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Objectives: Pregnant women are among the priority groups to receive influenza vaccines in the Czech Republic since 2011, data on vaccination coverage are not yet available. The aim of the study was to determine the influenza vaccination coverage (IVC) and provide source data for further activities., Methods: A prospective observational study was performed in a large maternity hospital in Prague. The self-completed questionnaire was distributed to 5,475 pregnant women between September 1, 2020 and August 31, 2021. Questions included maternal sociodemographic characteristics, influenza vaccination status and sources of maternal vaccination recommendations during pregnancy., Results: A total of 4,617 completed questionnaires have been analysed. The median age of study participants (N = 4,592) was 33 years (range: 18-51 years). The majority (69.7%) of women had completed their university education, most women were childless (58.5%) or had one child (32.5%) before the start of the study. Less than 2% of women reported being vaccinated against influenza during their pregnancy (1.5%; 95% CI, 1.1-1.9%). Only 21% of women knew that it's possible to get vaccinated against influenza during pregnancy. Participants considered influenza vaccination in pregnancy as important (3.3%), useful (41.1%) and useless (44.4%). Out of 959 pregnant women who had information about influenza vaccination during pregnancy, only 6.9% were vaccinated, while among those who did not have this information, 0.1% were vaccinated during pregnancy (p < 0.001). The most frequent source of information was Internet, then media and a general practitioner., Conclusions: The IVC during pregnancy in our study was extremely low. In order to improve IVC among pregnant women, it is necessary to increase awareness of recommendations and vaccination options among the public and professionals and incorporating vaccination recommendation in routine antenatal practice., (© 2023. The Author(s).)

Objectives: For the proper diagnosis of toxoplasmosis it is essential to determine the stage of the infection, for which the most preferred method is IgG avidity test. The avidity index (AI) should initially be low (AI≤0.3) in the acute phase and increase during the infection. However, persistent low avidity can occur in patients with latent toxoplasmosis, which can complicate the interpretation of the results. The aim of the study is to explain the causes of this phenomenon., Methodology: A retrospective study was carried out with 717 serum samples collected from 442 patients from the categories of pregnant and non-pregnant women, men, and newborns + infants (age < 0.5 year). The trends of AI kinetics were evaluated in repeatedly examined patients. The frequency of cases with low avidity was compared in individual categories of patients and in groups of people with acute and non-acute toxoplasmosis., Results: The proportion of patients with initially low avidity was 42.1% in the acute toxoplasmosis group while it was 13.0% in the non-acute groups. In uninfected newborns with anti-Toxoplasma antibodies transmitted from the mother, a decrease in IgG avidity levels over time was observed, resulting in 29.2% of samples showing low (improper) avidity. While the dynamics of IgG avidity and the frequency of cases of improperly low avidity were similar in men and pregnant and non-pregnant women, the category of newborns and infants differed substantially for these indicators., Conclusions: Due to acceptable specificity and negative predictive value, high avidity can rule out acute toxoplasmosis, but moderate sensitivity complicates the possibility of its confirmation. The results of the avidity test must be interpreted in the context of the results of other methods., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kodym et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: The completeness and timeliness of the pertussis questionnaire-based enhanced surveillance system (ESS) among infants and reported pertussis data within the electronic nationwide notification system (NNS) in the years 2015, 2017 and 2019 were evaluated in a pilot study., Methods: The completeness of the variables for demographic characteristics, date of symptom onset, hospitalisation and vaccination status were assessed in both systems. Timeliness of reporting in the NNS was analysed as the interval between symptom onset and a) the date of first specimen collection (diagnostic delay), and b) the date of the Regional Public Health Authority receiving notification (notification delay)., Results: A total of 121 confirmed pertussis cases were reported to the NNS in the study years, while in the ESS a total of 104 confirmed cases were reported in infants. In both systems most cases were in the age group of one completed month of life (20% versus 23%) and males (55% versus 55%). The majority of cases were hospitalised (81% versus 85%) and unvaccinated (77% versus 78%). Within the NNS, the first dose of vaccine was reported in 13 cases, the second dose in 11, and third dose in three cases. Within the NNS, 100% completeness of following variables was found: symptom onset, week and region of reporting, age, gender and place of isolation. Median diagnostic delay was nine days. Median notification delay was 18 days., Conclusions: Data completeness was high in the NNS, except for lack of vaccination data in those eligible by age. Efforts to improve the completeness of laboratory-related variables and timeliness are essential. Based on the study results, the project of improving the ESS for infants will continue with regular evaluation., (© 2023 Monika LIPTÁKOVÁ et al., published by Sciendo.)

Background: Predicting stroke risk in patients with carotid artery stenosis (CS) remains challenging. Circulating biomarkers seem to provide improvements with respect to risk stratification. Methods: Study patients who underwent carotid endarterectomy were categorized into four groups according to symptomatology and compared as follows: symptomatic with asymptomatic patients; and asymptomatic patients including amaurosis fugax (AF) (asymptomatic + AF group) with patients with a transient ischemic attack (TIA) or brain stroke (BS) (hemispheric brain stroke group). Carotid specimens were histologically analyzed and classified based on the American Heart Classification (AHA) standard. As a marker of OS, the plasma levels of malondialdehyde (MDA) were measured. Comparisons of MDA plasma levels between groups were analyzed. Results : In total, 35 patients were included in the study. There were 22 (63%) patients in the asymptomatic group and 13 (37%) in the symptomatic group. Atheromatous plaque ( p = 0.03) and old hemorrhage ( p = 0.05), fibrous plaque ( p = 0.04), myxoid changes ( p = 0.02), plaques without hemorrhage ( p = 0.04), significant neovascularization ( p = 0.04) and AHA classification ( p = 0.006) had significant correlations with clinical presentation. There were 26 (74%) patients in the asymptomatic group and 9 (26%) in the hemispheric brain stroke group. Atheromatous plaque ( p = 0.02), old hemorrhage ( p = 0.05) and plaques without neovascularization ( p = 0.02), fibrous plaque ( p = 0.03), plaques without hemorrhage ( p = 0.02) and AHA classification ( p = 0.01) had significant correlations with clinical presentation. There was no significant difference between symptomatic and asymptomatic groups with respect to MDA plasma levels ( p = 0.232). A significant difference was observed when MDA plasma levels were compared to asymptomatic + AF and the hemispheric stroke group ( p = 0.002). Conclusions : MDA plasma level correlates with the risk of hemispheric stroke (TIA or BS) and is a reliable marker of plaque vulnerability in carotid artery stenosis.

Background: Comparative data regarding the effect of percutaneous and thoracoscopic ablation of atrial fibrillation (AF) on cognitive function are very limited. The aim of the study was to determine and compare the effect of both types of ablations on patient cognitive functions in the mid-term., Methods: Patients with AF indicated for ablation procedure were included. Forty-six patients underwent thoracoscopic, off-pump ablation using the COBRA Fusion radiofrequency system, followed by a catheter ablation three months afterward (Hybrid group). A comparative cohort of 53 AF patients underwent pulmonary vein isolation only (PVI group). Neuropsychological examinations were done before and nine months after the surgical or catheter ablation procedure. Neuropsychological testing comprised 13 subtests of seven domains, and the results were expressed as post-operative cognitive dysfunction (POCD) nine months after the procedure., Results: Patients in both groups were similar with respect to the baseline clinical characteristics; only non-paroxysmal AF was more common in the hybrid group (98% vs. 34%). Major POCD was present in eight (17.4%) of hybrid patients versus three (5.7%) of PVI patients (p = 0.11), combined (major/minor) worsened cognitive decline was present in 10 (21.7%) hybrid patients versus three (5.6%) PVI patients (p = 0.034). On the other hand, combined (major/minor) improvement was present in 15 (32.6%) hybrid patients versus nine (16.9%) patients in the PVI group (p = 0.099)., Conclusion: Hybrid ablation, a combination of thoracoscopic and percutaneous ablation, is associated with a higher risk of cognitive decline compared to sole percutaneous ablation., (© 2022 Wiley Periodicals LLC.)

