Genetic Characterization of Three Distinct Mechanisms Supporting RNA-Driven DNA Repair and Modification Reveals Major Role of DNA Polymerase ζ.

DNA double-stranded breaks (DSBs) are dangerous lesions threatening genomic stability. Fidelity of DSB repair is best achieved by recombination with a homologous template sequence. In yeast, transcript RNA was shown to template DSB repair of DNA. However, molecular pathways of RNA-driven repair processes remain obscure. Utilizing assays of RNA-DNA recombination with and without an induced DSB in yeast DNA, we characterize three forms of RNA-mediated genomic modifications: RNA- and cDNA-templated DSB repair (R-TDR and c-TDR) using an RNA transcript or a DNA copy of the RNA transcript for DSB repair, respectively, and a new mechanism of RNA-templated DNA modification (R-TDM) induced by spontaneous or mutagen-induced breaks. While c-TDR requires reverse transcriptase, translesion DNA polymerase ζ (Pol ζ) plays a major role in R-TDR, and it is essential for R-TDM. This study characterizes mechanisms of RNA-DNA recombination, uncovering a role of Pol ζ in transferring genetic information from transcript RNA to DNA.

Investigating the knowledge of and public attitudes towards genetic testing within the Visegrad countries: a cross-sectional study.

BACKGROUND: Previous studies have investigated various factors that can determine the attitudes of the citizens considering genetic testing. However, none of them investigated how these attitudes may differ between the Visegrad countries. METHODS: In this cross-sectional study a questionnaire developed by Dutch researchers was translated and used in Hungary, Slovakia, Czechia and Poland. In each country 1000 adult citizens were asked on the topics of personal benefits regarding genetic tests, genetic determinism, and finally, the availability and usage of genetic testing. Multivariate robust regression model was created including several possible influencing factors (such as age, sex, education, marital status, religiousness, and having a genetic test within the nuclear family) to identify the possible differences between the four countries. RESULTS: The Hungarian citizens had the most positive opinion on the personal benefits of genetic testing followed by the Czech, Slovak and Polish. All differences were significant in this regard. Considering genetic determinism, the Slovak citizens had a significantly firmer belief in this issue compared to the Hungarians. No other significant differences were observed in this domain. On the topic of the availability and use of genetic testing the Hungarian citizens had the most accepting opinion among the four countries, followed by the Czech citizens. In this domain the Polish and Slovak answers did not differ significantly from each other. CONCLUSIONS: Significant differences were observed even when considering various confounding effects. As the underlying reasons for these discrepancies are unknown, future studies should investigate this enigma among the four countries.

[Comparative analysis of the full and shortened versions of the Oldenburg Burnout Inventory]. / Az Oldenburg Kiégés Kérdôív és rövidített változatának összehasonlító elemzése.

Background - The two free-to-use versions of the Oldenburg Burnout Inventory (OLBI) have been increasingly utilised to assess the prevalence of burnout among human service workers. The OLBI has been developed to overcome some of the psychometric and conceptual limitations of the Maslach Burnout Inventory, the gold standard of burnout measures. There is a lack of data on the structural validity of the Mini Oldenburg Burnout Inventory and the Oldenburg Burnout Inventory in Hungary. Purpose - To assess the structural validity of the Hungarian versions of the Oldenburg Burnout Inventory and the Mini-Oldenburg Burnout Inventory. Methods - We enrolled 564 participants (196 healthcare workers, 104 nurses and 264 clinicians) in three cross-sectional surveys. In our analysis we assessed the construct validity of the instruments using confirmatory factor analysis and internal consistency using coefficient Cronbach's α. Results - We confirmed the two-dimensional structure (exhaustion and disengagement) of the Mini-Oldenburg Inventory and a shortened version of the Oldenburg Burnout Inventory Internal consistency coefficient confirmed the reliability of the instruments. The burnout appeared more than a 50 percent of the participants in every subsample. The prevalence of exhaustion was above 54.5% in each of the subsamples and the proportion of disengaged clinicians was particularly high (92%). Conclusions - Our findings provide support for the construct validity and reliability of the Hungarian versions of the Mini-Oldenburg Burnout Inventory and a shortened version of the Oldenburg Burnout Inventory in the assessment of burnout among clinicians and nurses in Hungary.

