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Abstract Background Bacteria of the candidate phyla radiation (CPR), constituting about 25% of the bacterial biodiversity, are characterized by small cell size and patchy genomes without complete key metabolic pathways, suggesting a symbiotic lifestyle. Gracilibacteria (BD1-5), which are part of the CPR branch, possess alternate coded genomes and have not yet been cultivated. The lifestyle of Gracilibacteria, their temporal dynamics, and activity in natural ecosystems, particularly in groundwater, has remained largely unexplored. Here, we aimed to investigate Gracilibacteria activity in situ and to discern their lifestyle based on expressed genes, using the metaproteogenome of Gracilibacteria as a function of time in the cold-water geyser Wallender Born in the Volcanic Eifel region in Germany. Results We coupled genome-resolved metagenomics and metaproteomics to investigate a cold-water geyser microbial community enriched in Gracilibacteria across a 12-day time-series. Groundwater was collected and sequentially filtered to fraction CPR and other bacteria. Based on 725 Gbps of metagenomic data, 1129 different ribosomal protein S3 marker genes, and 751 high-quality genomes (123 population genomes after dereplication), we identified dominant bacteria belonging to Gallionellales and Gracilibacteria along with keystone microbes, which were low in genomic abundance but substantially contributing to proteomic abundance. Seven high-quality Gracilibacteria genomes showed typical limitations, such as limited amino acid or nucleotide synthesis, in their central metabolism but no co-occurrence with potential hosts. The genomes of these Gracilibacteria were encoded for a high number of proteins involved in cell to cell interaction, supporting the previously surmised host-dependent lifestyle, e.g., type IV and type II secretion system subunits, transporters, and features related to cell motility, which were also detected on protein level. Conclusions We here identified microbial keystone taxa in a high-CO2 aquifer, and revealed microbial dynamics of Gracilibacteria. Although Gracilibacteria in this ecosystem did not appear to target specific organisms in this ecosystem due to lack of co-occurrence despite enrichment on 0.2-µm filter fraction, we provide proteomic evidence for the complex machinery behind the host-dependent lifestyle of groundwater Gracilibacteria. Video Abstract

CRISPR-Cas systems defend prokaryotic cells from invasive DNA of viruses, plasmids and other mobile genetic elements. Here, we show using metagenomics, metatranscriptomics and single-cell genomics that CRISPR systems of widespread, uncultivated archaea can also target chromosomal DNA of archaeal episymbionts of the DPANN superphylum. Using meta-omics datasets from Crystal Geyser and Horonobe Underground Research Laboratory, we find that CRISPR spacers of the hosts Candidatus Altiarchaeum crystalense and Ca. A. horonobense, respectively, match putative essential genes in their episymbionts' genomes of the genus Ca. Huberiarchaeum and that some of these spacers are expressed in situ. Metabolic interaction modelling also reveals complementation between host-episymbiont systems, on the basis of which we propose that episymbionts are either parasitic or mutualistic depending on the genotype of the host. By expanding our analysis to 7,012 archaeal genomes, we suggest that CRISPR-Cas targeting of genomes associated with symbiotic archaea evolved independently in various archaeal lineages.

Background This study aimed to compare the stage of coronary heart disease in patients who underwent invasive cardiac diagnostics during the COVID-19 pandemic and before, based on the number of medical devices used and the number of complex coronary angioplasty procedures performed. Material and Methods A retrospective, single-center study was conducted, which included 187 successive patients with diagnosed coronary heart disease, who were divided into 2 groups: group I (N = 92, pre-COVID-19 pandemic) and group II (N = 95, during COVID-19 pandemic). Results Despite a comparable number of invasive procedures in both groups, stent length and contrast use per procedure were significantly higher during the COVID-19 pandemic. Similarly, a higher number of stents was used per patient in 2021, however, the difference was not statistically significant (p = 0.0817). Similarly, fluoroscopy time per procedure and procedure duration were significantly longer in the 2021 group. Among patients treated during the COVID-19 pandemic, higher glucose concentration, blood pressure parameters, low-density lipoprotein and total cholesterol were observed; however, these differences were not statistically significant. Conclusions During the COVID-19 pandemic, coronary atherosclerosis progression were found, requiring a higher number of complex coronary angioplasty procedures, which contributed to a statistically significant increase in the number of medical devices used (angioplasty guidewires, angioplasty balloons) and procedures duration. Med Pr Work Health Saf. 2024;75(4):333–342

