Detection of Small-Molecule Interactions with Fibrillar Tau Protein Aggregates Using Microscale Thermophoresis.

Aggregated fibrillar tau protein is a pathological hallmark of several neurodegenerative diseases. Small molecules that bind to tau fibrils may be applied for their detection and quantification. This is of great importance as they can potentially be used for earlier diagnosis of disease and disease progression. Microscale thermophoresis (MST) enables the detection of biomolecular interactions in an aqueous environment in which no immobilization of either reaction partner is required. Here, an MST assay methodology is described for the detection of the interaction between a variety of small molecules and tau fibrils. The results of this study demonstrate that MST is a practical methodology to quantify interactions between small molecules and tau fibrillar aggregates.

Examination of the Virulence of Actinobacillus pleuropneumoniae Serovar 16 in Pigs.

Different virulence variants of A. pleuropneumoniae are involved in the etiology of porcine pleuropneumonia. The purpose of the present trial was examination of the virulence of the Actinobacillus pleuropneumoniae A-85/14 strain, the type strain of serovar 16, in an animal challenge experiment. Thirty 12-week-old piglets seronegative for A. pleuropneumoniae were allocated into three trial groups each of 10 animals, and they were infected intranasally with 106, 107, or 108 colony forming units (cfu) of the strain, respectively. Clinical signs were recorded twice a day, and the animals were euthanized 6 days after the infection. Typical clinical signs and postmortem lesions of porcine pleuropneumonia were seen in the animals of each trial group; however, they were generally mild, and no significant differences could be seen between the three groups. Even 106 colony forming units of A. pleuropneumoniae A-85/14 strain could induce clinical signs and lesions. Based on these results, the type strain of serovar 16 of A. pleuropneumoniae must be regarded as a typical pathogenic strain of the species.

Mannich-type modifications of (-)-cannabidiol and (-)-cannabigerol leading to new, bioactive derivatives.

(-)-Cannabidiol (CBD) and (-)-cannabigerol (CBG) are two major non-psychotropic phytocannabinoids that have many beneficial biological properties. However, due to their low water solubility and prominent first-pass metabolism, their oral bioavailability is moderate, which is unfavorable for medicinal use. Therefore, there is a great need for appropriate chemical modifications to improve their physicochemical and biological properties. In this study, Mannich-type reaction was used for the synthetic modification of CBD and CBG for the first time, and thus fifteen new cannabinoid derivatives containing one or two tertiary amino groups were prepared. Thereafter the antiviral, antiproliferative and antibacterial properties of the derivatives and their effects on certain skin cells were investigated. Some modified CBD derivatives showed remarkable antiviral activity against SARS-CoV-2 without cytotoxic effect, while synthetic modifications on CBG resulted in a significant increase in antiproliferative activity in some cases compared to the parent compound.

Patch seriation to visualize data and model parameters.

We developed a new seriation merit function for enhancing the visual information of data matrices. A local similarity matrix is calculated, where the average similarity of neighbouring objects is calculated in a limited variable space and a global function is constructed to maximize the local similarities and cluster them into patches by simple row and column ordering. The method identifies data clusters in a powerful way, if the similarity of objects is caused by some variables and these variables differ for the distinct clusters. The method can be used in the presence of missing data and also on more than two-dimensional data arrays. We show the feasibility of the method on different data sets: on QSAR, chemical, material science, food science, cheminformatics and environmental data in two- and three-dimensional cases. The method can be used during the development and the interpretation of artificial neural network models by seriating different features of the models. It helps to identify interpretable models by elucidating clusters of objects, variables and hidden layer neurons.

Estimated versus field measured soil gas radon concentration and soil gas permeability.