Influenza Vaccination Coverage among People with Self-Reported Cardiovascular Diseases-Findings from the Hungarian Implementation of the European Health Interview Survey.

Worldwide, cardiovascular diseases are the leading cause of mortality. This has significant implications for public health. Influenza, a common infectious disease, poses an increased risk for individuals with chronic conditions, such as cardiovascular diseases. However, little is known about influenza vaccination coverage in this group. This study utilized data from the Hungarian implementation of the European Health Interview Survey to assess influenza vaccination coverage and its determinants among cardiovascular respondents from 2009 to 2019. The findings reveal a downward trend in the vaccination rates over the years (from 24% to 21%), despite the availability of free vaccination in Hungary for this high-risk population. The main factors influencing low influenza vaccine uptake were identified, as follows: young age, a lower level of education, good self-perceived health status, smoking, a lower frequency of medical visits, and not suffering from respiratory diseases. Addressing these disparities necessitates targeted vaccination strategies supported by enhanced education, better access to healthcare services, and the promotion of preventive healthcare measures. Improving vaccination coverage among patients with cardiovascular diseases is imperative for reducing influenza-related morbidity and mortality. This highlights the importance of comprehensive public health interventions and healthcare provider engagement in promoting vaccination among groups at increased risk.

Changes in Antibiotic Redemption Related to Hungarian Dental Care During COVID-19.

INTRODUCTION AND AIMS: Antibiotic resistance is a global health problem that can affect individuals, health systems, and the economy. Our aim was to investigate how the redemption of antibiotics and the associated dental care changed during COVID-19 in Hungary. METHODS: The ICD codes for the infections for which antibiotics should be prescribed in dental care were identified based on the professional recommendation. The database was provided by the National Health Insurance Fund Manager of Hungary. Pearson's chi-squared test with Bonferroni correction was used to compare both the redemption of antibiotics and the associated dental care based on the recommendations in the year 2020 of the COVID-19 and the 2 preceding years. RESULTS: There were fewer antibiotic redemptions and associated dental care in 2020 than in 2018 and 2019. The ratio of professionally relevant and nonrelevant antibiotics redeemed did not change significantly between 2018 and 2019, but the proportion of antibiotics redeemed following recommendations increased significantly during the COVID-19 year of 2020 compared to the previous 2 years. The proportion of all dental care involving antibiotic redemptions increased significantly from 2018 to 2019, and then showed a further increase in 2020. The sex and age of the patient did not affect these findings, while the type of dental care did alter the trends in some cases. CONCLUSIONS: Antibiotics in dentistry are required only for very low proportion of cases, due to the nature of dental care where most infections such as caries and periodontal disease could be managed without resorting to antibiotics. Antibiotic redemptions during COVID-19 pandemic period increased compared to the pre-pandemic period in Hungary. Therefore, Hungarian policy makers need to develop control systems to promote the correct use of antibiotics in dental care. CLINICAL RELEVANCE: It is important to reduce unjustified use of antibiotics so as to curb the emergence of antibiotic resistance organisms.

A global view of aging and Alzheimer's pathogenesis-associated cell population dynamics and molecular signatures in human and mouse brains.

Conventional methods fall short in unraveling the dynamics of rare cell types related to aging and diseases. Here we introduce EasySci, an advanced single-cell combinatorial indexing strategy for exploring age-dependent cellular dynamics in the mammalian brain. Profiling approximately 1.5 million single-cell transcriptomes and 400,000 chromatin accessibility profiles across diverse mouse brains, we identified over 300 cell subtypes, uncovering their molecular characteristics and spatial locations. This comprehensive view elucidates rare cell types expanded or depleted upon aging. We also investigated cell-type-specific responses to genetic alterations linked to Alzheimer's disease, identifying associated rare cell types. Additionally, by profiling 118,240 human brain single-cell transcriptomes, we discerned cell- and region-specific transcriptomic changes tied to Alzheimer's pathogenesis. In conclusion, this research offers a valuable resource for probing cell-type-specific dynamics in both normal and pathological aging.

