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Primary (non-motile) cilia represent structurally and functionally diverse organelles whose roles as specialized cellular antenna are central to animal cell signaling pathways, sensory physiology and development. An ever-growing number of ciliary proteins, including those found in vertebrate photoreceptors, have been uncovered and linked to human disorders termed ciliopathies. Here, we demonstrate that an evolutionarily-conserved PPEF-family serine-threonine phosphatase, not functionally linked to cilia in any organism but associated with rhabdomeric (non-ciliary) photoreceptor degeneration in the Drosophila rdgC (retinal degeneration C) mutant, is a bona fide ciliary protein in C.   elegans . The nematode protein, PEF-1, depends on transition zone proteins, which make up a ‘ciliary gate’ in the proximal-most region of the cilium, for its compartmentalization within cilia. Animals lacking PEF-1 protein function display structural defects to several types of cilia, including potential degeneration of microtubules. They also exhibit anomalies to cilium-dependent behaviors, including impaired responses to chemical, temperature, light, and noxious CO 2 stimuli. Lastly, we demonstrate that PEF-1 function depends on conserved myristoylation and palmitoylation signals. Collectively, our findings broaden the role of PPEF proteins to include cilia, and suggest that the poorly-characterized mammalian PPEF1 and PPEF2 orthologs may also have ciliary functions and thus represent ciliopathy candidates.

Northern British Columbia has been identified as an important habitat for several coastal cetacean species, including humpback whales (Megaptera novaeangliae). This species is listed as being of “Special Concern” under Canada's Species at Risk Act, partly due to data deficiencies concerning genetic population structure and demographics in British Columbia. Anthropogenic activities threaten North Coast humpback whale populations, with particular concern for the impact of vessel noise, entanglement, and ship strikes. Current methodology (i.e., biopsy sampling) for obtaining cetacean genetic data is invasive, challenging, and costly; therefore, there is an urgency to develop effective and minimally invasive methodologies for efficiently collecting this data. Environmental DNA (eDNA) has been identified as an ideal tool for monitoring the presence and distribution of numerous species within marine ecosystems; however, the feasibility for cetaceans is not yet well established. In this study, we opportunistically collected targeted 1 L seawater eDNA samples from flukeprints when individual humpback whales were observed diving between the years of 2020 and 2022. A total of 93 samples were collected from individual humpback whales identified using a photographic identification catalogue. We successfully detected humpback whale eDNA in 28 samples using novel species‐specific qPCR primers (~500 mL of sample), with relatively equal successful detection between immediate (0 days) and delayed (up to 10 days) sample filtration. Here, we have validated a qPCR assay for detecting humpback whale DNA from flukeprints and highlighted the future optimizations required to improve the potential application of flukeprint eDNA for conservation management. We developed and employed a species‐specific eDNA assay to detect humpback whale DNA from flukeprints. We detected humpback whales in ~30% of samples and determined there was no difference between instant and delayed filtration on detection.

Anthropogenic pressures on marine ecosystems are intensifying, highlighting the need to bridge knowledge gaps for species at risk. Data deficiencies, particularly for species recovering from historic declines, such as marine megafauna like whales, hinder effective management. Understanding long‐term population viability and identifying any sex‐biased threats is important for cetacean conservation. Killer whales (Orcinus orca) and humpback whales (Megaptera novaeangliae) are well‐studied in the Northeast Pacific, making them ideal for optimizing noninvasive environmental DNA (eDNA) techniques for sex determination. We collected eDNA flukeprint samples from killer whales (n = 67) and humpback whales (n = 18), analyzing ZFX/ZFY gene amplification using conventional PCR to compare results against known sexes. Samples from killer whales exhibited higher ZFX/ZFY PCR amplification success (53%) compared with humpback whales (44%). However, the close social structure of this species likely contributed to only 54% of samples matching the known sex of whales sampled. Conversely, humpback whale samples accurately matched the known sexes of individuals (100%). These findings demonstrate eDNA's potential to replace more invasive biopsies for sex determination but highlight the need for further optimization regarding sampling protocols and species‐specific ZFX/ZFY amplification approaches. Additionally, eDNA flukeprint sampling also shows promise for other solitary cetaceans such as large rorquals (Balaenoptera spp.), which remain among the most data‐deficient species. This study used environmental DNA (eDNA) flukeprint sampling and ZFX/ZFY gene amplification to determine the sex of killer whales and humpback whales, finding higher PCR success in killer whales but higher sex‐matching success in humpbacks. While eDNA shows promise as a noninvasive alternative to biopsies, further refinement is needed to optimize sampling techniques for broader application, especially for data‐deficient rorquals.

