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The Escherichia coli protein RecA catalyzes DNA strand exchange necessary for DNA repair and homologous genetic recombination. The binding of adenosine triphosphate (ATP) to RecA induces a conformation with a high affinity for DNA. However, adenosine diphosphate (ADP) binding results in a RecA conformation with a lower affinity for DNA. In an attempt to identify and compare the specific structural changes induced by the binding of the two nucleotides, we have utilized Fourier transform infrared difference spectroscopy in conjunction with the photolytic release of caged nucleotides. RecA-ADP minus RecA and RecA-ATP minus RecA difference spectra contain distinct, reproducible changes in the 1800-1300 cm -1 region associated with nucleotide binding to RecA. In addition, the first difference FT-IR evidence suggests that small structural perturbations in the RecA and/or nucleotide are responsible for differences between the low and high DNA affinity states of the protein. The results presented here are consistent with studies that predict the involvement of key amino acid side chains and also reveal secondary structural changes may be associated with nucleotide binding to RecA in the absence of DNA.

Cell lines are the mainstay in understanding the biology of COVID-19 infection but do not recapitulate many of the complexities of human infection. The use of human lung tissue is one solution for the study of such novel respiratory pathogens. We hypothesized that a cryopreserved bank of human lung tissue would allow for the ex vivo study of the interindividual heterogeneity of host response to SARS-CoV-2, thus providing a bridge between studies with cell lines and studies in animal models. We generated a cryobank of tissues from 21 donors, many of whom had clinical risk factors for severe COVID-19. Cryopreserved tissues preserved 90% cell viability and contained heterogenous populations of metabolically active epithelial, endothelial, and immune cell subsets of the human lung. Samples were readily infected with HCoV-OC43 and SARS-CoV-2 and demonstrated comparable susceptibility to infection. In contrast, we observed a marked donor-dependent heterogeneity in the expression of IL6 , CXCL8 , and IFNB1 in response to SARS-CoV-2. Treatment of tissues with dexamethasone and the experimental drug N-hydroxycytidine suppressed viral growth in all samples, whereas chloroquine and remdesivir had no detectable effect. Metformin and sirolimus, molecules with predicted but unproven antiviral activity, each suppressed viral replication in tissues from a subset of donors. In summary, we developed a system for the ex vivo study of human SARS-CoV-2 infection using primary human lung tissue from a library of donor tissues. This model may be useful for drug screening and for understanding basic mechanisms of COVID-19 pathogenesis.

Chloramphenicol (CAP) is commonly employed in veterinary clinics, but illegal and uncontrollable consumption can result in its potential contamination in environmental soil, and aquatic matrix, and thereby, regenerating microbial resistance, and antibiotic-resistant genes. Adsorption by efficient, and recyclable adsorbents such as mesoporous carbons (MPCs) is commonly regarded as a “green and sustainable” approach. Herein, the MPCs were facilely synthesized via the pyrolysis of the metal–organic framework Fe3O(BDC)3 with calcination temperatures (x °C) between 600 and 900 °C under nitrogen atmosphere. The characterization results pointed out mesoporous carbon matrix (MPC700) coating zero-valent iron particles with high surface area (~225 m2/g). Also, significant investigations including fabrication condition, CAP concentration, effect of pH, dosage, and ionic strength on the absorptive removal of CAP were systematically studied. The optimal conditions consisted of pH = 6, concentration 10 mg/L and dose 0.5 g/L for the highest chloramphenicol removal efficiency at nearly 100% after 4 h. Furthermore, the nonlinear kinetic and isotherm adsorption studies revealed the monolayer adsorption behavior of CAP onto MPC700 and Fe3O(BDC)3 materials via chemisorption, while the thermodynamic studies implied that the adsorption of CAP was a spontaneous process. Finally, adsorption mechanism including H-bonding, electrostatic attraction, π–π interaction, and metal–bridging interaction was proposed to elucidate how chloramphenicol molecules were adsorbed on the surface of materials. With excellent maximum adsorption capacity (96.3 mg/g), high stability, and good recyclability (4 cycles), the MPC700 nanocomposite could be utilized as a promising alternative for decontamination of chloramphenicol antibiotic from wastewater.

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