Search

Abstract Despite annual vaccination, influenza B viruses (IBV) continue to cause significant morbidity and mortality in humans. We have found that IBV infection resulted in a weaker innate and adaptive immune response than influenza A viruses (IAV) in ferrets. To understand and overcome the weak immune responses to IBV in ferrets, we administered type-I or type-III interferon (IFN) to ferrets following infection or vaccination and evaluated their effects on the immune response. IFN signaling following viral infection plays an important role in the initial innate immune response and affects subsequent adaptive immune responses. In the respiratory tract, IFN lambda (IFNL) has regulatory effects on adaptive immunity indirectly through thymic stromal lymphopoietin (TSLP), which then acts on immune cells to stimulate the adaptive response. Following IBV infection or vaccination, IFN treatment (IFN-Tx) upregulated gene expression of early inflammatory responses in the upper respiratory tract and robust IFN, TSLP, and inflammatory responses in peripheral blood cells. These responses were sustained following challenge or vaccination in IFN-Tx animals. Serum IFNL and TSLP levels were enhanced in IFN-Tx animals following challenge/rechallenge over mock-Tx; however, this difference was not observed following vaccination. Antibody responses in serum of IFN-Tx animals following IBV infection or vaccination increased more quickly and to higher titers and were sustained longer than mock-Tx animals over 3 months. Following rechallenge of infected animals 3 months post treatment, antibody levels remained higher than mock-Tx. However, IFN-Tx did not have an effect on antibody responses following challenge of vaccinated animals. A strong direct correlation was found between TSLP levels and antibody responses following challenge-rechallenge and vaccination-challenge indicating it as a useful tool for predicting adaptive immune responses following IBV infection or vaccination. The effects of IFN on strengthening both innate and adaptive responses to IBV may aid in development of more effective treatments following infection and improved influenza vaccines.

Abstract Climate change and population growth are putting increasing pressure on global food security. The development of high-yielding varieties for important crops such as wheat is crucial to meet these challenges. The basis for this is extensive exploitation of beneficial genetic variation resting in genebanks around the world. Selecting suitable donor genotypes from the vast number of wheat accessions stored in genebanks is a difficult task and depends critically on the density of information on the performance of individual accessions. Therefore, this study aimed to access phenotypic data from the Czech genebank, storing over 13,000 wheat accessions. We curated and analyzed data on heading date, plant height, and thousand grain weight for more than one-third of all available accessions regenerated across 70 years. The data underwent analysis using a linear mixed model, revealing high quality of curated data with heritability reaching 99%. The raw data, but also derived data such as the best linear unbiased estimations, are now available for the wheat collection of the Czech genebank for research and breeding.

Summary: Flaviviruses such as dengue virus (DENV), Zika virus (ZIKV), and yellow fever virus (YFV) are spread by mosquitoes and cause human disease and mortality in tropical areas. In contrast, Powassan virus (POWV), which causes severe neurologic illness, is a flavivirus transmitted by ticks in temperate regions of the Northern hemisphere. We find serologic neutralizing activity against POWV in individuals living in Mexico and Brazil. Monoclonal antibodies P002 and P003, which were derived from a resident of Mexico (where POWV is not reported), neutralize POWV lineage I by recognizing an epitope on the virus envelope domain III (EDIII) that is shared with a broad range of tick- and mosquito-borne flaviviruses. Our findings raise the possibility that POWV, or a flavivirus closely related to it, infects humans in the tropics.

This article provides background legal information on the problems of judicial review of EU external sanctions before the CJEU. It does so from two perspectives. First, the article discusses the general problems of judicial review of international (external) sanctions, hiding under the ambivalent label of “restrictive measures”; here, the legal problems arise mainly from the fact that EU secondary sanctions acts fall under both the CFSP and other EU policies, the two EU policy areas being relatively autonomous in terms of the types of secondary acts and their adoption. The article then analyses the human rights dimension of this judicial review; this is because certain fundamental rights (in particular the right to a defence and the right to effective remedy) place considerable demands on the procedural and evidentiary aspects of this judicial review in a situation where much of the evidence relies on classified intelligence information. The scope of the article allows the author to provide only an introduction to the issues discussed.

This paper discusses the development of rapid, reliable, and accurate polymerase chain reaction (PCR) assays for detecting opium poppy (Papaver somniferum L.) in food. Endpoint, quantitative, and digital PCRs were compared based on the amplification of a newly developed DNA marker targeting the NADPH-dependent codeinone reductase (COR) gene. Designed assays were shown to be highly specific and sensitive in discriminating opium poppy from other plant species, even in heat-treated and food samples. Digital PCR was the most sensitive, with a detection limit of up to 5 copies, i.e., approximately 14 pg of target DNA per reaction. Quantitative and digital PCR further allowed the quantification of opium poppy in up to 1.5 ng and 42 pg (15 copies) of target DNA in a sample, respectively. In addition, two duplex PCRs have been developed for the simultaneous detection of opium poppy DNA and representatives of (i) the Papaveraceae family or (ii) the Plantae kingdom. Finally, all designed assays were successfully applied for analysis of 15 commercial foodstuffs; two were suspected of being adulterated. The study results have an important impact on addressing food fraud and ensuring the safety and authenticity of food products. Beyond food adulteration, the study may also have significant implications for forensics and law enforcement.