Determining the sub-cellular localization of a protein within a cell is often an essential step towards understanding its function. In Caenorhabditis elegans, the relatively large size of the body wall muscle cells and the exquisite organization of their sarcomeres offer an opportunity to identify the precise position of proteins within cell substructures. Our goal in this study is to generate a comprehensive \"localizome\" for C. elegans body wall muscle by GFP-tagging proteins expressed in muscle and determining their location within the cell. For this project, we focused on proteins that we know are expressed in muscle and are orthologs or at least homologs of human proteins. To date we have analyzed the expression of about 227 GFP-tagged proteins that show localized expression in the body wall muscle of this nematode (e.g. dense bodies, M-lines, myofilaments, mitochondria, cell membrane, nucleus or nucleolus). For most proteins analyzed in this study no prior data on sub-cellular localization was available. In addition to discrete sub-cellular localization we observe overlapping patterns of localization including the presence of a protein in the dense body and the nucleus, or the dense body and the M-lines. In total we discern more than 14 sub-cellular localization patterns within nematode body wall muscle. The localization of this large set of proteins within a muscle cell will serve as an invaluable resource in our investigation of muscle sarcomere assembly and function.

A crucial step in the development of muscle cells in all metazoan animals is the assembly and anchorage of the sarcomere, the essential repeat unit responsible for muscle contraction. In Caenorhabditis elegans, many of the critical proteins involved in this process have been uncovered through mutational screens focusing on uncoordinated movement and embryonic arrest phenotypes. We propose that additional sarcomeric proteins exist for which there is a less severe, or entirely different, mutant phenotype produced in their absence. We have used Serial Analysis of Gene Expression (SAGE) to generate a comprehensive profile of late embryonic muscle gene expression. We generated two replicate long SAGE libraries for sorted embryonic muscle cells, identifying 7,974 protein-coding genes. A refined list of 3,577 genes expressed in muscle cells was compiled from the overlap between our SAGE data and available microarray data. Using the genes in our refined list, we have performed two separate RNA interference (RNAi) screens to identify novel genes that play a role in sarcomere assembly and/or maintenance in either embryonic or adult muscle. To identify muscle defects in embryos, we screened specifically for the Pat embryonic arrest phenotype. To visualize muscle defects in adult animals, we fed dsRNA to worms producing a GFP-tagged myosin protein, thus allowing us to analyze their myofilament organization under gene knockdown conditions using fluorescence microscopy. By eliminating or severely reducing the expression of 3,300 genes using RNAi, we identified 122 genes necessary for proper myofilament organization, 108 of which are genes without a previously characterized role in muscle. Many of the genes affecting sarcomere integrity have human homologs for which little or nothing is known.

Health disparities between Indigenous and non-Indigenous peoples persist globally. Northern interior British Columbia, where many Indigenous people live on Indian 1 reserves allocated in the late nineteenth century, is no exception. This article reviews findings from fifty-eight interviews with members of thirteen First Nations communities in Carrier, Sekani, Wet'suwet'en, and Babine territories. The results suggest that colonial geographies, both physical and social, along with extant anti-Indigenous racism, are significant determinants of the health and well-being (or lack thereof) of many First Nations in the region.

This article provides a practical guide and insights into developing a focus group research center or a similar \"learning lab\" for the purposes of experiential learning, by describing a project to remodel a set of existing contiguous spaces within a school of business into a focus group research facility that enhances teaching, fosters faculty collaboration, and serves the marketing research needs of the community. The article presents some of the key trade-off design issues and connects these decisions to cost impact. A schematic for the facility and summary budgeting model are presented to aid in the development of similar projects.

This project explores John’s portrait of Jesus in the Gospel of John and argues that John presents Jesus as the Servant of the Lord from Isaiah primarily through his allusions to the Servant passages. The Servant is not the predominant Christological title in John, but it is an important aspect of Johannine Christology. Chapter 1 surveys various works on John’s use of the Old Testament in general, and John’s use of Isaiah specifically. While Isaiah is a major emphasis in John, there have only been two other monographs on John’s use of Isaiah. Furthermore, other works have noted a few allusions to the Isaianic Servant, but no other works have systematically studied the Isaianic Servant in the Gospel of John. Chapter 2 proposes several criteria necessary to identitying allusions to the Isaianic Servant, with allusions classified as clear, probable, and possible. Chapter 3 contains an exegetical discussion of the relevant Servant passages in Isaiah, along with a discussion of the literary context of each Servant song. There is an excursus on the Second Temple Literature, which indicates there is no evidence of a Suffering Messiah in that period. Chapter 4 analyzes two quotations to Isaiah and other clear allusions to the Servant songs in Isaiah. This chapter, along with subsequent chapters, explores the meaning and significance of each allusion for the literary context of John. Chapter 5 identifies probable allusions to the Servant, including the Greeks coming to see Jesus, light and darkness, and Jesus obeying the will of God. Chapter 6 analyzes possible allusions, which have some overlap with the Servant songs, but lack the criterion of uniqueness that would assist an interpreter in connecting John to Isaiah. Chapter 7 discusses the theme of the Isaianic New Exodus and the Servant’s role in initiating this new exodus. John’s description of Jesus utilizes the same characteristics of the Servant to portray Jesus as the Servant in Isaiah. Chapter 8 summarizes the previous chapters and analyzes the implications of the study along with areas for further research.