Condensed ECM-based nanofilms on highly permeable PET membranes for robust cell-to-cell communications with improved optical clarity

The properties of a semipermeable porous membrane, including pore size, pore density, and thickness, play a crucial role in creating a tissue interface in a microphysiological system (MPS) because it dictates multicellular interactions between different compartments. The small pore-sized membrane has been preferentially used in an MPS for stable cell adhesion and the formation of tissue barriers on the membrane. However, it limited the applicability of the MPS because of the hindered cell transmigration via sparse through-holes and the optical translucence caused by light scattering through pores. Thus, there remain unmet challenges to construct a compartmentalized MPS without those drawbacks. Here we report a submicrometer-thickness (similar to 500 nm) fibrous extracellular matrix (ECM) film selectively condensed on a large pore-sized track-etched (TE) membrane (10 mu m-pores) in an MPS device, which enables the generation of functional tissue barriers simultaneously achieving optical transparency, intercellular interactions, and transmigration of cells across the membrane. The condensed ECM fibers uniformly covering the surface and 10 mu m-pores of the TE membrane permitted sufficient surface areas where a monolayer of the human induced pluripotent stem cell-derived brain endothelial cells is formed in the MPS device. The functional maturation of the blood-brain barrier (BBB) was proficiently achieved due to astrocytic endfeet sheathing the brain endothelial cells through 10 mu m pores of the condensed-ECM-coated TE (cECMTE) membrane. We also demonstrated the extravasation of human metastatic breast tumor cells through the human BBB on the cECMTE membrane. Thus, the cECMTE membrane integrated with an MPS can be used as a versatile platform for studying various intercellular communications and migration, mimicking the physiological barriers of an organ compartment

Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey▿

In this study, an enzyme-linked immunosorbent assay (ELISA) using glycoprotein and a monoclonal antibody (MAb) was developed for the detection of antibodies to vesicular stomatitis virus (VSV) serotype New Jersey (NJ). The glycoprotein to be used as a diagnostic antigen was extracted from partially purified VSV-NJ, and a neutralizing MAb specific to VSV-NJ was incorporated to compete with antibodies in a blocking ELISA using glycoprotein (GP ELISA). The cutoff of the GP ELISA was set at 40% inhibition, which corresponded to a virus neutralization test (VNT) titer of 32. With this threshold, the GP ELISA exhibited 99.6% specificity for naïve sera (n = 3,005) from cattle (n = 1,040), pigs (n = 1,120), and horses (n = 845) from domestic farms. The GP ELISA did not cross-react with sera positive for foot-and-mouth disease virus, swine vesicular disease virus, or VSV serotype Indiana. The GP ELISA was more compatible with the VNT than was the nucleocapsid-based ELISA for VSV-NJ-positive sera (n = 19). Taken together, this GP ELISA could be a useful tool as an alternative to the VNT for detecting antibodies specific to VSV-NJ

Accuracy of the qSOFA Score and RED Sign in Predicting Critical Care Requirements in Patients with Suspected Infection in the Emergency Department: A Retrospective Observational Study

Background and objectives: We aimed to compare the accuracy of positive quick sequential organ failure assessment (qSOFA) scores and the RED sign in predicting critical care requirements (CCRs) in patients with suspected infection who presented to the emergency department (ED). Materials and Methods: In this retrospective observational study, we examined adult patients with suspected infection in the ED from June 2018 to September 2018. A positive qSOFA (qSOFA+) was defined as the presence of ≥2 of the following criteria: altered mental status (AMS), systolic blood pressure (SBP) < 100 mmHg, and respiratory rate (RR) ≥ 22 breaths/min. A positive RED sign (RED sign+) was defined as the presence of at least one of the RED sign criteria: AMS, skin mottling, SBP < 90 mmHg, heart rate >130 beats/min, or RR > 30 breaths/min. A qSOFA/RED+ was defined as the presence of qSOFA+ or RED+. We applied these tools twice using the initial values upon ED arrival and all values within 2 h after ED arrival. The accuracy of qSOFA+, RED+, and qSOFA/RED+ in predicting CCR was assessed. Results: Data from 5353 patients with suspected infection were analyzed. The area under the receiver operating characteristic curve (AUC) of RED+ (0.67, 95% confidence interval [CI]: 0.65–0.70) and that of qSOFA/RED+ (0.68, 95% CI: 0.66–0.70, p < 0.01) were higher than the AUC of qSOFA+ (0.59, 95% CI: 0.57–0.60) in predicting CCR on ED arrival. The qSOFA/RED+ within 2 h showed the highest accuracy (AUC 0.72, 95% CI: 0.70–0.75, p < 0.001). Conclusions: The accuracy of the RED sign in predicting CCR in patients with suspected infection who presented at ED was better than that of qSOFA. The combined use of the RED sign and qSOFA (positive qSOFA or RED sign) showed the highest accuracy

Use of a Baculovirus-Expressed Structural Protein for the Detection of Antibodies to Foot-and-Mouth Disease Virus Type A by a Blocking Enzyme-Linked Immunosorbent Assay▿

A blocking enzyme-linked immunosorbent assay (ELISA) with a baculovirus-expressed structural protein was developed for the detection of antibodies to foot-and-mouth disease virus type A. It exhibited 99% specificity with a cutoff of 53% inhibition. Its sensitivity was comparable to the sensitivities of the virus neutralization test and the liquid-phase blocking ELISA, indicating its potential as an alternative assay