Molecular Characterization of 'Candidatus Phytoplasma prunorum' in the Czech Republic and Susceptibility of Apricot Rootstocks to the Two Most Abundant Haplotypes.

'Candidatus Phytoplasma prunorum' is one of the most destructive pathogens of Prunus species, where susceptible species render unproductive several years after infection. In epidemiology, the molecular characterization of phytoplasmas is based on sequence analysis of variable nonribosomal genes. In this study aceF, pnp, imp and secY genes were used for characterization of the 'Ca. P. prunorum' genotypes present in the Czech Republic. In total, 56 plant and 33 vector (Cacopsylla pruni) samples positive to 'Ca. P. prunorum' collected in seven localities were used in the study. Based on sequence analysis, four aceF, two pnp, six imp, and three secY genotypes were identified in analyzed samples. The most abundant in both plant and insect samples were the A6, P2, I4, and S2 genotypes. Most of the Czech 'Ca. P. prunorum' haplotypes clustered together in the haplotype network analysis. Next, two isolates representing the two most abundant Czech haplotypes (A6-P2-I4-S2 and A5-P2-I4-S2) were used in the susceptibility test of three apricot rootstock types (St. Julien A, M-VA-1, GF-305). Susceptibility was analyzed by phytoplasma quantification using quantitative real-time PCR and evaluation of symptom manifestation. Based on the results, the influence of the rootstock type on the phytoplasma titer and symptom manifestation was greater than of the phytoplasma isolate, while the year of analysis had no influence on the results. The results also showed that the phytoplasma titer is increasing in plant tissues during the vegetation period.

Evaluation of eHealth assistance in-hospital care for improved quality of life in patients.

Health conditions contribute significantly to patients' quality of life. Healthcare infrastructure and healthcare services, including their accessibility, belong to objective factors influencing their perception of their health. The growing disparity between supply and demand for specialized inpatient facilities due to the aging population calls for new solutions, including eHealth technologies. Automatized activities could be taken over by eHealth technologies that do not require a constant presence of staff. We tested whether eHealth technical solutions reduce patients' health risks on a sample of 61 patients on the covid-19 unit in Tomas Bata hospital in Zlin. We have applied the randomized control trial to select patients for the treatment and the control groups. Moreover, we tested eHealth technologies and their help to staff in the hospital. Due to the severity of the covid-19 disease and its rapid course and the size of the sample in our research, we did not demonstrate a statistically significant impact of eHealth technologies on patient health. The evaluation results confirm that even the limited number of technologies deployed proves to be an effective help for staff in critical situations like the pandemic. The main issue is psychological support to staff in hospitals and relieving stressful work.

The Expanding Menagerie of Prunus-Infecting Luteoviruses.

Members of the genus Luteovirus are responsible for economically destructive plant diseases worldwide. Over the past few years, three luteoviruses infecting Prunus trees have been characterized. However, the biological properties, prevalence, and genetic diversity of those viruses have not yet been studied. High-throughput sequencing of samples of various wild, cultivated, and ornamental Prunus species enabled the identification of four novel species in the genus Luteovirus for which we obtained complete or nearly complete genomes. Additionally, we identified another new putative species recovered from Sequence Read Archive data. Furthermore, we conducted a survey on peach-infecting luteoviruses in eight European countries. Analyses of 350 leaf samples collected from germplasm, production orchards, and private gardens showed that peach-associated luteovirus (PaLV), nectarine stem pitting-associated virus (NSPaV), and a novel luteovirus, peach-associated luteovirus 2 (PaLV2), are present in all countries; the most prevalent virus was NSPaV, followed by PaLV. The genetic diversity of these viruses was also analyzed. Moreover, the biological indexing on GF305 peach indicator plants demonstrated that PaLV and PaLV2, like NSPaV, are transmitted by graft at relatively low rates. No clear viral symptoms have been observed in either graft-inoculated GF305 indicators or different peach tree varieties observed in an orchard. The data generated during this study provide a broader overview of the genetic diversity, geographical distribution, and prevalence of peach-infecting luteoviruses and suggest that these viruses are likely asymptomatic in peach under most circumstances.

Movement disorders, cerebral palsy and vaccination.

