MOESM7 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 7. Uncorrected and corrected cell dry weight results for multicomponent fed-batch cultivations. (a) Direct cell dry weight measurements of culture medium; (b) Cell dry weight measurements corrected for evaporation and water back-diffusion; Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate, reservoir: 250/375/500/750 g/L glucose, 40 g/L ammonium carbonate, blue dextran concentration: 1 g/L), temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP

MOESM1 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 1. Set-up of the membrane-based fed-batch shake flask. (a) The offline and (b) The online membrane-based fed-batch shake flask

MOESM9 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 9. Average values and standard deviations for validation of reproducibility. Depiction of (a) Eight replicates within one experiment; (b) Five replicates from independent experiments; Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate, initial reservoir glucose concentrations: 250 g/L, blue dextran concentration: 4 g/L, temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP

MOESM8 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 8. Effect of dipotassium phosphate concentration variation on OTR in cultivations with additional glucose and ammonium carbonate in the reservoir. Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate, reservoir: 500 g/L glucose, 40 g/L ammonium carbonate, varied dipotassium phosphate concentration, blue dextran concentration: 1 g/L), temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP

MOESM3 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 3. Comparison of calculated and measured values at varied initial reservoir glucose concentrations with the membrane-based fed-batch shake flask. Depiction of (a) + (b) Measured OTR; (c) + (d) Measured and calculated amount of metabolized glucose; (e) + (f) Measured and calculated amount of cell dry weight; (g) + (h) Measured and calculated amount of metabolized ammonium and accumulated glucose. For calculations the stoichiometric equation C6H12O6 + 0.57 NH3 + 2.34 O2 → 3.38 CH1.7O0.43N0.17 + 2.62 CO2 + 3.99 H2O was used. Calculations until the abrupt OTR increases are based on measured total oxygen consumed values, calculations after the abrupt OTR increase are based on the calculated glucose release; Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate, reservoir: 250/375/500/750 g/L glucose, blue dextran concentration: 1 g/L), temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP; Concentration values have been volume-corrected

MOESM5 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 5. Depiction of cultivations with 750 g/L glucose and 40 g/L ammonium carbonate in the reservoir for the multi-component release with the membrane-based fed-batch shake flask. Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate in culture medium, reservoir: 750 g/L glucose, 40 g/L ammonium carbonate concentration and blue dextran concentration: 1 g/L), temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP; The arrow points to the occurrence of an abrupt OTR increase

MOESM4 of Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask

Additional file 4. Depiction of ammonium concentrations in the culture medium and reservoir of the multi-component fed-batch cultivations at varied initial glucose concentrations and 40 g/L ammonium carbonate in the reservoir. Depiction of (a) Oxygen transfer rate; (b) Ammonium concentration in the culture medium; (c) Ammonium concentration in the reservoir; Cultivation conditions: Wilms-MOPS-mineral medium (culture medium: 200 mM MOPS with 7 g/L initial ammonium sulfate, initial reservoir glucose concentrations: 250/375/500/750 g/L, 40 g/L ammonium carbonate, blue dextran concentration: 1 g/L), temperature: 37 °C, shaking frequency: 350 rpm, shaking diameter: 50 mm, initial culture medium volume: 10 mL, inoculation OD600: 0.5, reservoir filling volume: 2 mL, dialysis membrane: Reichelt 10-20 kDa, membrane area: 18.1 mm2; Strain: E. coli BL21 (DE3) pRhotHi-2-EcFbFP; Concentration values have been volume-corrected

Additional file 1 of Midregional proatrial naturetic peptide (MRproANP) and copeptin (COPAVP) as predictors of all-cause mortality in recently diagnosed mild to moderate COPD—results from COSYCONET

Additional file 1: Table S1. Results of consecutive Cox regression analyses in all patients of grades GOLD 0-4. Hazard ratios are given for the respective biomarkers in the upper quartile versus the three lower quartiles. * Age, BMI, FEV1% predicted, TLCO % predicted, SGRQ total score, 6-minute walking distance, ≥1 moderate/severe exacerbations in the previous year, smoking status, pack years, hypercapnia (PaCO2 >55 mmHg), hypoxemia (PaO2 <60 mmHg). ** All variables of model 2 plus sex, hypertension, diabetes, hyperlipidemia, log HbA1c, and log CRP. *** All variables of model 3 plus heart failure, coronary artery disease, history of myocardial infarction, history of stroke, ankle-brachial index ≤0.90, log NT-proBNP, left-ventricular ejection fraction from echocardiography