Abstract Background The transformation of a pharmacist’s role from that of a drug dispenser to an advisor and patient educator, partially accelerated by the COVID-19 pandemic, requires a thorough change in the pharmacy curriculum. Preparation for the provision of modern pharmaceutical services requires the use of the most advanced teaching methods, such as pharmaceutical simulation. Knowledge alone does not guarantee students’ readiness and motivation to take on new challenges in their professional work, but it seems crucial that graduates of medical faculties have the ability to practically apply their knowledge, including in new and nonstandard situations. Therefore, in our study, we proposed an intervention using a simulation method (peer role play) in teaching pharmaceutical care, and we assessed its impact on students’ levels of self-perceived confidence and self-efficacy in accordance with Bandura’s theory. The aim of the study was to verify whether the introduction of these types of classes could be a useful element of pharmacy curriculum renewal. Methods The questionnaire-based study was conducted during pharmaceutical care peer role-playing classes in a simulation environment with a debriefing session among 85 final-year pharmacy students at Poznan University of Medical Sciences, Poland. The questionnaire consisted of two surveys: the General Self-Efficacy Scale (GSE) and a pre-post self-assessment and self-efficacy questionnaire. Results There was a positive correlation between the GSE score and self-efficacy (R = 0.52, p

Background: In many European countries, flu vaccination coverage rates are below the 75% target. During the COVID-19 pandemic, many pharmacists around Europe were involved as vaccine administrators and demonstrated positive results in improving vaccine uptake. This paper explores the challenges, accomplishments, and best practices of various European pharmacists' associations in administering vaccines and positively contributing to public health. Methods: Eight pharmacists representing various associations from different countries across Europe (Italy, Belgium, Poland, Portugal, France, and Germany) convened to discuss their role as vaccination providers, the advantages, and strategies for improvement, and to identify barriers and gaps in the vaccination administration process, especially focusing on the administration of seasonal flu vaccines. Results: Currently, 15 European countries allow community pharmacists to dispense and administer flu vaccines. Among the ones that attended the meeting, Portugal initiated the flu immunization program at the pharmacy earliest, before the COVID era, but in other countries, the process started only in the last couple of years. Initial hesitancy and reluctance by other HCPs or institutions were overcome as the pilot projects showed positive and cost-effective public health results. Today, pharmacists are considered crucial professional figures to provide immunization services against COVID-19, the flu, and other vaccine-preventable diseases, and pursue important public health goals.Key takeaways to enhance the pharmacist's role in providing immunization services against vaccine-preventable diseases include improving interaction with policymakers and the public, generating real-world evidence highlighting public health benefits, and ensuring ongoing professional education and training for pharmacists. Conclusion: Vaccinating pharmacists are gaining recognition of their role and the benefits derived from their broader involvement in the healthcare system, including immunization programs. Further efforts are needed in each country for an adequate recognition of the profession and a broader utilization of pharmacy services to exploit the benefit of immunization, especially against the flu.

Abstract Background Patients with asthma and chronic obstructive pulmonary disease could benefit from education on using inhalers provided by pharmacists. However, pharmacists may have limited competencies, indicating the necessity to implement appropriate postgraduate courses. The study aimed to evaluate an inhaler use course for pharmacists, including its impact on participants’ knowledge and satisfaction. Methods The study involved 261 pharmacists from community pharmacies and was conducted between September 2019 and March 2021. A pre-post analysis of their knowledge of the topic was applied. Additionally, at the beginning of the course, participants were asked about their educational needs, and at the end, they completed a satisfaction survey. The preferred learning formats indicated by participants were interactive workshops and lectures. Results As a result of the course, both their actual and self-assessed level of knowledge significantly increased. The percentage of correct answers in the test before the training was 24.4%, while after, it was 84.3% (p

Abstract Marine viruses in seawater have frequently been studied, yet their dispersal from neuston ecosystems at the air-sea interface towards the atmosphere remains a knowledge gap. Here, we show that 6.2% of the studied virus population were shared between air-sea interface ecosystems and rainwater. Virus enrichment in the 1-mm thin surface microlayer and sea foams happened selectively, and variant analysis proved virus transfer to aerosols collected at ~2 m height above sea level and rain. Viruses detected in rain and these aerosols showed a significantly higher percent G/C base content compared to marine viruses. CRISPR spacer matches of marine prokaryotes to foreign viruses from rainwater prove regular virus-host encounters at the air-sea interface. Our findings on aerosolization, adaptations, and dispersal support transmission of viruses along the natural water cycle.

