Autoclave sterilization and ethanol treatment of re-used surgical masks and N95 respirators during COVID-19: impact on their performance and integrity.

BACKGROUND: An exceptionally high demand for surgical masks and N95 filtering facepiece respirators (FFRs) during the COVID-19 pandemic has considerably exceeded their supply. These disposable devices are generally not approved for routine decontamination and re-use as a standard of care, while this practice has widely occurred in hospitals. The US Centers for Disease Control and Prevention allowed it "as a crisis capacity strategy". However, limited testing was conducted on the impact of specific decontamination methods on the performance of N95 FFRs and no data was presented for surgical masks. AIM: We evaluated common surgical masks and N95 respirators with respect to the changes in their performance and integrity resulting from autoclave sterilization and a 70% ethanol treatment; these methods are frequently utilized for re-used filtering facepieces in hospitals. METHODS: The filter collection efficiency and pressure drop were determined for unused masks and N95 FFRs, and for those subjected to the treatments in a variety of ways. The collection efficiency was measured for particles of approximately 0.037-3.2 µm to represent aerosolized single viruses, their agglomerates, bacteria and larger particle carriers. FINDINGS: The initial collection efficiency and the filter breathability may be compromised by sterilization in an autoclave and ethanol treatment. The effect depends on a protective device, particle size, breathing flow rate, type of treatment and other factors. Additionally, physical damages were observed in N95 respirators after autoclaving. CONCLUSION: Strategies advocating decontamination and re-use of filtering facepieces in hospitals should be re-assessed considering the data obtained in this study.

DNA-methyltransferase SsoII interaction with own promoter region binding site.

The investigation of Sso II DNA-methyltransferase (M.Sso II) interaction with the intergenic region of Sso II restriction-modification system was carried out. Seven guanine residues protected by M. Sso II from methylation with dimethylsulfate and thus probably involved in enzyme-DNA recognition were identified. Six of them are located symmetrically within the 15 bp inverted repeat inside the Sso II promoter region. The crosslinking of Sso II methyltransferase with DNA duplexes containing 5-bromo-2'-deoxyuridine (br5dU) instead of thymidine was performed. The crosslinked products were obtained in all cases, thus proving that tested thymines were in proximity with enzyme. The ability to produce the crosslinked products in one case was 2-5-fold higher than in other ones. This allowed us to imply that thymine residue in this position of the inverted repeat could be in contact with M. Sso II. Based on the experimental data, two symmetrical 4 bp clusters (GGAC), which could be involved in the interaction with M. Sso II in the DNA-protein complex, were identified. The model of M. Sso II interaction with its own promoter region was proposed.

Specific binding of sso II DNA methyltransferase to its promoter region provides the regulation of sso II restriction-modification gene expression.

The regulation of the Sso II restriction-modification system from Shigella sonnei was studied in vivo and in vitro . In lacZ fusion experiments, Sso II methyltransferase (M. Sso II) was found to repress its own synthesis but stimulate expression of the cognate restriction endonuclease (ENase). The N-terminal 72 amino acids of M. Sso II, predicted to form a helix-turn-helix (HTH) motif, was found to be responsible for the specific DNA-binding and regulatory function of M. Sso II. Similar HTH motifs are predicted in the N-terminus of a number of 5-methylcytosine methyltransferases, particularly M. Eco RII, M.dcm and M. Msp I, of which the ability to regulate autogenously has been proposed. In vitro, the binding of M. Sso II to its target DNA was investigated using a mobility shift assay. M. Sso II forms a specific and stable complex with a 140 bp DNA fragment containing the promoter region of Sso II R-M system. The dissociation constant (Kd) was determined to be 1.5x10(-8) M. DNaseI footprinting experiments demonstrated that M. Sso II protects a 48-52 bp region immediately upstream of the M. Sso II coding sequence which includes the predicted -10 promoter sequence of M. Sso II and the -10 and -35 sequences of R. Sso II.