Phylogeny of Parasitic Parabasalia and Free-Living Relatives Inferred from Conventional Markers vs. Rpb1, a Single-Copy Gene

Parabasalia are single-celled eukaryotes (protists) that are mainly comprised of endosymbionts of termites and wood roaches, intestinal commensals, human or veterinary parasites, and free-living species. Phylogenetic comparisons of parabasalids are typically based upon morphological characters and 18S ribosomal RNA gene sequence data (rDNA), while biochemical or molecular studies of parabasalids are limited to a few axenically cultivable parasites. These previous analyses and other studies based on PCR amplification of duplicated protein-coding genes are unable to fully resolve the evolutionary relationships of parabasalids. As a result, genetic studies of Parabasalia lag behind other organisms.Comparing parabasalid EF1α, α-tubulin, enolase and MDH protein-coding genes with information from the Trichomonas vaginalis genome reveals difficulty in resolving the history of species or isolates apart from duplicated genes. A conserved single-copy gene encodes the largest subunit of RNA polymerase II (Rpb1) in T. vaginalis and other eukaryotes. Here we directly sequenced Rpb1 degenerate PCR products from 10 parabasalid genera, including several T. vaginalis isolates and avian isolates, and compared these data by phylogenetic analyses. Rpb1 genes from parabasalids, diplomonads, Parabodo, Diplonema and Percolomonas were all intronless, unlike intron-rich homologs in Naegleria, Jakoba and Malawimonas.The phylogeny of Rpb1 from parasitic and free-living parabasalids, and conserved Rpb1 insertions, support Trichomonadea, Tritrichomonadea, and Hypotrichomonadea as monophyletic groups. These results are consistent with prior analyses of rDNA and GAPDH sequences and ultrastructural data. The Rpb1 phylogenetic tree also resolves species- and isolate-level relationships. These findings, together with the relative ease of Rpb1 isolation, make it an attractive tool for evaluating more extensive relationships within Parabasalia

Werewolf Arena: A Case Study in LLM Evaluation via Social Deduction

This paper introduces Werewolf Arena, a novel framework for evaluating large language models (LLMs) through the lens of the classic social deduction game, Werewolf. In Werewolf Arena, LLMs compete against each other, navigating the game's complex dynamics of deception, deduction, and persuasion. The framework introduces a dynamic turn-taking system based on bidding, mirroring real-world discussions where individuals strategically choose when to speak. We demonstrate the framework's utility through an arena-style tournament featuring Gemini and GPT models. Our results reveal distinct strengths and weaknesses in the models' strategic reasoning and communication. These findings highlight Werewolf Arena's potential as a challenging and scalable LLM benchmark.Comment: 13 pages, 10 figure

Just Because It\u27S Gay? : Transgressive Design In Queer Coming Of Age Visual Novels

While queer narratives have recently gained greater prominence and attention [35][51][48] both in games and game studies, they still represent a tiny portion of the playable romances available. Even fewer among these represent the potential of what Edmond Chang [12] has referred to as queergaming, or games which refuse to conform in their narrative, mechanics, and modes of play to a cisgender, white, heteronormative gaze. We argue that meaningful queergaming is impossible within the current norms and genres of so-called AAA, or mainstream. It thus requires space and platforms dedicated to transgressive game design, building on Aarseth\u27s [1] framework of transgressive play. To substantiate our argument, we use an examination of recent queer visual novels built using Ren\u27Py and released on Itch.io, with a case study of most popular current exemplar Butterfly Soup [32] alongside previous exemplars of queer romance in games. 1