Introduction: Imprinting broadly neutralizing antibody (bNAb) paratopes by shape complementary protein mimotopes represents a potential alternative for developing vaccine immunogens. This approach, designated as a Non-Cognate Ligand Strategy (NCLS), has recently been used for the identification of protein variants mimicking CD4 binding region epitope or membrane proximal external region (MPER) epitope of HIV-1 envelope (Env) glycoprotein. However, the potential of small binding proteins to mimic viral glycan-containing epitopes has not yet been verified., Methods: In this work, we employed a highly complex combinatorial Myomedin scaffold library to identify variants recognizing paratopes of super candidate bNAbs, PGT121 and PGT126, specific for HIV-1 V3 loop epitopes., Results: In the collection of Myomedins called MLD variants targeted to PGT121, three candidates competed with gp120 for binding to this bNAb in ELISA, thus suggesting an overlapping binding site and epitope-mimicking potential. Myomedins targeted to PGT126 designated MLB also provided variants that competed with gp120. Immunization of mice with MLB or MLD binders resulted in the production of anti-gp120 and -Env serum antibodies. Mouse hyper-immune sera elicited with MLB036, MLB041, MLB049, and MLD108 moderately neutralized 8-to-10 of 22 tested HIV-1-pseudotyped viruses of A, B, and C clades in vitro ., Discussion: Our data demonstrate that Myomedin-derived variants can mimic particular V3 glycan epitopes of prominent anti-HIV-1 bNAbs, ascertain the potential of particular glycans controlling neutralizing sensitivity of individual HIV-1 pseudoviruses, and represent promising prophylactic candidates for HIV-1 vaccine development., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Daniel Lišková, Kosztyu, Kuchař, Černý, Bharadwaj, Petroková, Vroblová, Křupka, Malý, Zosinčuková, Šulc, Rašková Kafková, Raška and Malý.)

S1 nuclease from Aspergillus oryzae is a single-strand-specific nuclease from the S1/P1 family that is utilized in biochemistry and biotechnology. S1 nuclease is active on both RNA and DNA but with differing catalytic efficiencies. This study clarifies its catalytic properties using a thorough comparison of differences in the binding of RNA and DNA in the active site of S1 nuclease based on X-ray structures, including two newly solved complexes of S1 nuclease with the products of RNA cleavage at atomic resolution. Conclusions derived from this comparison are valid for the whole S1/P1 nuclease family. For proper model building and refinement, multiple lattice-translocation defects present in the measured diffraction data needed to be solved. Two different approaches were tested and compared. Correction of the measured intensities proved to be superior to the use of the dislocation model of asymmetric units with partial occupancy of individual chains. As the crystals suffered from multiple lattice translocations, equations for their correction were derived de novo. The presented approach to the correction of multiple lattice-translocation defects may help to solve similar problems in the field of protein X-ray crystallography.

One of the main limitations of laser powder bed fusion technology is the residual stress (RS) introduced into the material by the local heating of the laser beam. RS restricts the processability of some materials and causes shape distortions in the process. Powder bed preheating is a commonly used technique for RS mitigation. Therefore, the objective of this study was to investigate the effect of powder bed preheating in the range of room temperature to 400 °C on RS, macrostructure, microstructure, mechanical properties, and properties of the unfused powder of the nickel-based superalloy Inconel 939. The effect of base plate preheating on RS was determined by an indirect method using deformation of the bridge-shaped specimens. Inconel 939 behaved differently than titanium and aluminum alloys when preheated at high temperatures. Preheating at high temperatures resulted in higher RS, higher 0.2% proof stress and ultimate strength, lower elongation at brake, and higher material hardness. The increased RSs and the change in mechanical properties are attributed to changes in the microstructure. Preheating resulted in a larger melt pool, increased the width of columnar grains, and led to evolution of the carbide phase. The most significant microstructure change was in the increase of the size and occurrence of the carbide phase when higher preheating was applied. Furthermore, it was detected that the evolution of the carbide phase strongly corresponds to the build time when high-temperature preheating is applied. Rapid oxidation of the unfused powder was not detected by EDX or XRD analyses.

Background: We estimated what risk factors affect hospitalisation for confirmed pertussis cases among infants (child up to 1 year) in the Czech Republic based on data from the questionnaire-based enhanced surveillance system (ESS) in years 2015, 2017 and 2019., Methods: Retrospective cohort study was conducted in which we assessed demographic, clinical data, vaccination status and risk/protective factors. Vaccination status was extracted from the electronic nationwide notification system (NNS). We performed descriptive, univariable and multivariable analysis using risk ratio (RR) and logistic regression with odds ratio (OR)., Results: A total of 104 confirmed cases (27, 13, 64) were reported in the ESS during 2015, 2017 and 2019. Most cases were in age group 1 month (24), more males (57). Fifteen cases were vaccinated and 89 unvaccinated. Of 88 hospitalised cases, 31 cases reported stay in Intensive Care Unit (ICU). The median length of hospitalisation was 8 days. Although the variable vaccinated in infants was statistically significant in a univariable analysis for outcome hospitalisation, RR 0.76 (95% CI 0.53-1.10), it was not in multivariable. Hospitalisation was strongly associated with the younger age group of 0-3 months adjusted by a smoking family member in a household (OR = 9.72; 95% CI: 2.97-31.81). Stay in ICU was strongly correlated with the younger age group of 0-3 months (OR = 5.07; 95% CI: 1.44-17.87) and with a contact with confirmed or probable pertussis (OR = 7.05; 95% CI: 1.36-36.52)., Conclusions: Our study demonstrated younger age and contact with other pertussis case as risk factors for hospitalisation of infants with pertussis. It is necessary to consider adolescent and adult boosters, including vaccination during pregnancy. We suggest integrating the variables from the enhanced surveillance system into the nationwide notification system, in order to simplify the data reporting and evaluation. Further studies are needed to evaluate the ESS and to monitor the vaccination of pregnant women against pertussis.

Water plays an important role in stabilizing the structure of DNA and mediating its interactions. Here, the hydration of DNA was analyzed in terms of dinucleotide fragments from an ensemble of 2727 nonredundant DNA chains containing 41 853 dinucleotides and 316 265 associated first-shell water molecules. The dinucleotides were classified into categories based on their 16 sequences and the previously determined structural classes known as nucleotide conformers (NtCs). The construction of hydrated dinucleotide building blocks allowed dinucleotide hydration to be calculated as the probability of water density distributions. Peaks in the water densities, known as hydration sites (HSs), uncovered the interplay between base and sugar-phosphate hydration in the context of sequence and structure. To demonstrate the predictive power of hydrated DNA building blocks, they were then used to predict hydration in an independent set of crystal and NMR structures. In ten tested crystal structures, the positions of predicted HSs and experimental waters were in good agreement (more than 40% were within 0.5 Å) and correctly reproduced the known features of DNA hydration, for example the `spine of hydration' in B-DNA. Therefore, it is proposed that hydrated building blocks can be used to predict DNA hydration in structures solved by NMR and cryo-EM, thus providing a guide to the interpretation of experimental data and computer models. The data for the hydrated building blocks and the predictions are available for browsing and visualization at the website https://watlas.datmos.org/watna/., (open access.)

Aim: The aim is to characterize in more detail the group of HIV-positive persons in the Czech Republic diagnosed with tuberculosis (TB) in 2000-2020., Material and Methods: Data sources were mainly the national online TB register (RTBC), which is part of the information system of the Public Health Service, and the national electronic register of HIV-positive persons (RHIV) maintained by the National Reference Laboratory for HIV/AIDS of the National Institute of Public Health., Results: Of 3,763 TB cases reported to the RHIV since 1985 and 16,212 TB cases reported to the RTBC since 2000, 91 occurred in 88 HIV-positive persons (69 males and 22 females) between 2000 and 2020. Sixty-five (74%) of the 88 HIV-positive persons were foreign born. Twenty-six per cent of TB cases were screened for HIV. The mean age of patients with TB/HIV coinfection was 35 years. The largest number of coinfected persons (35 cases) were from the capital city of Prague. Pulmonary TB was detected in 84 cases. Ninety-two per cent of the TB cases were bacteriologically confirmed, and 10 cases were multidrug-resistant TB. At the time of TB diagnosis, the median CD4+ lymphocyte count was 91.5 cells per mm3 of blood. TB was the most common reason for HIV testing in the analysed cohort (23 cases). The most common mode of HIV transmission was sexual intercourse (heterosexual in 39 cases and homosexual in 13 cases). Treatment success at 12-month follow-up was only recorded in 32% of cases of culture-positive pulmonary TB in HIV-positive patients., Conclusions: TB/HIV co-infection remains a serious health concern, especially in the foreign-born residents of the Czech Republic. Of foreign-born persons with TB, 42% were tested for HIV over the 21-year study period, with their percentage increasing over the years. Almost 6% of them tested HIV positive. The most relevant finding is that treatment success was only recorded for less than one third of HIV-positive persons with culture-positive pulmonary TB and that every fourth patient with TB/HIV died before or during TB treatment.

