Automated and rapid identification of multidrug resistant Escherichia coli against the lead drugs of acylureidopenicillins, cephalosporins, and fluoroquinolones using specific Raman marker bands

A Raman-based, strain-independent, semi-automated method is presented that allows the rapid (<3 hours) determination of antibiotic susceptibility of bacterial pathogens isolated from clinical samples. Applying a priori knowledge about the mode of action of the respective antibiotic, we identified characteristic Raman marker bands in the spectrum and calculated batch-wise weighted sum scores from standardized Raman intensity differences between spectra of antibiotic exposed and nonexposed samples of the same strains. The lead substances for three relevant antibiotic classes (fluoroquinolone ciprofloxacin, third-generation cephalosporin cefotaxime, ureidopenicillin piperacillin) against multidrug-resistant Gram-negative bacteria (MRGN) revealed a high sensitivity and specificity for the susceptibility testing of two Escherichia coli laboratory strains and 12 clinical isolates. The method benefits from the parallel incubation of control and treated samples, which reduces the variance due to alterations in cultivation conditions and the standardization of differences between batches leading to long-term comparability of Raman measurements. © 2020 The Authors. Journal of Biophotonics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinhei

Framework and baseline examination of the German National Cohort (NAKO)

The German National Cohort (NAKO) is a multidisciplinary, population-based prospective cohort study that aims to investigate the causes of widespread diseases, identify risk factors and improve early detection and prevention of disease. Specifically, NAKO is designed to identify novel and better characterize established risk and protection factors for the development of cardiovascular diseases, cancer, diabetes, neurodegenerative and psychiatric diseases, musculoskeletal diseases, respiratory and infectious diseases in a random sample of the general population. Between 2014 and 2019, a total of 205,415 men and women aged 19–74 years were recruited and examined in 18 study centres in Germany. The baseline assessment included a face-to-face interview, self-administered questionnaires and a wide range of biomedical examinations. Biomaterials were collected from all participants including serum, EDTA plasma, buffy coats, RNA and erythrocytes, urine, saliva, nasal swabs and stool. In 56,971 participants, an intensified examination programme was implemented. Whole-body 3T magnetic resonance imaging was performed in 30,861 participants on dedicated scanners. NAKO collects follow-up information on incident diseases through a combination of active follow-up using self-report via written questionnaires at 2–3 year intervals and passive follow-up via record linkages. All study participants are invited for re-examinations at the study centres in 4–5 year intervals. Thereby, longitudinal information on changes in risk factor profiles and in vascular, cardiac, metabolic, neurocognitive, pulmonary and sensory function is collected. NAKO is a major resource for population-based epidemiology to identify new and tailored strategies for early detection, prediction, prevention and treatment of major diseases for the next 30 years. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10654-022-00890-5

Pom1 regulates the assembly of Cdr2-Mid1 cortical nodes for robust spatial control of cytokinesis

Proper division plane positioning is essential to achieve faithful DNA segregation and to control daughter cell size, positioning, or fate within tissues. In Schizosaccharomyces pombe, division plane positioning is controlled positively by export of the division plane positioning factor Mid1/anillin from the nucleus and negatively by the Pom1/DYRK (dual-specificity tyrosine-regulated kinase) gradients emanating from cell tips. Pom1 restricts to the cell middle cortical cytokinetic ring precursor nodes organized by the SAD-like kinase Cdr2 and Mid1/anillin through an unknown mechanism. In this study, we show that Pom1 modulates Cdr2 association with membranes by phosphorylation of a basic region cooperating with the lipid-binding KA-1 domain. Pom1 also inhibits Cdr2 interaction with Mid1, reducing its clustering ability, possibly by down-regulation of Cdr2 kinase activity. We propose that the dual regulation exerted by Pom1 on Cdr2 prevents Cdr2 assembly into stable nodes in the cell tip region where Pom1 concentration is high, which ensures proper positioning of cytokinetic ring precursors at the cell geometrical center and robust and accurate division plane positioning

Aquaporin regulation under salt and osmotic stress in the halophyte mesembryanthemum crystallinum L.