Prediction of areas with elevated natural radiation is fundamental for the prevention of human exposure. Soil gas radon activity concentration and soil gas permeability are predictive parameters for the radon potential, which has great importance in areas where future urban development is planned. In this study, the soil gas radon equilibrium concentration (C∞) and soil gas permeability (K) were estimated through the application of theoretical and empirical models found in the literature. These models apply soil properties as input parameters. Using already existing soil parameters to predict the radon potential of an area would be useful in avoiding direct field measurements. Therefore, in this study, we examined whether the estimated soil gas radon activity concentration and soil gas permeability values match the values measured in the field. The soil gas radon activity concentration estimated by two theoretical models is about 50% of the measured value in the studied area. This underestimation can be attributed to the assumption that the radon activity concentration measured in the field depends only on soil parameters and the models do not take into account the underlying bedrock. Additionally, these models neglect the radon transport by advection and consider only the radon availability and migration in homogeneous media. Furthermore, they do not count certain characteristics of the soil that can be relevant, e.g. organic matter and clay content in the soil. To investigate more in detail such soil characteristics, seven samples located roughly along the slope, were selected to determine the soil chemical composition by ICP-MS. Evaluating the physical and chemical properties of the soil, it was found that the sampling sites with pH < 8 (low calcium content) the preferential adsorption was a dominant process. This causes radium enrichment in organic matter and clay, which directly influence the soil gas radon activity concentration. At pH > 8, radium is no longer preferentially adsorbed on organic matter but continues to be adsorbed on clays albeit this process is weak because radium competes with calcium cations. Also, there are other factors that may affect radon emanation in soil such as radium concentration and distribution, porosity and water content. In contrast, empirical model of soil gas permeability overestimates the measured value in the study area by an order of magnitude. A new model was made by modifying the previously proposed one, which can be used as a guide for the estimation of the median value of soil gas permeability in granitic areas, but not as an accurate predictor due to the lack of correlation between the estimated and measured values.

Current Practice in Pediatric Cow's Milk Protein Allergy-Immunological Features and Beyond.

Cow's milk protein allergy is one of the most common pediatric food allergies. It poses a significant socioeconomic burden in industrialized countries and has a profound effect on the quality of life of affected individuals and their families. Diverse immunologic pathways can lead to the clinical symptoms of cow's milk protein allergy; some of the pathomechanisms are known in detail, but others need further elucidation. A comprehensive understanding of the development of food allergies and the features of oral tolerance could have the potential to unlock more precise diagnostic tools and novel therapeutic approaches for patients with cow's milk protein allergy.

Fecal calprotectin levels in pediatric cow's milk protein allergy.

Introduction: The most prevalent food allergy in younger children is cow's milk protein allergy (CMPA), a hypersensitivity reaction to cow's milk protein and its most common clinical manifestation is allergic colitis. The goal of our recent study was to assess somatic symptoms of CMPA and to prospectively observe the effects of a dairy elimination diet using objective parameters and questionnaires. Methods: The County Hospital in Szekszárd, Hungary, investigated children aged 1 to 18 who had clinical signs that might indicate CMPA. Stool samples were taken and analyzed using a fecal calprotectin (FC) rapid test (Quantum Blue fCAL, Bühlmann Laboratories, Switzerland) at the time of the diagnosis and following 3 months of an elimination diet. At the baseline visit as well as the first and second follow-up, questionnaires were filled out. Patients were divided into two subgroups according to dietary guidelines based on the results of the questionnaires. Results: A total of 47 patients participated in the study [42.55% female, mean age: 7.36 (SD 4.22) years]. There was no significant difference in FC levels between baseline and after 3-month elimination diet [73.98 (71.12) µg/g and 68.11 (74.4) µg/g, respectively, p = 0.331]. After three months, there was a significant decrease in FC levels among patients who adhered to the strict diet [84.06 (79.48) µg/g and 41.11 (34.24) µg/g, respectively, p = 0.001]. Conclusion: The findings of our study suggest that FC can be an objective marker in confirming the diagnosis of CMPA. Significant improvement in clinical symptoms and in FC levels can only be expected after a strictly followed elimination diet.

The Relevance of Goodness-of-fit, Robustness and Prediction Validation Categories of OECD-QSAR Principles with Respect to Sample Size and Model Type.