This review focused on vaccination in children with movement disorders, including cerebral palsy and the movement disorders triggered by vaccination in children with and without neurological disabilities. The following clinical questions were addressed: 1) Can children with movement disorders be vaccinated? 2) Can vaccination trigger movement disorders in children without neurological disabilities? 3) Can vaccination trigger movement disorders in children with neurological disabilities? and 4) Is there any consensus of care concerning vaccination in children with movement disorders? Following the PRISMA reporting guidelines, 1096 records were identified and 34 relevant papers were included. No evidence that vaccinations are contraindicated for children with movement disorders was noticed. Several reports of neurological adverse events, including movement disorders in children without neurological disabilities after various types of vaccination, were found. The reporting rates were low, the causality was controversial, and patient outcomes were mostly favourable. There was limited (if any) evidence in our search that any vaccination leads to any movement disorder exacerbation. Finally, no generally accepted consensus or standards of care concerning vaccination in patients with movement disorders were found. In summary, we found few precautions for vaccination in this group of patients and concluded that general best practice guidelines for immunization should be followed. In addition, influenza and pneumococcal vaccines are recommended because they can reduce morbidity and mortality in individuals severely affected by movement restrictions.

Quality assessment of clinical practice guidelines for perioperative care and use of GRADE: a systematic review protocol.

INTRODUCTION: Perioperative care is a broad field covering an array of elective and emergency procedures. Clinical practice guidelines (CPGs) for perioperative care exist with various degrees of methodological quality. We intend to critically appraise them using AGREE II instrument and investigate the use of Grading of Recommendations, Assessment, Development and Evaluations (GRADE). METHODS AND ANALYSIS: We searched MEDLINE (Ovid), Epistemonikos, Cochrane Database of Systematic Reviews and PROSPERO and did not identify any similar systematic review in this area. We will search databases, repositories and websites of guideline developers and medical societies, including MEDLINE (Ovid), Embase (Ovid), DynaMed, the GIN international guideline library and registry of guidelines in development, BIGG international database of GRADE guidelines, ECRI Guideline Trust or National Institute for Clinical Evidence to identify all CPGs for perioperative care in an adult population in a general clinical setting. We will include CPGs, expert guidance, position papers, guidance documents and consensus statements published in the last 5 years by experts or international organisations that provide guidance or recommendations in the available full text with no geographical or language limitation. Excluded will be those containing only good practice statements. Two independent reviewers will perform critical appraisal using the AGREE II tool. The data presented in a narrative and tabular form will include the results of the critical appraisal for all identified CPGs for all AGREE II domains and an assessment of the use of the GRADE approach. ETHICS AND DISSEMINATION: Ethics approval is not required. We will disseminate the findings through professional networks and conference presentations and will publish the results.

A novel coronavirus (SARS-CoV-2) and COVID-19.

By the end of 2019 the first cases of severe pneumonia of unknown origin were reported in Wuhan, China. The causative agent was identified as a novel b-coronavirus SARS-CoV-2 and the disease was named COVID-19. Since the beginning of 2020, the infection has spread worldwide, which led the WHO to declare COVID-19 a public health emergency of international concern and to characterize the current situation as a pandemic. The transmission occurs mainly via respiratory droplets and the incubation period ranges from 2 to 14 days. Most cases are mild, but some patients develop severe pneumonia with acute respiratory distress, septic shock and multi-organ failure. The most common symptoms include fever, dry cough, myalgia and shortness of breath. Characteristic laboratory findings are normal white blood cell count or mild leukopenia, marked lymphopenia, in severe cases elevated CRP, procalcitonin, LDH, and D-dimer are commonly found. Typical imaging findings include multifocal peripherally distributed ground-glass opacities or consolidations, interlobular septal thickening, crazy paving appearance and cystic changes. The overall case fatality rate is estimated to range from 1 to 3 %, however, it is dependent on age and underlying medical comorbidities. Current potential treatment options include hydroxychloroquine, remdesivir, lopinavir/ritonavir and convalescent plasma.

Assessment of Antioxidants in Selected Plant Rootstocks.

The service tree (Sorbus domestica) is a wild fruit tree with immense medicinal and industrial value. This study aimed at determining the four major groups of antioxidants (flavonoids, phenolic acids and aldehydes, catechin and procyanidin) in rootstocks of Crataegus laevigata (genotypes O-LE-14 and O-LE-21), Aronia melanocarpa (genotypes O-LE-14 and O-LE-21), Chaenomeles japonica (genotype O-LE-9) and Cydonia oblonga (BA 29) (genotypes O-LE-14 and O-LE-21). Hyperoside (Quercetin 3-D-galactoside) was the most abundant flavonoid compound, since its average content in the rootstocks of Crataegus laevigata (O-LE-21) was 180.68 ± 0.04 µg·g-1. Dihydrokaempherol was the least frequently found flavonoid compound, with an average concentration of 0.43 ± 0.01 µg·g-1 in all the rootstocks of plants considered in this study. Among the phenolic compounds, the most represented one was protocatechuic acid, with 955.92 ± 10.25 µg·g-1 in the rootstocks of Aronia melanocarpa (O-LE-14). On the other hand, the least represented p-Coumaric acid exhibited the average concentration of 0.34 ± 0.01 µg·g-1 in the plant rootstocks. Epicatechin was the most abundant catechin compound, with a content of 3196.37 ± 50.10 µg·g-1 in the rootstocks of Aronia melanocarpa (O-LE-14). The lowest represented catechin compound was epigallocatechin, with the average concentration of 0.95 ± 0.08 µg·g-1 in the screened plant rootstocks. From the procyanidin compounds, the most abundant one was procyanidin b2 in the rootstocks of Crataegus laevigata (O-LE-14), with a concentration of 5550.40 ± 99.56 µg·g-1. On the contrary, procyanidin a2, with an average concentration of 40.35 ± 1.61 µg·g-1, represented the least frequent procyanidin compound in all the plant rootstocks screened herein.