Aim: Homelessness is a social, societal, economic, and health problem. The aim of the study was to describe the characteristics of hospitalized homeless persons in the Czech Republic in relation to their health status and to propose targeted measures for improvement., Material and Methods: The study used anonymous data from the National Registry of Hospitalized Patients from 2006-2015, including the primary diagnosis at hospital admission and up to five additional diagnoses according to the 10th revision of the International Classification of Diseases (ICD-10). The basic statistical unit is the completed case of hospitalization of a homeless person in an inpatient health care facility such as hospital or nursing home., Results: Over the 10-year study period (2006-2015), 3387 homeless persons, 77.5% of them male, were admitted to inpatient health care facilities in the Czech Republic. The length of hospital stay tended to decrease over the years, with a median of six days. The majority of hospitalized homeless persons were aged between 40 and 69 years. Their distribution by type of health care facility: 2,710 (80.0%) were admitted to hospitals, 468 (13.8%) to psychiatric hospitals, 180 (5.3%) to long-term care facilities, and 29 (0.9%) to tuberculosis and respiratory disease hospitals. According to the mode of termination of hospitalization, 2189 (64.6%) homeless persons were discharged to their original environment, 280 (8.3%) were transferred to an after-care facility, 277 (8.2%) were transferred to an acute care facility, 222 (6.6%) left against medical advice, 180 (5.3%) were transferred to another ward, and 80 (2.4%) were transferred to a social care facility. A total of 159 (4.7%) in-hospital deaths occurred. The underlying causes of hospitalization according to ICD-10 were mostly those under Chapter V - Mental and behavioural disorders (19.9%) and Chapter XIX - Injury, poisoning and certain other consequences of external causes (18.1%). Among the 175 cases with a primary diagnosis from Chapter I - Certain infectious and parasitic diseases, the most common conditions were scabies, pediculosis, mycosis, bacterial infections, and tuberculosis., Conclusion: Data on hospital admissions is one of the important tools for monitoring the health of homeless people. They illustrate the seriousness of the situation of homeless people and point to the need to obtain more information on this issue in order to improve the availability of specific health care and social care for sick homeless people. Prevention programs, residential services, and specific outreach health and social activities can contribute to reducing the negative impacts.

Supramolecular structures, such as micelles, liposomes, polymerosomes or dendrimerosomes, are widely studied and used as drug delivery systems. The behavior of amphiphilic building blocks strongly depends on their spatial distribution and shape of polar and nonpolar component. This report is focused on the development of new versatile synthetic protocols for amphiphilic carbosilane dendrons (amp-CS-DDNs) capable of self-assembly to regular micelles and other supramolecular objects. The presented strategy enables the fine modification of amphiphilic structure in several ways and also enables the facile connection of a desired functionality. DLS experiments demonstrated correlations between structural parameters of amp-CS-DDNs and the size of formed nanoparticles. For detailed information about the organization and spatial distribution of amp-CS-DDNs assemblies, computer simulation models were studied by using molecular dynamics in explicit water.

Fog is an important atmospheric phenomenon highly relevant to ecosystems and/or the environment. Two essential prerequisites of fog formation are the presence of fog condensation nuclei and water in the atmosphere. The aim of our study was to examine in detail how fog occurrence is influenced by water areas in the immediate vicinity of the fog observation site. We have used as input data long-term observations on fog occurrence measured at 56 professional meteorological stations in Romania in 1981-2017 and GIS-derived information on water areas and on two topographical indices, TWI and TPI, in the neighbourhood of these stations. We formulated three alternative models of different complexity based on a semiparametric generalised additive logistic model for the probability of fog occurrence with potentially nonlinear, smooth effects modelled via penalised splines. A radius of 9 km appeared to be the most influential when considering the water area in a circle around the fog observation station. Based on our results, we concluded that (i) the water area in the vicinity of the station is a factor influencing fog occurrence, (ii) the water's effect differs according to water type (freshwater or seawater proximity), and (iii) GIS-derived topographical indices are informative for the explanation of fog occurrence and their inclusion enhanced the fit of the models substantially. Our findings, based on a reliable long-term data set of fog occurrence and recent GIS-derived data, explored by a relevant statistical approach will enhance further considerations related to fog formation and its environmental consequences., Competing Interests: Declaration of competing interest None., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Lipids modified by oxidative stress are key players in atherosclerosis progression. Superimposed thrombosis with subsequent closure of the coronary artery leads to the clinical manifestation of acute coronary syndrome (ACS). While several studies focusing on alterations in lipid metabolism in the acute phase have been conducted, no information is available on patients' lipidome alterations over longer time periods. In the current follow-up study, we analyzed plasma samples obtained from 17 patients three years after their ACS event (group AC). Originally, these patients were sampled 3-5 days after an index event (group B). Lipidome stability over time was studied by untargeted lipidomics using high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS). Multi-dimensional statistics used for data processing indicated that plasmalogen lipids were the most prominent lipids separating the above patient groups and that they increased in the follow-up AC group. A similar trend was observed for lysophosphatidylethanolamine (LPE) and phosphatidylethanolamine (PE). The opposite trend was observed for two fatty acyls of hydroxy fatty acid (FAHFAs) lipids and free stearic acid. In addition, a decrease in the "classic" oxitadive stress marker, malondialdehyde (MDA), occurred during the follow-up period. Our findings present unique information about long-term lipidome changes in patients after ACS.

The complexity of drug delivery mechanisms calls for the development of new transport system designs. Here, we report a robust synthetic procedure toward stable glycodendrimer (glyco-DDM) series bearing glucose, galactose, and oligo(ethylene glycol)-modified galactose peripheral units. In vitro cytotoxicity assays showed exceptional biocompatibility of the glyco-DDMs. To demonstrate applicability in drug delivery, the anticancer agent doxorubicin (DOX) was encapsulated in the glyco-DDM structure. The anticancer activity of the resulting glyco-DDM/DOX complexes was evaluated on the noncancerous (BJ) and cancerous (MCF-7 and A2780) cell lines, revealing their promising generation- and concentration-dependent effect. The glyco-DDM/DOX complexes show gradual and pH-dependent DOX release profiles. Fluorescence spectra elucidated the encapsulation process. Confocal fluorescence microscopy demonstrated preferential cancer cell internalization of the glyco-DDM/DOX complexes. The conclusions were supported by computer modeling. Overall, our results are consistent with the assumption that novel glyco-DDMs and their drug complexes are very promising in drug delivery and related applications.

One of the proposed strategies for the development of a more efficient HIV-1 vaccine is based on the identification of proteins binding to a paratope of chosen broadly neutralizing antibody (bNAb) that will mimic cognate HIV-1 Env (glyco)protein epitope and could be used as potent immunogens for induction of protective virus-neutralizing antibodies in the immunized individuals. To verify this "non-cognate ligand" concept, we developed a highly complex combinatorial library designed on a scaffold of human myomesin-1 protein domain and selected proteins called Myomedins specifically binding to variable regions of HIV-1 broadly neutralizing antibody 10E8. Immunization of mice with these Myomedin variants elicited the production of HIV-1 Env-specific antibodies. Hyperimmune sera bound to Env pseudotyped viruses and weakly/moderately neutralized 54% of tested clade A, B, C, and AE pseudotyped viruses variants in vitro . These results demonstrate that Myomedin variants have the potential to mimic Env epitopes and could be used as potential HIV-1 vaccine components.

In macromolecular crystallography, paired refinement is generally accepted to be the optimal approach for the determination of the high-resolution cutoff. The software tool PAIREF provides automation of the protocol and associated analysis. Support for phenix.refine as a refinement engine has recently been implemented in the program. This feature is presented here using previously published data for thermolysin. The results demonstrate the importance of the complete cross-validation procedure to obtain a thorough and unbiased insight into the quality of high-resolution data.

Alterations in lipid metabolism mediated by oxidative stress play a key role in the process of atherosclerosis and superimposed thrombosis; these can lead to acute coronary syndrome (ACS) and acute ischemic stroke (AIS). Multiple studies have shown that the formation of atheromatous lesions is initiated by oxidation of low-density lipoproteins incorporated into the intima of the vessel wall. Here, we studied lipids in plasma samples from three cohorts: 61 patients with ACS (group A), 49 patients with AIS (group D), and 82 controls (group K). Untargeted lipidomics based on high-performance liquid chromatography coupled to mass spectrometry (UHPLC-HRMS) was employed to obtain comprehensive information on whether relationships exist between these patient categories based on lipid patterns. In addition, malondialdehyde (MDA) as a standard marker of oxidative stress was monitored. The most characteristic lipids in group K were fatty acyls of hydroxyfatty acids (FAHFAs). As expected, MDA concentrations were the lowest in group K. Our findings can better explain ongoing pathologies, both acute and chronic, with the potential for future diagnosis and treatment.

