Exploiting the anti-biofilm effect of the engineered phage endolysin PM-477 to disrupt in vitro single- and dual-species biofilms of vaginal pathogens associated with bacterial vaginosis

Supplementary Materials: The following are available online at: https://www.mdpi.com/article/10.3390/antibiotics11050558/s1, Figure S1: An example data set on the organization of the dual-species BV-associated biofilms by epifluorescence microscopy, Table S1: Bacterial strains used in this study, Table S2: Specific primers for the quantification of the bacterial species present in the dual-species biofilm.Bacterial vaginosis (BV) is the most frequent vaginal infection in women of reproductive age. It is caused by the overgrowth of anaerobic vaginal pathogens, such as Gardnerella vaginalis, Fannyhessea vaginae, and Prevotella bivia, which are vaginal pathogens detected during the early stages of incident BV and have been found to form multi-species biofilms. Treatment of biofilm-associated infections, such as BV, is challenging. In this study, we tested the role of an investigational engineered phage endolysin, PM-477, in the eradication of dual-species biofilms composed of G. vaginalis–F. vaginae or G. vaginalis–P. bivia. Single-species biofilms formed by these species were also analysed as controls. The effect of PM-477 on biomass and culturability of single- and dual-species biofilms was assessed in vitro using a microtiter plate assay, epifluorescence microscopy, confocal laser scanning microscopy, and quantitative PCR. The results showed that PM-477 was particularly effective in the disruption and reduction of culturability of G. vaginalis biofilms. In dual-species biofilms, PM-477 exhibited lower efficiency but was still able to selectively and significantly eliminate G. vaginalis. Since polymicrobial interactions have been shown to strongly affect the activity of various antibiotics, the activity of PM-477 in dual-species biofilms is a potentially promising result that should be further explored, aiming to completely eradicate multi-species biofilms associated with BV.This research was partially funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of unit (UIDB/04469/2020). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.info:eu-repo/semantics/publishedVersio

Extended Thromboprophylaxis with Betrixaban in Acutely Ill Medical Patients

Background Patients with acute medical illnesses are at prolonged risk for venous thrombosis. However, the appropriate duration of thromboprophylaxis remains unknown. Methods Patients who were hospitalized for acute medical illnesses were randomly assigned to receive subcutaneous enoxaparin (at a dose of 40 mg once daily) for 10±4 days plus oral betrixaban placebo for 35 to 42 days or subcutaneous enoxaparin placebo for 10±4 days plus oral betrixaban (at a dose of 80 mg once daily) for 35 to 42 days. We performed sequential analyses in three prespecified, progressively inclusive cohorts: patients with an elevated d-dimer level (cohort 1), patients with an elevated d-dimer level or an age of at least 75 years (cohort 2), and all the enrolled patients (overall population cohort). The statistical analysis plan specified that if the between-group difference in any analysis in this sequence was not significant, the other analyses would be considered exploratory. The primary efficacy outcome was a composite of asymptomatic proximal deep-vein thrombosis and symptomatic venous thromboembolism. The principal safety outcome was major bleeding. Results A total of 7513 patients underwent randomization. In cohort 1, the primary efficacy outcome occurred in 6.9% of patients receiving betrixaban and 8.5% receiving enoxaparin (relative risk in the betrixaban group, 0.81; 95% confidence interval [CI], 0.65 to 1.00; P=0.054). The rates were 5.6% and 7.1%, respectively (relative risk, 0.80; 95% CI, 0.66 to 0.98; P=0.03) in cohort 2 and 5.3% and 7.0% (relative risk, 0.76; 95% CI, 0.63 to 0.92; P=0.006) in the overall population. (The last two analyses were considered to be exploratory owing to the result in cohort 1.) In the overall population, major bleeding occurred in 0.7% of the betrixaban group and 0.6% of the enoxaparin group (relative risk, 1.19; 95% CI, 0.67 to 2.12; P=0.55). Conclusions Among acutely ill medical patients with an elevated d-dimer level, there was no significant difference between extended-duration betrixaban and a standard regimen of enoxaparin in the prespecified primary efficacy outcome. However, prespecified exploratory analyses provided evidence suggesting a benefit for betrixaban in the two larger cohorts. (Funded by Portola Pharmaceuticals; APEX ClinicalTrials.gov number, NCT01583218. opens in new tab.

