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Smoking, a risk factor for periodontitis and peri-implantitis, is associated with shifts in the oral microbiome (OM) composition. Although smoking habits are almost always established before adulthood, data on effects of smoking on the OM in adolescents is rare. The aim of this study was to investigate the early impact of smoking on the OM composition in pupils. The adolescent cohort, aged 14–20, comprised 98 smokers and 98 non-smokers matched for several physiological co-variates. Buccal swabs were analysed for OM composition using high-throughput sequencing of the full-length 16 S rRNA gene targeting species-level resolution. Parameters of bacterial diversity and abundance of individual bacterial taxa were related to information on smoking. The microbiome dataset contained 733 species-level taxa. Streptococcus , Rothia , and Haemophilus dominated both groups, smokers and non-smokers. Smoking exerted a discernible influence on the overall microbial composition as measured by weighted UniFrac distances. The number of species-level bacterial taxa was significantly higher in individual smokers compared to non-smokers. Furthermore, several taxa, including known pathogens, exhibited significant differences in abundance between the two groups. The genera Veillonella , and Actinomyces , as well as and multiple Actinomyces species, Dialister invisus , Atopobium parvulum , Streptococcus mutans and Prevotella melaninogenica were significantly more abundant in smokers. Our findings indicated an early onset of smoking-related changes already in the oral microbiome of adolescents.

Endoprosthesis loosening is the major cause of arthroplasty failure. Currently, radiography, histo-pathological classification of the periprosthetic membrane, and microbiological examination are used retrospectively for diagnosis. Prospective options for early diagnosis are not available. The study presented here aimed to identify (prospective) molecular biomarkers of implant loosening in tissue samples from patients. Four patient cohorts (primary and revision arthroplasty due to aseptic loosening of hip or knee) were defined. Aseptic loosening of implants was assessed by the standardised approach of the Knee Society Total Knee Arthroplasty Roentgenographic Evaluation and Scoring System. Synovial fluid, bone marrow, and blood were collected from 96 patients. Mesenchymal stromal cells (BM-MSCs) were isolated from the bone marrow. Bulk RNA-sequencing of 28 samples (including a fifth patient cohort with knee arthrofibrosis as aseptic cause for revision surgery) showed that Glycoprotein Non-Metastatic Melanoma Protein B mRNA ( GPNMB ) was significantly upregulated in BM-MSCs derived from revision patients compared to patients with primary implantations. Elevated GPNMB mRNA levels were confirmed with qRT-PCR. Synovial fluid plasma study by ELISA revealed increased levels of GPNMB protein in patients undergoing revision surgery compared to patients undergoing primary arthroplasty. GPNMB concentration in synovial fluid plasma, which can be obtained non-invasively, could be a potential biomarker for early detection of implant loosening, ahead of current diagnostic procedures.

This is the first study to quantify the dependence on wind velocity of airborne bacterial emission fluxes from soil. It demonstrates that manure bacteria get aerosolized from fertilized soil more easily than soil bacteria, and it applies bacterial genomic sequencing for the first time to trace environmental fecal contamination back to its source in the chicken barn. Summary This is the first study to quantify the dependence on wind velocity of airborne bacterial emission fluxes from soil. It demonstrates that manure bacteria get aerosolized from fertilized soil more easily than soil bacteria, and it applies bacterial genomic sequencing for the first time to trace environmental faecal contamination back to its source in the chicken barn. We report quantitative, airborne emission fluxes of bacteria during and following the fertilization of agricultural soil with manure from broiler chickens. During the fertilization process, the concentration of airborne bacteria culturable on blood agar medium increased more than 600 000‐fold, and 1 m3 of air carried 2.9 × 105 viable enterococci, i.e. indicators of faecal contamination which had been undetectable in background air samples. Trajectory modelling suggested that atmospheric residence times and dispersion pathways were dependent on the time of day at which fertilization was performed. Measurements in a wind tunnel indicated that airborne bacterial emission fluxes from freshly fertilized soil under local climatic conditions on average were 100‐fold higher than a previous estimate of average emissions from land. Faecal bacteria collected from soil and dust up to seven weeks after fertilization could be traced to their origins in the poultry barn by genomic sequencing. Comparative analyses of 16S rRNA gene sequences from manure, soil and dust showed that manure bacteria got aerosolized preferably, likely due to their attachment to low‐density manure particles. Our data show that fertilization with manure may cause substantial increases of bacterial emissions from agricultural land. After mechanical incorporation of manure into soil, however, the associated risk of airborne infection is low.