A novel approach to inducing anion transport over the dialytic membrane was proposed and successfully tested using the dihydrogen phosphate anion. The anion receptor based on isophthalamide was anchored on a dendritic skeleton, resulting in a macromolecular structure with a limited possibility to cross the dialytic membrane. The dendritic receptor was placed in a compartment separated from a mother anion solution by a membrane. The resulting anion complexation reduced the actual concentration of the anion and induced the anion transfer across the membrane. The anion concentration in mother solution decreased, while it was found to be increased in the compartment with the dendritic receptor. This phenomenon was observed using dendritic receptors with four and eight complexation sites. A detailed analysis of a series of dialytic experiments by 1 H NMR spectroscopy enabled an assessment of the complexation behavior of both receptors and an evaluation of the dendritic effect on the anion complexation., Competing Interests: The authors declare no competing financial interest., (© 2021 The Authors. Published by American Chemical Society.)

Aim: Treatment with sacubitril/valsartan (S/V) significantly improves cardiovascular morbidity, mortality, quality of life and prolongs the survival of chronic heart failure patients with reduced ejection fraction. The aim of the study was to evaluate changes in ejection fraction, NT-proBNP and glomerular filtration after 12 months of sacubitril/valsartan treatment., Methods: 30 patients (28 men) with chronic heart failure with reduced ejection fraction, functional classes NYHA II-III, EF LK &lt; 40%, NT-proBNP (&gt; 450 ng/l), with glomerular filtration &gt; 0.5 ml/s/1.73 m2, with a potassium &lt; 5.4 mmol/l were classified in the study. S/V treatment was started at systolic blood pressure &gt; 100 mmHg. Ejection fraction, glomerular filtration rate and NT-proBNP values were compared before treatment and after 12 months of S/V treatment. The number of hospitalizations and deaths was also monitored., Results: During 12 months of S/V treatment there was a significant improvement in left ventricular ejection fraction (median initial 26.3%, after treatment 36.3%, difference 7.5%, p.

Fog is a very complex phenomenon, relevant to both atmospheric physics and chemistry, contributing to the atmospheric inputs of both nutrients and pollutants to the environment. Fog occurrence is affected by numerous factors. The aim of this study is to examine the effects of terrain on fog occurrence. Namely, we studied in detail how altitude, slope and landform influence the probability of fog occurrence using the generalized additive model. In particular, we investigated how different explanatory variables might modify (deform) the trend and the seasonal component of the probability of fog occurrence. We used long-term records of daily fog occurrence measured in 1981-2017 at 56 professional meteorological stations in Romania, reflecting different environments and geographical areas. The altitude of the sites under review ranged between 13 and 2504 m above sea level, the coverage of localities at different altitudes being highly uneven. Out of the terrain variables considered, the most decisive influence was found to be altitude. We have included information on slope and landform, which refined and bettered the basic model. Our model results indicated a significant decrease in the probability of fog occurrence over the examined period. The behaviour of fog differed according to the altitude, the most profound effects being observed for ground-level fog and fog above flat terrain. The probability of fog occurrence at different altitudes varied mostly in summer and autumn, whereas it was very similar in winter., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Reports of SARS-CoV-2 reinfections are on the rise. This study focused on reinfections in patients with confirmed COVID-19 in the Czech Republic. Between 1 March 2020 and 9 November 2020, 362 084 cases with the onset of symptoms before 31 October 2020 were reported. Overall, 28 cases of symptomatic SARS-CoV-2 reinfections were identified, 11 in males and 17 in females, age range 25-80 years, median age 46 years. The interval between the first and second episodes of the disease ranged from 101 to 231 days, and the median interval was 201.5 days. During both symptomatic episodes, all patients have been tested by RT-PCR. Altogether 26 patients (92.9%) have been tested negative after recovery from the first episode of COVID-19. Symptomatic reinfections occurred in nearly 0.2% of all patients at risk. Most patients with reinfection had mild symptoms in both episodes, and only three episodes were moderate to severe. Thus, reinfections may have been underdiagnosed. In summary, COVID-19 reinfections are possible and not exceptional.

Background: The aim of our study was to analyse the influence of air temperature and implemented veterinary measures on salmonellosis incidence in the Czech Republic (CZ)., Methods: We conducted a descriptive analysis of salmonellosis as reported to the Czech national surveillance system during 1998-2017 and evaluated the influence of applied veterinary measures (started in January 2008) on salmonellosis incidence by comparing two 9-year periods (1998-2006, 2009-2017). Using a generalized additive model, we analysed association between monthly mean air temperature and log-transformed salmonellosis incidence over the entire twenty-year period., Results: A total of 410,533 salmonellosis cases were reported during the study period in the CZ. Annual mean incidences of salmonellosis were 313.0/100,000 inhabitants before and 99.0/100,000 inhabitants after implementation of the veterinary measures. The time course of incidence was non-linear, with a sharp decline during 2006-2010. Significant association was found between disease incidence and air temperature. On average, the data indicated that within a common temperature range every 1 °C rise in air temperature contributed to a significant 6.2% increase in salmonellosis cases., Conclusions: Significant non-linear effects of annual trend, within-year seasonality, and air temperature on the incidence of salmonellosis during 1998-2017 were found. Our study also demonstrates significant direct effect of preventive veterinary measures taken in poultry in reducing incidence of human salmonellosis in the CZ. The annual mean number of salmonellosis cases in the period after introducing the veterinary measures was only 32.5% of what it had been in the previous period.

The Fish Embryo Acute Toxicity (FET) Test was adopted by the Organisation for Economic Co-operation and Development as OECD TG 236 in 2013. The test has been designed to determine acute toxicity of chemicals on embryonic stages of fish and proposed as an alternative method to the Fish Acute Toxicity Test performed according to OECD TG 203. In recent years fish embryos were used not only in the assessment of toxicity of chemicals but also for environmental and wastewater samples. In our study we investigated the acute toxicity of treated wastewater from seven hospitals in the Czech Republic. Our main purpose was to compare the suitability and sensitivity of zebrafish embryos with the sensitivity of two other aquatic organisms commonly used for wastewater testing - Daphnia magna and Aliivibrio fischeri. For the aim of this study, in addition to the lethal endpoints of the FET test, sublethal effects such as delayed heartbeat, lack of blood circulation, pericardial and yolk sac edema, spinal curvature and pigmentation failures were evaluated. The comparison of three species demonstrated that the sensitivity of zebrafish embryos is comparable or in some cases higher than the sensitivity of D. magna and A. fischeri. The inclusion of sublethal endpoints caused statistically significant increase of the FET test efficiency in the range of 1-12 %. Based on our results, the FET test, especially with the addition of sublethal effects evaluation, can be considered as a sufficiently sensitive and useful additional tool for ecotoxicity testing of the acute toxicity potential of hospital effluents.

Ambient or ground-level ozone (O 3 ), a key player in atmospheric chemistry, has been studied extensively throughout the world in recent decades due to its harmful effects on human health and the environment. The aim of this study is to examine in detail the time patterns of O 3 in the Czech Republic, a Central European country with a long-term history of ambient air pollution. We have examined the interplay between annual trends and within-a-year seasonality in daily O 3 concentrations at twelve stations representing different environments, geographical regions and altitudes in the Czech Republic in 1993-2018. We base our analysis on a generalised additive model (GAM) framework as a modern regression approach suitable for addressing non-linear trend shapes in a formalised and unified way. In particular, we use penalised splines. The inconsistent behaviour of the stations prior to 1998, two local maxima around 2003 and 2006 and a steady increase in O 3 at all sites from 2014 to the present are the main features of our study. A seasonality O 3 pattern with a clear maximum in May-June is similar for all stations. In parallel with the O 3 concentration changes over the years, the deformation of seasonality profiles over a long period of time occurs, resulting in a shift of the peak O 3 concentrations towards later days in the year. Our statistical modelling offers a detailed view of both long-term trend and seasonality components separately. Additionally, in a model with trend*seasonality interaction, we are able to study the gradual deformation of the seasonality profile over the years. Capsule: Our analysis of daily mean O 3 concentrations at twelve Czech sites in 1993-2018 revealed a steady increase in O 3 from 2014 and deformation of the seasonality O 3 profiles over the year., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Perfluoralkylated substances (PFASs) are persistent and bioaccumulative environmental contaminants. They are included on the list of emergent compounds monitored in the frame of HBM4EU project., Objectives: To analyze PFASs levels in human milk samples collected in the period 2006 through 2017, to follow their time trends, to assess the PFASs exposure in breastfed infants, to calculate the daily intake of PFASs and to compare it with the tolerable daily/weekly) intakes and to quantify risk from exposure using the hazard quotient and hazard index approach., Material and Methods: A broad spectrum of PFASs were analyzed by means of UHPLC-MS/MS in primipara human milk samples collected in four consecutive time periods 2006, 2010/11, 2014, and 2017; N = 46, 183, 164 and 232, respectively. Mothers living in urban and suburban residences were recruited after their delivery at maternity hospitals, and milk samples were taken within 2 and 8 weeks after delivery. The questionnaire was focused on possible sources of exposure, dietary habits and lifestyle., Results: Only perfluorooctane sulfonate (PFOS) and perfluorooctanoid acid PFOA (in 2017, also perfluorononanoic acid (PFNA)) were quantified in more than 90% of analyzed human milk samples. In all sampling periods, the levels of PFOA were higher than those of PFOS (p < 0.05). A significant downward temporal trend (p < 0.001) was observed for both PFOA and PFOS levels. The median concentrations in sampling years 2006, 2010/11, 2014, and 2017 were 0.075, 0.059, 0.035, and 0.023 ng/mL for PFOA and 0.045, 0.031, 0.029, and 0.020 ng/mL for PFOS, respectively. In 2017, PFNA was also quantified in 99% of samples with the median concentration of 0.007 ng/mL. The levels of PFASs correlated with maternal sea fish consumption. No maternal age-related relationship was observed. Using the tolerably daily intake (TDI) values for PFOS and PFOA set by the European Food Safety Authority (EFSA) in 2008, the calculated daily intakes from breastfeeding were clearly below these limits. Using the new, more conservative EFSA Provisional Tolerably Weekly Intake (PTWI) values set in 2018, we demonstrated a considerable exceedance of PTWI, with a hazard index above 1., Conclusion: Significant time-related decreasing trends in the PFOS and PFOA levels in human milk were observed. Nevertheless, the body burden of infants from breastfeeding might pose an enhanced health risk to infants when the current PTWI values are applied. These findings strongly support the present EU efforts to phase out PFOA, its salts and PFOA related compounds. Since PFOS exposure there has still been widely detected despite PFOS usage reduction measures, the major exposure routes should be further monitored and, if possible, eliminated., (Copyright © 2020 Elsevier Inc. All rights reserved.)

