Direct Quantitation of SARS-CoV-2 Virus in Urban Ambient Air via a Continuous-Flow Electrochemical Bioassay.

Airborne SARS-CoV-2 virus surveillance faces challenges in complicated biomarker enrichment, interferences from various non-specific matters and extremely low viral load in the urban ambient air, leading to difficulties in detecting SARS-CoV-2 bioaerosols. This work reports a highly specific bioanalysis platform, with an exceptionally low limit-of-detection (≤1 copy m-3 ) and good analytical accordance with RT-qPCR, relying on surface-mediated electrochemical signaling and enzyme-assisted signal amplification, enabling gene and signal amplification for accurate identification and quantitation of low doses human coronavirus 229E (HCoV-229E) and SARS-CoV-2 viruses in urban ambient air. This work provides a laboratory test using cultivated coronavirus to simulate the airborne spread of SARS-CoV-2, and validate that the platform could reliably detect airborne coronavirus and reveal the transmission characteristics. This bioassay conducts the quantitation of real-world HCoV-229E and SARS-CoV-2 in airborne particulate matters collected from road-side and residential areas in Bern and Zurich (Switzerland) and Wuhan (China), with resultant concentrations verified by RT-qPCR.

On-Site Quantification and Infection Risk Assessment of Airborne SARS-CoV-2 Virus Via a Nanoplasmonic Bioaerosol Sensing System in Healthcare Settings.

On-site quantification and early-stage infection risk assessment of airborne severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with high spatiotemporal resolution is a promising approach for mitigating the spread of coronavirus disease 2019 (COVID-19) pandemic and informing life-saving decisions. Here, a condensation (hygroscopic growth)-assisted bioaerosol collection and plasmonic photothermal sensing (CAPS) system for on-site quantitative risk analysis of SARS-CoV-2 virus-laden aerosols is presented. The CAPS system provided rapid thermoplasmonic biosensing results after an aerosol-to-hydrosol sampling process in COVID-19-related environments including a hospital and a nursing home. The detection limit reached 0.25 copies/µL in the complex aerosol background without further purification. More importantly, the CAPS system enabled direct measurement of the SARS-CoV-2 virus exposures with high spatiotemporal resolution. Measurement and feedback of the results to healthcare workers and patients via a QR-code are completed within two hours. Based on a dose-responseµ model, it is used the plasmonic biosensing signal to calculate probabilities of SARS-CoV-2 infection risk and estimate maximum exposure durations to an acceptable risk threshold in different environmental settings.

SARS-CoV-2 and other airborne respiratory viruses in outdoor aerosols in three Swiss cities before and during the first wave of the COVID-19 pandemic.

Caused by the SARS-CoV-2 virus, Coronavirus disease 2019 (COVID-19) has been affecting the world since the end of 2019. While virus-laden particles have been commonly detected and studied in the aerosol samples from indoor healthcare settings, studies are scarce on air surveillance of the virus in outdoor non-healthcare environments, including the correlations between SARS-CoV-2 and other respiratory viruses, between viruses and environmental factors, and between viruses and human behavior changes due to the public health measures against COVID-19. Therefore, in this study, we collected airborne particulate matter (PM) samples from November 2019 to April 2020 in Bern, Lugano, and Zurich. Among 14 detected viruses, influenza A, HCoV-NL63, HCoV-HKU1, and HCoV-229E were abundant in air. SARS-CoV-2 and enterovirus were moderately common, while the remaining viruses occurred only in low concentrations. SARS-CoV-2 was detected in PM10 (PM below 10 µm) samples of Bern and Zurich, and PM2.5 (PM below 2.5 µm) samples of Bern which exhibited a concentration positively correlated with the local COVID-19 case number. The concentration was also correlated with the concentration of enterovirus which raised the concern of coinfection. The estimated COVID-19 infection risks of an hour exposure at these two sites were generally low but still cannot be neglected. Our study demonstrated the potential functionality of outdoor air surveillance of airborne respiratory viruses, especially at transportation hubs and traffic arteries.

Exploring the signaling space of a GPCR using bivalent ligands with a rigid oligoproline backbone.

G protein-coupled receptors (GPCRs) are one of the most important drug-target classes in pharmaceutical industry. Their diversity in signaling, which can be modulated with drugs, permits the design of more effective and better-tolerated therapeutics. In this work, we have used rigid oligoproline backbones to generate bivalent ligands for the gastrin-releasing peptide receptor (GRPR) with a fixed distance between their recognition motifs. This allows the stabilization of GPCR dimers irrespective of their physiological occurrence and relevance, thus expanding the space for medicinal chemistry. Specifically, we observed that compounds presenting agonists or antagonists at 20- and 30-Å distance induce GRPR dimerization. Furthermore, we found that 1) compounds with two agonists at 20- and 30-Å distance that induce dimer formation show bias toward Gq efficacy, 2) dimers with 20- and 30-Å distance have different potencies toward ß-arrestin-1 and ß-arrestin-2, and 3) the divalent agonistic ligand with 10-Å distance specifically reduces Gq potency without affecting ß-arrestin recruitment, pointing toward an allosteric effect. In summary, we show that rigid oligoproline backbones represent a tool to develop ligands with biased GPCR signaling.

Pharmacological inhibition of mTORC1 increases CCKBR-specific tumor uptake of radiolabeled minigastrin analogue [177Lu]Lu-PP-F11N.