Summary: Tick-borne encephalitis virus (TBEV) is a flavivirus that causes human neuroinfections and represents a growing health problem. The human monoclonal antibody T025 targets envelope protein domain III (EDIII) of TBEV and related tick-borne flaviviruses, potently neutralizing TBEV in vitro and in preclinical models, representing a promising candidate for clinical development. We demonstrate that TBEV escape in the presence of T025 or T028 (another EDIII-targeting human monoclonal antibody) results in virus variants of reduced pathogenicity, characterized by distinct sets of amino acid changes in EDII and EDIII that are jointly needed to confer resistance. EDIII substitution K311N impairs formation of a salt bridge critical for T025-epitope interaction. EDII substitution E230K is not on the T025 epitope but likely induces quaternary rearrangements of the virus surface because of repulsion of positively charged residues on the adjacent EDI. A combination of T025 and T028 prevents virus escape and improves neutralization.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has prompted great interest in novel broad-spectrum antivirals, including perylene-related compounds. In the present study, we performed a structure–activity relationship analysis of a series of perylene derivatives, which comprised a large planar perylene residue, and structurally divergent polar groups connected to the perylene core by a rigid ethynyl or thiophene linker. Most of the tested compounds did not exhibit significant cytotoxicity towards multiple cell types susceptible to SARS-CoV-2 infection, and did not change the expressions of cellular stress-related genes under normal light conditions. These compounds showed nanomolar or sub-micromolar dose-dependent anti-SARS-CoV-2 activity, and also suppressed the in vitro replication of feline coronavirus (FCoV), also termed feline infectious peritonitis virus (FIPV). Perylene compounds exhibited high affinity for liposomal and cellular membranes, and efficiently intercalated into the envelopes of SARS-CoV-2 virions, thereby blocking the viral–cell fusion machinery. Furthermore, the studied compounds were demonstrated to be potent photosensitizers, generating reactive oxygen species (ROS), and their anti-SARS-CoV-2 activities were considerably enhanced after irradiation with blue light. Our results indicated that photosensitization is the major mechanism underlying the anti-SARS-CoV-2 activity of perylene derivatives, with these compounds completely losing their antiviral potency under red light. Overall, perylene-based compounds are broad-spectrum antivirals against multiple enveloped viruses, with antiviral action based on light-induced photochemical damage (ROS-mediated, likely singlet oxygen-mediated), causing impairment of viral membrane rheology.

The article concerns the problematics of unburned earth used as material for moulding formwork or clay plaster with lowered contraction, especially about interior unburned earth plaster with the heightened percentage of clay and therefore improved sorption abilities. By adding a plasticizer, it is possible to modify the fluidity of the mixture. With lessening the amount of water it is possible to lower the shrinkage. Furthermore, we can increase the ratio of clay to sand and thus we can control the indoor microclimate. This is because the clay provides adsorption properties. Unburnt clay has been widely used as a building material in the past around the world. In the last century, this material was rather despised. Nowadays, however, this trend is reversing and the unburnt clay is experiencing a renaissance. Increasingly people emphasize ecology, a healthy and balanced indoor climate, and demand buildings with low operational and energy intensity. All this can be offered by unburnt clay, because it is energy-efficient in production, easily accessible in almost all parts of the world, harmless, and beneficial to human health, as clay can contribute to the improvement of the internal microclimate to a much greater extent than other building materials, as confirmed by many scientific findings.

The carbon (C) fixed by crops, which is exported with harvest and retained as postharvest residues in a field, is important for calculating the C balance. The aim of this study was to determine the effect of water availability on the C content in whole wheat plants. In a three-year field trial, the weights of grain, straw, chaff, stubble, and roots of two cultivars of winter wheat (Triticum aestivum L.) and one cultivar of einkorn wheat (Triticum monococcum L.) and their carbon contents were determined in water stress, irrigation, and rain-fed control treatments. The water availability, year, and cultivar had a significant influence on the C content in aboveground plant parts, but the effect of water on grain C was weak. The C content decreased with irrigation and increased with drought, but the differences were small (at most, 3.39% in chaff). On average, the C contents of grain, straw, chaff, and roots reached 45.0, 45.7, 42.6, and 34.9%, respectively. The amount of C exported with grain and left on the field in the form of postharvest residues depended on the weight of the total biomass and the ratio of grain to straw and residue. Whole plant C yield reached 8.99, 7.46, and 9.65 t ha−1 in rain-fed control, stressed, and irrigated treatments, respectively, and 8.91, 9.45, and 7.47 t ha−1 in Artix, Butterfly, and Rumona, respectively. Irrigation significantly increased the C content in grain and straw (but not in chaff, stubble, and roots) in comparison with water shortage conditions. On average, a grain yield of 1 t ha−1 corresponded to an average export of 0.447–0.454 t C ha−1 in the grain of all cultivars and inputs of 0.721, 0.832, and 2.207 t C ha−1 of residue to the soil in the form of straw and postharvest residue in the two cultivars of common wheat and one of einkorn. The results of the study provided reliable data for the calculation of the C balance of wheat under conditions of different water availability.

Crop Wild Relatives (CWRs), as potential sources of new genetic variants, are being extensively studied to identify genotypes that will be able to confer resistance to biotic stresses. In this study, a collection of barley wild relatives was assessed in the field, and their phenotypic variability was evaluated using a Barley Description List, reflecting the identified ecosites. Overall, the CWRs showed significant field resistance to various fungal diseases. To further investigate their resistance, greenhouse tests were performed, revealing that several CWRs exhibited resistance against Fusarium culmorum, Pyrenophora teres, and Puccinia hordei G.H. Otth. Additionally, to characterize the genetic diversity within the collection, DNA polymorphisms at 21 loci were examined. We successfully employed barley-specific SSR markers, confirming their suitability for identifying H. spontaneum and even H. marinum, i.e., perennial species. The SSR markers efficiently clustered the investigated collection according to species and ecotypes, similarly to the phenotypic assessment. Moreover, SSR markers associated with disease resistance revealed different alleles in comparison to those found in resistant barley cultivars. Overall, our findings highlight that this evaluated collection of CWRs represents a valuable reservoir of genetic variability and resistance genes that can be effectively utilized in breeding programs.

Perylenylethynyl derivatives have been recognized as broad-spectrum antivirals that target the lipid envelope of enveloped viruses. In this study, we present novel perylenylethynylphenols that exhibit nanomolar or submicromolar antiviral activity against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) and feline infectious peritonitis virus (FIPV) in vitro. Perylenylethynylphenols incorporate into viral and cellular membranes and block the entry of the virus into the host cell. Furthermore, these compounds demonstrate an ability to generate singlet oxygen when exposed to visible light. The rate of singlet oxygen production is positively correlated with antiviral activity, confirming that the inhibition of fusion is primarily due to singlet-oxygen-induced damage to the viral envelope. The unique combination of a shape that affords affinity to the lipid bilayer and the capacity to generate singlet oxygen makes perylenylethynylphenols highly effective scaffolds against enveloped viruses. The anticoronaviral activity of perylenylethynylphenols is strictly light-dependent and disappears in the absence of daylight (under red light). Moreover, these compounds exhibit negligible cytotoxicity, highlighting their significant potential for further exploration of the precise antiviral mechanism and the broader scope and limitations of this compound class.