A detailed description of the dnatco.datmos.org web server implementing the universal structural alphabet of nucleic acids is presented. It is capable of processing any mmCIF- or PDB-formatted files containing DNA or RNA molecules; these can either be uploaded by the user or supplied as the wwPDB or PDB-REDO structural database access code. The web server performs an assignment of the nucleic acid conformations and presents the results for the intuitive annotation, validation, modeling and refinement of nucleic acids., (open access.)

Crystallographic resolution is a key characteristic of diffraction data and represents one of the first decisions an experimenter has to make in data evaluation. Conservative approaches to the high-resolution cutoff determination are based on a number of criteria applied to the processed X-ray diffraction data only. However, high-resolution data that are weaker than arbitrary cutoffs can still result in the improvement of electron-density maps and refined structure models. Therefore, the impact of reflections from resolution shells higher than those previously used in conservative structure refinement should be analysed by the paired refinement protocol. For this purpose, a tool called PAIREF was developed to provide automation of this protocol. As a new feature, a complete cross-validation procedure has also been implemented. Here, the design, usage and control of the program are described, and its application is demonstrated on six data sets. The results prove that the inclusion of high-resolution data beyond the conventional criteria can lead to more accurate structure models., (© Martin Malý et al. 2020.)

Thermodynamic acidity constants (acid or acid-base dissociation constants, sometimes called also as ionization constants) and limiting ionic mobilities (both of them at defined temperature, usually 25°C) are the fundamental physicochemical characteristics of a weak electrolyte, that is, weak acid or weak base or ampholyte. We introduce a novel method for determining the data of a weak electrolyte by the nonlinear regression of effective electrophoretic mobility versus buffer composition dependence when measured in a set of BGEs with various pH. To correct the experimental data for zero ionic strength we use the extended Debye-Hückel model and Onsager-Fuoss law with no simplifications. Contrary to contemporary approaches, the nonlinear regression is performed on limiting mobility data calculated by PeakMaster's correction engine, not on the raw experimental mobility data. Therefore, there is no requirement to perform all measurements at a constant ionic strength of the set of BGEs. We devised the computer program AnglerFish that performs the necessary calculations in a user-friendly fashion. All thermodynamic pKa values and limiting electrophoretic mobilities for arbitrarily charged substances having any number of ionic forms are calculated by one fit. The user input consists of the buffer composition of the set of BGEs and experimentally measured effective mobilities of the inspected weak electrolyte., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Fog is a very important and complex atmospheric phenomenon of the utmost importance for the environment and for human society. For practical reasons, fog occurrence is observed regularly at meteorological stations worldwide. Decreasing trends in fog frequency reported from numerous regions have been often associated with either decreasing pollution or climate change, including increasing temperature and changes in atmospheric circulation. We have examined the data on fog occurrence from twelve Czech sites representing different environments (urban, rural, mountain), geographical areas, and altitudes across the country. For our analysis we used long-term records from the time period of 1961-2018, covering both the ambient air's heavily polluted periods of the 1970s and 1980s and the cleaner period, following the adoption of new, more stringent legislation and effective countermeasures after the 1990s. We applied a generalised additive model (GAM) framework as a flexible, semiparametric regression approach to address nonlinear trend shapes in a formalised and unified way. In particular, we employed a penalised spline approach with cross-validated penalty coefficient estimation. Our study confirmed non-linear behaviour for both year-to-year trends and annual seasonality. Our results showed further that over the analysed, almost sixty-year period, fog occurrence has decreased significantly at all the examined sites, though the pattern of the long-term change differed among individual sites. Moreover, we have found significant seasonality in fog occurrence, though it is different at individual sites. Furthermore, apart from the overall annual fog probability change over the years, at some sites the fog's seasonal profile has also deformed substantially over the long term., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Fourteen low molecular mass UV absorbing ampholytes containing 1 or 2 weakly acidic and 1 or 2 weakly basic functional groups that best satisfy Rilbe's requirement for being good carrier ampholytes (ΔpK a = pKa monoanion - pKa monocation < 2) were selected from a large group of commercially readily available ampholytes in a computational study using two software packages (ChemSketch and SPARC). Their electrophoretic mobilities were measured in 10 mM ionic strength BGEs covering the 2 < pH < 12 range. Using our Debye-Hückel and Onsager-Fuoss laws-based new software, AnglerFish (freeware, https://echmet.natur.cuni.cz/software/download), the effective mobilities were recalculated to zero ionic strength from which the thermodynamic pK a values and limiting ionic mobilities of the ampholytes were directly calculated by Henderson-Hasselbalch equation-type nonlinear regression. The tabulated thermodynamic pK a values and limiting ionic mobilities of these ampholytes (pI markers) facilitate both the overall and the narrow-segment characterization of the pH gradients obtained in IEF in order to mitigate the errors of analyte ampholyte pI assignments caused by the usual (but rarely proven) assumption of pH gradient linearity. These thermodynamic pK a and limiting mobility values also enable the reality-based numeric simulation of the IEF process using, for example, Simul (freeware, https://echmet.natur.cuni.cz/software/download)., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Bioresorbable scaffold (BRS) Absorb™ clinical use has been stopped due to higher rate of device thrombosis. Scaffold struts persist longer than 2 years in the vessel wall. Second generation devices are being developed. This study evaluates long-term invasive imaging in STEMI patients., Methods: PRAGUE-19 study is an academic study enrolling consecutive STEMI patients with intention to implant Absorb™ BRS. A total of 83 STEMI patients between December 2012 and March 2014 fulfilled entry criteria. Coronary angiography and optical coherence tomography at 5 year follow-up was performed in 25 patients., Results: Primary combined clinical endpoint (death, myocardial infarction or target vessel revascularization) occurred in 12.6% during the five-year follow-up with overall mortality 6.3%. Definite scaffold thrombosis occurred in 2 patients in the early phase after BRS implantation. Quantitative coronary angiography after 5 years demonstrated low late lumen loss of 0.11 ± 0.35 mm with binary restenosis rate of 0%. Optical coherence tomography demonstrated complete resorption of scaffold struts and mean lumen diameter of 3.25 ± 0.30 and 3.22 ± 0.49 (P = 0.73) at baseline and after 5 years, respectively. Three patients developed small coronary artery aneurysm in the treated segment., Conclusion: Invasive imaging results 5 years after BRS implantation in STEMI showed complete resorption of scaffold struts and stable lumen vessel diameter. Trial registration ISRCTN43696201 (retrospectivelly registred, June 7th, 2019). https://www.isrctn.com/ISRCTN43696201.