It is evident that plant aquaporins should play a dynamic role in maintaining cellular water homeostasis under conditions that necessitate modifications in water flux, including drought- and salt-stress. Changes in water uptake and allocation would be required to balance alterations in cellular osmotic potential and therefore, both plasma membrane and vacuolar membrane (tonoplast) aquaporin activity and/or expression must be tightly regulated

Automated and rapid identification of multidrug resistant Escherichia coli against the lead drugs of acylureidopenicillins, cephalosporins, and fluoroquinolones using specific Raman marker bands

A Raman-based, strain-independent, semi-automated method is presented that allows the rapid (<3 hours) determination of antibiotic susceptibility of bacterial pathogens isolated from clinical samples. Applying a priori knowledge about the mode of action of the respective antibiotic, we identified characteristic Raman marker bands in the spectrum and calculated batch-wise weighted sum scores from standardized Raman intensity differences between spectra of antibiotic exposed and nonexposed samples of the same strains. The lead substances for three relevant antibiotic classes (fluoroquinolone ciprofloxacin, third-generation cephalosporin cefotaxime, ureidopenicillin piperacillin) against multidrug-resistant Gram-negative bacteria (MRGN) revealed a high sensitivity and specificity for the susceptibility testing of two Escherichia coli laboratory strains and 12 clinical isolates. The method benefits from the parallel incubation of control and treated samples, which reduces the variance due to alterations in cultivation conditions and the standardization of differences between batches leading to long-term comparability of Raman measurements. © 2020 The Authors. Journal of Biophotonics published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinhei

Three Dimensional Checkerboard Synergy Analysis of Colistin, Meropenem, Tigecycline against Multidrug-Resistant Clinical <i>Klebsiella pneumonia</i> Isolates

<div><p>The spread of carbapenem-non-susceptible <i>Klebsiella pneumoniae</i> strains bearing different resistance determinants is a rising problem worldwide. Especially infections with KPC (<i>Klebsiella pneumoniae</i> carbapenemase) - producers are associated with high mortality rates due to limited treatment options. Recent clinical studies of KPC-blood stream infections revealed that colistin-based combination therapy with a carbapenem and/or tigecycline was associated with significantly decreased mortality rates when compared to colistin monotherapy. However, it remains unclear if these observations can be transferred to <i>K</i>. <i>pneumoniae</i> harboring other mechanisms of carbapenem resistance. A three-dimensional synergy analysis was performed to evaluate the benefits of a triple combination with meropenem, tigecycline and colistin against 20 <i>K</i>. <i>pneumoniae</i> isolates harboring different β-lactamases. To examine the mechanism behind the clinically observed synergistic effect, efflux properties and outer membrane porin (Omp) genes (<i>ompK35</i> and <i>ompK36</i>) were also analyzed. Synergism was found for colistin-based double combinations for strains exhibiting high minimal inhibition concentrations against all of the three antibiotics. Adding a third antibiotic did not result in further increased synergistic effect in these strains. Antagonism did not occur. These results support the idea that colistin-based double combinations might be sufficient and the most effective combination partner for colistin should be chosen according to its MIC.</p></div

Results of susceptibility testing (MICs), the molecular analyses of β-lactamases and porins, the efflux properties (EHT), and the synergism testing (FICIs).

<p>The EHT and the FICI values are given as median ± standard deviation. Synergistic FICI values are indicated in bold letters. The FICI values indicated in brackets represent the lowest values for the given combination.</p><p>*Strains were obtained from: NRZ = National Reference Laboratory for multidrug-resistant gram-negative bacteria, Ruhr-University Bochum, Germany, and RKI = Robert-Koch-Institut, Wernigerode, Germany; MEM = meropenem; CST = colistin; TCG = tigecycline; FICI = fractional inhibitory concentration index; EHT = efflux half-time; nm = not measureable; aa = amino acid;:: = gene dispuption by insertion sequence; del = deletion; ins insertion; sub = substitution;</p><p>**non-functional (STOP mutation in <i>bla</i>_OXA-9 gene)</p><p>Results of susceptibility testing (MICs), the molecular analyses of β-lactamases and porins, the efflux properties (EHT), and the synergism testing (FICIs).</p

Inhibition zones of the antibiotics in disc diffusion test of selected strains.

<p>Inhibition zones of the antibiotics in disc diffusion test of selected strains.</p

Exemplary isoboles of the double combinations of the antibiotics.

<p>A-C: Isoboles of strain NRZ 00246 (OXA-48 producer) exhibiting synergistic effects. D-F: Isoboles of strain RKI 318/11 (KPC producer) yielding no synergism. FIC = fractional inhibitory concentration.</p

Exemplary plots of the checkerboard assays for the double combinations of the antibiotics.

<p>A-C: Strain NRZ 00246 (OXA-48 producer) exhibiting FICI <0.8. D-F: Strain RKI 318/11 (KPC producer) exhibiting FICI >0.8. Grey lines indicate the breakpoints of the respective antibiotic (according to EUCAST).</p