13C discrimination (Δ13C) has been used in research as an indicator of water availability in crops; however, few data are available concerning fruit trees. The aim of this study was to examine the effect of irrigation on the Δ13C values of apple leaves. We assumed that Δ13C would increase with irrigation intensity. The Δ13C of apple trees (Malus domestica) cv. ‘Red Jonaprince’ was determined in the years 2019–2022. Leaf samples were collected in spring, in June, at the beginning of the irrigation campaign, and in autumn, in September, following the harvest. The irrigation doses were applied to replenish the water consumption, 0% (ET0), 50% (ET50), 75% (ET75), and 100% (ET100), of the calculated evapotranspiration (ET) levels. In November, the leaves collected from different positions on the shoots were sampled, assuming the Δ13C signature would reflect the changes occurring in the water supply during their growth. The irrigation rates had a significant effect on the Δ13C of the leaves when the data for the spring and summer months were pooled. On average, Δ13C increased from 20.77‰ and 20.73‰ for ET0 and ET50, respectively, to 20.80‰ and 20.95‰ for ET75 and ET100, respectively. When the data obtained for the spring and summer months were analysed separately, the effect of irrigation was weak (p < 0.043). The Δ13C value was always higher for treatment ET100 than treatment ET0, for individual experimental years and terms; however, the differences were minor and mostly insignificant. The leaf position had a strong significant effect on Δ13C; the values gradually decreased from the leaves growing from two-years-old branches (22.50‰) to the youngest leaves growing at the top of the current year’s shoots (21.07‰). This order was similar for all the experimental years. The results of the experiment suggest that 13C discrimination in apples is affected not only by water availability during growth, but also by the use of C absorbed in previous years.

Abstract Opium poppy (Papaver somniferum L.) is a versatile plant exploited by the pharmaceutical and food industries. Unfortunately, it is also infamously known as a source of highly addictive narcotics, primarily heroin. Drug abuse has devastating consequences for users and also has many direct or indirect negative impacts on human society as a whole. Therefore, developing a molecular genetic tool for the individualization of opium poppy, raw opium or heroin samples could help in the fight against the drug trade by retrieving more information about the source of narcotics and linking isolated criminal cases. Bioinformatic analysis provided insight into the distribution, density and other characteristics of roughly 150 thousand microsatellite loci within the poppy genome and indicated underrepresentation of microsatellites with the desired attributes. Despite this fact, 27 polymorphic STR markers, divided into three multiplexed assays, were developed in this work. Internal validation confirmed species-specific amplification, showed that the optimal amount of DNA is within the range of 0.625–1.25 ng per reaction, and indicate relatively well balanced assays according to the metrics used. Moreover, the stutter ratio (mean + 3 SD 2.28–15.59%) and allele-specific stutters were described. The analysis of 187 individual samples led to the identification of 158 alleles in total, with a mean of 5.85 alleles and a range of 3–14 alleles per locus. Most of the alleles (151) were sequenced by the Sanger method, which enabled us to propose standardized nomenclature and create three allelic ladders. The OpiumPlex system discriminates most of the varieties from each other and pharmaceutical varieties from the others (culinary, dual and ornamental).

Quinoa displays huge genetic variability and adaptability to distinct climatic conditions. Quinoa seeds are a good source of nutrients; however, the overall nutritional composition and nutrient content is influenced by numerous factors. This study focused on the nutritional and morphologic evaluation of various quinoa genotypes grown in the Czech Republic. Significant differences between years were observed for morphological traits (plant height, inflorescence length, weight of thousand seeds). The weather conditions in the year 2018 were favorable for all the morphological traits. The protein content of quinoa accessions ranged between 13.44 and 20.01% and it was positively correlated to mauritianin. Total phenolic content varied greatly from year to year, while the antioxidant activity remained relatively stable. The most abundant phenolic compounds were the flavonoids miquelianin, rutin, and isoquercetin. Isoquercetin, quercetin, and N-feruoloyl octopamine showed the highest stability under variable weather conditions in the analyzed years. A total of six compounds were detected and quantified in quinoa for the first time. Most varieties performed well under Central European conditions and can be considered a good source of nutrients and bioactive compounds. These data can be used as a source of information for plant breeders aiming to improve the quality traits of quinoa.

In a three-year experiment (2019–2021), the roots of 7-year-old apple trees (Malus domestica cv. ‘Red Jonaprince’) grown under drip irrigation were studied. The aim of the study was to determine the effect of irrigation on root density at different depths and distances from the trunk. The working hypothesis assumed that irrigation significantly affects the total length of apple roots. The irrigation treatments corresponding to the calculated water evapotranspiration (ET100), 50% of the calculated ET (ET50), a control (ET0, no irrigation, under rainfed conditions), and a treatment using double-drip lines (2Drops) were monitored. Soil cores were collected in spring and autumn. The total length of the roots (TRLt) and the length of new vital roots (TRLv) to a depth of 80 cm were evaluated. The effects of treatments were mostly insignificant for the TRLt; only in the dry season in 2019 were the TRLt values of the irrigated treatments (ET50 and ET100) significantly higher, 18.67 km·m−2 and 17.45 km·m−2, in comparison to 11.16 km·m−2 for the ET0, at a 10 cm distance from the tree trunk. The irrigation treatments had a statistically significant effect on the TRLv values near the trunk in 2019 and 2020, while in autumn 2020 and 2021, irrigation significantly affected the TRLv at greater distances from the tree trunk. In summary, the irrigation treatments mostly had no significant effect on the total root length. However, an effect of irrigation on the root length of new vital roots was observed at certain sampling dates and distances from the trunk.

In this article, an innovative new method of non-destructive testing for estimating the sensitivity of limestone is presented. An advantage of the laser ultrasonic structuroscopy method is the time and the quality, due to which less tests are necessary for an analysis of rock properties. Limestone is measured with an innovative structural control method, which shows a good qualitative and quantitative agreement with the basic structural analysis.

