zebrafish individuals originating from the SiO2@ZrO2 NP exposed treatments compared to the groups supplemented by the same concentration of ZrOCl2, which did not result in the elevation of Zr in fish tissue. The trace element homeostasis of D. rerio was not affected by the acute SiO2@ZrO2 NP exposition and no lethality occurred.

At the beginning of the 20th century, the Croatian-Hungarian palaeontologist Ottokar Kadic discovered a rich Late Cretaceous vertebrate assemblage around Valioara in the Haţeg Basin, including fossils of several dinosaurs and the proposed lectotype of the iconic basal eusuchian crocodyliform Allodaposuchus. These fossils were collected from seven main localities and have been housed in the collections of the Mining and Geological Survey of Hungary. However, the collection was mixed after World War II, and this unprovenanced material currently cannot be used for palaeoecological investigations. Nevertheless, the map marking the location of Kadic’s sites has been recently uncovered, showing the positions of the fossiliferous localities, which allows matching these with the historically collected specimens using geochemistry. Based on Kadic’s map, we georeferenced, relocated, and re-excavated these vertebrate-bearing outcrops, and documented their sedimentological context. Detailed stratigraphical investigations of the exposed successions indicate that this Valioara material represents one of the oldest (= earliest Maastrichtian) Late Cretaceous faunal assemblages known from the Haţeg Basin. The vertebrate remains collected during our new excavations around Valioara represent turtles (including Kallokibotion), crocodyliforms (Allodaposuchus, Doratodon, ‘Theriosuchus’, Acynodon), dinosaurs (Zalmoxes, Telmatosaurus, titanosaurs, theropods), and mammals. In order to determine potential geochemical differences among the sites, we selected several specimens with recorded stratigraphic position, measured their trace element compositions, and used these as independent proxies to assess the probable stratigraphic origin of the historical vertebrate fossils. Our detailed sedimentological, geochemical and palaeontological investigations around Valioara contribute to a deeper understanding of the distribution, evolution and palaeoecology of the Haţeg vertebrate faunas during the latest Cretaceous.

The composition of fungal biota and air quality of five traditional subterranean wine cellars and one store building of a modern wine production facility were examined in the Tokaj wine region (northeastern Hungary). Air samples were collected with SAS IAQ sampler onto PDA, MEA and RBA. Strains representing morphotypes were isolated from colonies formed on agar plates from either air or surface samples. The internal transcribed spacer (ITS) region of the rRNA gene cluster was amplified with primers ITS1 and ITS4. Altogether 90 morphotypes were isolated, 48 and 12 strains (43 species) from the air and surfaces, respectively. The number of spore-forming species generated high diversity of indoor fungi and differences between the cellars’ fungal compositions; however, their dominant species were proved to be the same. Among the isolated strains Penicillium spp. were the most frequent. The walls of cellars were covered by colonies of Zasmidium (Cladosporium) cellare often referred to as a noble mold. Even so, this mold has been found only at a small concentration in the air samples (10–30 CFU/m3). The walls of the modern store were free of molds. Diversity of fungi of the examined wine cellars was influenced by environmental conditions to a certain degree, such as elevation (height above sea level), age, reconstruction time of cellars, indoor ethanol concentration and the number of chimneys. The location of cellars poorly influenced the concentration of fungi of the air inside cellars, contrary to outdoors where the air of the municipal area contained more CFUs than that of rural spaces.