Investigation of the relationship between incidence of mental disorders and economic growth among the Visegrad countries.

Prevention and care for mental disorders represent an important public health task in achieving global development goals. Proper access to adequate healthcare and social services is an important step related to care for mental disorders, which is presumably strongly related to economic growth. The main aim of the study was to investigate the relationship between the economic growth and the incidence of mental disorders in the V4 countries. An ecological correlation study was conducted regarding the four Visegrad countries. Indicators were derived from the World Health Organization (WHO) 'Health for All' (HFA) online database and Penn World Table version 10. The incidence of mental disorders increased in the V4 countries throughout the years between 2000-2018 except in Hungary, where a decreasing trend was observed. The prevalence of mental disorders increased in all countries as well, but it stagnated in Hungary. At the same time standardized death rate due to mental disorders increased in all Visegrad countries. According to the Hungarian data, while the prevalence of the disease did not change remarkably, the incidence decreased and the mortality increased as well as the prescription of drugs used in the treatment of mental disorders. This could indicate a serious hidden morbidity.

The Emergency Performance of the Hungarian Ambulance Service during the COVID-19 Pandemic.

The COVID-19 pandemic had a considerable impact on the whole health sector, particularly on emergency services. Our aim was to examine the performance of the Hungarian National Ambulance Service during the first four waves of the pandemic. We defined the 2019 performance of the service as the baseline and compared it with the activity during the pandemic years of 2020 and 2021. The data contained deliveries related to acute myocardial infarction, hemorrhagic stroke, ischemic stroke, overall non-COVID-related ambulance deliveries, COVID screenings performed by the ambulance service, and COVID-related ambulance deliveries. The data were aggregated for each week of the investigated time period and stratified by gender and age. Compared with the pre-pandemic era, we found a significant increase in all three medical conditions and overall deliveries (p < 0.001 in all cases). As a result of the increased burden, it is important for emergency services to prepare for the next global epidemic and to improve organizational performance and rescue activities. The Hungarian example highlights that in a pandemic, it can be beneficial to organize the emergency care of a country or a larger region under a single provider with a single decision maker supported by business intelligence.

Differences in Beliefs About Cholesterol-Lowering Medications Among the Visegrad Group Countries: A Cross-Sectional Study.

Background: New cholesterol guidelines highlight more personalized risk assessments and new cholesterol-lowering drugs for people at the highest risk for cardiovascular disease. Adherence due to fear of and lack of trust in medications prevents treatment to provide better health outcomes. Objectives: The aim of our study was to investigate the possible differences in the beliefs about the necessity and concerns regarding lipid-lowering drugs among the Visegrad Group countries. Methods: The Beliefs About Medicines Questionnaire (BMQ-Specific) was used in our research. The responses of 205 Hungarian, 200 Slovak, 235 Czech, and 200 Polish participants, all taking cholesterol-lowering medications, were compared to each other. Results: Hungarian participants' belief in the necessity of cholesterol-lowering drugs was significantly lower compared to the Slovak (P = 0.001), Czech (P = 0.037), and Polish (P < 0.001) participants. While no difference was observed between the Czech and Slovak responses (P = 0.154), both the Czech (P < 0.001) and Slovak (P = 0.006) respondents' belief regarding necessity was lower than that of the Polish. Regarding concerns, the only significant difference was observed between the Czech and the Polish respondents (P = 0.011). Conclusions: While the beliefs about benefits (necessity) are most prominent among the Polish participants, except in comparison to Czech responses, the Visegrad Group countries do not differ considerably regarding their beliefs about the fear (concerns) of the treatment.

Preference for patient-centered communication among the citizens of the Visegrad countries.