The article deals with an innovative method designed to check project documentation of buildings at the design stage, specifically exploring the possibility to implement FMEA and PDCA methodologies. Based on performed measurements and data collection, it theoretically determines the riskiest areas of the project documentation, which should be given special attention in order to reduce later costs for construction companies to fix the reported complaints. The research proves that the application of the FMEA and PDCA methodology can be very useful regarding the elimination of defects in the project documentation of constructions already in the phase of construction preparation.

Seeds of Czech poppies used for culinary purposes are frequently mixed with seeds of imported low-quality poppies with higher alkaloid content and poor taste properties. That not only decreases the quality of the product on the market, but it can also represent a potential health hazard for consumers. To prevent such adulteration, the method is needed that could clearly detect the presence of the low-quality poppies in the products on the market and which can also confirm the authenticity of the genetic material for growers. For that reason, length polymorphism of Simple Sequence Repeats was evaluated in the blue and non-blue poppy varieties of Czech and foreign origin. Used markers clearly distinguished 'Czech blue poppy' cultivars from blue poppies of another origin as well as from poppies of non-blue seed colour. We conclude that used markers can be applied to a commodity verification on the market to detect/exclude the presence of other genotypes.

Fluctuations in precipitation and higher evapotranspiration due to rising temperatures are reflected in reduced wheat yields, even in areas with a low historical incidence of drought. In this study, the effects of drought (S) and irrigation (IR) on spelt, einkorn wheat, and two common wheat cultivars were assessed in a field experiment in the years 2018–2021. Water availability was differentiated from the flowering stage using a mobile cover and drip irrigation. Grain yield, canopy temperature, and discrimination of 13C in grain (Δ 13C) were monitored. Drought reduced the average grain yield of common wheat to 5.24 t.ha−1, which was 67.00% of the rain-fed control (C) yield, and 62.09% of the irrigated wheat yield. For spelt and einkorn wheat, the average grain yield from stressed plants was 2.02 t.ha−1; this was 79.97% of the C-variant yield, and 70.82% of the IR-variant yield. Higher stand temperatures were an excellent indicator of water deficit in the stressed crops. The relationship between temperature and final grain yield in the monitored variants was always negative. In all years, discrimination of 13C in grain corresponded to water availability; in its effect on yields, the correlation was always positive. Between 2018 and 2020, spelt and einkorn exhibited lower Δ 13C in comparison with common wheat in all variants, suggesting a greater impact of differentiated water supply. The results of the experiment conclusively demonstrated systematic effects of drought after flowering upon yields and other studied characteristics.

Background: Vedolizumab demonstrated different placental pharmacokinetics than other immunoglobulin G1 antibodies, leading to lower drug levels in cord blood in contrast to maternal blood at the time of delivery. The placental transfer of ustekinumab seems to have a pattern similar to anti-tumour necrosis factor agents. Current evidence on the placental pharmacokinetics of vedolizumab and ustekinumab is limited. We aimed to assess the placental transfer of ustekinumab and vedolizumab in pregnant patients with inflammatory bowel disease. Methods: Consecutive women from a prospective observational study who were exposed to ustekinumab or vedolizumab within 2 months prior to conception or during pregnancy were included. Ustekinumab and vedolizumab levels were measured in maternal and cord blood at the time of delivery. Results: Drug levels were available in 31 infant-mother pairs (15 exposed to ustekinumab and 16 to vedolizumab). The median maternal and newborn ustekinumab levels were 5.3 mg/l and 10.3 mg/l, respectively (the median infant-to-maternal ratio was 1.7), while the median maternal and cord vedolizumab levels were 7.3 mg/l and 4.5 mg/l (the median infant-to-maternal ratio was 0.66). The ustekinumab levels in cord blood positively correlated with the maternal levels at delivery (ρ = 0.751, p = 0.001). However, no correlation with the timing of the last drug administration was found. In contrast, the vedolizumab levels in cord blood demonstrated significant positive correlation with the maternal levels (ρ = 0.831, p

Abstract Background Tick-borne encephalitis (TBE) is a severe neuropathological disorder caused by tick-borne encephalitis virus (TBEV). Brain TBEV infection is characterized by extensive pathological neuroinflammation. The mechanism by which TBEV causes CNS destruction remains unclear, but growing evidence suggests that it involves both direct neuronal damage by the virus infection and indirect damage caused by the immune response. Here, we aimed to examine the TBEV-infection-induced innate immune response in mice and in human neural cells. We also compared cytokine/chemokine communication between naïve and infected neuronal cells and astrocytes. Methods We used a multiplexed Luminex system to measure multiple cytokines/chemokines and growth factors in mouse serum samples and brain tissue, and in human neuroblastoma cells (SK-N-SH) and primary cortical astrocytes (HBCA), which were infected with the highly pathogenic TBEV strain Hypr. We also investigated changes in cytokine/chemokine production in naïve HBCA cells treated with virus-free supernatants from TBEV-infected SK-N-SH cells and in naïve SK-N-SH cells treated with virus-free supernatants from TBEV-infected HBCA cells. Additionally, a plaque assay was performed to assess how cytokine/chemokine treatment influenced viral growth following TBEV infection. Results TBEV-infected mice exhibited time-dependent increases in serum and brain tissue concentrations of multiple cytokines/chemokines (mainly CXCL10/IP-10, and also CXCL1, G-CSF, IL-6, and others). TBEV-infected SK-N-SH cells exhibited increased production of IL-8 and RANTES and downregulated MCP-1 and HGF. TBEV infection of HBCA cells activated production of a broad spectrum of pro-inflammatory cytokines, chemokines, and growth factors (mainly IL-6, IL-8, CXCL10, RANTES, and G-CSF) and downregulated the expression of VEGF. Treatment of SK-N-SH with supernatants from infected HBCA induced expression of a variety of chemokines and pro-inflammatory cytokines, reduced SK-N-SH mortality after TBEV infection, and decreased virus growth in these cells. Treatment of HBCA with supernatants from infected SK-N-SH had little effect on cytokine/chemokine/growth factor expression but reduced TBEV growth in these cells after infection. Conclusions Our results indicated that both neurons and astrocytes are potential sources of pro-inflammatory cytokines in TBEV-infected brain tissue. Infected/activated astrocytes produce cytokines/chemokines that stimulate the innate neuronal immune response, limiting virus replication, and increasing survival of infected neurons.