Genetic and morphological differentiation between Melica ciliata L. and M. transsilvanica Schur (Poaceae) in Europe reveals the non-presence of M. ciliata in the Polish flora

A good knowledge of species delimitation is crucial for the biodiversity protection and the conservation of wild species. We studied the efficiency of AFLP markers and morphological characters to assist species determination for Melica ciliata L. and M. transsilvanica Schur within European range of distribution, including isolated and range-limit populations of "M. ciliata" (i.e. M. cf. ciliata) from the Polish Sudetes, where it is regarded as critically endangered. AFLP markers were found to be more effective then morphological characters (more or less continuous) in distinguishing the both studied species. AMOVA revealed very low genetic diversity within populations and high differentiation among populations of M. ciliata and M. transsilvanica (FST = 0.89 and 0.95, respectively). The species-diagnostic AFLP markers of M. transsilvanica shared with "M. ciliata" from the Sudetes were detected. On the other hand, no species-diagnostic genetic markers of M. ciliata or hybrid-diagnostic markers of M. × thuringiaca were found within "M. ciliata". PCoA and NJ showed an overlapping genetic diversity of "M. ciliata" and M. transsilvanica. Hierar­chical AMOVA supported the absence of a significant genotypic distinction between "M. ciliata" and M. transsilvanica. ANOVA showed that the length ratio of lower to upper glumes was the best morphological character to discriminate between M. ciliata and M. transsilvanica. Combined morphological and genetic data show that M. ciliata is not currently present in Poland as its putative Polish populations represent M. transsilvanica. A significant decrease in genetic varia­bility that could influence viability was not observed the in Sudetian populations of M. transsilvanica. However, the population size changes significantly as a result of plant succession. Correction of the northern limit of the continuous distribution of M. ciliata L. in Central Europe is presented

Distribution and ecology of Carex secalina in the Czech Republic and Slovakia

Current and historical occurrence as well as coenotic affinity and ecology of Carex secalina in the Czech Republic and Slovakia are presented. The study is based on revision of herbarium specimens from 25 herbaria, literature data excerption and a field survey carried out during 2004-2010. Altogether, 168 sites were documented in the Czech Republic divided to two separate areas. Stabile occurrence trend was found in last decades, because 45 localities were documented in period 1975-1999 (27% of sites) and recent occurrence was confirmed at 42 localities (24% of sites). On the basis of our current knowledge the species was included into the IUCN threat category VU (vulnerable). In Slovakia, 19 localities of the species were recorded in total, but only 4 sites were confirmed recently (21% of sites recorded). However, only a small number of individuals was recorded there (less than 50 individuals in total) and half of this sites is acutely threatened by destruction (ploughing, secondary succession). Therefore, the C. secalina status in the Slovak Red List was changed to the IUCN category "critically endangered" (CR). Results of the study are summarised in the maps of historical and current species distribution. C. secalina usually occurred in halophytic grassland communities, especially in association Agrostio stoloniferae-Juncetum ranarii Vicherek 1962. However, it may occur in many other communities which suggest that the species had not well-defined coenotic relationships. On the basis of our knowledge, we consider C. secalina as an obligate halophyte

Effects of alirocumab on types of myocardial infarction: insights from the ODYSSEY OUTCOMES trial

Aims  The third Universal Definition of Myocardial Infarction (MI) Task Force classified MIs into five types: Type 1, spontaneous; Type 2, related to oxygen supply/demand imbalance; Type 3, fatal without ascertainment of cardiac biomarkers; Type 4, related to percutaneous coronary intervention; and Type 5, related to coronary artery bypass surgery. Low-density lipoprotein cholesterol (LDL-C) reduction with statins and proprotein convertase subtilisin–kexin Type 9 (PCSK9) inhibitors reduces risk of MI, but less is known about effects on types of MI. ODYSSEY OUTCOMES compared the PCSK9 inhibitor alirocumab with placebo in 18 924 patients with recent acute coronary syndrome (ACS) and elevated LDL-C (≥1.8 mmol/L) despite intensive statin therapy. In a pre-specified analysis, we assessed the effects of alirocumab on types of MI. Methods and results  Median follow-up was 2.8 years. Myocardial infarction types were prospectively adjudicated and classified. Of 1860 total MIs, 1223 (65.8%) were adjudicated as Type 1, 386 (20.8%) as Type 2, and 244 (13.1%) as Type 4. Few events were Type 3 (n = 2) or Type 5 (n = 5). Alirocumab reduced first MIs [hazard ratio (HR) 0.85, 95% confidence interval (CI) 0.77–0.95; P = 0.003], with reductions in both Type 1 (HR 0.87, 95% CI 0.77–0.99; P = 0.032) and Type 2 (0.77, 0.61–0.97; P = 0.025), but not Type 4 MI. Conclusion  After ACS, alirocumab added to intensive statin therapy favourably impacted on Type 1 and 2 MIs. The data indicate for the first time that a lipid-lowering therapy can attenuate the risk of Type 2 MI. Low-density lipoprotein cholesterol reduction below levels achievable with statins is an effective preventive strategy for both MI types.For complete list of authors see http://dx.doi.org/10.1093/eurheartj/ehz299</p