We investigated the relevance of the validation principles on the Quantitative Structure Activity Relationship models issued by Organization for Economic and Co-operation and Development. We checked the goodness-of-fit, robustness and predictivity categories in linear and nonlinear models using benchmark datasets. Most of our conclusions are drawn using the sample size dependence of the different validation parameters. We found that the goodness-of-fit parameters misleadingly overestimate the models on small samples. In the case of neural network and support vector models, the feasibility of the goodness-of-fit parameters often might be questioned. We propose to use the simplest y-scrambling method to estimate chance correlation. We found that the leave-one-out and leave-many-out cross-validation parameters can be rescaled to each other in all models and the computationally feasible method should be chosen depending on the model type. We assessed the interdependence of the validation parameters by calculating their rank correlations. Goodness of fit and robustness correlate quite well over a sample size for linear models and one of the approaches might be redundant. In the rank correlation between internal and external validation parameters, we found that the assignment of good and bad modellable data to the training or the test causes negative correlations.

The epidemiology of smoking and e-cigarette use in the Hungarian adult population in 2018 / A dohányzás és az e-cigaretta-használat epidemiológiája a felnott magyar népesség körében 2018-ban.

Összefoglaló. Bevezetés: Nagyszámú kutatás igazolta, hogy a dohányzás növeli a legjelentosebb krónikus betegségek kockázatát. Habár 2009 óta csökkeno tendenciát mutat Magyarországon a hagyományos dohányzók aránya, az e-cigarettát kipróbálóké az utóbbi években folyamatosan növekszik. Célkituzés: A 2018-ban a felnott lakosság körében végzett Népegészségügyi Felmérés dohányzásra és e-cigaretta-használatra vonatkozó eredményeinek bemutatása az elozo vizsgálatok tükrében. Módszer: A kérdoíves felmérésben 1586 fo került személyesen lekérdezésre. Az iteratív súlyozás a többlépcsos mintavételi designhatást és a 2016-os mikrocenzus adatait vette figyelembe. Eredmények: 2018-ban a dohányzók aránya a magyar felnott lakosság körében 28,7% (95% MT: 26,3-31,1%), az e-cigarettát használók aránya pedig 1,7% (95% MT: 1,1-2,5%) volt. Az iskolai végzettség a 65 év alattiak esetében a dohányzást befolyásoló tényezo volt (EH: 3,32; 95% MT: 2,53-4,34), de a 65 éves és annál idosebb korcsoportban már nem (EH: 1,11; 95% MT: 0,59-2,09). Az e-cigarettát kipróbálók és használók között a leginkább említett (54,3% 95% MT: 44,0-64,5%) motivációs tényezocsoport a dohányzásról való leszokással, az ártalomcsökkentéssel és a visszaesés megelozésével volt kapcsolatos. A 65 éves és idosebb korcsoportban a dohányzók aránya 2015-höz képest emelkedett. 2018-ban az alapfokú iskolai végzettséguek körében volt a legmagasabb a dohányzók aránya, míg 2014-ben az érettségivel nem rendelkezo középfokú végzettséguek körében. Következtetés: Bár összességében csökkent, az alacsony iskolai végzettséguek és az idosek körében emelkedett a dohányzók aránya Magyarországon. Az e-cigarettát kipróbálók és használók száma növekvo tendenciát mutat hazánkban. Eredményeink az alacsony iskolai végzettséguekre kiemelten fókuszáló, megelozo és leszokást támogató népegészségügyi alprogramokat is tartalmazó komplex beavatkozást sürgetnek. Orv Hetil. 2022; 163(1): 31-38. INTRODUCTION: The body of evidence suggests that smoking increases the risk of the most prevalent chronic diseases. Although the proportion of traditional smokers in Hungary has been on a declining trend since 2009, the proportion of those who tried e-cigarette has been steadily increasing in recent years. OBJECTIVE: To present - in the light of previous studies - the results of the Public Health Survey among adults in 2018 on smoking and e-cigarette use. METHOD: 1586 persons were personally interviewed in a survey. The iterative weighting algorithm considered both the design effect of multistaged sampling and the 2016 Hungarian microcensus. RESULTS: In 2018, the proportion of smokers in the Hungarian adult population was 28.7% (95% CI 26.3-31.1%), and the proportion of e-cigarette users was 1.7% (95% CI 1.1-2.5%). Educational level was a predictor of smoking among respondents younger than 65 years old (OR 3.32; 95% CI 2.53-4.34), but not for those aged 65 years or older (OR 1.11; 95% CI 0.59-2.09). Among e-cigarette ever or current users, the most commonly mentioned (54.3% 95% CI 44.0-64.5%) motivational factor-group to try or use e-cigarettes included motivations to quit smoking, to reduce harm, and to avoid relapsing. In the population aged 65 years old or older, the proportion of smokers increased compared to 2015. The proportion of smokers was the highest among those with primary education in 2018, while in 2014, it was the highest among those with secondary education without a graduation certificate. CONCLUSION: In Hungary, although overall smoking rates are declining, the smoking rate in the low educational group and among the elderly increased. The number of people trying or using e-cigarettes is showing an increasing trend in our country. Our results call for a complex public health intervention program including prevention and smoking cessation supporting subprograms with high focus on those with primary education. Orv Hetil. 2022; 163(1): 31-38.