The effect of leaf-applied fertilizers on nutritional status and yields of apple cv. Rubinola grown on two soil types (Luvisol, Cambisol) differing in yield levels was studied in a three-year experiment at the orchard in Vanovice, Czech Republic. Two fertilizers, A (containing N, Mg, Ca, B and Zn) and B (N, B, Zn, Mn and Fe) were applied after blooming, leaves and fruit were analyzed. Zn and B concentrations increased significantly in leaves and fruit in both soils after the application of solution A compared to the control. Foliar fertilizer B increased the concentration of Zn and Mn significantly both in leaves and fruit compared to the control. The content of Mn in dry leaf matter increased almost 4 times (from the control level about 30 mg/kg), of B up to 1.5 times (from 28 mg/kg) and the content of Zn even more than 10 times (from 15 mg/kg). The application of foliar fertilizers was more efficient in the Cambisol orchard section with worse soil conditions; however, the enhanced nutritional status did not significantly increase fruit yields in either of the experimental soil types.

Human rhinoviruses (HRV) are the most common cause of viral respiratory tract infections. While normally mild and self-limiting in healthy adults, HRV infections are associated with bronchiolitis in infants, pneumonia in immunocompromised patients, and exacerbations of asthma and COPD. The human cathelicidin LL-37 is a host defense peptide (HDP) with broad immunomodulatory and antimicrobial activities that has direct antiviral effects against HRV. However, LL-37 is known to be susceptible to the enzymatic activity of peptidyl arginine deiminases (PAD), and exposure of the peptide to these enzymes results in the conversion of positively charged arginines to neutral citrullines (citrullination). Here, we demonstrate that citrullination of LL-37 reduced its direct antiviral activity against HRV. Furthermore, while the anti-rhinovirus activity of LL-37 results in dampened epithelial cell inflammatory responses, citrullination of the peptide, and a loss in antiviral activity, ameliorates this effect. This study also demonstrates that HRV infection upregulates PAD2 protein expression, and increases levels of protein citrullination, including histone H3, in human bronchial epithelial cells. Increased PADI gene expression and HDP citrullination during infection may represent a novel viral evasion mechanism, likely applicable to a wide range of pathogens, and should therefore be considered in the design of therapeutic peptide derivatives.

Abstract Background The wild relatives of crop species represent a potentially valuable source of novel genetic variation, particularly in the context of improving the crop’s level of tolerance to abiotic stress. The mechanistic basis of these tolerances remains largely unexplored. Here, the focus was to characterize the transcriptomic response of the nodes (meristematic tissue) of couch grass (a relative of barley) to dehydration stress, and to compare it to that of the barley crown formed by both a drought tolerant and a drought sensitive barley cultivar. Results Many of the genes up-regulated in the nodes by the stress were homologs of genes known to be mediated by abscisic acid during the response to drought, or were linked to either development or lipid metabolism. Transporters also featured prominently, as did genes acting on root architecture. The resilience of the couch grass node arise from both their capacity to develop an altered, more effective root architecture, but also from their formation of a lipid barrier on their outer surface and their ability to modify both their lipid metabolism and transporter activity when challenged by dehydration stress. Conclusions Our analysis revealed the nature of dehydration stress response in couch grass. We suggested the tolerance is associated with lipid metabolism, the induction of transporters and the re-programming of development coordinated by ABA. We also proved the applicability of barley microarray for couch grass stress-response analysis.

Dogs are frequently infected with the tick-borne encephalitis virus (TBEV). However, to date, only a few clinically manifest cases of tick-borne encephalitis (TBE) have been reported in dogs. In this study, three-month-old beagle dogs were infected with TBEV through a subcutaneous injection. Body temperature, clinical signs, blood haematology, blood biochemistry, and immune responses were monitored for up to 28 days postinfection (p.i.). No changes in body temperature or clinical signs were observed in the infected dogs. Most haematology and blood biochemistry parameters were unchanged after the infection, except for a slight reduction in blood lymphocyte counts, but they were within the physiological range. Low-titre viraemia was detected in 2/4 infected dogs between days 1 and 3 p.i. All infected dogs developed a robust immune response, in terms of neutralising antibodies. Thus, TBEV infections lead to effective seroconversion in dogs. Next, to assess TBEV exposure in dogs in the TBEV-endemic region of the Czech Republic, we conducted a serosurvey. Virus neutralisation tests revealed TBEV-specific antibodies in 17 of 130 (13.07%) healthy dogs, which confirmed a high, but clinically inappreciable TBEV exposure rate in the endemic area. The seropositivity rate was similar (12.7%; 41 positives out of 323) in a subgroup of dogs with various clinical disorders, and it was 13.4% (23 out of 171) in a subgroup of dogs with signs of acute neurological disease. Two dogs with fatal acute meningoencephalitis showed positive results for TBEV-specific IgM and IgG antibodies. These data extended our understanding of the clinical presentation of TBEV infections.

SARS-CoV-2 has caused an extensive pandemic of COVID-19 all around the world. Key viral enzymes are suitable molecular targets for the development of new antivirals against SARS-CoV-2 which could represent potential treatments of the corresponding disease. With respect to its essential role in the replication of viral RNA, RNA-dependent RNA polymerase (RdRp) is one of the prime targets. HeE1-2Tyr and related derivatives were originally discovered as inhibitors of the RdRp of flaviviruses. Here, we present that these pyridobenzothiazole derivatives also significantly inhibit SARS-CoV-2 RdRp, as demonstrated using both polymerase- and cell-based antiviral assays.

