The source control effect of personal protection equipment and physical barrier on short-range airborne transmission.

In order to control the spread of Covid-19, authorities provide various prevention guidelines and recommendations for health workers and the public. Personal protection equipment (PPE) and physical barrier are the most widely applied prevention measures in practice due to their affordability and ease of implementation. This study aims to investigate the effect of PPE and physical barriers on mitigating the short-range airborne transmission between two people in a ventilated environment. Four types of PPE (surgical mask, two types of face shield, and mouth visor), and two different sizes of the physical barrier were tested in a controlled environment with two life-size breathing thermal manikins. The PPE was worn by the source manikin to test the efficiency of source control. The measurement results revealed that the principles of PPE on preventing short-range droplet and airborne transmission are different. Instead of filtering the fine droplet nuclei, they mainly redirect the virus-laden exhalation jet and avoid the exhaled flow entering the target's inhalation region. Physical barriers can block the spreading of droplet nuclei and create a good micro environment at short distances between persons. However, special attention should be paid to arranging the physical barrier and operating the ventilation system to avoid the stagnant zone where the contaminant accumulates.

Human exhalation characterization with the aid of schlieren imaging technique.

The purpose of this paper is to determine the dispersion and distribution characteristics of exhaled airflow for accurate prediction of disease transmission. The development of airflow dynamics of human exhalation was characterized using nonhazardous schlieren photography technique, providing a visualization and quantification of turbulent exhaled airflow from 18 healthy human subjects whilst standing and lying. The flow shape of each breathing pattern was characterized by two angles and averaged values of 18 subjects. Two exhaled air velocities, u m and u p , were measured and compared. The mean peak centerline velocity, u m was found to decay correspondingly with increasing horizontal distance x in a form of power function. The mean propagation velocity, u p was found to correlate with physiological parameters of human subjects. This was always lower than u m at the mouth/nose opening, due to a vortex like airflow in front of a single exhalation cycle. When examining the talking and breathing process between two persons, the potential infectious risk was found to depend on their breathing patterns and spatial distribution of their exhaled air. Our study when combined with information on generation and distributions of pathogens could provide a prediction method and control strategy to minimize infection risk between persons in indoor environments.

Solvent dependent photosensitized singlet oxygen production from an Ir(III) complex: pointing to problems in studies of singlet-oxygen-mediated cell death.

A cationic cyclometallated Ir(III) complex with 1,10-phenanthroline and 2-phenylpyridine ligands photosensitizes the production of singlet oxygen, O2(a(1)Δ(g)), with yields that depend appreciably on the solvent. In water, the quantum yield of photosensitized O2(a(1)Δ(g)) production is small (ϕ(Δ) = 0.036 ± 0.008), whereas in less polar solvents, the quantum yield is much larger (ϕ(Δ) = 0.54 ± 0.05 in octan-1-ol). A solvent effect on ϕ(Δ) of this magnitude is rarely observed and, in this case, is attributed to charge-transfer-mediated processes of non-radiative excited state deactivation that are more pronounced in polar solvents and that kinetically compete with energy transfer to produce O2(a(1)Δ(g)). A key component of this non-radiative deactivation process, electronic-to-vibrational energy transfer, is also manifested in pronounced H2O/D2O isotope effects that indicate appreciable coupling between the Ir(III) complex and water. This Ir(III) complex is readily incorporated into HeLa cells and, upon irradiation, is cytotoxic as a consequence of the O2(a(1)Δ(g)) thus produced. The data reported herein point to a pervasive problem in mechanistic studies of photosensitized O2(a(1)Δ(g))-mediated cell death: care must be exercised when interpreting the effective cytotoxicity of O2(a(1)Δ(g)) photosensitizers whose photophysical properties depend strongly on the local environment. Specifically, the photophysics of the sensitizer in bulk solutions may not accurately reflect its intracellular behavior, and the control and quantification of the O2(a(1)Δ(g)) "dose" can be difficult in vivo.

Effect of chromophore encapsulation on linear and nonlinear optical properties: the case of "miniSOG", a protein-encased flavin.