Vaping-Related Adverse Events and Perceived Health Improvements: A Cross-Sectional Survey among Daily E-Cigarette Users.

Web-based samples of e-cigarette users commonly report significant vaping-related health improvements (HIs) and mild adverse events (AEs). This cross-sectional study with in-person interviewing data collection examined self-reported AEs and perceived HIs among Hungarian adult current daily exclusive e-cigarette (n = 65) and dual users (n = 127), and former daily e-cigarette users (n = 91) in 2018. Logistic regression was used to evaluate associations between reporting any AEs/HIs, vaping status, and covariates. More former users (52.7%) reported AEs than current users (39.6%; p = 0.038). Exclusive and dual daily users reported similar rates of AEs (44.6% and 37.0%, respectively; p = 0.308). More current users (46.9%) experienced HIs than former users (35.2%; p = 0.064). Exclusive daily users were more likely to report HIs than dual users (63.1% versus 38.6%; p = 0.001). Former user status and smoking cessation/reduction reasons increased the odds of reporting AEs, whereas nicotine-containing e-liquid use and older age decreased the odds of reporting AEs. Exclusive vaper status, using advanced generation devices, and smoking cessation/reduction reasons increased the odds of experiencing HIs. This study, which used a traditional data collection methodology, found a higher rate of AEs and a lower rate of HIs compared to web-based surveys. Our results highlight that experiencing AEs and HIs is affected by users' characteristics, in addition to the device and e-liquid type.

Understanding Potential Heavy Metal Contamination, Absorption, Translocation and Accumulation in Rice and Human Health Risks.

Rice is a worldwide staple food and heavy metal contamination is often reported in rice production. Heavy metal can originate from natural sources or be present through anthropogenic contamination. Therefore, this review summarizes the current status of heavy metal contamination in paddy soil and plants, highlighting the mechanism of uptake, bioaccumulation, and health risk assessment. A scoping search employing Google Scholar, Science Direct, Research Gate, Scopus, and Wiley Online was carried out to build up the review using the following keywords: heavy metals, absorption, translocation, accumulation, uptake, biotransformation, rice, and human risk with no restrictions being placed on the year of study. Cadmium (Cd), arsenic (As), and lead (Pb) have been identified as the most prevalent metals in rice cultivation. Mining and irrigation activities are primary sources, but chemical fertilizer and pesticide usage also contribute to heavy metal contamination of paddy soil worldwide. Further to their adverse effect on the paddy ecosystem by reducing the soil fertility and grain yield, heavy metal contamination represents a risk to human health. An in-depth discussion is further offered on health risk assessments by quantitative measurement to identify potential risk towards heavy metal exposure via rice consumption, which consisted of in vitro digestion models through a vital ingestion portion of rice.

The promise of crowdfunding to finance R&D of novel diagnostics and therapeutics for incurable diseases.

The level of funding available for research and development (R&D) of diagnostics (D) and therapeutics (T) for incurable diseases varies and is not associated with the extent of their disease burden. Crowdfunding is a promising way to increase funding for R&D of D&T for underfunded incurable diseases, such as Alzheimer's and Parkinson's disease, which has not been exploited to its full capacity. Investing into efforts to educate patients and researchers about its prospective is a worthwhile endeavor, which could lead to the generation of substantial new capital to finance the development of novel therapeutics for these diseases.

Copper Content and Export in European Vineyard Soils Influenced by Climate and Soil Properties.