Buckwheat is a nutritionally valuable crop, an alternative to common cereals also usable in gluten-free diets. The selection of buckwheat genotypes suitable for further breeding requires the characterization and evaluation of genetic resources. The main objective of this work was to evaluate selected phenotypic and morphological traits using international buckwheat descriptors, including total phenolic content and antioxidant activity, on a unique set of 136 common buckwheat accessions grown in 2019–2020 under Czech Republic conditions. In addition, UHPLC-ESI- MS/MS was used to analyze a wide spectrum of 20 phenolic compounds in buckwheat seeds, including four flavanols, three phenolic acids, seven flavonols, four flavones, and two flavanones. Significant differences among years and genotypes were observed for morphological traits (plant height and 1000-seed weight) and antioxidant activity, as well as levels of observed chemical compounds. Antioxidant activity, crude protein content, plant height and rutin content were characterized by higher mean values in 2020 than in 2019 and vice versa for total polyphenol content and 1000-seed weight. Crude protein content was the most stable across years, while total polyphenol content and rutin content varied greatly from year to year. The most abundant phenolic compounds were rutin, hyperoside, epicatechin, catechin, vitexin, isovitexin, orientin and isoorientin. Protein content was negatively correlated with plant height, catechin and epicatechin content. On the other hand, AA and TPC were positively correlated with rutin, hyperoside and chlorogenic acid. Five accessions showed high stability of the evaluated traits under changing conditions within both years of observation. These materials can be used in breeding programmes aimed at improving buckwheat genotypes with emphasis on quality traits.

The monitoring of trace metals in microbial cells is relevant for diagnosis of inflammatory bowel disease (IBD). Sulfate-reducing bacteria (SRB) represent an important factor in the IBD development. The content of trace metals in bacterial cells may reflect the functioning of the enzyme systems and the environmental impact on the occurrence of SRB. The aim of our research was to compare the content of trace elements in the cells of SRB cultures isolated from fecal samples of patients with IBD and healthy people. The contents of 11 chemical elements in the bacterial cells of SRB were analyzed by the inductively coupled plasma-mass-spectrometry (ICP-MS) method. Significant changes in the content of calcium, zinc, magnesium, potassium, and iron were observed in patients with IBD compared to healthy individuals. Through a principal component analysis (PCA), a total variability of 67.3% in the difference between the samples was explained. The main factors influencing the total variability in the bacterial cells of SRB isolated from patients suffering from IBD were the content of the micro- and trace elements, such as manganese (with power 0.87), magnesium and cobalt (0.86), calcium (0.84), molybdenum (0.81), and iron (0.78). Such changes in the elemental composition of SRB under different conditions of existence in the host may indicate adaptive responses of the microorganisms, including the inclusion of oxidative stress systems, which can lead to changes in SRB metabolism and the manifestation of parameters of IBD in humans. The use of PCA might make it possible in the future to predict the development and ratio of SRB in patients with various diseases.

During the operation of engineering structures made of natural stone, for industrial and civil purposes, an important parameter in monitoring their technical condition is the assessment of their reliability and safety under the influence of various external influences. In this case, high-quality monitoring of the stress–strain state of natural stone structures, its physical, mechanical and filtration properties, as well as internal structural features is necessary to study the possibility of replacing individual elements of objects that have lost their original characteristics. To assess the state of geomaterials, this article proposes using a complex of introscopic methods, including infrared radiometry and laser-ultrasound structuroscopy. An important aspect is the calculation based on the Green–Christoffel equation of the velocity of a quasi-longitudinal wave in limestone consisting of densely packed, chaotically oriented calcite grains with a small quartz content. For the first time, using laser-ultrasonic structuroscopy and standard methods for determining open porosity, both total and closed porosity were determined. This allowed us to find the values of specific heat capacities of dry and water-saturated samples. The obtained values are used to find the ratio of changes in the temperature of dry and water-saturated samples at the same stress values. The results obtained demonstrate the need to take into account changes in the intensity of thermal radiation of limestone with different moisture content under conditions of uniaxial compression, when identifying changes in the stress state of elements of stone structures in real conditions.

Six onion cultivars Bingo, Dormo, Elenka, Elbrus, Spirit, and Sturon grown in the Czech Republic for commercial purposes were analysed to investigate the content of health-promoting compounds. The results showed that at harvest time, cysteine sulfoxide content varied from 32.38 to 44.16 g/kg of dry weight, polyphenol content was between 2.66 and 3.37 g/kg of dry weight, and antioxidant activity ranged from 0.75 to 0.83 g/kg. Cv. Bingo had the highest level of the analysed compounds. The cultivars were concurrently analysed by DNA (microsatellite) markers. Dendrograms based on the chemical composition and DNA analysis were almost identical. This finding confirms the dependence of the secondary metabolite content on onion genotype.

Determination of the water supply available in soils for crops is important for both the calculation of water balance and the prediction of water stress. An approach to calculations of available water content in layers of the root zone, depletion of water during growth, and water balance, with limited access to data on farms, is presented. Soil water retention was calculated with simple pedotransfer functions from the texture of soil layers, root depth, and depletion function were derived from observed data; and the potential evapotranspiration was calculated from the temperature. A comparison of the calculated and experimental soil water contents showed a reasonable fit.

LL-37, the only member of the mammalian cathelicidin in humans, plays an essential role in innate immunity by killing pathogens and regulating the inflammatory response. However, at an inflammatory focus, arginine residues in LL-37 can be converted to citrulline via a reaction catalyzed by peptidyl-arginine deiminases (PAD2 and PAD4), which are expressed in neutrophils and are highly active during the formation of neutrophil extracellular traps (NETs). Citrullination impairs the bactericidal activity of LL-37 and abrogates its immunomodulatory functions. Therefore, we hypothesized that citrullination-resistant LL-37 variants would retain the functionality of the native peptide in the presence of PADs. To test this hypothesis, we synthetized LL-37 in which arginine residues were substituted by homoarginine (hArg-LL-37). Bactericidal activity of hArg-LL-37 was comparable with that of native LL-37, but neither treatment with PAD4 nor exposure to NETs affected the antibacterial and immunomodulatory activities of hArg-LL-37. Importantly, the susceptibilities of LL-37 and hArg-LL-37 to degradation by proteases did not significantly differ. Collectively, we demonstrated that citrullination-resistant hArg-LL-37 is an attractive lead compound for the generation of new agents to treat bacterial infections and other inflammatory diseases associated with enhanced PAD activity. Moreover, our results provide a proof-of-concept for synthesis of therapeutic peptides using homoarginine.