Abstract Recent advances in immuno-oncology have opened up new and impressive treatment options for cancer. Notwithstanding, overcoming the limitations of the current FDA-approved therapies with monoclonal antibodies (mAbs) that block the PD-1/PD-L1 pathway continues to lead to the testing of multiple approaches and optimizations. Recently, a series of macrocyclic peptides have been developed that exhibit binding strengths to PD-L1 ranging from sub-micromolar to micromolar. In this study, we present the most potent non-antibody-based PD-1/PD-L1 interaction inhibitor reported to date. The structural and biological characterization of this macrocyclic PD-L1 targeting peptide provides the rationale for inhibition of both PD-1/PD-L1 and CD80/PD-L1 complexes. The IC50 and EC50 values obtained in PD-L1 binding assays indicate that the pAC65 peptide has potency equivalent to the current FDA-approved mAbs and may have similar activity to the BMS986189 peptide, which entered the clinical trial and has favorable safety and pharmacokinetic data. The data presented here delineate the generation of similar peptides with improved biological activities and applications not only in the field of cancer immunotherapy but also in other disorders related to the immune system.

Abstract FUS is a multifunctional protein involved in many aspects of RNA metabolism, including transcription, splicing, translation, miRNA processing, and replication-dependent histone gene expression. In this work, we show that FUS depletion results in the differential expression of numerous small nucleolar RNAs (snoRNAs) that guide 2’-O methylation (2’-O-Me) and pseudouridylation of specific positions in ribosomal RNAs (rRNAs) and small nuclear RNAs (snRNAs). Using RiboMeth-seq and HydraPsiSeq for the profiling of 2’-O-Me and pseudouridylation status of rRNA species, we demonstrated considerable hypermodification at several sites in HEK293T and SH-SY5Y cells with FUS knockout (FUS KO) compared to wild-type cells. We observed a similar direction of changes in rRNA modification in differentiated SH-SY5Y cells with the FUS mutation (R495X) related to the severe disease phenotype of amyotrophic lateral sclerosis (ALS). Furthermore, the pattern of modification of some rRNA positions was correlated with the abundance of corresponding guide snoRNAs in FUS KO and FUS R495X cells. Our findings reveal a new role for FUS in modulating the modification pattern of rRNA molecules, that in turn might generate ribosome heterogeneity and constitute a fine-tuning mechanism for translation efficiency/fidelity. Therefore, we suggest that increased levels of 2’-O-Me and pseudouridylation at particular positions in rRNAs from cells with the ALS-linked FUS mutation may represent a possible new translation-related mechanism that underlies disease development and progression.

Comparative analyses of fleshy red algae mats and seagrass meadows highlight their value in fostering sessile invertebrate biodiversity.

Abstract Microbial communities in freshwater streams play an essential role in ecosystem functioning via biogeochemical cycling. Yet, the impacts of treated wastewater influx into stream ecosystems on microbial strain diversity remain mostly unexplored. Here, we coupled full‐length 16S ribosomal RNA gene Nanopore sequencing and strain‐resolved metagenomics to investigate the impact of treated wastewater on a mesocosm system (AquaFlow) run with restored river water. Over 10 days, community Bray–Curtis dissimilarities between treated and control mesocosm decreased (0.57 ± 0.058 to 0.26 ± 0.046) based on ribosomal protein S3 gene clustering, finally converging to nearly identical communities. Similarly, strain‐resolved metagenomics revealed a high diversity of bacteria and viruses after the introduction of treated wastewater; these microbes also decreased over time resulting in the same strain clusters in control and treatment at the end of the experiment. Specifically, 39.2% of viral strains detected in all samples were present after the introduction of treated wastewater only. Although bacteria present at low abundance in the treated wastewater introduced additional antibiotic resistance genes, signals of naturally occurring ARG‐encoding organisms resembled the resistome at the endpoint. Our results suggest that the previously stressed freshwater stream and its microbial community are resilient to a substantial introduction of treated wastewater.