Oxidative stress in humans is related to various pathophysiological processes, which can manifest in numerous diseases including cancer, cardiovascular diseases, and Alzheimer's disease. On the atomistic level, oxidative stress causes posttranslational modifications, thus inducing structural and functional changes into the proteins structure. This study focuses on fibrinogen, a blood plasma protein that is frequently targeted by reagents causing posttranslational modifications in proteins. Fibrinogen was in vitro modified by three reagents, namely sodium hypochlorite, malondialdehyde, and 3-morpholinosydnonimine that mimic the oxidative stress in diseases. Newly induced posttranslational modifications were detected via mass spectrometry. Electron microscopy was used to visualize changes in the fibrin networks, which highlight the extent of disturbances in fibrinogen behavior after exposure to reagents. We used molecular dynamics simulations to observe the impact of selected posttranslational modifications on the fibrinogen structure at the atomistic level. In total, 154 posttranslational modifications were identified, 84 of them were in fibrinogen treated with hypochlorite, 51 resulted from a reaction of fibrinogen with malondialdehyde, and 19 were caused by 3-morpholinosydnonimine. Our data reveal that the stronger reagents induce more posttranslational modifications in the fibrinogen structure than the weaker ones, and they extensively alter the architecture of the fibrin network. Molecular dynamics simulations revealed that the effect of posttranslational modifications on fibrinogen secondary structure varies from negligible alternations to serious disruptions. Among the serious disruptions is the oxidation of γR375 resulting in the release of Ca2+ ion that is necessary for appropriate fibrin fiber formation. Folding of amino acids γE72-γN77 into a short α-helix is a result of oxidation of γP76 to glutamic acid. The study describes behaviour of fibrinogen coiled-coil connecter in the vicinity of plasmin and hementin cleavage sites., Competing Interests: NO authors have competing interests.

We discuss several possible phenomena in electrophoretic systems with complexing agents present in the background electrolyte. In our previous work, we extended the linear theory of electromigration with the first-order nonlinear term, which originally applied to acid-base equilibria only, by generalizing it to any fast chemical equilibria. This extension provides us with a fresh insight into the well-established technique of elecktrokinetic chromatography (EKC). We combine mathematical analysis of the generalized model with its solution by means of the new version of our software PeakMaster 6, and experimental data. We re-examine the fundamental equations by Wren and Rowe and Tiselius in the frame of the generalized linear theory of electromigration. Besides, we show that selector concentration can increase inside the interacting-analyte zone due to its complexation with the analyte, which contradicts the generally accepted idea of a consumption of a portion of the selector inside the zone. Next, we focus our discussion on interacting buffers (i.e., buffer constituents that form a complex with the selector). We demonstrate how such side-interaction of the selector with another buffer constituent can influence measuring analyte-selector interactions. Finally, we describe occurrence and mobilities of system peaks in these EKC systems. We investigate systems with fully charged analytes and neutral cyclodextrins as selectors. Although the theory is not limited in terms of the charge and/or the degree of (de)protonation of any constituent, this setup allows us to find analytical solutions to generalized model under approximate, yet realistic, conditions and to demonstrate all important phenomena that may occur in EKC systems. An occurrence of system peaks in a system with fully charged selector is also investigated., (Copyright © 2019. Published by Elsevier B.V.)

Background: Entry of human immunodeficiency virus type 1 (HIV-1) in target cells is enabled by CD4 receptor and one of two co-receptors, CXCR4 or CCR5. Deletion of 32 bp in CCR5 gene (CCR5Δ32) in both alleles provides strong but not absolute resistance to HIV-1 infection and deletion in one allele slows disease progression to AIDS. Here, we analyzed the prevalence and the role of CCR5Δ32 heterozygosity on the disease progression in HIV positive patients in the Czech Republic., Patients and Methods: A total of 92 HIV-1 seropositive subjects that included 80 Czech individuals from the AIDS center in the Hospital Na Bulovce in Prague were enrolled in CCR5 genotyping as a part of a study of the role of HIV fitness on disease progression. DNA was extracted from patient’s peripheral blood mononuclear cells and subjected to real-time PCR with specific probes detecting wild-type and 32 bp-deleted CCR5 variants. A subgroup of 74 antiretroviral therapy-naive patients with more than one year of follow-up was used to determine the role of the CCR5Δ32 heterozygous phenotype in disease progression., Results: CCR5Δ32 was found heterozygous in 23.8% of 80 Czech HIV-1 seropositive individuals which is very similar to 21% and 24% prevalence reported in HIV negative Czech population. Homozygous mutant variant was not detected. In CCR5Δ32 heterozygous group we observed slightly higher mean CD4+ T-cell count and lower mean plasma viremia levels., Conclusions: Overall, our study indicates no obvious benefit of CCR5Δ32 heterozygosity on HIV transmission and only small benefit on disease progression in the Czech HIV-1 cohort.

A simple, sensitive, specific, and cost-effective analytical methodology was developed for the analysis of human plasma samples spiked with imatinib by CZE with on-line UV detection in the context of Therapeutic Drug Monitoring. Several analytical conditions such as the ionic strength (I) and the pH of the BGE composed of citric acid and ε-amino caproic acid were studied in regards of the presence of sodium chloride (NaCl) in plasma samples (1% m/v). Computer simulations (Simul software) were used to confirm the experimental results and to understand imatinib electrophoretic behavior in the presence of NaCl. Furthermore, the advantages of adding ACN to the sample containing NaCl to combine efficient protein precipitation and on-line CZE stacking of imatinib were demonstrated. LOD and LOQ values of 48 and 191 ng/mL were obtained from plasma sample supernatant after protein precipitation with ACN, which is much lower than mean imatinib plasma level observed for patients treated by imatinib mesylate (about 1000 ng/mL). Good linearity was obtained in the concentration range 191-5000 ng/mL (R 2  > 0.997). RSD of less than 1.68% and 2.60% (n = 6) for migration times and corrected peak areas, respectively, were observed at the LOQ., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A survey was conducted in the Czech Republic to determine whether serological responses to the 15/17-kDa and 27-kDa Cryptosporidium antigens had changed since the end of the communist era and if these responses were associated with drinking water sources. Sera from 301 blood donors residing in six areas served by various sources of drinking water were analysed by Western Blot (mini-immunoblots) to measure the IgG response. The intensity of response and percentage of persons with a strong response to the 27-kDa, but not the 15/17-kDa, antigen were higher than found 20 years earlier. A strong response to both the 15/17- and 27-kDa-antigens was higher than reported in other countries, and the probability of persons having a strong response was greater in areas with surface water sources than river-bank infiltration. Few cases of cryptosporidiosis were reported in spite of these high responses to Cryptosporidium antigens. These responses suggest a chronic low-level exposure from several sources that may be affording protection against symptoms and illness. Although strong serological responses were associated with surface water sources, drinking water is not likely to be the most important exposure for Cryptosporidium in the Czech Republic.

Objectives: Individuals with HIV infection are at an increased risk for a number of infectious diseases, some of which are preventable by vaccination. Unfortunately, little is known about the attitudes of this population group to vaccination, therefore, we decided to find out vaccination coverage against 5 infections among newly diagnosed HIV-infected patients in the Czech Republic., Methods: This cross-sectional study was conducted on newly diagnosed patients who started their follow-up care at the HIV Clinic of Na Bulovce Hospital during the two following years. Vaccination history data and results of serological tests were collected from all participants., Results: Enrolled were 269 HIV-positive subjects (94.1% males) with a mean age of 34.4 years, 64 subjects (23.8%) had tertiary education, 229 (85.1%) were men having sex with men, 32 (11.9%) were heterosexual, and 8 (3.0%) were injection drug users. The mean CD4+ T-lymphocyte count was 556.2/µL, with 149 persons (55.4%) who had a CD4+ T-lymphocyte count &gt; 500/µL, and 68 (25.3%) individuals were late presenters with CD4+ T-lymphocyte count &lt; 350/µL. A vaccination against tetanus was reported by 262 subjects (97.4%), against influenza by 18 subjects (6.7%), against tick-borne encephalitis by 18 subjects (6.7%), against viral hepatitis A by 78 persons (29.0%), and against hepatitis B by 104 subjects (38.7%). For influenza, tick-borne encephalitis and hepatitis A, a significant positive impact of tertiary education was found (p-values &lt; 0.001-0.044). Vaccination coverage against both types of hepatitis was significantly lower in late presenters (p = 0.044 and p = 0.004, respectively)., Conclusions: Vaccination rates found in our cohort were except tetanus and hepatitis B in young people low, especially for influenza and tick-borne encephalitis. Higher level of education and less advanced HIV infection were associated with higher vaccination rates. To improve this unsatisfactory situation, more attention should be paid to vaccination.

