Protein display, secretion, and export in prokaryotes are essential for utilizing microbial systems as engineered living materials, medicines, biocatalysts, and protein factories. To select for improved signal peptides for Escherichia coli protein display, we utilized error-prone polymerase chain reaction (epPCR) coupled with single-cell sorting and microplate titer to generate, select, and detect improved Ag43 signal peptides. Through just three rounds of mutagenesis and selection using green fluorescence from the 56 kDa sfGFP-beta-lactamase, we isolated clones that modestly increased surface display from 1.4- to 3-fold as detected by the microplate plate-reader and native SDS-PAGE assays. To establish that the functional protein was displayed extracellularly, we trypsinized the bacterial cells to release the surface displayed proteins for analysis. This workflow demonstrated a fast and high-throughput method leveraging epPCR and single-cell sorting to augment bacterial surface display rapidly that could be applied to other bacterial proteins.

Workplace bureaucracies impact many areas of an employee’s working life. High levels of perceived bureaucracy can diminish employees’ job motivation, satisfaction, and performance. During the COVID-19 pandemic, infection containment measures likely contributed to the factors underlying the “great resignation” via the lack of self-efficacy from perceived bureaucracy. Recruiting 240 full-time employees between late 2021 to early 2022 across organizations in both private and public sectors, the Work Extrinsic Intrinsic Motivation Scale, the Workplace Bureaucracy Questionnaire, and the General Self-Efficacy Scale were employed along with demographic parameters for analysis, showing that salary range, education levels, self-efficacy and job motivation were negatively associated with perceived workplace bureaucracies, but not gender, age, and length of employment. The findings of the study provide insight into the effect of increased perceived bureaucracy during the COVID-19 pandemic on employee experience that would be relevant to the workplace environment even in normal times.

The belief in the supernatural and paranormal is deeply intertwined in human history and culture with the recent inclusion of science fiction entity types e.g., extra-terrestrials. Investigating for the association between the beliefs in formal religions, pagan, and science fiction with worldview elements, 255 participants were recruited and surveyed. The demographic of the participants and their declared religious affiliation were found to be associated but not with their theistic beliefs using the non-parametric Kruskal-Wallis and Dunn’s tests. Non-monotheistic beliefs, particularly polytheistic and agnostic beliefs were significantly positively associated with beliefs in ghosts (H = 34.51, p < 0.01) and other pagan entities such as nature spirits (H = 36.41, p < 0.01) and luck (H = 28.85, p < 0.01). Monotheists tend to have more significant exclusive beliefs in supernatural entities e.g., angels and demons compared to the other theists including atheists, agnostics, polytheists, and pantheists (H = 53.58, p < 0.01). By gender, females were found to significantly believe in the paranormal (W = 9308, p < 0.01) and ghosts (W = 9308, p < 0.01) more than males. The findings here serve to uncover associations of religious beliefs with supernatural and paranormal beliefs such as extra-terrestrials, luck, and objective morality, with demographic background that includes education and professional vocations. The findings here would inform the basis for future studies on beliefs in supernatural and paranormal entities.

Protein display, secretion and export in prokaryotes are essential for utilizing microbial systems as engineered living materials for medicines, biocatalysts, and protein factories. To select for improved signal peptides for Escherichia coli protein display, we utilized error-prone polymerase chain reaction (epPCR) coupled with single-cell sorting and microplate titer to generate, select, and detect improved Ag43 signal peptides. Through three rounds of mutagenesis and selection using green fluorescence from the 56 kDa sfGFP-beta-lactamase, we isolated clones that increased surface display from 1.4 to 3 folds as detected by the microplate plate-reader and native SDS-PAGE assays. To establish that the protein was displayed extracellularly, we trypsinised the bacterial cells to release the surface displayed proteins for analysis. This workflow demonstrated a fast and high-throughput method leveraging on epPCR and single-cell sorting to rapidly augment bacterial surface display, a method that could be applied to other bacterial proteins.