Copper-based fungicides (Cuf) are used in European (EU) vineyards to prevent fungal diseases. Soil physicochemical properties locally govern the variation of the total copper content (Cut) in EU vineyards. However, variables controlling Cut distribution at a larger scale are poorly known. Here, machine learning techniques were used to identify governing variables and to predict the Cut distribution in EU vineyards. Precipitation, aridity and soil organic carbon are key variables explaining together 45% of Cut distribution across EU vineyards. This underlines the effect of both climate and soil properties on Cut distribution. The average net export of Cu at the EU scale is 0.29 kg Cu ha-1, which is 2 orders of magnitude less than the net accumulation of Cu (24.8 kg Cu ha-1). Four scenarios of Cuf application were compared. The current EU regulation with a maximum of 4 kg Cu ha-1 year-1 may increase by 2% of the EU vineyard area, exceeding the predicted no-effect concentration (PNEC) in soil in the next 100 years. Overall, our results highlight the vineyard areas requiring specific remediation measures and strategies of Cuf use to manage a trade-off between pest control and soil and water contamination.

Age-associated insolubility of parkin in human midbrain is linked to redox balance and sequestration of reactive dopamine metabolites.

The mechanisms by which parkin protects the adult human brain from Parkinson disease remain incompletely understood. We hypothesized that parkin cysteines participate in redox reactions and that these are reflected in its posttranslational modifications. We found that in post mortem human brain, including in the Substantia nigra, parkin is largely insoluble after age 40 years; this transition is linked to its oxidation, such as at residues Cys95 and Cys253. In mice, oxidative stress induces posttranslational modifications of parkin cysteines that lower its solubility in vivo. Similarly, oxidation of recombinant parkin by hydrogen peroxide (H2O2) promotes its insolubility and aggregate formation, and in exchange leads to the reduction of H2O2. This thiol-based redox activity is diminished by parkin point mutants, e.g., p.C431F and p.G328E. In prkn-null mice, H2O2 levels are increased under oxidative stress conditions, such as acutely by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxin exposure or chronically due to a second, genetic hit; H2O2 levels are also significantly increased in parkin-deficient human brain. In dopamine toxicity studies, wild-type parkin, but not disease-linked mutants, protects human dopaminergic cells, in part through lowering H2O2. Parkin also neutralizes reactive, electrophilic dopamine metabolites via adduct formation, which occurs foremost at the primate-specific residue Cys95. Further, wild-type but not p.C95A-mutant parkin augments melanin formation in vitro. By probing sections of adult, human midbrain from control individuals with epitope-mapped, monoclonal antibodies, we found specific and robust parkin reactivity that co-localizes with neuromelanin pigment, frequently within LAMP-3/CD63+ lysosomes. We conclude that oxidative modifications of parkin cysteines are associated with protective outcomes, which include the reduction of H2O2, conjugation of reactive dopamine metabolites, sequestration of radicals within insoluble aggregates, and increased melanin formation. The loss of these complementary redox effects may augment oxidative stress during ageing in dopamine-producing cells of mutant PRKN allele carriers, thereby enhancing the risk of Parkinson's-linked neurodegeneration.

Current trends in and future potential of crowdfunding to finance R&D of treatments for neglected tropical diseases.

A serious lack of funding exists for the research and development (R&D) of therapeutics, diagnostics, and preventive measures for neglected tropical diseases (NTDs). Hence, crowdfunding to finance R&D for NTDs has high importance, because it is a new and alternate source of capital. This study explores current trends of crowdfunding for R&D for NTDs. Our study showed that, although the number of crowdfunding campaigns for NTDs has been increasing since 2010, crowdfunding overall has not reached its full potential. Several factors contributing positively to the success of crowdfunding campaigns were identified. These and the promotion of the crowdfunding ecosystem could aid the unlocking of its potential as a complementary financing source to conventional funding practices of R&D for NTDs.

Defining healthstyles to plan behavior change interventions in representative samples of children and adults.