Linear and nonlinear spectroscopic parameters of flavin mononucleotide, FMN, have been examined both experimentally and computationally under conditions in which FMN is (1) solvated in a buffered aqueous solution, and (2) encased in a protein that is likewise solvated in a buffered aqueous solution. The latter was achieved using "miniSOG" which is an FMN-containing protein engineered from Arabidopsis thaliana phototropin 2. Although it is reasonable to expect that the encasing protein could have an appreciable effect, certainly on the nonlinear two-photon absorption cross section, we find that replacing the dynamic aqueous environment with the more static protein environment does little to influence the spectroscopic properties of FMN. The experimental and computational studies are consistent in this regard, and this agreement indicates that comparatively high-level computational methods can indeed be used with success on large chromophores with a complicated local environment. The results of the present study facilitate the much-needed development of well-characterized and readily-controlled chromophores suitable for use as intracellular sensitizers and fluorophores.

Oxygen-dependent photochemistry and photophysics of "miniSOG," a protein-encased flavin.

Selected photochemical and photophysical parameters of flavin mononucleotide (FMN) have been examined under conditions in which FMN is (1) solvated in a buffered aqueous solution, and (2) encased in a protein likewise solvated in a buffered aqueous solution. The latter was achieved using the so-called "mini Singlet Oxygen Generator" (miniSOG), an FMN-containing flavoprotein engineered from Arabidopsis thaliana phototropin 2. Although FMN is a reasonably good singlet oxygen photosensitizer in bulk water (Ï•Δ = 0.65 ± 0.04), enclosing FMN in this protein facilitates photoinitiated electron-transfer reactions (Type-I chemistry) at the expense of photosensitized singlet oxygen production (Type-II chemistry) and results in a comparatively poor yield of singlet oxygen (Ï•Δ = 0.030 ± 0.002). This observation on the effect of the local environment surrounding FMN is supported by a host of spectroscopic and chemical trapping experiments. The results of this study not only elucidate the behavior of miniSOG but also provide useful information for the further development of well-characterized chromophores suitable for use as intracellular sensitizers in mechanistic studies of reactive oxygen species.

Singlet-oxygen-mediated cell death using spatially-localized two-photon excitation of an extracellular sensitizer.

Controlling and quantifying the photosensitized production of singlet oxygen are key aspects in mechanistic studies of oxygen-dependent photoinitiated cell death. In this regard, the commonly accepted practice of using intracellular photosensitizers is, unfortunately, plagued by problems that include the inability to accurately (1) quantify the sensitizer concentration in the irradiated domain and (2) control the local environment that influences light delivery and sensitizer photophysics. However, capitalizing on the fact that singlet oxygen produced outside a cell is also cytotoxic, many of these problems can be avoided with the use of an extracellular sensitizer. For the present study, a hydrophilic dendrimer-encased membrane-impermeable sensitizer was used to generate an extracellular population of singlet oxygen upon spatially localized two-photon irradiation. Through the use of this sensitizer and this approach, it is now possible to better control the singlet oxygen dose in microscope-based time- and space-resolved single cell experiments. Thus, we provide a solution to a limiting problem in mechanistic studies of singlet-oxygen-mediated cell death.

Direct site-directed photocoupling of proteins onto surfaces coated with beta-cyclodextrins.

A method called Dock'n'Flash was developed to offer site-specific capture and direct UVA-induced photocoupling of recombinant proteins. The method involves the tagging of recombinant proteins with photoreactive p-benzoyl-L-phenylalanine (pBpa) by genetic engineering. The photoreactive pBpa tag is used for affinity capture of the recombinant protein by beta-cyclodextrin (beta-CD), which provides hydrogen atoms to be abstracted in the photocoupling process. To exemplify the method, a recombinant, folded, and active N27pBpa mutant of cutinase from Fusarium solani pisi was produced in E. coli. Insertion of pBpa was verified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy. A molecular dynamic simulation, with water as solvent, showed high solvent accessibility of the pBpa benzophenone group in N27pBpa-cutinase mutant. The formation of an inclusion complex between the benzophenone group of N27pBpa-cutinase and beta-CD was shown, and an apparent K(d) of 1.65 mM was determined using (1)H NMR. Photocoupling of beta-CD to N27pBpa-cutinase in a 1:1 ratio, upon UVA irradiation at 360 +/- 20 nm, was shown by MALDI-TOF mass spectroscopy. UVA photoimmobilization of N27pBpa-cutinase on quartz slides coated with beta-CD was achieved from liquid or dry films by total internal reflection fluorescence (TIRF). The Dock'n'Flash method offers a solution for direct photocoupling and patterning of recombinant proteins onto surfaces with site-specific attachment.