Colloids

All lipids were purchased from Avanti Polar Lipids. For binding of NeutrAvidin, 5% 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) was introduced. Vesicles were prepared by drying the lipids onto the interior of a flask for 30 min, followed by hydration in buffer and extrusion 11 times through 30 nm pores (1 bar). The I-BAR domain of IRSp53 was produced as described previously (Saarikangas et al., 2009 (link)). Standard chemicals for buffer preparation were from Sigma. NeutrAvidin was from ThermoFisher.
Nanowells were prepared as described previously (Junesch et al., 2012 (link), 2015 (link); Malekian et al., 2017 (link); Ferhan et al., 2018 (link)) using 107 nm polystyrene colloids on Nb₂O₅ and 158 nm on SiO₂ (Microparticles). Nanowells in SiO₂ were prepared on fused silica to enable direct etching of the solid support (Malekian et al., 2017 (link)). Nanowells in Nb₂O₅ were prepared on borosilicate glass (which cannot be easily etched) onto which Nb₂O₅ was first deposited by reactive sputter coating with O₂ and Ar (Junesch et al., 2012 (link)) (Nordiko). A 20 nm thick SiO₂ layer was deposited by plasma enhanced chemical vapor deposition (Surface Technology Systems). Recipes aiming for stochiometric SiO₂ or Si₃N₄ were used.
For bilayer formation with negative lipids, a 20 mM citric acid buffer was used with 150 mM KCl at a pH of 4.8. IRSp53 binding was performed in a buffer with 20 mM tris(hydroxymethyl) aminomethane and 150 mM NaCl with pH adjusted to 7.4 unless stated otherwise. The pH values were adjusted with concentrated HCl and NaOH.
The setup for extinction spectroscopy with high resolution and tracking of multiple resonance features has been described previously (Junesch et al., 2015 (link); Ferhan et al., 2018 (link)). Extinction is presented using the natural logarithm of the ratio between reference and measured intensities.

This study was a restrictive observation study from the Medical Information Mart for Intensive Care IV (MIMIC-IV version 0.4) database from 2008 to 2019 [24 ]. An individual who has finished the Collaborative Institutional Training Initiative examination (Certification number 35931520 for author Zhou) can access the database. This is a longitudinal, single-center database including 257,366 individuals and 196,527 adults, and 11,263 patients with sepsis (Defined by sepsis-3 criteria [1 (link)]). In our study, we extracted patients’ parameters containing age, gender, ethnic group, admission type, insurance condition, the first 24-h Sequential Organ Failure Assessment (SOFA) score, Simplified Acute Physiology Score II (SAPS) score, mean arterial blood pressure (MAP), heart rate, respiratory rate, temperature, SpO₂, total urine output during the first 24 h after ICU admission, lactate level, the use of vasopressors, weight, mechanical ventilation, renal replacement therapy (RRT), the stage of acute kidney injury (AKI), anamnesis (myocardial infarction, cancer, renal disease, cirrhosis and diabetes) and the type and volume of their fluid administration during the whole ICU stay. Vasopressors included norepinephrine, phenylephrine, epinephrine, vasopressin, dopamine, and dobutamine. For the antibiotics, Carbapenems (meropenem), Glycopeptide (vancomycin), β-lactams (ceftriaxone, cefotaxime, and cefepime), and Aminoglycosides (gentamicin and amikacin) were extracted into our analysis. In this study, types of administration for crystalloids and albumin including normal saline and lactated Ringer’s (LR) solution, while 5% and 25% HSA for colloids. The code of data extraction is available on Github (https://github.com/MIT-LCP/mimic-iv).
Adults patients (≥ 18 years) with sepsis and complete fluid administration records were screened in the analysis. The following exclusion criteria were used: (1) patients who have not received any crystalloids administration; (2) patients who received albumin longer than 24 h after the initiation of crystalloids administration or preceded the crystalloids. For patients who had ICU admission more than once, only data of the first ICU admission of the first hospital stay were included.