Spatial variability of crop growth and yields is the result of many interacting factors. The contribution of the factors to variable yields is often difficult to separate. This work studied the relationships between the 13C discrimination (Δ13C) of plants and the spatial variability of field soil conditions related to impacts of water shortage on crop yield. The 13C discrimination, the indicator of water shortage in plants, 15N (δ15N) discrimination, and nitrogen (N) content were determined in grains of winter wheat, spring barley, and pea. The traits were observed at several dozens of grid spots in seven fields situated in two regions with different soil and climate conditions between the years 2017 and 2019. The principles of precision agriculture were implemented in some of the studied fields and years by variable rate nitrogen fertilization. The Δ13C significantly correlated with grain yields (correlation coefficient from 0.66 to 0.94), with the exception of data from the wetter year 2019 at the site with higher soil water capacity. The effect of drought was demonstrated by statistically significant relationships between Δ13C in dry years and soil water capacity (r from 0.46 to 0.97). The significant correlations between Δ13C and N content of seeds and soil water capacity agreed with the expected impact of water shortage on plants. The 13C discrimination of crop seeds was confirmed as a reliable indicator of soil spatial variability related to water shortage. Stronger relationships were found in variably fertilized areas.

The purpose of this study was to determine the effect of sixty years of contrasting fertilization treatments on the roots of winter wheat (Triticum aestivum L.) at sites with different soil and climate conditions. The depth and length density distribution of the wheat roots were determined between 2014 and 2016 in a crop rotation experiment established in 1955 at three sites: Lukavec, Čáslav, and Ivanovice (Czech Republic). Three fertilization treatments were examined: Zero fertilization (N0), organic (ORG) fertilization, and mineral (MIN) fertilization. The fertilization, site, and year all had a significant effect on the total root length (TRL). The average TRL per square meter reached 30.2, 37.0, and 46.1 km with the N0, ORG, and MIN treatments at Lukavec, respectively, which was the site with the lightest soil and the coldest climate. At Čáslav and Ivanovice (warmer sites with silt and loamy soils), the average TRL per square meter reached 41.2, 42.4, and 47.7 km at Čáslav and 49.2, 55.3, and 62.9 km at Ivanovice with the N0, MIN, and ORG treatments, respectively. The effect of fertilization on the effective root depth (EfRD), the depth at which the root length density dropped below 2.0 cm cm−3, was significant, while the maximum root depth (RMD) was only marginally affected. With the sites and years averaged, the MIN-treated plants showed a greater EfRD (102.2 cm) in comparison to the N0 (81.8 cm) and ORG (93.5 cm) treatments. The N0 treatment showed no signs of an adaptive reaction to the root system, with potential improvement for nutrient acquisition, while optimal fertilization contributed to the potential for resource depletion from the soil profile.

The paper is focused on the examination of the internal quality of joints created in a multi-material additive manufacturing process. The main part of the work focuses on experimental production and non-destructive testing of restrained joints of modified PLA (polylactic acid) and ABS (Acrylonitrile butadiene styrene) three-dimensional (3D)-printed on RepRap 3D device that works on the “open source” principle. The article presents the outcomes of a non-destructive materials test in the form of the data from the Laser Amplified Ultrasonography, microscopic observations of the joints area and tensile tests of the specially designed samples. The samples with designed joints were additively manufactured of two materials: Specially blended PLA (Market name—PLA Tough) and conventionally made ABS. The tests are mainly focused on the determination of the quality of material connection in the joints area. Based on the results obtained, the samples made of two materials were compared in the end to establish which produced material joint is stronger and have a lower amount of defects.

The onion (Allium cepa L.) constitutes an important part of the human diet and is of economic importance on the vegetable market. Methods that allow unambiguous differentiation of cultivars and breeding lines are required for market control and the protection of plant breeders' rights. DNA analyses, such as single sequence repeat polymorphism (SSRs), represent such a tool. Based on the evaluation of polymorphism in 21 SSR loci, we established a panel of 15 easy to score SSR markers that differentiated 16 commercial onion cultivars in the Czech Republic. The polymorphism ranged from 2-3 alleles per locus, indicating a low level of diversity among the cultivars. The panel can also be used to evaluate genetic resources.

Protegrins are porcine antimicrobial peptides (AMPs) that belong to the cathelicidin family of host defense peptides. Protegrin-1 (PG-1), the most investigated member of the protegrin family, is an arginine-rich peptide consisting of 18 amino acid residues, its main chain adopting a β-hairpin structure that is linked by two disulfide bridges. We report on the immune modulatory activity of PG-1 and its analogs in neutralizing bacterial endotoxin and capsular polysaccharides, consequently inhibiting inflammatory mediators’ release from macrophages. We demonstrate that the β-hairpin structure motif stabilized with at least one disulfide bridge is a prerequisite for the immune modulatory activity of this type of AMP.

The content of nitrate or mineral nitrogen (Nmin = N-NO-3 + N-NH +4) in soil in autumn is recognized as the indicator of potential risk of N leaching during winter. In this contribution, the apparent changes of Nmin in the 0-60 cm soil layer, during winter, on farm fields in the Czech Republic were calculated. A significant positive relationship between Nmin in autumn and the change during winter was observed in eight out of the nine farms. Nitrate N data produced similar relationships as Nmin. The regression analysis suggested that 40-90% of Nmin above a specific amount, 14-35 kg N/ha (interception of regression line, I1), on farms was apparently lost from the soil zone. Corresponding results for pooled data (n = 187) were 74% and 25 kg N/ha (r = 0.90, P < 0.001). The proportion of N leached from the 0-60 cm layer, calculated with a simple leaching equation was significantly correlated (n = 187, r = 0.92, P < 0.001) with observed Nminchange during winter, with the intercept (I2) significantly different from zero (-30.9 kg/ha). When the average value of regression intercept I1 of farms, or of pooled data, were introduced to the leaching equation as a constant correction parameter, the fit was satisfactory (r = 0.93 and 0.92, resp.) and the intercepts (-3.1 kg and -5.4 kg N/ha, resp.) were not significantly different from zero (at P < 0.01). The results of the study support the use of autumn Nmin within the leaching equation as a robust indicator of the risk of N leaching.