OBJECTIVES: The aim of the study was to investigate and compare the preferences that citizens of four Visegrad countries hold concerning the communication between patient and provider. METHODS: The patient-practitioner orientation scale was used on a general population in our research, which consists of the Sharing and Caring subscales and assesses patient-centered or doctor-centered orientation toward communication. The statistical analysis included 4000 respondents of citizens from the Czech Republic, Slovakia, Poland and Hungary. When comparing the various demographic data and the four countries with each other univariate and multivariate logistic regressions were performed. RESULTS: Being female, middle aged, having a higher education and a poor health status were associated with significantly higher Sharing and Caring scores. Also, Hungarian citizens had a significantly higher Caring score compared to the other three countries. CONCLUSIONS: Key demographic variables were identified that affect how citizens perceive the communication between patient and provider. With the exception of Caring among Hungarian citizens, no differences were observed among the Visegrad countries. PRACTICE IMPLICATIONS: These findings have potential implications for understanding the preferences of the citizens and thus better promote a more patient-centered communication.

Expanding the binding specificity for RNA recognition by a PUF domain.

The ability to design a protein to bind specifically to a target RNA enables numerous applications, with the modular architecture of the PUF domain lending itself to new RNA-binding specificities. For each repeat of the Pumilio-1 PUF domain, we generate a library that contains the 8,000 possible combinations of amino acid substitutions at residues critical for RNA contact. We carry out yeast three-hybrid selections with each library against the RNA recognition sequence for Pumilio-1, with any possible base present at the position recognized by the randomized repeat. We use sequencing to score the binding of each variant, identifying many variants with highly repeat-specific interactions. From these data, we generate an RNA binding code specific to each repeat and base. We use this code to design PUF domains against 16 RNAs, and find that some of these domains recognize RNAs with two, three or four changes from the wild type sequence.

Cross-Cultural Adaptation and Lingual Validation of the Beliefs about Medicines Questionnaire (BMQ)-Specific for Cholesterol Lowering Drugs in the Visegrad Countries.

The goal of this study was to translate the Beliefs about Medicines Questionnaire-Specific (BMQ-Specific) for cholesterol-lowering drugs, into the Hungarian, Slovak, Czech and Polish languages and test their reliability with statistical methods. For this purpose, Cronbach's alpha, confirmatory and exploratory factor analyses were conducted. The analyses included 235 Czech, 205 Hungarian, 200 Polish, and 200 Slovak respondents, all of whom were taking cholesterol-lowering drugs. The translations from English into the target languages were always done by two independent translators. As part of the validation process these translations were pilot tested and after the necessary alterations, they were translated back into English by a third translator. After the approval by the creator of the questionnaire, nationwide surveys were conducted in all four countries. The results of the confirmatory factor analysis were exceptionally good for the Czech and Slovak translations, while the Polish and Hungarian translations marginally crossed the predetermined thresholds. With the exception of a single Polish question, the results of the exploratory factor analysis were deemed acceptable. The translated versions of BMQ-Specific are reliable and valid tools to assess patients' beliefs about medication, especially medication adherence among patients taking cholesterol-lowering medication. A comparison between the four countries with this questionnaire is now possible.

Variations in maternal vitamin A intake modifies phenotypes in a mouse model of 22q11.2 deletion syndrome.

BACKGROUND: Vitamin A regulates patterning of the pharyngeal arches, cranial nerves, and hindbrain that are essential for feeding and swallowing. In the LgDel mouse model of 22q11.2 deletion syndrome (22q11DS), morphogenesis of multiple structures involved in feeding and swallowing are dysmorphic. We asked whether changes in maternal dietary Vitamin A intake can modify cranial nerve, hindbrain and pharyngeal arch artery development in the embryo as well as lung pathology that can be a sign of aspiration dysphagia in LgDel pups. METHODS: Three defined amounts of vitamin A (4, 10, and 16 IU/g) were provided in the maternal diet. Cranial nerve, hindbrain and pharyngeal arch artery development was evaluated in embryos and inflammation in the lungs of pups to determine the impact of altering maternal diet on these phenotypes. RESULTS: Reduced maternal vitamin A intake improved whereas increased intake exacerbated lung inflammation in LgDel pups. These changes were accompanied by increased incidence and/or severity of pharyngeal arch artery and cranial nerve V (CN V) abnormalities in LgDel embryos as well as altered expression of Cyp26b1 in the hindbrain. CONCLUSIONS: Our studies demonstrate that variations in maternal vitamin A intake can influence the incidence and severity of phenotypes in a mouse model 22q11.2 deletion syndrome.

