A novel live-attenuated vaccine candidate for mayaro Fever.

Mayaro virus (MAYV) is an emerging, mosquito-borne alphavirus that causes a dengue-like illness in many regions of South America, and which has the potential to urbanize. Because no specific treatment or vaccine is available for MAYV infection, we capitalized on an IRES-based approach to develop a live-attenuated MAYV vaccine candidate. Testing in infant, immunocompetent as well as interferon receptor-deficient mice demonstrated a high degree of attenuation, strong induction of neutralizing antibodies, and efficacy against lethal challenge. This vaccine strain was also unable to infect mosquito cells, a major safety feature for a live vaccine derived from a mosquito-borne virus. Further preclinical development of this vaccine candidate is warranted to protect against this important emerging disease

Casimir effect due to a single boundary as a manifestation of the Weyl problem

The Casimir self-energy of a boundary is ultraviolet-divergent. In many cases the divergences can be eliminated by methods such as zeta-function regularization or through physical arguments (ultraviolet transparency of the boundary would provide a cutoff). Using the example of a massless scalar field theory with a single Dirichlet boundary we explore the relationship between such approaches, with the goal of better understanding the origin of the divergences. We are guided by the insight due to Dowker and Kennedy (1978) and Deutsch and Candelas (1979), that the divergences represent measurable effects that can be interpreted with the aid of the theory of the asymptotic distribution of eigenvalues of the Laplacian discussed by Weyl. In many cases the Casimir self-energy is the sum of cutoff-dependent (Weyl) terms having geometrical origin, and an "intrinsic" term that is independent of the cutoff. The Weyl terms make a measurable contribution to the physical situation even when regularization methods succeed in isolating the intrinsic part. Regularization methods fail when the Weyl terms and intrinsic parts of the Casimir effect cannot be clearly separated. Specifically, we demonstrate that the Casimir self-energy of a smooth boundary in two dimensions is a sum of two Weyl terms (exhibiting quadratic and logarithmic cutoff dependence), a geometrical term that is independent of cutoff, and a non-geometrical intrinsic term. As by-products we resolve the puzzle of the divergent Casimir force on a ring and correct the sign of the coefficient of linear tension of the Dirichlet line predicted in earlier treatments.Comment: 13 pages, 1 figure, minor changes to the text, extra references added, version to be published in J. Phys.

Differential Alphavirus Defective RNA Diversity between Intracellular and Extracellular Compartments Is Driven by Subgenomic Recombination Events

Our understanding of viral defective RNAs (D-RNAs), or truncated viral genomes, comes largely from passaging studies in tissue culture under artificial conditions and/or packaged viral RNAs. Here, we show that specific populations of alphavirus D-RNAs arise de novo and that they are not packaged into virions, thus imposing a transmission bottleneck and impeding their prior detection. This raises important questions about the roles of D-RNAs, both in nature and in tissue culture, during viral infection and whether their influence is constrained by packaging requirements. Further, during the course of these studies, we found a novel type of alphavirus D-RNA that is enriched intracellularly; dubbed subgenomic D-RNAs (sgD-RNAs), they are defined by deletion boundaries between the capsid-E3 region and the E1-3′ untranslated region (UTR) and are common to chikungunya, Mayaro, Sindbis, and Aura viruses. These sgD-RNAs are enriched intracellularly and do not appear to be selectively packaged, and additionally, they may exist as subgenome-derived transcripts.Alphaviruses are positive-sense RNA arboviruses that can cause either a chronic arthritis or a potentially lethal encephalitis. Like other RNA viruses, alphaviruses produce truncated, defective viral RNAs featuring large deletions during replication. These defective RNAs (D-RNAs) have primarily been isolated from virions after high-multiplicity-of-infection passaging. Here, we aimed to characterize both intracellular and packaged viral D-RNA populations during early-passage infections under the hypothesis that D-RNAs arise de novo intracellularly that may not be packaged and thus have remained undetected. To this end, we generated next-generation sequencing libraries using RNA derived from passage 1 (P1) stock chikungunya virus (CHIKV) 181/clone 25, intracellular virus, and P2 virions and analyzed samples for D-RNA expression, followed by diversity and differential expression analyses. We found that the diversity of D-RNA species is significantly higher for intracellular D-RNA populations than P2 virions and that specific populations of D-RNAs are differentially expressed between intracellular and extracellular compartments. Importantly, these trends were likewise observed in a murine model of CHIKV AF15561 infection, as well as in vitro studies using related Mayaro, Sindbis, and Aura viruses. Additionally, we identified a novel subtype of subgenomic D-RNA that is conserved across arthritogenic alphaviruses. D-RNAs specific to intracellular populations were defined by recombination events specifically in the subgenomic region, which were confirmed by direct RNA nanopore sequencing of intracellular CHIKV RNAs. Together, these studies show that only a portion of D-RNAs generated intracellularly are packaged and D-RNAs readily arise de novo in the absence of transmitted template

Question Formulation Skills Training Using a Rubric with First-Year Medical Students. Dataset

The attached dataset consists of survey responses organized in the following manner. Column 1: De-identified student number Column 2: Cohort Class of 2022 Column 3: Pre-Test Score