We recently reported that HIV-1 infection can be inhibited by innate antimicrobial components of human seminal plasma (SP). Conversely, naturally occurring peptidic fragments from the SP-derived prostatic acid phosphatase (PAP) have been reported to form amyloid fibrils called "SEVI" and enhance HIV-1 infection in vitro. In order to understand the biological consequence of this proviral effect, we extended these studies in the presence of human SP. PAP-derived peptides were agitated to form SEVI and incubated in the presence or absence of SP. While PAP-derived peptides and SEVI alone were proviral, the presence of 1% SP ablated their proviral activity in several different anti-HIV-1 assays. The anti-HIV-1 activity of SP was concentration dependent and was reduced following filtration. Supraphysiological concentrations of PAP peptides and SEVI incubated with diluted SP were degraded within hours, with SP exhibiting proteolytic activity at dilutions as high as 1:200. Sub-physiological concentrations of two prominent proteases of SP, prostate-specific antigen (PSA) and matriptase, could degrade physiological and supraphysiological concentrations of PAP peptides and SEVI. While human SP is a complex biological fluid, containing both antiviral and proviral factors, our results suggest that PAP peptides and SEVI may be subject to naturally occurring proteolytic components capable of reducing their proviral activity.

The extensive world-wide morbidity and mortality caused by influenza A viruses highlights the need for new insights into the host immune response and novel treatment approaches. Cationic Host Defense Peptides (CHDP, also known as antimicrobial peptides), which include cathelicidins and defensins, are key components of the innate immune system that are upregulated during infection and inflammation. Cathelicidins have immunomodulatory and anti-viral effects, but their impact on influenza virus infection has not been previously assessed. We therefore evaluated the effect of cathelicidin peptides on disease caused by influenza A virus in mice. The human cathelicidin, LL-37, and the murine cathelicidin, mCRAMP, demonstrated significant anti-viral activity in vivo, reducing disease severity and viral replication in infected mice to a similar extent as the well-characterized influenza virus-specific antiviral drug zanamivir. In vitro and in vivo experiments suggested that the peptides may act directly on the influenza virion rather than via receptor-based mechanisms. Influenza virus-infected mice treated with LL-37 had lower concentrations of pro-inflammatory cytokines in the lung than did infected animals that had not been treated with cathelicidin peptides. These data suggest that treatment of influenza-infected individuals with cathelicidin-derived therapeutics, or modulation of endogenous cathelicidin production may provide significant protection against disease.

BackgroundThe emergence and rapid spread of the 2009 H1N1 pandemic influenza A virus (H1N1pdm) in humans highlights the importance of enhancing the capability of existing influenza surveillance systems with tools for rapid identification of emerging and re-emerging viruses. One of the new approaches is the RT-PCR electrospray ionization mass spectrometry (RT-PCR/ESI-MS) technology, which is based on analysis of base composition (BC) of RT-PCR amplicons from influenza "core" genes. Combination of the BC signatures represents a "genomic print" of an influenza A virus.Methodology/principal findingsHere, 757 samples collected between 2006 and 2009 were tested, including 302 seasonal H1N1, 171 H3N2, 7 swine triple reassortants, and 277 H1N1pdm viruses. Of the 277 H1N1pdm samples, 209 were clinical specimens (throat, nasal and nasopharyngeal swabs, nasal washes, blood and sputum). BC signatures for the clinical specimen from one of the first cases of the 2009 pandemic, A/California/04/2009, confirmed it as an unusual, previously unrecognized influenza A virus, with "core" genes related to viruses of avian, human and swine origins. Subsequent analysis of additional 276 H1N1pdm samples revealed that they shared the genomic print of A/California/04/2009, which differed from those of North American swine triple reassortant viruses, seasonal H1N1 and H3N2 and other viruses tested. Moreover, this assay allowed distinction between "core" genes of co-circulating groups of seasonal H1N1, such as clades 2B, 2C, and their reassortants with dual antiviral resistance to adamantanes and oseltamivir.Conclusions/significanceThe RT-PCR/ESI-MS assay is a broad range influenza identification tool that can be used directly on clinical specimens for rapid and accurate detection of influenza virus genes. The assay differentiates the H1N1pdm from seasonal and other nonhuman hosts viruses. Although not a diagnostic tool, this assay demonstrates its usefulness and robustness in influenza virus surveillance and detection of novel and unusual viruses with previously unseen genomic prints.

Capsular polysaccharides (CPS) are a major virulence factor in meningococcal infections and form the basis for serogroup designation and protective vaccines. Our work has identified meningococcal CPS as a pro-inflammatory ligand that functions through TLR2 and TLR4-MD2-dependent activation. We hypothesized that human cationic host defense peptides interact with CPS and influence its biologic activity. Accordingly, the interaction of meningococcal CPS with the human-derived cationic peptide LL-37, which is expressed by phagocytic and epithelial cells that interface with meningococci during infection, was investigated. LL-37 neutralized the pro-inflammatory activity of endotoxin-free CPS as assessed by TLR2 and TLR4-MD-2-dependent release of TNFα, IL-6 and IL-8 from human and murine macrophages. The cationic and hydrophobic properties of LL-37 were crucial for this inhibition, which was due to binding of LL-37 to CPS. LL-37 also inhibited the ability of meningococcal CPS to induce nitric oxide release, as well as TNFα and CXCL10 (IP-10) release from TLR4-sufficient and TLR4-deficient murine macrophages. Truncated LL-37 analogs, especially those that retained the antibacterial domain, inhibited vaccine grade CPS and meningococcal CPS prepared from the major serogroups (A, B C, Y and W135). Thus, LL-37 interaction with CPS was independent of specific glucan structure. We conclude that the capacity of meningococcal CPS to activate macrophages via TLR2 and TLR4-MD-2 can be inhibited by the human cationic host defense peptide LL-37 and propose that this impacts CPS-based vaccine responses.