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SUMMARY Confocal microscopy has greatly aided our understanding of the major cellular processes and trafficking pathways responsible for plant growth and development. However, a drawback of these studies is that they often rely on the manual analysis of a vast number of images, which is time‐consuming, error‐prone, and subject to bias. To overcome these limitations, we developed Dot Scanner, a Python program for analyzing the densities, lifetimes, and displacements of fluorescently tagged particles in an unbiased, automated, and efficient manner. Dot Scanner was validated by performing side‐by‐side analysis in Fiji‐ImageJ of particles involved in cellulose biosynthesis. We found that the particle densities and lifetimes were comparable in both Dot Scanner and Fiji‐ImageJ, verifying the accuracy of Dot Scanner. Dot Scanner largely outperforms Fiji‐ImageJ, since it suffers far less selection bias when calculating particle lifetimes and is much more efficient at distinguishing between weak signals and background signal caused by bleaching. Not only does Dot Scanner obtain much more robust results, but it is a highly efficient program, since it automates much of the analyses, shortening workflow durations from weeks to minutes. This free and accessible program will be a highly advantageous tool for analyzing live‐cell imaging in plants. 

Abstract Transmissible cancers are infectious parasitic clones that metastasize to new hosts, living past the death of the founder animal in which the cancer initiated. We investigated the evolutionary history of a cancer lineage that has spread though the soft-shell clam (Mya arenaria) population by assembling a chromosome-scale soft-shell clam reference genome and characterizing somatic mutations in transmissible cancer. We observe high mutation density, widespread copy-number gain, structural rearrangement, loss of heterozygosity, variable telomere lengths, mitochondrial genome expansion and transposable element activity, all indicative of an unstable cancer genome. We also discover a previously unreported mutational signature associated with overexpression of an error-prone polymerase and use this to estimate the lineage to be >200 years old. Our study reveals the ability for an invertebrate cancer lineage to survive for centuries while its genome continues to structurally mutate, likely contributing to the evolution of this lineage as a parasitic cancer. 

Abstract We use the results of a survey for low-surface-gravity stars in Galactic (and LMC) globular clusters to show that “yellow” post-asymptotic-branch (yPAGB) stars are likely to be excellent extragalactic standard candles, capable of producing distances to early-type galaxies that are accurate to a couple of percent. We show that the mean bolometric magnitude of the 10 yPAGB stars in globular clusters is 〈 M bol 〉 = −3.38 ± 0.03, a value that is ∼0.2 mag brighter than that predicted from the latest post-horizontal-branch evolutionary tracks. More importantly, we show that the observed dispersion in the distribution is only 0.10 mag, i.e., better than the scatter for individual Cepheids. We describe the physics that can produce such a small dispersion and show that, if one restricts surveys to the color range 0.0 ≲ ( B − V ) 0 ≲ 0.5, then samples of nonvariable yPAGB stars can be identified quite easily with a minimum of contamination. The extremely bright absolute V magnitudes of these stars (〈 M V 〉 = −3.37) make them, by far, the visually brightest objects in old stellar populations and ideal Population II standard candles for measurements out to ∼10 Mpc with current instrumentation. A Hubble Space Telescope survey in the halos of galaxies in the M81 and Sculptor groups could therefore serve as an effective cross-check on both the Cepheid and tip-of-the-red-giant-branch distance scales. 

Abstract We have carried out a search for above-horizontal-branch (AHB) stars—objects lying above the horizontal branch (HB) and blueward of the asymptotic giant branch (AGB) in the color–magnitude diagram—in 97 Galactic and seven Magellanic Cloud globular clusters (GCs). We selected AHB candidates based on photometry in the uBVI system, which is optimized for detection of low-surface-gravity stars with large Balmer jumps, in the color range −0.05 ≤ ( B − V ) 0 ≤1.0. We then used Gaia astrometry and Gaussian-mixture modeling to confirm cluster membership and remove field interlopers. Our final catalog contains 438 AHB stars, classified and interpreted in the context of post-HB evolution as follows: (1) AHB1: 280 stars fainter than M V = −0.8, evolving redward from the blue HB (BHB) toward the base of the AGB. (2) Post-AGB (PAGB): 13 stars brighter than M V ≃ −2.75, departing from the top of the AGB and evolving rapidly blueward. (3) AHB2: 145 stars, with absolute magnitudes between those of the AHB1 and PAGB groups. This last category includes a mixture of objects leaving the extreme BHB and evolving toward the AGB, and brighter ones moving back from the AGB toward higher temperatures. Among the AHB1 stars are 59 RR Lyrae interlopers, observed by chance in our survey near maximum light. PAGB and AHB2 stars (including W Virginis Cepheids) overwhelmingly belong to GCs containing BHB stars, in accordance with predictions of post-HB evolutionary tracks. We suggest that most W Vir variables are evolving toward lower temperatures and are in their first crossings of the instability strip. Nonvariable yellow PAGB stars show promise as a Population II standard candle for distance measurement. 

Canine distemper virus (CDV) is a multi-host pathogen with variable clinical outcomes of infection across and within species. We used whole-genome sequencing (WGS) to search for viral markers correlated with clinical distemper in African lions. To identify candidate markers, we first documented single-nucleotide polymorphisms (SNPs) differentiating CDV strains associated with different clinical outcomes in lions in East Africa. We then conducted evolutionary analyses on WGS from all global CDV lineages to identify loci subject to selection. SNPs that both differentiated East African strains and were under selection were mapped to a phylogenetic tree representing global CDV diversity to assess if candidate markers correlated with documented outbreaks of clinical distemper in lions (n = 3). Of 54 SNPs differentiating East African strains, ten were under positive or episodic diversifying selection and 20 occurred in the clinical strain despite strong purifying selection at those loci. Candidate markers were in functional domains of the RNP complex (n = 19), the matrix protein (n = 4), on CDV glycoproteins (n = 5), and on the V protein (n = 1). We found mutations at two loci in common between sequences from three CDV outbreaks of clinical distemper in African lions; one in the signaling lymphocytic activation molecule receptor (SLAM)-binding region of the hemagglutinin protein and another in the catalytic center of phosphodiester bond formation on the large polymerase protein. These results suggest convergent evolution at these sites may have a functional role in clinical distemper outbreaks in African lions and uncover potential novel barriers to pathogenicity in this species. 

Numerous pairs of evolutionarily divergent mammalian species have been shown to produce hybrid offspring. In some cases, F 1 hybrids are able to produce F 2 s through matings with F 1 s. In other instances, the hybrids are only able to produce offspring themselves through backcrosses with a parent species owing to unisexual sterility (Haldane's Rule). Here, we explicitly tested whether genetic distance, computed from mitochondrial and nuclear genes, can be used as a proxy to predict the relative fertility of the hybrid offspring resulting from matings between species of terrestrial mammals. We assessed the proxy's predictive power using a well-characterized felid hybrid system, and applied it to modern and ancient hominins. Our results revealed a small overlap in mitochondrial genetic distance values that distinguish species pairs whose calculated distances fall within two categories: those whose hybrid offspring follow Haldane's Rule, and those whose hybrid F 1 offspring can produce F 2 s. The strong correlation between genetic distance and hybrid fertility demonstrated here suggests that this proxy can be employed to predict whether the hybrid offspring of two mammalian species will follow Haldane's Rule.