Peri-implantitis is a severe biofilm-associated infection affecting millions worldwide. This cross-sectional study aimed to identify taxonomic and functional biomarkers that reliably indicate peri-implantitis by utilizing paired data from full length 16S rRNA gene amplicon sequencing (full-16S) and metatranscriptomics (RNAseq) in 48 biofilm samples from 32 patients. Both full-16S and RNAseq analyses revealed significant differences between healthy and peri-implantitis samples, with a shift toward anaerobic Gram-negative bacteria in peri-implantitis. Metatranscriptomics identified enzymatic activities and metabolic pathways associated with peri-implantitis and uncovered complex peri-implant biofilm ecology related to amino acid metabolism. Integrating taxonomic and functional data enhanced predictive accuracy (AUC = 0.85) and revealed diagnostic biomarkers including health-associated Streptococcus and Rothia species and peri-implantitis-associated enzymes (urocanate hydratase, tripeptide aminopeptidase, NADH:ubiquinone reductase, phosphoenolpyruvate carboxykinase and polyribonucleotide nucleotidyltransferase). Thus, biofilm profiling at taxonomic and functional levels provides highly predictive disease biomarkers, laying the foundation for novel diagnostic and personalized treatment approaches for peri-implant disease.

Changes in bacterial species composition within oral biofilms, known as biofilm dysbiosis, are associated with the development of severe oral diseases. To better understand this process and help establish early detection systems, models are needed which replicate oral biofilm dysbiosis - ideally by also mimicking natural salivary flow conditions. For this purpose, the present study cultivated two different combinations of oral commensal and pathogenic strains - and - comparatively within an established flow chamber model on the implant material titanium, and statically in 6-well plates for 21 days. Biofilm morphology, species distribution, and bacterial metabolism were analyzed by fluorescence microscopy, molecular biological methods, and metabolic interaction prediction. Biofilm growth and composition were strongly influenced by bacterial species selection, and to a more minor extent, by cultivation conditions. Within the model containing and a laboratory strain, a diversification of commensal species was observed over time along with a significantly reduced pH-value. In contrast, the model containing and the clinical isolate W83, a dysbiotic shift with increased pathogen levels, pH-value, and virulence factors was achieved. Within the present study, different oral multispecies biofilm models were successfully developed. Depending on bacterial species selection, these models were able to depict the infection-associated dysbiotic shift in species composition under flow conditions solely by intrinsic interactions and without the use of external stimuli.

The Helicobacter pylori energy sensor TlpD determines tactic behaviour under low energy conditions and is important in vivo . We explored protein-protein interactions of TlpD and their impact on TlpD localisation and function. Pull-down of tagged TlpD identified protein interaction partners of TlpD, which included the chemotaxis histidine kinase CheAY2, the central metabolic enzyme aconitase (AcnB) and the detoxifying enzyme catalase (KatA). We confirmed that KatA and AcnB physically interact with TlpD. While the TlpD-dependent behavioural response appeared not influenced in the interactor mutants katA and acnB in steady-state behavioural assays, acetone carboxylase subunit ( acxC ) mutant behaviour was altered. TlpD was localised in a bipolar subcellular pattern in media of high energy. We observed a significant change in TlpD localisation towards the cell body in cheAY2 -, catalase- or aconitase-deficient bacteria or in bacteria incubated under low energy conditions, including oxidative stress or respiratory inhibition. Inactivation of tlpD resulted in an increased sensitivity to iron limitation and oxidative stress and influenced the H. pylori transcriptome. Oxidative stress, iron limitation and overexpressing the iron-sulfur repair system nifSU altered TlpD-dependent behaviour. We propose that TlpD localisation is instructed by metabolic activity and protein interactions, and its sensory activity is linked to iron-sulfur cluster integrity.

Demographic change and demographic development meet digitalization, which reveals the significant need for revolutionary measures regarding the handling of demographic changes in the workforce. In addition to the rising integration of immigrants into the education and employment system and the skills shortage, the identification of a new course of action is one of the most profound challenges of demographic change (Jeschke et al, 2013). Since the implementation of such revolutionary actions is tightly linked to technological advances whose introduction cannot always be predicted, it is necessary to concurrently refine and improve existing approaches. In order to improve existing measures and define future models, a demography management evaluation focusing a holistic lifecycle management has been developed and tested in the automotive industry. Therefore, a concept has been developed for evaluating the current demographic situation in a company, which can, in the long term, help in reducing and counteracting the negative consequences of demographic change. The potential outcomes of demographic change are manifold, so countermeasures have to include different fields of action regarding personnel commitment, staff recruitment, personnel development and personnel activation like health management (Bauer, Cernavin, 2015). The concept is based on three elements: (1) evaluation of the current demographic situation, (2) definition of strategic goals and (3) derivation of suitable measures for action. The concept consists of workshops and semi-standardized interviews with leaders and employees as well as an audit of assessment measures with the human resource department. Through choosing the participants out of the principle of maximum contrast - from leaders of different hierarchies, members of the works council to employees with different educational and work background - different perspectives are taken into consideration. This analysis results in a rating, which has been validated by developing and assessing semi-standardized interviews. The development of this concept, its prototypical application and resulting methodological adjustments are discussed in this paper. Concluding, an outlook on the large-scale role out is given.