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure &lt; 100 mmHg (n = 1127), estimated glomerular filtration rate &lt; 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation

Effect of alirocumab on mortality after acute coronary syndromes. An analysis of the ODYSSEY OUTCOMES randomized clinical trial

Background: Previous trials of PCSK9 (proprotein convertase subtilisin-kexin type 9) inhibitors demonstrated reductions in major adverse cardiovascular events, but not death. We assessed the effects of alirocumab on death after index acute coronary syndrome. Methods: ODYSSEY OUTCOMES (Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab) was a double-blind, randomized comparison of alirocumab or placebo in 18 924 patients who had an ACS 1 to 12 months previously and elevated atherogenic lipoproteins despite intensive statin therapy. Alirocumab dose was blindly titrated to target achieved low-density lipoprotein cholesterol (LDL-C) between 25 and 50 mg/dL. We examined the effects of treatment on all-cause death and its components, cardiovascular and noncardiovascular death, with log-rank testing. Joint semiparametric models tested associations between nonfatal cardiovascular events and cardiovascular or noncardiovascular death. Results: Median follow-up was 2.8 years. Death occurred in 334 (3.5%) and 392 (4.1%) patients, respectively, in the alirocumab and placebo groups (hazard ratio [HR], 0.85; 95% CI, 0.73 to 0.98; P=0.03, nominal P value). This resulted from nonsignificantly fewer cardiovascular (240 [2.5%] vs 271 [2.9%]; HR, 0.88; 95% CI, 0.74 to 1.05; P=0.15) and noncardiovascular (94 [1.0%] vs 121 [1.3%]; HR, 0.77; 95% CI, 0.59 to 1.01; P=0.06) deaths with alirocumab. In a prespecified analysis of 8242 patients eligible for ≥3 years follow-up, alirocumab reduced death (HR, 0.78; 95% CI, 0.65 to 0.94; P=0.01). Patients with nonfatal cardiovascular events were at increased risk for cardiovascular and noncardiovascular deaths (P<0.0001 for the associations). Alirocumab reduced total nonfatal cardiovascular events (P<0.001) and thereby may have attenuated the number of cardiovascular and noncardiovascular deaths. A post hoc analysis found that, compared to patients with lower LDL-C, patients with baseline LDL-C ≥100 mg/dL (2.59 mmol/L) had a greater absolute risk of death and a larger mortality benefit from alirocumab (HR, 0.71; 95% CI, 0.56 to 0.90; Pinteraction=0.007). In the alirocumab group, all-cause death declined wit h achieved LDL-C at 4 months of treatment, to a level of approximately 30 mg/dL (adjusted P=0.017 for linear trend). Conclusions: Alirocumab added to intensive statin therapy has the potential to reduce death after acute coronary syndrome, particularly if treatment is maintained for ≥3 years, if baseline LDL-C is ≥100 mg/dL, or if achieved LDL-C is low. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT01663402

Initial Education Of Early Years Teachers And The Profession's Needs

The thesis deals with undergraduate training of kindergarten teachers. The aim is to compare the issue of preparation for the profession of kindergarten teacher from different perspectives in historical and international comparisons and reflect and analyze the needs of the profession towards initial training kindergarten teachers on the basis of theoretical concepts and statements from practice. The reaserch aims to analyze the curriculum of initial training kindergarten teachers at selected faculties of education in the Czech Republic, analyze and reflect on meeting the needs of the profession in these faculties and learn how to evaluate their students' level of development of their professional competence at the beginning of undergraduate training and at the end. Research design is a multi-case study. Working with reaserch respondents has a longitudinal character. Methods were used quantitative and qualitative methodology - questionnaire, rating scales, content analysis of documents, interview and focus group. The thesis analyzes the curriculum of undergraduate training at Faculty of Education Charles University in Prague, Faculty of Education University of West Bohemia in Pilsen and Faculty of Education University of South Bohemia in České Budějovice in the context of the model of the..