Existing drugs are slow to eradicate Mycobacterium tuberculosis (Mtb) in patients and have failed to control tuberculosis globally. One reason may be that host conditions impair Mtb's replication, reducing its sensitivity to most antiinfectives. We devised a highthroughput screen for compounds that kill Mtb when its replication has been halted by reactive nitrogen intermediates (RNIs), acid, hypoxia, and a fatty acid carbon source. At concentrations routinely achieved in human blood, oxyphenbutazone (OPB), an inexpensive anti-inflammatory drug, was selectively mycobactericidal to nonreplicating (NR) Mtb. Its cidal activity depended on mild acid and was augmented by RNIs and fatty acid. Acid and RNIs fostered OPB's 4-hydroxylation. The resultant 4-butyl-4-hydroxy-1-(4-hydroxyphenyl)-2-phenylpyrazolidine-3,5-dione (4-OH-OPB) killed both replicating and NR Mtb, including Mtb resistant to standard drugs. 4-OH-OPB depleted flavins and formed covalent adducts with N-acetyl-cysteine and mycothiol. 4-OH-OPB killed Mtb synergistically with oxidants and several antituberculosis drugs. Thus, conditions that block Mtb's replication modify OPB and enhance its cidal action. Modified OPB kills both replicating and NR Mtb and sensitizes both to host-derived and medicinal antimycobacterial agents.

We report two series of novel cephalosporins that are bactericidal to Mycobacterium tuberculosis alone of the pathogens tested, which only kill M. tuberculosis when its replication is halted by conditions resembling those believed to pertain in the host, and whose bactericidal activity is not dependent upon or enhanced by clavulanate, a β-lactamase inhibitor. The two classes of cephalosporins bear an ester or alternatively an oxadiazole isostere at C-2 of the cephalosporin ring system, a position that is almost exclusively a carboxylic acid in clinically used agents in the class. Representatives of the series kill M. tuberculosis within macrophages without toxicity to the macrophages or other mammalian cells.

Herein we provide a living summary of the data generated during the COVID Moonshot project focused on the development of SARS-CoV-2 main protease (Mpro) inhibitors. Our approach uniquely combines crowdsourced medicinal chemistry insights with high throughput crystallography, exascale computational chemistry infrastructure for simulations, and machine learning in triaging designs and predicting synthetic routes. This manuscript describes our methodologies leading to both covalent and non-covalent inhibitors displaying protease IC50 values under 150 nM and viral inhibition under 5 uM in multiple different viral replication assays. Furthermore, we provide over 200 crystal structures of fragment-like and lead-like molecules in complex with the main protease. Over 1000 synthesized and ordered compounds are also reported with the corresponding activity in Mpro enzymatic assays using two different experimental setups. The data referenced in this document will be continually updated to reflect the current experimental progress of the COVID Moonshot project, and serves as a citable reference for ensuing publications. All of the generated data is open to other researchers who may find it of use.

The detection and successful typing of dengue virus (DENV) from patients with suspected dengue fever is important both for the diagnosis of the disease and for the implementation of epidemiologic control measures. A technique for the multiplex detection and typing of DENV serotypes 1 to 4 (DENV-1 to DENV-4) from clinical samples by PCR-ligase detection reaction (LDR) has been developed. A serotype-specific PCR amplifies the regions of genes C and E simultaneously. The two amplicons are targeted in a multiplex LDR, and the resultant fluorescently labeled ligation products are detected on a universal array. The assay was optimized using 38 DENV strains and was evaluated with 350 archived acute-phase serum samples. The sensitivity of the assay was 98.7%, and its specificity was The dengue virus (DENV), a mosquito-borne flavivirus, consists of four closely related but genetically distinct antigenic serotypes: DENV serotype 1 (DENV-1), DENV-2, DENV-3, and DENV-4. It is tropical and subtropical in distribution and is prevalent in Asia, Africa, and Central and South America (45). Infection with any of the four serotypes of DENV may cause a mild febrile illness, dengue fever (DF). In some cases, however, more-severe manifestations, such as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), occur; these may prove fatal without proper early intervention (15).Geographic spread of both the mosquito vector and the virus over the past 25 years has led to the increased occurrence of epidemic DF/DHF/DSS, making dengue a major global health problem. The disease is endemic in more than 100 countries, with an estimated 2.5 billion people at risk of infection. It is estimated that 50 million DENV infections occur each year, with 500,000 cases of DHF and at least 22,000 deaths, mainly in children (31,32,45; WHO/WPRO/SEARO meeting on DengueNet implementation in Southeast Asia and the Western Pacific, Kuala Lumpur, Malaysia, 11 to 13 December 2003).DENV infection confers lifelong serotype-specific immunity. Multiple infections with different DENV serotypes occur in regions of hyperendemicity (31, 35). Secondary infections with a different DENV serotype are major risk factors for DHF and DSS (13, 14, 39) due to antibody-dependent enhancement of disease (35). Serotype identification and the differentiation of primary and secondary infections are therefore important both for patient management and for the implementation of public health measures (26,33).The diagnosis of DENV infection and the typing of DENV serotypes can be confirmed using viral isolation techniques, serology, or molecular methods. Virus isolation is the gold standard for detection but requires 7 to 10 days and is often insensitive (26). Serological tests for the detection of viral antibodies, such as immunoglobulin M and immunoglobulin G antibody capture enzyme-linked immunosorbent assays, require the demonstration of a rise in antibody titer from an acute-phase to a convalescent-phase serum sample and therefore have little impact on patient management (24,...

We have developed a novel high-throughput PCR-ligase detection reaction-capillary electrophoresis (PCR-LDR