Summary During a field experiment applying broiler manure for fertilization of agricultural land, we detected viable Clostridioides (formerly, Clostridium) difficile in broiler feces, manure, dust, and fertilized soil. A large diversity of toxigenic C. difficile isolates was recovered, including PCR ribotypes common from human disease. Genomic relatedness of C. difficile isolates from dust and from soil, recovered more than two years after fertilization, traced their origins to the specific chicken farm that had delivered the manure. We present evidence of long-term contamination of agricultural soil with manure-derived C. difficile and demonstrate the potential for airborne dispersal of C. difficile through dust emissions during manure application. Clostridioides genome sequences virtually identical to those from manure had been recovered from chicken meat and from human infections in previous studies, suggesting broiler-associated C. difficile are capable of zoonotic transmission. Competing Interest Statement The authors have declared no competing interest.

Background The gastric microbiome and inflammation play a key role in gastric cancer (GC) by regulating the immune response in a complex manner and by inflammatory events supporting carcinogenesis. Meprin β is a zinc endopeptidase and participates in tissue homeostasis, intestinal barrier function and immunological processes. It influences local inflammatory processes, dysbiosis and the microbiome. Here, we tested the hypothesis that meprin β is expressed in GC and of tumor biological significance. Patients and methods Four hundred forty whole mount tissue sections of patients with therapy-naive GC were stained with an anti-meprin β antibody. The histoscore and staining pattern were analyzed for each case. Following dichotomization at the median histoscore into a “low” and “high” group, the expression was correlated with numerous clinicopathological patient characteristics. Results Meprin β was found intracellularly and at the cell membrane of GC. Cytoplasmic expression correlated with the phenotype according to Lauren, microsatellite instability and PD-L1 status. Membranous expression correlated with intestinal phenotype, mucin-1-, E-cadherin-, β-catenin status, mucin typus, microsatellite instability, KRAS mutation and PD-L1-positivity. Patients with cytoplasmic expression of meprin β showed a better overall and tumor-specific survival. Conclusions Meprin β is differentially expressed in GC and has potential tumor biological relevance. It might function as a tumor suppressor or promotor depending on histoanatomical site and context.

Aflatoxin B 1 (AFB 1 ) is a highly hepatotoxic and carcinogenic mycotoxin produced by Aspergillus species. The compound is mainly metabolized in the liver and its metabolism varies between species. The present study quantified relevant AFB 1 - metabolites formed by mouse, rat, and human primary hepatocytes after treatment with 1 µM and 10 µM AFB 1 . The use of liquid chromatographic separation coupled with tandem mass spectrometric detection enabled the selective and sensitive determination of phase I and phase II metabolites of AFB 1 over incubation times of up to 24 h. The binding of AFB 1 to macromolecules was also considered. The fastest metabolism of AFB 1 was observed in mouse hepatocytes which formed aflatoxin P 1 as a major metabolite and also its glucuronidated form, while AFP 1 occurred only in traces in the other species. Aflatoxin M 1 was formed in all species and was, together with aflatoxin Q 1 and aflatoxicol, the main metabolite in human cells. Effective epoxidation led to high amounts of DNA adducts already 30 min post-treatment, especially in rat hepatocytes. Lower levels of DNA adducts and fast DNA repair were found in mouse hepatocytes. Also, protein adducts arising from reactive intermediates were formed rapidly in all three species. Detoxification via glutathione conjugation and subsequent formation of the N -acetylcysteine derivative appeared to be similar in mice and in rats and strongly differed from human hepatocytes which did not form these metabolites at all. The use of qualitative reference material of a multitude of metabolites and the comparison of hepatocyte metabolism in three species using advanced methods enabled considerations on toxification and detoxification mechanisms of AFB 1 . In addition to glutathione conjugation, phase I metabolism is strongly involved in the detoxification of AFB 1 .