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76 records · Page 5

Engineered Cell Line Imaging Assay Differentiates Pathogenic from Non-Pathogenic Bacteria

Cell culture systems have greatly expanded our understanding of how bacterial pathogens target signaling pathways to manipulate the host and cause infection. Advances in genetic engineering have allowed for the creation of fluorescent protein readouts within signaling pathways, but these techniques have been underutilized in pathogen biology. Here, we genetically engineered a lung cell line with fluorescent reporters for extracellular signal-related kinase (ERK) and the downstream transcription factor FOS-related antigen 1 (Fra1) and evaluated signaling after inoculation with pathogenic and non-pathogenic bacteria. Cells were inoculated with 100 colony-forming units of Acinetobacter baylyi, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus agalactiae, or Staphylococcus epidermidis and imaged in a multi-mode reader. The alamarBlue cell viability assay was used as a reference test and showed that pathogenic P. aeruginosa induced significant (p < 0.05) cell death after 8 h in both wild-type and engineered cell lines compared to non-pathogenic S. epidermidis. In engineered cells, we found that Fra1 signaling was disrupted in as little as 4 h after inoculation with bacterial pathogens compared to delayed disruption in signaling by non-pathogenic S. epidermidis. Overall, we demonstrate that low levels of pathogenic versus non-pathogenic bacteria can be rapidly and sensitively screened based on ERK-Fra1 signaling

59 BASIC BIOLOGICAL SCIENCES↗

Testing convolutional neural network based deep learning systems: a statistical metamorphic approach

Machine learning technology spans many areas and today plays a significant role in addressing a wide range of problems in critical domains,i.e., healthcare, autonomous driving, finance, manufacturing, cybersecurity,etc. Metamorphic testing (MT) is considered a simple but very powerful approach in testing such computationally complex systems for which either an oracle is not available or is available but difficult to apply. Conventional metamorphic testing techniques have certain limitations in verifying deep learning-based models (i.e., convolutional neural networks (CNNs)) that have a stochastic nature (because of randomly initializing the network weights) in their training. In this article, we attempt to address this problem by using a statistical metamorphic testing (SMT) technique that does not require software testers to worry about fixing the random seeds (to get deterministic results) to verify the metamorphic relations (MRs). We propose seven MRs combined with different statistical methods to statistically verify whether the program under test adheres to the relation(s) specified in the MR(s). We further use mutation testing techniques to show the usefulness of the proposed approach in the healthcare space and test two CNN-based deep learning models (used for pneumonia detection among patients). The empirical results show that our proposed approach uncovers 85.71% of the implementation faults in the classifiers under test (CUT). Furthermore, we also propose an MRs minimization algorithm for the CUT, thus saving computational costs and organizational testing resources.

Computer Science↗

Ventilation in Residential Care Environments

The purpose of this study was to evaluate the effect of ventilation rate, directional airflow (e.g. pressure relationships) and airflow barriers on bioaerosol concentration and movement within assisted-living and residential care environments. Included within this report is a comprehensive literature review, field data collection test plan and, an evaluation of commercially available mitigation technologies related to the transmission of COVID-19 in LTC environments. Additionally, this report includes aerosol testing in an actual LTC facility using the test plan developed herein. Nearly 1.5 million people live in 16,000 nursing homes in the U.S. As of December 20, 2020 the Centers for Medicare and Medicaid Services (CMS) identified a total of 662,549 confirmed and suspected COVID-19 cases in these and other long-term care (LTC) facilities resulting in 92,373 deaths. At this time, COVID-19 cases in LTC facilities accounted for only 3% of the U.S. total of 21M confirmed cases yet nearly a third of the 350,000 total U.S. deaths. Preliminary studies have found little correlation between the quality of care and transmission of COVID-19 in these facilities, suggesting that even the best infection control practices may not be effective in containing the spread of this potentially airborne disease. As a result, many LTC facilities have implemented quarantine procedures and other measures to isolate infectious residents from the general population. Unfortunately, most LTC facilities were not designed for airborne infection control and guidance for retrofitting existing LTC spaces for airborne isolation is limited.

42 ENGINEERING↗