Corynebacterium diphtheriae invasion-associated protein (DIP1281) is involved in cell surface organization, adhesion and internalization in epithelial cells

<p>Abstract</p> <p>Background</p> <p><it>Corynebacterium diphtheriae</it>, the causative agent of diphtheria, is well-investigated in respect to toxin production, while little is known about <it>C. diphtheriae </it>factors crucial for colonization of the host. In this study, we investigated the function of surface-associated protein DIP1281, previously annotated as hypothetical invasion-associated protein.</p> <p>Results</p> <p>Microscopic inspection of DIP1281 mutant strains revealed an increased size of the single cells in combination with an altered less club-like shape and formation of chains of cells rather than the typical V-like division forms or palisades of growing <it>C. diphtheriae </it>cells. Cell viability was not impaired. Immuno-fluorescence microscopy, SDS-PAGE and 2-D PAGE of surface proteins revealed clear differences of wild-type and mutant protein patterns, which were verified by atomic force microscopy. DIP1281 mutant cells were not only altered in shape and surface structure but completely lack the ability to adhere to host cells and consequently invade these.</p> <p>Conclusions</p> <p>Our data indicate that DIP1281 is predominantly involved in the organization of the outer surface protein layer rather than in the separation of the peptidoglycan cell wall of dividing bacteria. The adhesion- and invasion-negative phenotype of corresponding mutant strains is an effect of rearrangements of the outer surface.</p

Application of next-generation sequencing to Enterobacter hormaechei subspecies analysis during a neonatal intensive care unit outbreak

Introduction: The Enterobacter cloacae complex (ECC) species are potential neonatal pathogens, and ECC strains are among the most commonly encountered Enterobacter spp. associated with nosocomial bloodstream infections. Outbreaks caused by ECC can lead to significant morbidity and mortality in susceptible neonates. At the molecular level, ECC exhibits genomic heterogeneity, with six closely related species and subspecies. Genetic variability poses a challenge in accurately identifying outbreaks by determining the clonality of ECC isolates. This difficulty is further compounded by the limitations of the commonly used molecular typing methods, such as pulsed field gel electrophoresis, which do not provide reliable accuracy in distinguishing between ECC strains and can lead to incorrect conclusions. Next-generation sequencing (NGS) offers superior resolution in determining strain relatedness. Therefore, we investigated the clinical pertinence of incorporating NGS into existing bundle measures to enhance patient management during an outbreak of ECC in a level-3 neonatal intensive care unit (NICU) in Germany. Methods: As the standard of care, all neonates on the NICU received weekly microbiological swabs (nasopharyngeal and rectal) and analysis of endotracheal secretion, where feasible. During the 2.5-month outbreak, colonisation with ECC was detected in n = 10 neonates. The phylogenetic relationship and potential antimicrobial resistance genes as well as mobile genetic elements were identified via bacterial whole-genome sequencing (WGS) using Illumina MiSeq followed by in silico data analysis. Results: Although all ECC isolates exhibited almost identical antimicrobial susceptibility patterns, the WGS data revealed the involvement of four different ECC clones. The isolates could be characterised as Enterobacter hormaechei subspecies steigerwaltii (n = 6, clonal), subsp. hoffmannii (n = 3, two clones) and subsp. oharae (n = 1). Despite the collection of environmental samples, no source of this diffuse outbreak could be identified. A new standardised operating procedure was implemented to enhance the management of neonates colonised with MRGN. This collaborative approach involved both parents and medical professionals and successfully prevented further transmission of ECC. Conclusions: Initially, it was believed that the NICU outbreak was caused by a single ECC clone due to the similarity in antibiotic resistance. However, our findings show that antibiotic susceptibility patterns can be misleading in investigating outbreaks of multi-drug-resistant ECC. In contrast, bacterial WGS accurately identified ECC at the clonal level, which significantly helped to delineate the nature of the observed outbreak

Increased zinc levels facilitate phenotypic detection of ceftazidime-avibactam resistance in metallo-β-lactamase-producing Gram-negative bacteria.