Rationale: A high tumor-to-healthy-tissue uptake ratio of radiolabeled ligands is an essential prerequisite for safe and effective peptide receptor radionuclide therapy (PRRT). In the present study, we searched for novel opportunities to increase tumor-specific uptake of the radiolabeled minigastrin analogue [177Lu]Lu-DOTA-(DGlu)6-Ala-Tyr-Gly-Trp-Nle-Asp-Phe-NH2 ([177Lu]Lu-PP-F11N), that targets the cholecystokinin B receptor (CCKBR) in human cancers. Methods: A kinase inhibitor library screen followed by proliferation and internalization assays were employed to identify compounds which can increase uptake of [177Lu]Lu-PP-F11N in CCKBR-transfected human epidermoid carcinoma A431 cells and natural CCKBR-expressing rat pancreatic acinar AR42J cells. Western blot (WB) analysis verified the inhibition of the signaling pathways and the CCKBR level, whereas the cell-based assay analyzed arrestin recruitment. Biodistribution and SPECT imaging of the A431/CCKBR xenograft mouse model as well as histological analysis of the dissected tumors were used for in vivo validation. Results: Our screen identified the inhibitors of mammalian target of rapamycin complex 1 (mTORC1), which increased cell uptake of [177Lu]Lu-PP-F11N. Pharmacological mTORC1 inhibition by RAD001 and metformin increased internalization of [177Lu]Lu-PP-F11N in A431/CCKBR and in AR42J cells. Analysis of protein lysates from RAD001-treated cells revealed increased levels of CCKBR (2.2-fold) and inhibition of S6 phosphorylation. PP-F11N induced recruitment of ß-arrestin1/2 and ERK1/2 phosphorylation. In A431/CCKBR-tumor bearing nude mice, 3 or 5 days of RAD001 pretreatment significantly enhanced tumor-specific uptake of [177Lu]Lu-PP-F11N (ratio [RAD001/Control] of 1.56 or 1.79, respectively), whereas metformin treatment did not show a significant difference. Quantification of SPECT/CT images confirmed higher uptake of [177Lu]Lu-PP-F11N in RAD001-treated tumors with ratios [RAD001/Control] of average and maximum concentration reaching 3.11 and 3.17, respectively. HE staining and IHC of RAD001-treated tumors showed a significant increase in necrosis (1.4% control vs.10.6% of necrotic area) and the reduction of proliferative (80% control vs. 61% of Ki67 positive cells) and mitotically active cells (1.08% control vs. 0.75% of mitotic figures). No significant difference in the tumor vascularization was observed after five-day RAD001 or metformin treatment. Conclusions: Our data demonstrates, that increased CCKBR protein level by RAD001 pretreatment has the potential to improve tumor uptake of [177Lu]Lu-PP-F11N and provides proof-of-concept for the development of molecular strategies aimed at enhancing the level of the targeted receptor, to increase the efficacy of PRRT and nuclear imaging.

New Insights into Arrestin Recruitment to GPCRs.

G protein-coupled receptors (GPCRs) are cellular master regulators that translate extracellular stimuli such as light, small molecules or peptides into a cellular response. Upon ligand binding, they bind intracellular proteins such as G proteins or arrestins, modulating intracellular signaling cascades. Here, we use a protein-fragment complementation approach based on nanoluciferase (split luciferase assay) to assess interaction of all four known human arrestins with four different GPCRs (two class A and two class B receptors) in live cells. Besides directly tagging the 11S split-luciferase subunit to the receptor, we also could demonstrate that membrane localization of the 11S subunit with a CAAX-tag allowed us to probe arrestin recruitment by endogenously expressed GPCRs. Varying the expression levels of our reporter constructs changed the dynamic behavior of our assay, which we addressed with an advanced baculovirus-based multigene expression system. Our detection assay allowed us to probe the relevance of each of the two arrestin binding sites in the different GPCRs for arrestin binding. We observed remarkable differences between the roles of each arresting binding site in the tested GPCRs and propose that the distinct advantages of our system for probing receptor interaction with effector proteins will help elucidate the molecular basis of GPCR signaling efficacy and specificity in different cell types.

A comparative cross-sectional study of personality traits in internists and surgeons.

BACKGROUND: A stereotype of surgeons' personality persists in the general public and among health-care professionals. Only a few studies have attempted to describe this "surgical personality" in detail. The aim of this study was to investigate the personality traits of surgeons compared with internists and to prove the existence of a stereotype among health-care professionals concerning surgeons. METHODS: To investigate the existence of a stereotype, nursing staff members in a public tertiary referral 900-bed hospital rated the personality traits of internists and surgeons. Simultaneously, all internists and surgeons in the same hospital were asked to complete the Freiburg Personality Inventory-the most frequently used German self-report form. RESULTS: Three hundred and thirty-four of 543 (62%) eligible nursing staff members participated; their responses confirmed the existence of a stereotype. A total of 253 of 284 eligible doctors completed the self-report form for a response rate of 89%. Compared with the general population, internists differed in most of 12 personality domains, whereas surgeons differed in 6 of 12 personality traits. The self-assessment revealed a statistically significant excess of achievement orientation (P = .00005) and extraversion (P < .00001) among surgeons and decreased aggressiveness (P = .00012) among internists. No significant difference was found between board-certified surgeons and internists in any of the 12 personality domains. CONCLUSION: This study identified a clear discrepancy between the self- and external assessment of personality but only among surgeons. This outcome provides an opportunity for surgeons to reflect on any potential lack of self-awareness and its impact on interdisciplinary patient care.