Czech national project 'Clinical Practice Guidelines' methodology and current results.

Currently in the Czech Republic, there does not exist such an institution as a 'National Centre for Clinical Practice Guidelines'. In 2017, there were about 123 professional medical organizations which developed about 1909 'guidelines' until 2017. However, the majority of these guidelines are 'expert opinion' or 'consensual' based 'guidance' or rather recommendations in the most cases missing a systematic approach that reflects evidence-based medicine principles and methods. The project is led by the Czech Health Research Council, the first partner is the Ministry of Health of the Czech Republic and the second partner is the Institute of Health Information and Statistics of the Czech Republic with support from policy makers, academics, clinicians and members of the Czech National Centre for Evidence-Based Healthcare and Knowledge Translations. This centre is an umbrella for three very important international collaborations which play a key role in Evidence-Based Healthcare, Evidence Synthesis, Evidence Implementation and trustworthy guidelines development. These are Cochrane Czech Republic, Masaryk University Grade Centre and the Czech Republic Centre for Evidence-Based Healthcare: The Joanna Briggs Institute Centre of Excellence.The main aim of this article is to present the Czech National Methodology of the Trustworthy Clinical Practice Guideline (CPG) development and the first results of the project 'Clinical Practice Guidelines'.A pilot phase of the project was realized during the first year of the project from January to December 2018. As the first step, there were established managing authorities including a Guarantee Committee and an Appraisal (Methodological) Committee. The Members of the Appraisal Committee developed a pilot version of the National Methodology of CPG development based on the best available approaches to Trustworthy CPGs development followed by testing on the first five pilot CPGs.

Development of Czech National Stroke Guidelines.

Despite a significant transformation of ischaemic stroke management, it remains one of the leading causes of death, disability, functional impairment and cognitive deficits. With the advent of intravenous thrombolysis and mechanical thrombectomy, stroke is not an untreatable disease, mostly occurring in older people, any more. The most common cause of cardioembolic strokes is atrial fibrillation or flutter and atrial fibrillation is the most prevalent arrhythmia.The aim of this short communication is to describe the methodology of the Czech stroke guidelines development.High-quality 2017 Australian Stroke Foundation stroke guideline was considered for adaptation and a new guideline was developed and disseminated.

Epidemiological analyses for preparation of Clinical Practice Guidelines related to acute coronary syndromes in the Czech Republic.

Coronary heart disease, sometimes also referred to as ischemic heart disease, remains the leading condition causing most deaths and disability-adjusted life years worldwide. Acute coronary syndrome (ACS) represents a subset that is defined by sudden reduction of blood supply in the coronary arteries. ACS consists of unstable angina, non-ST-segment elevation myocardial infarction (NSTEMI), and ST-segment elevation myocardial infarction (STEMI).The current short communication aims to provide current ACS prevalence and incidence data analysis to inform development of clinical practice guidelines in the Czech Republic.The Institute of Health Information and Statistics of the Czech Republic has provided the data that are collected by the National Health Information System with the National Register of Reimbursed Health Services as a primary source providing data for the period from 2015 to 2017.There has been a slight decrease in the number of hospitalized patients for ACS in the Czech Republic from 2015 to 2017. Sex difference remains large, with majority (two thirds) of those hospitalized for unstable angina, NSTEMI, or STEMI being men. Hospitalization with STEMI is reported in younger age with no sex difference compared with NSTEMI and unstable angina.

Use of epidemiological analyses in Clinical Practice Guideline development focused on the diabetic patients treated with insulin.

