A High-Avidity, Thermostable, and Low-Cost Synthetic Capture for Ultrasensitive Detection and Quantification of Viral Antigens and Aerosols.

Lateral flow assays (LFAs) are currently the most popular point-of-care diagnostics, rapidly transforming disease diagnosis from expensive doctor checkups and laboratory-based tests to potential on-the-shelf commodities. Yet, their sensitive element, a monoclonal antibody, is expensive to formulate, and their long-term storage depends on refrigeration technology that cannot be met in resource-limited areas. In this work, LCB1 affibodies (antibody mimetic miniproteins) were conjugated to bovine serum albumin (BSA) to afford a high-avidity synthetic capture (LCB1-BSA) capable of detecting the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and virus like particles (VLPs). Substituting the monoclonal antibody 2B04 for LCB1-BSA (stable up to 60 °C) significantly improved the thermal stability, shelf life, and affordability of plasmonic-fluor-based LFAs (p-LFAs). Furthermore, this substitution significantly improved the sensitivity of p-LFAs toward the spike protein and VLPs with precise quantitative ability over 2 and 3 orders of magnitude, respectively. LCB1-BSA sensors could detect VLPs at 100-fold lower concentrations, and this improvement, combined with their robust nature, enabled us to develop an aerosol sampling technology to detect aerosolized viral particles. Synthetic captures like LCB1-BSA can increase the ultrasensitivity, availability, sustainability, and long-term accuracy of LFAs while also decreasing their manufacturing costs.

Major source categories of PM2.5 oxidative potential in wintertime Beijing and surroundings based on online dithiothreitol-based field measurements.

Fine particulate matter (PM2.5) causes millions of premature deaths each year worldwide. Oxidative potential (OP) has been proposed as a better metric for aerosol health effects than PM2.5 mass concentration alone. In this study, we report for the first time online measurements of PM2.5 OP in wintertime Beijing and surroundings based on a dithiothreitol (DTT) assay. These measurements were combined with co-located PM chemical composition measurements to identify the main source categories of aerosol OP. In addition, we highlight the influence of two distinct pollution events on aerosol OP (spring festival celebrations including fireworks and a severe regional dust storm). Source apportionment coupled with multilinear regression revealed that primary PM and oxygenated organic aerosol (OOA) were both important sources of OP, accounting for 41 ± 12 % and 39 ± 10 % of the OPvDTT (OP normalized by the sampled air volume), respectively. The small remainder was attributed to fireworks and dust, mainly resulting from the two distinct pollution events. During the 3.5-day spring festival period, OPvDTT spiked to 4.9 nmol min-1 m-3 with slightly more contribution from OOA (42 ± 11 %) and less from primary PM (31 ± 15 %). During the dust storm, hourly-averaged PM2.5 peaked at a very high value of 548 µg m-3 due to the dominant presence of dust-laden particles (88 % of total PM2.5). In contrast, only mildly elevated OPvDTT values (up to 1.5 nmol min-1 m-3) were observed during this dust event. This observation indicates that variations in OPvDTT cannot be fully explained using PM2.5 alone; one must also consider the chemical composition of PM2.5 when studying aerosol health effects. Our study highlights the need for continued pollution control strategies to reduce primary PM emissions, and more in-depth investigations into the source origins of OOA, to minimize the health risks associated with PM exposure in Beijing.

Rapid Direct Detection of SARS-CoV-2 Aerosols in Exhaled Breath at the Point of Care.

Airborne transmission via virus-laden aerosols is a dominant route for the transmission of respiratory diseases, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Direct, non-invasive screening of respiratory virus aerosols in patients has been a long-standing technical challenge. Here, we introduce a point-of-care testing platform that directly detects SARS-CoV-2 aerosols in as little as two exhaled breaths of patients and provides results in under 60 s. It integrates a hand-held breath aerosol collector and a llama-derived, SARS-CoV-2 spike-protein specific nanobody bound to an ultrasensitive micro-immunoelectrode biosensor, which detects the oxidation of tyrosine amino acids present in SARS-CoV-2 viral particles. Laboratory and clinical trial results were within 20% of those obtained using standard testing methods. Importantly, the electrochemical biosensor directly detects the virus itself, as opposed to a surrogate or signature of the virus, and is sensitive to as little as 10 viral particles in a sample. Our platform holds the potential to be adapted for multiplexed detection of different respiratory viruses. It provides a rapid and non-invasive alternative to conventional viral diagnostics.

