Structural and functional analysis of the minimal orthomyxovirus-like polymerase of Tilapia Lake Virus from the highly diverged Amnoonviridae family.

Tilapia Lake Virus (TiLV), a recently discovered pathogen of tilapia fish, belongs to the Amnoonviridae family from the Articulavirales order. Its ten genome segments have characteristic conserved ends and encode proteins with no known homologues, apart from the segment 1, which encodes an orthomyxo-like RNA-dependent-RNA polymerase core subunit. Here we show that segments 1-3 encode respectively the PB1, PB2 and PA-like subunits of an active heterotrimeric polymerase that maintains all domains found in the distantly related influenza polymerase, despite an unprecedented overall size reduction of 40%. Multiple high-resolution cryo-EM structures of TiLV polymerase in pre-initiation, initiation and active elongation states, show how it binds the vRNA and cRNA promoters and performs RNA synthesis, with both transcriptase and replicase configurations being characterised. However, the highly truncated endonuclease-like domain appears inactive and the putative cap-binding domain is autoinhibited, emphasising that many functional aspects of TiLV polymerase remain to be elucidated.

Tandem mass spectrometry and density functional theory of RDX fragmentation pathways: Role of ion-molecule complexes in loss of NO3 and lack of molecular ion peak.

RATIONALE: Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is an explosive compound that finds a wide range of military and civilian applications. RDX has been a target in environmental matrices by gas chromatography/tandem mass spectrometry (GC/MS/MS). MS/MS in negative chemical ionization (NCI) mode of RDX provides important fragmentation patterns that are useful for structural elucidation. The fragmentation patterns are needed for proper identification of precursor and product ions in analytical methods that depend on MS/MS approaches for a reliable identification of RDX. METHODS: This study focuses on the MS fragmentation mechanisms of RDX in NCI mode using both MS/MS and density functional theory (DFT). The DFT studies were performed at the B3LYP/6-311G(d,p) level of theory. RESULTS: The DFT results showed that NCI of RDX leads to the formation of an anion-molecule complex that was energetically more stable than the RDX anion. The fragmentation proceeds through two pathways, leading to the loss of NO(2) and NO(3). The loss of NO(3) takes place in an anion-molecule complex leading to the formation of characteristic nitroso group fragment ions. Using the fragmentation schemes, important ion structures are proposed including structures for m/z 160, 129, 102, and 86. CONCLUSIONS: The results demonstrate the importance of both charge-induced and charge-remote dissociations in RDX pathways. The ion structures identified along the pathways could be used as targets in analytical methods for reliable identification purposes.

Creating an environment for diversity in dental schools: one school's approach.

Recent reports have indicated the need to improve the diversity in the dental profession's workforce. The enrollment of underrepresented minority students in the nation's dental schools must increase to accomplish this goal. A complex change process within the dental schools is required to prepare schools to enroll a more diverse student body. While each dental school in the United States is unique, a product of its history and institutional culture, and will, therefore, create an environment for diversity in different ways, it is appropriate to describe lessons learned in individual schools as they strive for diversity. The purpose of this paper is to describe how one dental school, the Columbia University School of Dental and Oral Surgery, approached diversity, so that appropriate strategies can be shared among schools.