Artificial intelligence tool for the study of COVID-19 microdroplet spread across the human diameter and airborne space.

The 2019 novel coronavirus (SARS-CoV-2 / COVID-19), with a point of origin in Wuhan, China, has spread rapidly all over the world. It turned into a raging pandemic wrecking havoc on health care facilities, world economy and affecting everyone's life to date. With every new variant, rate of transmission, spread of infections and the number of cases continues to rise at an international level and scale. There are limited reliable researches that study microdroplets spread and transmissions from human sneeze or cough in the airborne space. In this paper, we propose an intelligent technique to visualize, detect, measure the distance of spread in a real-world settings of microdroplet transmissions in airborne space, called "COVNET45". In this paper, we investigate the microdroplet transmission and validate the measurements accuracy compared to published researches, by examining several microscopic and visual images taken to investigate the novel coronavirus (SARS-CoV-2 / COVID-19). The ultimate contribution is to calculate the spread of the microdroplets, measure it precisely and provide a graphical presentation. Additionally, the work employs machine learning and five algorithms for image optimization, detection and measurement.

Multiplexed profiling of RNA and protein expression signatures in individual cells using flow or mass cytometry.

Advances in single-cell analysis technologies are providing novel insights into phenotypic and functional heterogeneity within seemingly identical cell populations. RNA within single cells can be analyzed using unbiased sequencing protocols or through more targeted approaches using in situ hybridization (ISH). The proximity ligation assay for RNA (PLAYR) approach is a sensitive and high-throughput technique that relies on in situ and proximal ligation to measure at least 27 specific RNAs by flow or mass cytometry. We provide detailed instructions for combining this technique with antibody-based detection of surface/internal protein, allowing simultaneous highly multiplexed profiling of RNA and protein expression at single-cell resolution. PLAYR overcomes limitations on multiplexing seen in previous branching DNA-based RNA detection techniques by integration of a transcript-specific oligonucleotide sequence within a rolling-circle amplification (RCA). This unique transcript-associated sequence can then be detected by heavy metal (for mass cytometry)- or fluorophore (for flow cytometry)-conjugated complementary detection oligonucleotides. Included in this protocol is methodology to label oligonucleotides with lanthanide metals for use in mass cytometry. When analyzed by mass cytometry, up to 40 variables (with scope for future expansion) can be measured simultaneously. We used the described protocol to demonstrate intraclonal heterogeneity within primary cells from chronic lymphocytic leukemia patients, but it can be adapted to other primary cells or cell lines in suspension. This robust, reliable and reproducible protocol can be completed in 2-3 d and can be paused at several stages for convenience.

High incidence of New Delhi metallo-beta-lactamase-producing Klebsiella pneumoniae isolates in Sharjah, United Arab Emirates.

The purpose of this study was to identify carbapenem-resistant Klebsiella pneumoniae in a tertiary care hospital in Sharjah Emirate, to identify the plasmids carrying the carbapenemase genes and to reveal clonal relationships among the isolates. Two hundred and two clinically relevant isolates collected between September 2011 and October 2012 at Al-Qassimi hospital, Sharjah, were investigated for meropenem resistance. Strains with decreased susceptibility were further tested with the modified Hodge test, by EDTA and phenylboronic acid synergy and by E-test. The genes of New Delhi metallo-ß-lactamase (NDM), IMP, VIM, OXA-48, and KPC beta-lactamases were targeted by polymerase chain reaction and the genes were located on plasmids by Southern blotting. Clusters of the isolates were revealed by macrorestriction analysis. Seven percent of the isolates were originally found to be meropenem resistant, one isolate have lost its resistance during storage. All of the 13 resistant isolates were positive for the NDM-1 gene located on plasmids of 125 to >170 kb, while three isolates also carried the blaOXA-48-like genes. Clusters having repeatedly been isolated over the study period were identified. Carbapenem-resistant Klebsiella pneumoniae carrying the blaNDM-1 gene is a fast emerging problem, emphasizing the potential role of the Middle East as a secondary reservoir for these organisms.