Real-time environmental surveillance of SARS-CoV-2 aerosols.

Real-time surveillance of airborne SARS-CoV-2 virus is a technological gap that has eluded the scientific community since the beginning of the COVID-19 pandemic. Offline air sampling techniques for SARS-CoV-2 detection suffer from longer turnaround times and require skilled labor. Here, we present a proof-of-concept pathogen Air Quality (pAQ) monitor for real-time (5 min time resolution) direct detection of SARS-CoV-2 aerosols. The system synergistically integrates a high flow (~1000 lpm) wet cyclone air sampler and a nanobody-based ultrasensitive micro-immunoelectrode biosensor. The wet cyclone showed comparable or better virus sampling performance than commercially available samplers. Laboratory experiments demonstrate a device sensitivity of 77-83% and a limit of detection of 7-35 viral RNA copies/m3 of air. Our pAQ monitor is suited for point-of-need surveillance of SARS-CoV-2 variants in indoor environments and can be adapted for multiplexed detection of other respiratory pathogens of interest. Widespread adoption of such technology could assist public health officials with implementing rapid disease control measures.

Effect of Biomass Burning, Diwali Fireworks, and Polluted Fog Events on the Oxidative Potential of Fine Ambient Particulate Matter in Delhi, India.

We investigated the influence of biomass burning (BURN), Diwali fireworks, and fog events on the ambient fine particulate matter (PM2.5) oxidative potential (OP) during the postmonsoon (PMON) and winter season in Delhi, India. The real-time hourly averaged OP (based on a dithiothreitol assay) and PM2.5 chemical composition were measured intermittently from October 2019 to January 2020. The peak extrinsic OP (OPv: normalized by the volume of air) was observed during the winter fog (WFOG) (5.23 ± 4.6 nmol·min-1·m-3), whereas the intrinsic OP (OPm; normalized by the PM2.5 mass) was the highest during the Diwali firework-influenced period (29.4 ± 18.48 pmol·min-1·µg-1). Source apportionment analysis using positive matrix factorization revealed that traffic + resuspended dust-related emissions (39%) and secondary sulfate + oxidized organic aerosols (38%) were driving the OPv during the PMON period, whereas BURN aerosols dominated (37%) the OPv during the WFOG period. Firework-related emissions became a significant contributor (∼32%) to the OPv during the Diwali period (4 day period from October 26 to 29), and its contribution peaked (72%) on the night of Diwali. Discerning the influence of seasonal and episodic sources on health-relevant properties of PM2.5, such as OP, could help better understand the causal relationships between PM2.5 and health effects in India.

Sources of cellular oxidative potential of water-soluble fine ambient particulate matter in the Midwestern United States.

We investigated the spatiotemporal distribution and sources of cellular oxidative potential (OP) in the Midwest US. Weekly samples were collected from three urban [Chicago (IL), Indianapolis (IN), and St. Louis (MO)], one rural [Bondville (IL], and one roadside site [Champaign (IL)] for a year (May 2018 to May 2019), and analyzed for water-soluble cellular OP using a macrophage reactive oxygen species (ROS) assay. Chemical composition of the samples including several carbonaceous components, inorganic ions, and water-soluble elementals, were also analyzed. The emission sources contributing to water-soluble cellular OP and PM2.5 mass were analyzed using positive matrix factorization. The secondary organic aerosols contributed substantially (≥54%) to PM2.5 cellular OP at urban sites, while the roadside and rural OP were dominated by road dust (54%) and agricultural activities (62%), respectively. However, none of these sources contributed substantially to the PM2.5 mass (≤21%). Other sources contributing significantly to the PM2.5 mass, i.e., secondary sulfate and nitrate, biomass burning and coal combustion (14-26%) contributed minimally to the cellular OP (≤13%). Such divergent profiles of the emission sources contributing to cellular OP vs. PM2.5 mass demonstrate the need of considering more health-relevant metrics such as OP in the design of air pollution control strategies.

Synergistic and antagonistic interactions among organic and metallic components of the ambient particulate matter (PM) for the cytotoxicity measured by Chinese hamster ovary cells.