Ceftazidime-avibactam is one of the last resort antimicrobial agents for the treatment of carbapenem-resistant, Gram-negative bacteria. Metallo-β-lactamase-producing bacteria are considered to be ceftazidime-avibactam resistant. Here, we evaluated a semi-automated antimicrobial susceptibility testing system regarding its capability to detect phenotypic ceftazidime-avibactam resistance in 176 carbapenem-resistant, metallo-β-lactamase-producing Enterobacterales and isolates. Nine clinical isolates displayed ceftazidime-avibactam susceptibility in the semi-automated system and six of these isolates were susceptible by broth microdilution, too. In all nine isolates, metallo-β-lactamase-mediated hydrolytic activity was demonstrated with the EDTA-modified carbapenemase inactivation method. As zinc is known to be an important co-factor for metallo-β-lactamase activity, test media of the semi-automated antimicrobial susceptibility testing system and broth microdilution were supplemented with zinc. Thereby, the detection of phenotypic resistance was improved in the semi-automated system and in broth microdilution. Currently, ceftazidime-avibactam is not approved as treatment option for infections by metallo-β-lactamase-producing, Gram-negative bacteria. In infections caused by carbapenem-resistant Gram-negatives, we therefore recommend to rule out the presence of metallo-β-lactamases with additional methods before initiating ceftazidime-avibactam treatment

High throughput method for analysis of repeat number for 28 phase variable loci of C. jejuni strain NCTC11168

Mutations in simple sequence repeat tracts are a major mechanism of phase variation in several bacterial species including Campylobacter jejuni. Changes in repeat number of tracts located within the reading frame can produce a high frequency of reversible switches in gene expression between ON and OFF states. The genome of C. jejuni strain NCTC11168 contains 29 loci with polyG/polyC tracts of seven or more repeats. This protocol outlines a method for rapidly determining ON/OFF states of these 28 phase-variable loci in a large number of individual colonies. The method combines a series of multiplex PCR assays with a GeneScan assay and automated extraction of tract length, repeat number and expression state. This high throughput, multiplex assay has utility for detecting shifts in phase variation states within and between populations over time and for exploring the effects of phase variation on adaptation to differing selective pressures. An important output of this assay is combinatorial expression states that cannot be determined by other methods. This method can be adapted to analysis of phase variation in other C. jejuni strains and in a diverse range of bacterial species

Етнологія релігії – актуальна сфера українського релігієзнавства

Background: Inhaled long-acting beta-2-adrenoceptor agonists (LABA) are frequently used in patients suffering from asthma and chronic obstructive pulmonary disease (COPD). For evaluation of real-life data, drug consumption studies are needed but results might be widely influenced due to methodological differences in particular regarding inter-country comparisons. Objectives: This study aims to compare the LABA prescribing in the general population and specifically in patients suffering from asthma and/or COPD in five European countries. Methods: Crude and age- and sex-standardized (European 2008 reference population) annual period prevalence rates per 10,000 persons were calculated for the period 2002-2009 based on seven European electronic health record databases (Denmark, Germany, Spain, the Netherlands (2), and the United Kingdom (2)). Stratification by sex, age, and indication were performed for the annual period prevalence rates. Results: In all databases, we observed an increase in LABA prescriptions during the study period for the general population and for patients suffering from asthma and/or COPD. In 2008, the highest standardized period prevalence was observed in the Dutch Mondriaan-AHC and the Spanish BIFAP database (443.3 and 395.5 per 10,000 persons), and the lowest in the German Bavarian Claims and Dutch Mondriaan- NPRCD database (278.7 and 290.6 per 10,000 persons). Prevalence rates for LABA increased with age and were highest in patients over 70 years. Patients with a combined diagnosis of asthma and COPD had higher prevalence rates of LABA compared to patients with a single diagnosis of asthma or COPD. The proportion of patients with one inhaled LABA prescription only ranged from 14% (UK databases) to 35% (Spanish BIFAP database) in 2008 in the general population. Conclusions: By using a standardized protocol, we demonstrated inter- and intra-country differences in LABA prescriptions. A general increase of LABA prescriptions during the study period was observed in all databases

HIF1A and NFAT5 coordinate Na+-boosted antibacterial defense via enhanced autophagy and autolysosomal targeting

Infection and inflammation are able to induce diet-independent Na+-accumulation without commensurate water retention in afflicted tissues, which favors the pro-inflammatory activation of mouse macrophages and augments their antibacterial and antiparasitic activity. While Na+-boosted host defense against the protozoan parasite Leishmania major is mediated by increased expression of the leishmanicidal NOS2 (nitric oxide synthase 2, inducible), the molecular mechanisms underpinning this enhanced antibacterial defense of mouse macrophages with high Na+ (HS) exposure are unknown. Here, we provide evidence that HS-increased antibacterial activity against E. coli was neither dependent on NOS2 nor on the phagocyte oxidase. In contrast, HS-augmented antibacterial defense hinged on HIF1A (hypoxia inducible factor 1, alpha subunit)-dependent increased autophagy, and NFAT5 (nuclear factor of activated T cells 5)-dependent targeting of intracellular E. coli to acidic autolysosomal compartments. Overall, these findings suggest that the autolysosomal compartment is a novel target of Na+- modulated cell autonomous innate immunity.This work was supported by the Deutsche Forschungsgemeinschaft [WA 2539/4-1, 5-1, 7-1]; Deutsche Forschungsgemeinschaft (DE) [JA 1993/ 4-1]; Universitätsklinikum Regensburg [Reform C]; NIHR Cambridge Blood and Transplant Research Unit Organ Donation