Cyclin C influences the timing of mitosis in fission yeast.

The multiprotein Mediator complex is required for the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator contains the Cdk8 regulatory subcomplex, which directs periodic transcription and influences cell cycle progression in fission yeast. Here we investigate the role of CycC, the cognate cyclin partner of Cdk8, in cell cycle control. Previous reports suggested that CycC interacts with other cellular Cdks, but a fusion of CycC to Cdk8 reported here did not cause any obvious cell cycle phenotypes. We find that Cdk8 and CycC interactions are stabilized within the Mediator complex and the activity of Cdk8-CycC is regulated by other Mediator components. Analysis of a mutant yeast strain reveals that CycC, together with Cdk8, primarily affects M-phase progression but mutations that release Cdk8 from CycC control also affect timing of entry into S phase.

Cdk1 activity acts as a quantitative platform for coordinating cell cycle progression with periodic transcription.

Cell proliferation is regulated by cyclin-dependent kinases (Cdks) and requires the periodic expression of particular gene clusters in different cell cycle phases. However, the interplay between the networks that generate these transcriptional oscillations and the core cell cycle machinery remains largely unexplored. In this work, we use a synthetic regulable Cdk1 module to demonstrate that periodic expression is governed by quantitative changes in Cdk1 activity, with different clusters directly responding to specific activity levels. We further establish that cell cycle events neither participate in nor interfere with the Cdk1-driven transcriptional program, provided that cells are exposed to the appropriate Cdk1 activities. These findings contrast with current models that propose self-sustained and Cdk1-independent transcriptional oscillations. Our work therefore supports a model in which Cdk1 activity serves as a quantitative platform for coordinating cell cycle transitions with the expression of critical genes to bring about proper cell cycle progression.

Mediator can regulate mitotic entry and direct periodic transcription in fission yeast.

Cdk8 is required for correct timing of mitotic progression in fission yeast. How the activity of Cdk8 is regulated is unclear, since the kinase is not activated by T-loop phosphorylation and its partner, CycC, does not oscillate. Cdk8 is, however, a component of the multiprotein Mediator complex, a conserved coregulator of eukaryotic transcription that is connected to a number of intracellular signaling pathways. We demonstrate here that other Mediator components regulate the activity of Cdk8 in vivo and thereby direct the timing of mitotic entry. Deletion of Mediator components Med12 and Med13 leads to higher cellular Cdk8 protein levels, premature phosphorylation of the Cdk8 target Fkh2, and earlier entry into mitosis. We also demonstrate that Mediator is recruited to clusters of mitotic genes in a periodic fashion and that the complex is required for the transcription of these genes. We suggest that Mediator functions as a hub for coordinated regulation of mitotic progression and cell cycle-dependent transcription. The many signaling pathways and activator proteins shown to function via Mediator may influence the timing of these cell cycle events.

Cyclin-dependent kinase 8 regulates mitotic commitment in fission yeast.

Temporal changes in transcription programs are coupled to control of cell growth and division. We here report that Mediator, a conserved coregulator of eukaryotic transcription, is part of a regulatory pathway that controls mitotic entry in fission yeast. The Mediator subunit cyclin-dependent kinase 8 (Cdk8) phosphorylates the forkhead 2 (Fkh2) protein in a periodic manner that coincides with gene activation during mitosis. Phosphorylation prevents degradation of the Fkh2 transcription factor by the proteasome, thus ensuring cell cycle-dependent variations in Fkh2 levels. Interestingly, Cdk8-dependent phosphorylation of Fkh2 controls mitotic entry, and mitotic entry is delayed by inactivation of the Cdk8 kinase activity or mutations replacing the phosphorylated serine residues of Fkh2. In addition, mutations in Fkh2, which mimic protein phosphorylation, lead to premature mitotic entry. Therefore, Fkh2 regulates not only the onset of mitotic transcription but also the correct timing of mitotic entry via effects on the Wee1 kinase. Our findings thus establish a new pathway linking the Mediator complex to control of mitotic transcription and regulation of mitotic entry in fission yeast.