The TSC22D, WNK, and NRBP gene families exhibit functional buffering and evolved with Metazoa for cell volume regulation.

The ability to sense and respond to osmotic fluctuations is critical for the maintenance of cellular integrity. We used gene co-essentiality analysis to identify an unappreciated relationship between TSC22D2, WNK1, and NRBP1 in regulating cell volume homeostasis. All of these genes have paralogs and are functionally buffered for osmo-sensing and cell volume control. Within seconds of hyperosmotic stress, TSC22D, WNK, and NRBP family members physically associate into biomolecular condensates, a process that is dependent on intrinsically disordered regions (IDRs). A close examination of these protein families across metazoans revealed that TSC22D genes evolved alongside a domain in NRBPs that specifically binds to TSC22D proteins, which we have termed NbrT (NRBP binding region with TSC22D), and this co-evolution is accompanied by rapid IDR length expansion in WNK-family kinases. Our study reveals that TSC22D, WNK, and NRBP genes evolved in metazoans to co-regulate rapid cell volume changes in response to osmolarity.

Multi-level cellular and functional annotation of single-cell transcriptomes using scPipeline.

Single-cell RNA-sequencing (scRNA-seq) offers functional insight into complex biology, allowing for the interrogation of cellular populations and gene expression programs at single-cell resolution. Here, we introduce scPipeline, a single-cell data analysis toolbox that builds on existing methods and offers modular workflows for multi-level cellular annotation and user-friendly analysis reports. Advances to scRNA-seq annotation include: (i) co-dependency index (CDI)-based differential expression, (ii) cluster resolution optimization using a marker-specificity criterion, (iii) marker-based cell-type annotation with Miko scoring, and (iv) gene program discovery using scale-free shared nearest neighbor network (SSN) analysis. Both unsupervised and supervised procedures were validated using a diverse collection of scRNA-seq datasets and illustrative examples of cellular transcriptomic annotation of developmental and immunological scRNA-seq atlases are provided herein. Overall, scPipeline offers a flexible computational framework for in-depth scRNA-seq analysis.

Phylogeny of the North-Central American clade of blood-sucking reduviid bugs of the tribe Triatomini (Hemiptera: Triatominae) based on the mitochondrial genome.

Triatominae is a subfamily of blood-sucking reduviid hemipterans of public health importance primarily in tropical and sub-tropical regions of the Americas, whose members possess various morphological adaptations closely associated to hematophagy. Despite their medical importance, the systematics of the subfamily is far from resolved, particularly within the tribe Triatomini. Here we employed mitochondrial genome DNA sequences to reconstruct the phylogenetic relationships among 19 species of the North-Central American (NCA) clade of Triatomini and to estimate the times of origin and diversification of its main clades. Twenty-nine mitogenomes were examined for representative specimens of 25 species, including the outgroup. Phylogenetic informativeness estimated for each protein-coding gene showed that cox1, cox2 and atp6 were the most informative markers, whereas atp8 and nad4 had high saturation levels. Phylogenetic analyses excluding the latter two protein-coding genes recovered an almost fully resolved topology. The NCA clade apparently originated shortly after emergence of an initial land bridge of the Panama Isthmus, ca. 15.05-20.05 Mya. An Asian/pantropical subclade with Linshcosteus costalis, Triatoma rubrofasciata and T. migrans was nested within the NCA clade, from which it diverged ca. 12.42-17.3Mya. Uncorrected cox1 and 13 protein-coding gene distances suggest the existence of additional species within the dimidiata complex. In contrast, T. phyllosoma, T. mazzottii and T. longipennis, from the phyllosoma complex, have considerably low cox1 and 13 PCG distances among them, suggesting mitochondrial introgression or conspecificity. Our study yielded a robust phylogeny for the group, which could be tested with further phylogenetic hypotheses based on nuclear genome-wide markers.

Three new species of Pambolus (Braconidae: Pambolinae) from Mexico, with comments on the variation of P. oblongispina Papp.

Three species of the braconid genus Pambolus (Braconidae) are described from Mexico: P. jarocho sp. n., P. chinanteco sp. n. and P. bizelab sp. n. The external morphological variation in males and females of P. oblongispina Papp, previously known only by two females from Honduras and northern Mexico, is described based on material from Jalisco and Oaxaca in central and southeast Mexico. Molecular characterisation of the examined species was carried out based on the 28S nuclear ribosomal and the COI mitochondrial DNA gene markers.