Similarities and differences in the historical records of lava dome-building volcanoes: implications for understanding magmatic processes and eruption forecasting

A key question for volcanic hazard assessment is the extent to which information can be exchanged between volcanoes. This question is particularly pertinent to hazard forecasting for dome-building volcanoes, where effusive activity may persist for years to decades, and may be punctuated by periods of repose, and sudden explosive activity. Here we review historical eruptive activity of fifteen lava dome-building volcanoes over the past two centuries, with the goal of creating a hierarchy of exchangeable (i.e., similar) behaviours. Eruptive behaviour is classified using empirical observations that include patterns of SO2 flux, eruption style, and magma composition. We identify two eruptive regimes: (i) an episodic regime where eruptions are much shorter than intervening periods of repose, and degassing is temporally correlated with lava effusion; and (ii) a persistent regime where eruptions are comparable in length to periods of repose and gas emissions do not correlate with eruption rates. A corollary to these two eruptive regimes is that there are also two different types of repose: (i) inter-eruptive repose separates episodic eruptions, and is characterised by negligible gas emissions and (ii) intra-eruptive repose is observed in persistently active volcanoes, and is characterised by continuous gas emissions. We suggest that these different patterns of can be used to infer vertical connectivity within mush-dominated magmatic systems. We also note that our recognition of two different types of repose raises questions about traditional definitions of historical volcanism as a point process. This is important, because the ontology of eruptive activity (that is, the definition of volcanic activity in time) influences both analysis of volcanic data and, by extension, interpretations of magmatic processes. Our analysis suggests that one identifying exchangeable traits or behaviours provides a starting point for developing robust ontologies of volcanic activity. Moreover, by linking eruptive regimes to conceptual models of magmatic processes, we illustrate a path towards developing a conceptual framework not only for comparing data between different volcanoes but also for improving forecasts of eruptive activity

Temporal changes in the epidemiology, management, and outcome from acute respiratory distress syndrome in European intensive care units: a comparison of two large cohorts

Background: Mortality rates for patients with ARDS remain high. We assessed temporal changes in the epidemiology and management of ARDS patients requiring invasive mechanical ventilation in European ICUs. We also investigated the association between ventilatory settings and outcome in these patients. Methods: This was a post hoc analysis of two cohorts of adult ICU patients admitted between May 1–15, 2002 (SOAP study, n = 3147), and May 8–18, 2012 (ICON audit, n = 4601 admitted to ICUs in the same 24 countries as the SOAP study). ARDS was defined retrospectively using the Berlin definitions. Values of tidal volume, PEEP, plateau pressure, and FiO2 corresponding to the most abnormal value of arterial PO2 were recorded prospectively every 24&nbsp;h. In both studies, patients were followed for outcome until death, hospital discharge or for 60&nbsp;days. Results: The frequency of ARDS requiring mechanical ventilation during the ICU stay was similar in SOAP and ICON (327[10.4%] vs. 494[10.7%], p = 0.793). The diagnosis of ARDS was established at a median of 3 (IQ: 1–7) days after admission in SOAP and 2 (1–6) days in ICON. Within 24&nbsp;h of diagnosis, ARDS was mild in 244 (29.7%), moderate in 388 (47.3%), and severe in 189 (23.0%) patients. In patients with ARDS, tidal volumes were lower in the later (ICON) than in the earlier (SOAP) cohort. Plateau and driving pressures were also lower in ICON than in SOAP. ICU (134[41.1%] vs 179[36.9%]) and hospital (151[46.2%] vs 212[44.4%]) mortality rates in patients with ARDS were similar in SOAP and ICON. High plateau pressure (&gt; 29 cmH2O) and driving pressure (&gt; 14 cmH2O) on the first day of mechanical ventilation but not tidal volume (&gt; 8&nbsp;ml/kg predicted body weight [PBW]) were independently associated with a higher risk of in-hospital death. Conclusion: The frequency of and outcome from ARDS remained relatively stable between 2002 and 2012. Plateau pressure &gt; 29 cmH2O and driving pressure &gt; 14 cmH2O on the first day of mechanical ventilation but not tidal volume &gt; 8&nbsp;ml/kg PBW were independently associated with a higher risk of death. These data highlight the continued burden of ARDS and provide hypothesis-generating data for the design of future studies

Surviving Sepsis Campaign: International guidelines for management of severe sepsis and septic shock: 2008

SCOPUS: ar.jinfo:eu-repo/semantics/publishe