Based on Michie's COM-B model, we developed a survey methodology and segmentation procedure to identify groups of Hungarian school children and adults with distinctive characteristics, named healthstyles. We aimed to find interventions fitting to each healthstyle to generate behavioural change. For the segmentation of data, the latent class analysis method was applied. The analysis resulted in 8 healthstyles for the school children and 13 for the adults. Each healthstyle possess distinctive 'traits' and, therefore, special behavioural change methods and prevention strategies can be fitted for them. For demonstrating the advantages of using healthstyles, we discuss possible approaches for selecting behavioural change interventions, one for school children and one for adults.

Farming by soil in Europe: status and outlook of cropping systems under different pedoclimatic conditions.

BACKGROUND: Despite of the importance of soils in agronomy, to date no comprehensive assessment of cropping in Europe has been performed from the viewpoint of the soil variability and its relationship to cropping patterns. In order to fill this knowledge gap, we studied the cropping patterns in different soils of European climate zones with regards to the shares of their crop types in a comparative manner. The study highlights the main features of farming by soil in Europe. Farming by soil in this context means the consideration of soil characteristics when selecting crop types and cropping patterns. METHODS: We first assessed the dissimilarity between the cropping compositions of different pedoclimatic zones in Europe. Next, we assessed the differences of crop distribution in the climate zones by soil types and main crop types by analyzing the degree of association of crops to soil types. A detailed country scale assessment was performed using crops-specific soil productivity maps and land use survey data from Hungary. RESULTS: Results suggest that, in general, farmers consciously take pedoclimatic condition of farming into account when selecting their cropping patterns. In other words, farming by soil is a common practice in the different climatic regions of Europe. However, we have strong reasons to believe that soil suitability-based cropping is not practiced to its full potential over the continent. For example, the findings of our European assessment suggest that production areas of legumes are not always optimized for the local pedoclimatic conditions in some zones. These findings also underline that economic drivers are decisive, when farmers adopt their cropping (eg. oil crops on Albeluvsiols in Europe). Win-win situations of economic considerations and soil suitability based management are observed in all pedoclimatic zones of Europe. The country analysis shows that cropping is progressively practiced on more suitable areas, depending also the crop tolerance to variable pedoclimatic conditions In conclusion, we can assume that pedoclimatic conditions of cropping are respected in most of Europe and farmers crops according to edaphic conditions whenever economic considerations do not override the ecological concerns of farming.

A fluorescent molecular imaging probe with selectivity for soluble tau aggregated protein.

Soluble forms of aggregated tau misfolded protein, generally termed oligomers, are considered to be the most toxic species of the different assembly states that are the pathological components of neurodegenerative disorders. Therefore, a critical biomedical need exists for imaging probes that can identify and quantify them. We have designed and synthesized a novel fluorescent probe, pTP-TFE for which binding and selectivity profiles towards aggregated tau and Aß proteins were assessed. Our results have shown pTP-TFE to be selective for early forms of soluble tau aggregates, with high affinity of dissociation constants (K d) = 66 nM, and tenfold selectivity over mature tau fibrils. Furthermore, we found that pTP-TFE is selective for tau over Aß aggregates and had good cell permeability. This selectivity of pTP-TFE towards early forms of aggregated tau protein ex vivo was also supported with studies on human brain tissue containing tau and Aß pathology. To the best of our knowledge, this is the first fluorescent molecule to be reported to have this form of selectivity profile, which suggests that pTP-TFE is a unique probe candidate for imaging-based detection of early stages of Alzheimer's disease and other tauopathies.

DJ-1 can form ß-sheet structured aggregates that co-localize with pathological amyloid deposits.

The loss of native function of the DJ-1 protein has been linked to the development of Parkinson's (PD) and other neurodegenerative diseases. Here we show that DJ-1 aggregates into ß-sheet structured soluble and fibrillar aggregates in vitro under physiological conditions and that this process is promoted by the oxidation of its catalytic Cys106 residue. This aggregation resulted in the loss of its native biochemical glyoxalase function and in addition oxidized DJ-1 aggregates were observed to localize within Lewy bodies, neurofibrillary tangles and amyloid plaques in human PD and Alzheimer's (AD) patients' post-mortem brain tissue. These findings suggest that the aggregation of DJ-1 may be a critical player in the development of the pathology of PD and AD and demonstrate that loss of DJ-1 function can happen through DJ-1 aggregation. This could then contribute to AD and PD disease onset and progression.