Over the last decades, multidrug-resistant bacteria have emerged and spread, increasing the number of bacteria, against which commonly used antibiotics are no longer effective. It has become a serious public health problem whose solution requires medical research in order to explore novel effective antimicrobial molecules. On the one hand, antimicrobial peptides (AMPs) are regarded as good alternatives because of their generally broad-spectrum activities, but sometimes they can be easily degraded by the organism or be toxic to animal cells. On the other hand, cationic carbosilane dendrons, whose focal point can be functionalized in many different ways, have also shown good antimicrobial activity. In this work, we synthetized first- and second-generation cationic carbosilane dendrons with a maleimide molecule on their focal point, enabling their functionalization with three different AMPs. After different microbiology studies, we found an additive effect between first-generation dendron and AMP3 whose study reveals three interesting effects: (i) bacteria aggregation due to AMP3, which could facilitate bacteria detection or even contribute to antibacterial activity by preventing host cell attack, (ii) bacteria disaggregation capability of second-generation cationic dendrons, and (iii) a higher AMP3 aggregation ability when dendrons were added previously to peptide treatment. These compounds and their different effects observed over bacteria constitute an interesting system for further mechanism studies.

The aim of this study is to observe the effect of process parameters on residual stresses and relative density of Ti6Al4V samples produced by Selective Laser Melting. The investigated parameters were hatch laser power, hatch laser velocity, border laser velocity, high-temperature preheating and time delay. Residual stresses were evaluated by the bridge curvature method and relative density by the optical method. The effect of the observed process parameters was estimated by the design of experiment and surface response methods. It was found that for an effective residual stress reduction, the high preheating temperature was the most significant parameter. High preheating temperature also increased the relative density but caused changes in the chemical composition of Ti6Al4V unmelted powder. Chemical analysis proved that after one build job with high preheating temperature, oxygen and hydrogen content exceeded the ASTM B348 limits for Grade 5 titanium., Competing Interests: The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

The interaction of neuropeptides, vasoactive intestinal peptide (VIP), or growth hormone-releasing hormone (GHRH), with a cationic carbosilane dendrimer forms dendriplexes with antitumoral behavior in advanced prostate cancer cells PC3. At the concentrations used for dendriplexes formation, the free peptides were protumoral and prometastatic in advanced prostate cancer, while dendrimer only showed low cytotoxicity, but did not avoid the metastatic behavior of PC3 cells. However, these nanoplexes favored also cell adhesion and avoided cell migration. Also, the dendriplexes were not toxic for no tumoral prostate cells (RPWE-1) or fibroblasts. The use of labeled GHRH peptide (rhodamine labeled) and a dendrimer (fluorescein labeled) allowed us to observe that both systems reach the intracellular milieu after dendriplex formation. The treatment of PC3 cells with the nanoplexes reduced expression of vascular endothelial growth factor (VEGF) and cyclic adenosine monophosphate (cAMP). Molecular modeling analysis highlights the important contribution of the carbosilane framework in the stabilization of the dendriplex, since dendrimer interacts with a peptide region where hydrophobic amino acids are presented.

The linear theory of electromigration, including the first-order nonlinear approximation, is generalized to systems with any equilibria fast enough to be considered instantaneous in comparison with the timescale of peak movement. For example, this theory is practically applied in the electrokinetic chromatography (EKC) mode of the CZE. The model enables the calculation of positions and shapes of analyte and system peaks without restricting the number of selectors, the complexation stoichiometry, or simultaneous acid-base equilibria. The latest version of our PeakMaster software, PeakMaster 6-Next Generation, implements the theory in a user-friendly way. It is a free and open-source software that performs all calculations and shows the properties of the background electrolyte and the expected electropherogram within a few seconds. In this paper, we mathematically derive the model, discuss its applicability to EKC systems, and introduce the PeakMaster 6 software., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

ÚVOD Populace je v sou&#269;asné vystavena r&#367;zným zevním vliv&#367;m, ze kterých mohou vyplývat i r&#367;zná zdravotní rizika. Jedním z mo&#382;ných je i riziko vzniku vrozené vady (VV). Z hlediska studia zdravotního stavu populace je d&#367;le&#382;itá znalost nejen pr&#367;m&#283;rných celkových incidencí VV, ale i p&#345;ípadných zm&#283;n t&#283;chto incidencí v pr&#367;b&#283;hu &#269;asu. Incidence VV je pova&#382;ována za jeden ze základních kvalitativních ukazatel&#367; popula&#269;ních i medicínských (1). Nelze opomenout ani hledání potenciálních rizik vedoucích ke zvý&#353;ené pravd&#283;podobnosti výskytu VV. T&#283;mito riziky mohou být ne&#382;ádoucí vlivy zevního prost&#345;edí nebo faktory biologicko-sociální (v&#283;k &#382;en, jejich onemocn&#283;ní aj). Podíl jednotlivých typ&#367; VV u narozených d&#283;tí se v &#269;ase m&#283;ní, obdobn&#283; jako struktura p&#345;í&#269;in úmrtí. Z hlediska epidemiologického je nutné hodnotit incidence VV a jejich zm&#283;ny p&#345;edev&#353;ím z pohledu jednotlivých diagnóz. B&#283;hem posledních let se intenzita VV v &#268;esku m&#283;nila. U n&#283;kterých typ&#367; se &#269;etnost v novorozenecké populaci sni&#382;uje díky úsp&#283;&#353;né prenatální diagnostice, u jiných diagnóz naopak &#269;etnost u narozených d&#283;tí stoupá (2). Krom&#283; prenatální diagnostiky se v&#353;ak na výsledné &#269;etnosti VV v populaci mohou uplat&#328;ovat i dal&#353;í vlivy. Jedním z nich m&#367;&#382;e být i zlep&#353;ení, zrychlení a zkvalitn&#283;ní postnatálních diagnostických mo&#382;ností - p&#345;edev&#353;ím zavedení a rozvoj ultrazvukové diagnostiky (3). Úsp&#283;&#353;nost prenatální diagnostiky a &#269;etnost VV u narozených jsou rovn&#283;&#382; významným ukazatelem prenatální a perinatální pé&#269;e, p&#345;e&#382;ívání d&#283;tí narozených s VV pak ukazatelem pé&#269;e postnatální (p&#345;edev&#353;ím neonatální a chirurgické). Registrace vrozených vad má v &#268;eské republice dlouhou tradici - Národní registr vrozených vad zahájil svou &#269;innost ji&#382; p&#345;ed více ne&#382; p&#367;lstoletím, v roce 1964 (4).   METODIKA Retrospektivní epidemiologická studie vyu&#382;ívá oficiální data z Národního registru vrozených vad vedeného v rámci Registru reproduk&#269;ního zdraví v Ústavu zdravotnických informací a statistiky &#268;R (ÚZIS). Analyzovány byly incidence jednotlivých diagnóz vrozených vad (kódy Q00-Q99) u narozených d&#283;tí z celého území &#268;eské republiky za &#269;asové období 1994-2015. Incidence VV byly analyzovány pro ob&#283; pohlaví, a to jak celkov&#283; (pro celou skupinu diagnóz VV), tak i dle základních diagnostických skupin Mezinárodní klasifikace nemocí (MKN).   VÝSLEDKY V období 1994-2015 se dle údaj&#367; ÚZIS &#268;R narodilo v &#268;eské republice celkem 87 359 d&#283;tí s vrozenou vadou zji&#353;t&#283;nou do 1 roku &#382;ivota. Z tohoto celkového po&#269;tu bylo 51 315 chlapc&#367; a 36 030 dívek. U 14 p&#345;ípad&#367; nebylo pohlaví známé / zji&#353;t&#283;no. Vývoj po&#269;tu diagnostikovaných p&#345;ípad&#367; ukazují p&#345;ehledn&#283; první t&#345;i grafy (obr. 1-3), zvlá&#353;&#357; pro chlapce, dívky a celkov&#283;. V relativních po&#269;tech to bylo za celé sledované období pr&#367;m&#283;rn&#283; 385,4 na 10 000 &#382;iv&#283; narozených d&#283;tí: nejmén&#283; 242,5 v roce 1994, nejvíce pak 448,3 v roce 2011. V p&#345;ípad&#283; &#382;iv&#283; narozených chlapc&#367; to bylo nejmén&#283; 263,6 v roce 1994, nejvíce pak 533,3 v roce 2011; pr&#367;m&#283;rná hodnota &#269;inila 440,6 na 10 000. U &#382;iv&#283; narozených dívek byla pr&#367;m&#283;rná hodnota za sledované období 327,1 na 10 000 ; nejni&#382;&#353;í hodnota 220,1 byla zaznamenána op&#283;t v roce 1994, nejvy&#353;&#353;í hodnota 380,9 v roce 2003. Grafy na obr. 4-6 ukazují relativní po&#269;ty na 10 000 &#382;iv&#283; narozených v pr&#367;b&#283;hu sledovaného období, op&#283;t zvlá&#353;&#357; pro chlapce, dívky a celkov&#283;. Dal&#353;í &#269;ást na&#353;í analýzy se v&#283;novala zm&#283;nám v pr&#367;b&#283;hu sledovaného období pro jednotlivé skupiny diagnóz dle rozd&#283;lení v Mezinárodní klasifikaci nemocí, 10. verze (tab. 1). various types of congenital anomalies are changing during the time according to different factors. Three main factors are methodical changes of the registration process, improvement of prenatal diagnostics and the real changes of incidences of selected diagnoses. While in the previous time period (till 1993) the registry included only selected diagnoses of congenital anomalies, in the new period (starting 1994) the registry includes all diagnoses of congenital anomalies from the ICD-10 classification. We can also see the difference in the incidences from 1994-1999 and 2000-2015 time period. The reason for this difference is the methodical change of registration, the Registry of congenital anomalies also receives the notifications about possible anomalies from the report of newborn (even when the report of congenital anomaly was not sent). As for the prenatal diagnostics - we may analyse possible changes of incidences - if the specific defect can be diagnosed prenatally.

