Microbial natural products are specialized metabolites that are sources of many bioactive compounds including antibiotics, antifungals, antiparasitics, anticancer agents, and probes of biology. The assembly of libraries of producers of natural products has traditionally been the province of the pharmaceutical industry. This sector has gathered significant historical collections of bacteria and fungi to identify new drug leads with outstanding outcomes-upwards of 60% of drug scaffolds originate from such libraries. Despite this success, the repeated rediscovery of known compounds and the resultant diminishing chemical novelty contributed to a pivot from this source of bioactive compounds toward more tractable synthetic compounds in the drug industry. The advent of advanced mass spectrometry tools, along with rapid whole genome sequencing and in silico identification of biosynthetic gene clusters that encode the machinery necessary for the synthesis of specialized metabolites, offers the opportunity to revisit microbial natural product libraries with renewed vigor. Assembling a suitable library of microbes and extracts for screening requires the investment of resources and the development of methods that have customarily been the proprietary purview of large pharmaceutical companies. Here, we report a perspective on our efforts to assemble a library of natural product-producing microbes and the establishment of methods to extract and fractionate bioactive compounds using resources available to most academic labs. We validate the library and approach through a series of screens for antimicrobial and cytotoxic agents. This work serves as a blueprint for establishing libraries of microbial natural product producers and bioactive extract fractions suitable for screens of bioactive compounds.

Obligate intracellular parasites must efficiently invade host cells in order to mature and be transmitted. For the malaria parasite Plasmodium falciparum, invasion of host red blood cells (RBCs) is essential. Here we describe a parasite-specific transcription factor PfAP2-I, belonging to the Apicomplexan AP2 (ApiAP2) family, that is responsible for regulating the expression of genes involved in RBC invasion. Our genome-wide analysis by ChIP-seq shows that PfAP2-I interacts with a specific DNA motif in the promoters of target genes. Although PfAP2-I contains three AP2 DNA-binding domains, only one is required for binding of the target genes during blood stage development. Furthermore, we find that PfAP2-I associates with several chromatin-associated proteins, including the Plasmodium bromodomain protein PfBDP1 and that complex formation is associated with transcriptional regulation. As a key regulator of red blood cell invasion, PfAP2-I represents a potential new antimalarial therapeutic target.

A gas-phase Advanced Oxidation Process (gAOP) was evaluated for decontaminating N95 and surgical masks. The continuous process was based on the generation of hydroxyl-radicals via the UV-C (254 nm) photo-degradation of hydrogen peroxide and ozone. The decontamination efficacy of the gAOP was dependent on the orientation of the N95 mask passing through the gAOP unit with those positioned horizontally enabling greater exposure to hydroxyl-radicals compared to when arranged vertically. The lethality of gAOP was independent of the applied hydrogen peroxide concentration (2-6% v/v) but was significantly (P<0.05) higher when H2O2 was introduced into the unit at 40 ml/min compared to 20 ml/min. A suitable treatment for N95 masks was identified as 3% v/v hydrogen peroxide delivered into the gAOP reactor at 40 ml/min with continuous introduction of ozone gas and a UV-C dose of 113 mJ/cm2 (30 s processing time). The treatment supported >6 log CFU decrease in Geobacillus stearothermophilus endospores, > 8 log reduction of human coronavirus 229E, and no detection of Escherichia coli K12 on the interior and exterior of masks. There was no negative effect on the N95 mask fitting or particulate efficacy after 20 passes through the gAOP system. No visual changes or hydrogen peroxide residues were detected (<1 ppm) in gAOP treated masks. The optimized gAOP treatment could also support >6 log CFU reduction of endospores inoculated on the interior or exterior of surgical masks. G. stearothermophilus Apex spore strips could be applied as a biological indicator to verify the performance of gAOP treatment. Also, a chemical indicator based on the oxidative polymerization of pyrrole was found suitable for reporting the generation of hydroxyl-radicals. In conclusion, gAOP is a verifiable treatment that can be applied to decontaminate N95 and surgical masks without any negative effects on functionality.