The prevalence of diabetes is on the rise worldwide especially in developed countries. The aim of glucose management in all types of diabetes is to minimize chronic and acute complications associated with diabetes. All patients with type 1 diabetes mellitus (T1DM) require insulin. Main areas of technology advances in diabetes are continuous subcutaneous insulin infusion (CSII) and also continuous glucose monitoring systems for the management of patients with both types of diabetes. It is very important to analyse the epidemiological situation within each country before and during the clinical practice guidelines (CPGs) development and implementation. The analyses will allow us to monitor the effect of the CPG after its implementation.The aim of this short communication is to analyse the epidemiology of prevalence and incidence of diabetes mellitus and use of CSII to inform development of CPGs in the Czech Republic.The analysis is developed based on the data managed by Institute of Health Information and Statistics of the Czech Republic. We used the National Register of Reimbursed Health Services 2015-2017 as primary source, and the annual report type A (Ministry of Health) 1-01: for Diabetology (A MH 004) 2007-2017 was used as validation source. The presented data are related to the year 2016 because we were able to validate them based on the data from 2015 to 2017 for this cohort of patients.The number of patients with T1DM is increasing in the Czech Republic with no significant sex difference. Life expectancy is about 11 years lower in the T1DM population. The majority of the patients are in older age; however, these are not treated with CSII compared with the younger population. From 61 533 patients with T1DM, 81% were reported with acute and chronic complications in 2016. Only 5011 of these patients were reported as using CSII.

Epidemiological analyses to inform stroke clinical practice guidelines development.

Stroke is one of the leading causes of mortality and the leading cause of functional impairment and cognitive deficits worldwide. It is important that clinical practice guidelines development is based on robust statistical and epidemiological data and their analysis throughout the whole process of guidelines development and implementation. The aim of this short communication is to analyse epidemiology of prevalence and incidence of ischaemic stroke, its main causes, brain imaging using MRI, recanalization therapies, secondary prevention with antiplatelet and anticoagulants, mortality data and to inform development of stroke clinical practice guidelines in the Czech Republic. The main analysed diagnosis was I63 (cerebral infarction) and secondary diagnoses were: I48 (atrial fibrillation and flutter), I35.9 (nonspecified aortic valve disease), Q21.1 (atrial septal defect) or I33.0 (acute and subacute endocarditis). We have also analysed use of brain imaging with MRI, recanalization treatment using intravenous thrombolysis and mechanical thrombectomy, stroke secondary prevention with antiplatelet drugs and anticoagulation as well as hospital admissions and mortality. In total, 159 344 patients were diagnosed with an ischaemic stroke from 2015 to 2017. Average prevalence of ischaemic stroke in the Czech Republic is 54.9 patients per 100 000. 22.2% of patients with stroke received intravenous thrombolysis or mechanical thrombectomy in 2017.

High-Throughput Sequencing Analysis of the Bacterial Community in Stone Fruit Phloem Tissues Infected by "Candidatus Phytoplasma prunorum".

"Candidatus Phytoplasma prunorum" (CPp) is a highly destructive phytopathogenic agent in many stone fruit-growing regions in Europe and the surrounding countries. In this work, we focused on documenting entire bacterial community in the phloem tissues of 60 stone fruit trees. Nested PCR and two real-time PCR assays were used to select CPp-positive (group A) and CPp-negative samples (group B). Afterwards, high-throughput amplicon sequencing was performed to assess bacterial community compositions in phloem tissues. The bacterial composition in phloem tissue consisted of 118 distinct genera, represented mainly by Pseudomonas, Acinetobacter, Methylobacterium, Sphingomonas, and Rhizobium. Statistics showed that CPp influenced the bacterial composition of infected plants (group A) and that the bacterial community depended on the geographical origin of the sample. This is the first work focusing on an analysis of the influence of CPp on the bacteria coexisting in the phloem tissues of stone fruit trees.

Use of Combined MSAP and NGS Techniques to Identify Differentially Methylated Regions in Somaclones: A Case Study of Two Stable Somatic Wheat Mutants.

The appearance of somaclonal variability induced by in vitro cultivation is relatively frequent and can, in some cases, provide a valuable source of new genetic variation for crop improvement. The cause of this phenomenon remains unknown; however, there are a number of reports suggesting that epigenetics, including DNA methylations, are an important factor. In addition to the non-heritable DNA methylation changes caused by transient and reversible stress-responsive gene regulation, recent evidence supports the existence of mitotically and meiotically inherited changes. The induction of phenotypes via stable DNA methylation changes has occasionally great economical value; however, very little is known about the genetic or molecular basis of these phenotypes. We used a novel approach consisting of a standard MSAP analysis followed by deep amplicon sequencing to better understand this phenomenon. Our models included two wheat genotypes, and their somaclones induced using in vitro cultivation with a changed heritable phenotype (shortened stem height and silenced high molecular weight glutenin). Using this novel procedure, we obtained information on the dissimilarity of DNA methylation landscapes between the standard cultivar and its respective somaclones, and we extracted the sequences and genome regions that were differentially methylated between subjects. Transposable elements were identified as the most likely factor for producing changes in somaclone properties. In summary, the novel approach of combining MSAP and NGS is relatively easy and widely applicable, which is a rather unique feature compared with the currently available techniques in the epigenetics field.