Although PM2.5 toxicity is known to be related to its chemical composition, the effect of interactions among various particles' components on the toxicity is not well explored. To understand these interactions, especially metals and organic compounds on PM2.5 cytotoxicity, we chose several redox-active substances known to be present in the ambient particles such as metals (Cu, Fe, and Mn) and quinones [9,10-phenanthraquinone (PQ), 1,2-naphthoquinone (1,2-NQ), 1,4-naphthoquinone (1,4-NQ), and 5-hydroxy-1,4-naphthoquinone (5,H-1,4-NQ)]. Cytotoxicity was assessed through a Chinese hamster ovary (CHO) cells assay and expressed by a median lethal concentration (LC50). Two methods were employed to assess the interactions. In the first method, we tested the impact of nontoxic level of a component on the LC50 of other components. In the second method, we mixed two components in different concentration ratios to expose the cells and calculated a mixture toxicity index (MTI). MTI is a composite value to quantify the nature of interactions such that the interactions are considered synergistic when MTI > 1, additive when 0 < MTI ≤ 1 and antagonistic when MTI < 0. The interactions between quinones and metals were largely synergistic by both methods. To further assess the environmental relevance of these mixtures, we extracted organic compounds termed as water-soluble Humic-like substances (HULIS) from real ambient PM samples and mixed them with individual metals. A similar pattern, as observed from the interaction of quinones and metals, was found. Moreover, the interactions became more synergistic as the relative concentration of metals with respect to water-soluble HULIS was decreased in these mixtures. With environmentally relevant mass concentration ratios of organics to metals (75-7500), the interactions were strongly synergistic (MTI = 1-115). These results indicate the importance of incorporating the interaction among various PM components for estimating the net toxicity of ambient PM2.5.

Dry Heat as a Decontamination Method for N95 Respirator Reuse.

A pandemic such as COVID-19 can cause a sudden depletion of the worldwide supply of respirators, forcing healthcare providers to reuse them. In this study, we systematically evaluated dry heat treatment as a viable option for the safe decontamination of N95 respirators (1860, 3M) before their reuse. We found that the dry heat generated by an electric cooker (100 °C, 5% relative humidity, 50 min) effectively inactivated Tulane virus (TV, >5.2-log10 reduction), rotavirus (RV, >6.6-log10 reduction), adenovirus (AdV, >4.0-log10 reduction), and transmissible gastroenteritis virus (TGEV, >4.7-log10 reduction). The respirator integrity (determined on the basis of the particle filtration efficiency and quantitative fit testing) was not compromised after 20 cycles of a 50 min dry heat treatment. On the basis of these results, dry heat decontamination generated by an electric cooker (e.g., rice cookers, instant pots, and ovens) could be an effective and accessible decontamination method for the safe reuse of N95 respirators. We recommend users measure the temperature during decontamination to ensure the respirator temperature can be maintained at 100 °C for 50 min.

Evaluation of recycled concrete aggregates for their suitability in construction activities: An experimental study.

Construction and demolition waste disposal is a major challenge in developing nations due to its ever increasing quantities. In this study, the recycling potential of waste concrete as aggregates in construction activities was studied. The metal leaching from the recycled concrete aggregates (RCA) collected from the demolition site of a 50year old building, was evaluated by performing three different leaching tests (compliance, availability and Toxic Characteristic Leaching Procedure). The metal leaching was found mostly within the permissible limit except for Hg. Several tests were performed to determine the physical and mechanical properties of the fine and coarse aggregates produced from recycled concrete. The properties of recycled aggregates were found to be satisfactory for their utilization in road construction activities. The suitability of using recycled fine and coarse aggregates with Portland pozzolanic cement to make a sustainable and environmental friendly concrete mix design was also analyzed. No significant difference was observed in the compressive strength of various concrete mixes prepared by natural and recycled aggregates. However, only the tensile strength of the mix prepared with 25% recycled fine aggregates was comparable to that of the control concrete. For other mixes, the tensile strength of the concrete was found to drop significantly. In summary, RCA should be considered seriously as a building material for road construction, mass concrete works, lightly reinforced sections, etc. The present work will be useful for the waste managers and policy makers particularly in developing nations where proper guidelines are still lacking.