Spondyloarthropathies (SpA) are a family of rheumatic disorders that could be divided into axial (axSpA) and peripheral (perSpA) sub-forms depending on the disease clinical presentation. The chronic inflammation is believed to be driven by innate immune cells such as monocytes, rather than self-reactive cells of adaptive immune system. The aim of the study was to investigate the micro-RNA (miRNA) profiles in monocyte subpopulations (classical, intermediate and non-classical subpopulations) acquired from SpA patients or healthy individuals in search for prospective disease specific and/or disease subtype differentiating miRNA markers. Several SpA-specific and axSpA/perSpA differentiating miRNAs have been identified that appear to be characteristic for specific monocyte subpopulation. For classical monocytes, upregulation of miR-567 and miR-943 was found to be SpA-specific, whereas downregulation of miR-1262 could serve as axSpA-differentiating, and the expression pattern of miR-23a, miR-34c, mi-591 and miR-630 as perSpA-differentiating markers. For intermediate monocytes, expression levels of miR-103, miR-125b, miR-140, miR-374, miR-376c and miR-1249 could be used to distinguish SpA patients from healthy donors, whereas the expression pattern of miR-155 was identified as characteristic for perSpA. For non-classical monocytes, differential expression of miR-195 was recognized as general SpA indicator, while upregulation of miR-454 and miR-487b could serve as axSpA-differentiating, and miR-1291 as perSpA-differentiating markers. Our data indicate for the first time that in different SpA subtypes, monocyte subpopulations bear disease-specific miRNA signatures that could be relevant for SpA diagnosis/differentiation process and may help to understand SpA etiopathology in the context of already known functions of monocyte subpopulations.

Background: Bacteria of the candidate phyla radiation (CPR), constituting about 25% of the bacterial biodiversity, are characterized by small cell size and patchy genomes without complete key metabolic pathways, suggesting a symbiotic lifestyle. Gracilibacteria (BD1-5), which are part of the CPR branch, possess alternate coded genomes and have not yet been cultivated. The lifestyle of Gracilibacteria, their temporal dynamics, and activity in natural ecosystems, particularly in groundwater, has remained largely unexplored. Here, we aimed to investigate Gracilibacteria activity in situ and to discern their lifestyle based on expressed genes, using the metaproteogenome of Gracilibacteria as a function of time in the cold-water geyser Wallender Born in the Volcanic Eifel region in Germany. Results: We coupled genome-resolved metagenomics and metaproteomics to investigate a cold-water geyser microbial community enriched in Gracilibacteria across a 12-day time-series. Groundwater was collected and sequentially filtered to fraction CPR and other bacteria. Based on 725 Gbps of metagenomic data, 1129 different ribosomal protein S3 marker genes, and 751 high-quality genomes (123 population genomes after dereplication), we identified dominant bacteria belonging to Gallionellales and Gracilibacteria along with keystone microbes, which were low in genomic abundance but substantially contributing to proteomic abundance. Seven high-quality Gracilibacteria genomes showed typical limitations, such as limited amino acid or nucleotide synthesis, in their central metabolism but no co-occurrence with potential hosts. The genomes of these Gracilibacteria were encoded for a high number of proteins involved in cell to cell interaction, supporting the previously surmised host-dependent lifestyle, e.g., type IV and type II secretion system subunits, transporters, and features related to cell motility, which were also detected on protein level. Conclusions: We here identified microbial keystone taxa in a high-CO2 aquifer, and revealed microbial dynamics of Gracilibacteria. Although Gracilibacteria in this ecosystem did not appear to target specific organisms in this ecosystem due to lack of co-occurrence despite enrichment on 0.2-µm filter fraction, we provide proteomic evidence for the complex machinery behind the host-dependent lifestyle of groundwater Gracilibacteria. 38kgAxaMce3Yn9ghud_S2p Video Abstract [ABSTRACT FROM AUTHOR]

Background: The pharmacists in a community pharmacies have already provided, or will provide in the near future advanced pharmaceutical care services. This requires modifying the approach to teaching pharmacy students as well as adapting the curriculum to the changing professional realities. It has been proven that in the field of medical and related sciences, learners-centered active teaching methods allow to achieve learning outcomes effectively, especially in the field of practical skills. Objectives: As the pharmaceutical services are only being introduced in many European countries, the question arises as to what active learning methods to use to prepare pharmacy graduates for this. Thus the review of worldwide literature occurred to be helpful in identifying what active learning methods are being used specifically in teaching aspects of pharmaceutical care. Methods: Three electronic databases: Pubmed, Scopus and Web of Science were searched using the keywords “active learning” and “pharmaceutical care”. Results: On the basis of the publications included in the review, 7 methods were distinguished. Case-study, role play and simulation exercises turned out to be the most popular. It was also possible to make preliminary conclusions on how to properly match the method to the learning outcomes. Moreover, a weak point of many studies was the lack of structured methods of assessing the skills acquired by the students. Conclusions: In conclusion, the curriculum renewal in pharmacy is necessary and requires taking many aspects into account, from the types of tasks assigned to pharmacists, through the selection of appropriate teaching methods, to the verification of assessment methods.