The study investigated the associations between the prevalence of the childhood respiratory diseases and the long-term exposure to air pollution in the burdened area of Moravian-Silesian Region in the Czech Republic. The health data were collected during 2014 in 7,239 children 5, 9, 13 and 17 years of age. Exposure to PM 10 and NO 2 in the residence addresses was based on dispersion models and GIS based traffic-related indicators. PM 10 levels were positively associated with both lifetime (OR 1.35; 95%CI 1.09-1.67) and current (OR 1.32; 95%CI 1.05-1.67) allergic rhinitis; current asthma was associated negatively. The associations between traffic indicator and respiratory health were not found. On the other hand, marked positive associations were found between the respiratory diseases and symptom severity structured into ordinal variables, and PM 10 and NO 2 . Modelled long-term exposure to air pollution was associated with childhood allergic rhinitis and deterioration of the respiratory symptoms.

Background: Prostate-specific membrane antigen (PSMA), also known as glutamate carboxypeptidase II (GCPII), is an important diagnostic and therapeutic target in prostate cancer. PSMA/GCPII is also expressed in many healthy tissues, but its function has only been established in the brain and small intestine. Several research groups have attempted to produce PSMA/GCPII-deficient mice to study the physiological role of PSMA/GCPII in detail. The outcomes of these studies differ dramatically, ranging from embryonic lethality to production of viable PSMA/GCPII-deficient mice without any obvious phenotype., Methods: We produced PSMA/GCPII-deficient mice (hereafter also referred as Folh1 -/- mice) by TALEN-mediated mutagenesis on a C57BL/6NCrl background. Using Western blot and an enzyme activity assay, we confirmed the absence of PSMA/GCPII in our Folh1 -/- mice. We performed anatomical and histopathological examination of selected tissues with a focus on urogenital system. We also examined the PSMA/GCPII expression profile within the mouse urogenital system using an enzyme activity assay and confirmed the presence of PSMA/GCPII in selected tissues by immunohistochemistry., Results: Our Folh1 -/- mice are viable, breed normally, and do not show any obvious phenotype. Nevertheless, aged Folh1 -/- mice of 69-72 weeks exhibit seminal vesicle dilation, which is caused by accumulation of luminal fluid. This phenotype was also observed in Folh1 +/- mice; the overall difference between our three cohorts (Folh1 -/- , Folh1 +/- , and Folh1 +/+ ) was highly significant (P < 0.002). Of all studied tissues of the mouse urogenital system, only the epididymis appeared to have a physiologically relevant level of PSMA/GCPII expression. Additional experiments demonstrated that PSMA/GCPII is also present in the human epididymis., Conclusions: In this study, we provide the first evidence characterizing the reproductive tissue phenotype of PSMA/GCPII-deficient mice. These findings will help lay the groundwork for future studies to reveal PSMA/GCPII function in human reproduction., (© 2018 Wiley Periodicals, Inc.)

The study compares diagnostic parameters of different commercial serological kits based on three different antigen types and correlates test results with the status of the patient's Borrelia infection. In total, 8 IgM and 8 IgG kits were tested, as follows: enzyme-linked immunosorbent assay (ELISA) (Euroimmun) based on whole-cell antigen, 3 species-specific enzyme immunoassays (EIAs) (TestLine), Liaison chemiluminescence (DiaSorin), ELISA-Viditest (Vidia), EIA, and Blot-Line (TestLine) using recombinant antigens. All tests were performed on a panel of 90 samples from patients with clinically characterized borreliosis (53 with neuroborreliosis, 32 with erythema migrans, and 5 with arthritis) plus 70 controls from blood donors and syphilis patients. ELISA based on whole-cell antigens has superior sensitivity and superior negative predictive value and serves as an excellent screening test, although its specificity and positive predictive values are low. Species-specific tests have volatile parameters. Their low sensitivity and low negative predictive value handicap them in routine diagnostics. Tests with recombinant antigens are characterized by high specificity and high positive predictive value and have a wide range of use in diagnostic practice. Diagnostic parameters of individual tests depend on the composition of the sample panel. Only a small proportion of contradictory samples giving both negative and positive results is responsible for discrepancies between test results. Correlation of test results with the patient's clinical state is limited, especially in the erythema migrans group with high proportions of negative and contradictory results. In contrast, IgG test results in the neuroborreliosis group, which are more concordant, show acceptable agreement with Borrelia status., (Copyright © 2018 Kodym et al.)

Interleukin 17 (IL-17) and its cognate receptor A (IL-17RA) play a crucial role in Th17 cells-mediated pro-inflammatory pathway and pathogenesis of several autoimmune disorders including psoriasis. IL-17 is mainly produced by activated Th-17 helper cells upon stimulation by IL-23 and, via binding to its receptors, mediates IL-17-driven cell signaling in keratinocytes. Hyper-proliferation of keratinocytes belongs to major clinical manifestations in psoriasis. To modulate IL-17-mediated inflammatory cascade, we generated a unique collection of IL-17RA-targeting protein binders that prevent from binding of human IL-17A cytokine to its cell-surface receptor. To this goal, we used a highly complex combinatorial library derived from scaffold of albumin-binding domain (ABD) of streptococcal protein G, and ribosome display selection, to yield a collection of ABD-derived high-affinity ligands of human IL-17RA, called ARS binders. From 67 analyzed ABD variants, 7 different sequence families were identified. Representatives of these groups competed with human IL-17A for binding to recombinant IL-17RA receptor as well as to IL-17RA-Immunoglobulin G chimera, as tested in enzyme-linked immunosorbent assay (ELISA). Five ARS variants bound to IL-17RA-expressing THP-1 cells and blocked binding of human IL-17 cytokine to the cell surface, as tested by flow cytometry. Three variants exhibited high-affinity binding with a nanomolar K d value to human keratinocyte HaCaT cells, as measured using Ligand Tracer Green Line. Upon IL-17-stimulated activation, ARS variants inhibited secretion of Gro-α (CXCL1) by normal human skin fibroblasts in vitro. Thus, we identified a novel class of inhibitory ligands that might serve as immunosuppressive IL-17RA-targeted non-IgG protein antagonists.

Glycodendrimers (Glyco-DDMs) represent a rapidly growing class of nanoparticles with promising properties for biomedical applications but concerns regarding the impact on human health and environment are still justified. Here we report, for the first time, the comparative study of in vivo developmental toxicity of carbosilane Glyco-DDMs and their cytotoxicity in vitro. Carbosilane Glyco-DDMs (generation 1-3) containing 4, 8, and 16 β-d-glucopyranosyl units at the periphery (DDM 1 Glu, DDM 2 Glu, and DDM 3 Glu) were synthesized and characterized by 1 H, 13 C and 29 Si NMR, mass spectrometry, dynamic light scattering, atomic force microscopy (AFM), and computer modeling. In vitro cytotoxicity assay (MTT) of DDM 1-3 Glu was performed on three different rodent cell lines (Cricetulus griseus) - B14 (ATCC, CCL-14.1), BRL 3A (ATCC, CRL-1442), and NRK 52E (ATCC, CRL-1571). Overall, very low cytotoxicity was observed with calculated IC 50 in mM range with slight difference between each cell line and DDM generation investigated. Modified fish embryo test (FET) was further used for DDM 3 Glu developmental toxicity testing on zebrafish (Danio rerio) embryos. While seemingly harmless to intact embryos, adverse effects of DDMs on the embryonic development become evident after chorion removal (LD 50 =2.78 µM at 96 hpe). We summarized that the modified FET test showed a two to three orders of magnitude difference between the in vitro cytotoxicity and in vivo developmental toxicity of DDM 3 Glu. While, in general, the Glyco-DDMs show great promises as efficient vectors in targeted drug delivery or as therapeutic molecules itself, we suggest that their developmental toxicity should be thoroughly investigated to exclude safety risks associated with their potential biomedical use.