Romero et al. perform NMR-based screening of 1500 fragments to identify fragments that bind at the oligomeric interface of malate dehydrogenase (MDH). Their study indicates an allosteric mechanism impacting enzymatic activity, paving the way for development of more selective molecules and a starting point for the future development of specific MDH inhibitors.

Objectives Cardiovascular and metabolic disorders constitute major health problems in the working populations in Europe. The aim of this project was to evaluate the health condition of workers employed in a Polish research and medical institution, and then to establish the necessary preventive actions by creating a modern model of occupational healthcare integrated with civilization disease (CivD) prevention. Material and Methods Overall, 100 workers voluntarily participated in a health program for CivD prevention during mandatory prophylactic examinations. Data from these examinations was collected in a system of electronic documentation to enable the analysis of the workers’ health condition and risk factors of CivDs. Results Women accounted for 72% of the employees who voluntarily participated in the prevention program, and 80% of the subjects had university education. As regards the health condition, 27% of the patients had elevated systolic, and 23% diastolic, blood pressure, and 21% had an abnormal fasting glucose level. Stressful job was an important factor correlated with an excessive body mass index, diastolic blood pressure, as well as total and low-density lipoprotein cholesterol levels. Smoking, sedentary work and a lack of physical activity were significant factors for abdominal obesity. Generally, 94% of the program participants required some further interventions in lifestyle, diagnostics or treatment. Conclusions Civilization disease prevention should focus on increasing physical activity both in leisure time and at the workplace as far as practicable. There is a need for implementing projects leading to occupational stress reduction and smoking cessation. Men as well as workers with vocational and elementary education need to be recruited for prevention programs dedicated to employees. Int J Occup Med Environ Health. 2021;34(3):403–13

The upside-down jellyfish Cassiopea increasingly occurs in many (sub-) tropical coastal habitats such as mangrove forests, seagrass meadows, and coral reefs. Its mixotrophic lifestyle and ecophysiological plasticity as well as a high regenerative capacity may be reasons for its success. While the regeneration of umbrella tissue and body structures (i.e. rhopalia and oral arms) was already demonstrated, it remains unclear whether a fully functioning medusa can regenerate from only umbrella tissue. In this study, we thus investigated the regeneration of umbrella fragments over time. We conducted a laboratory experiment for which we used 18 Cassiopea medusae of three different size classes that were cut into two pieces each, one fragment with oral arms and one without. Over a total observation period of 5 weeks, we regularly monitored survival, pulsation behavior, growth and the regeneration pattern of fragments. Findings revealed that 100% of the fragments with oral arms and 88% of the fragments without oral arms survived. Pulsation behavior occurred in all fragments and lasted until the end of the experiment in 94% of all fragments. The umbrella area of fragments without oral arms showed a significantly higher decrease in the first two weeks compared to fragments with oral arms. A complete regeneration of umbrella tissue was observed in all fragments, with and without oral arms alike, and 50% of all fragments even regenerated rhopalia or oral arms as body structures after 33 days. These results suggest an outstanding regenerative capacity of Cassiopea jellyfish after fragmentation. This may contribute to (i) explain the currently observed success of upside-down jellyfish and (ii) extend our knowledge about its regeneration process, which might even act as an asexual reproduction mode in Cassiopea.

In this study, we report a 4-month-old boy with T−B+NK− severe combined immunodeficiency (SCID) due to a novel mutation in exon 2 of IL2RG, the gene encoding the interleukin (IL) common gamma chain (γc) of the cytokine receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. The patient was born from a twin pregnancy. He manifested recurrent infections of the gastrointestinal tract, whereas his twin brother was asymptomatic with no immune defects. In order to evaluate the effect of this unreported variant on the protein structure, a structural modeling process was performed showing prominent biochemical alterations of the protein features, including molecular weight, isoelectric charge, and possible changes to its secondary and tertiary structure.