Search

The capability of synthesized zeolite A (SZ) to immobilize Ag ions (Ag-SZ) and Ag nanoparticles (AgNp-SZ) were comparatively studied. A novel approach of in situ biosynthesized AgNP-incorporated synthesized zeolite A (AgNp-SZ) was synthesized at an optimum volume of 0.4 mL of the Orthosiphon aristatus (O. aristatus) leaves plant extract (5%) using an in situ approach. In comparison, Ag-SZ was produced by loading the synthesized zeolite with Ag ions. All synthesized materials were characterized for their morphologies and physicochemical properties. The characterization analyses validate that the biosynthesized AgNP (

Worldwide, rising ocean temperatures are causing declines and range shifts in marine species. The direct effects of climate change on the biology of marine organisms are often well documented; yet, knowledge on the indirect effects, particularly through trophic interactions, is largely lacking. We provide evidence of ocean warming decoupling critical trophic interactions supporting a commercially important mollusc in a climate change hotspot. Dietary assessments of the Australian blacklip abalone (Haliotis rubra) indicate primary dependency on a widespread macroalgal species (Phyllospora comosa) which we show to be in state of decline due to ocean warming, resulting in abalone biomass reductions. Niche models suggest further declines in P. comosa over the coming decades and ongoing risks to H. rubra. This study highlights the importance of studies from climate change hotspots and understanding the interplay between climate and trophic interactions when determining the likely response of marine species to environmental changes.

The number of human–shark interactions has increased worldwide during the past decade resulting in injuries and fatalities. In Australia, the white shark (Carcharodon carcharias), tiger shark (Galeocerdo cuvier), and bull shark (Carcharhinus leucas) are responsible for the majority of fatal incidents. On the southeast coast of Australia, monitoring programs currently rely on SMART (Shark‐Management‐Alert‐in‐Real‐Time) drumlines and mesh nets to catch, tag, and monitor shark movement. However, these methods are laborious, costly, and involve the capture of only a fraction of the total shark population. Here, we develop a multiplex environmental DNA assay capable of detecting all three shark species simultaneously from water samples by targeting conserved but specific mitochondrial sequences that are characteristic of each species. The specificity of the assay was validated by testing for cross‐amplification across a range of non‐target but co‐occurring shark species from eastern Australia. We test the sensitivity of the assay on water samples collected from shark capture events and sites where these shark species are known to frequent, and undertake DNA sequencing on positive samples to confirm species haplotype authenticity. Samples collected from one of these sites also demonstrate that eDNA detections are dependent on shark activity in the area. This assay will allow for rapid detection of DNA from each shark species in water samples, providing a cost‐effective alternative for monitoring sharks along the east coast of Australia and potentially elsewhere.

Mesic grasslands in South Africa (> 650 mm a − 1 MAP) are rich in herbaceous forbs, which outnumber grass species by more than 5 to 1. Many of these forbs are geophytes with underground storage units (USOs), such as thickened rootstocks, rhizomes, bulbs, or corms, that provide resources (non-structural carbohydrates, minerals, and water) enabling them to resprout after dry, frosty winters, and fire. However, despite their extensive biomass and reserves ostensibly protected underground, mesic grassland geophytes can be severely depleted or extirpated by chronic trampling and grazing of their aerial parts by livestock. This study examined a possible explanation for the demise of grassland geophytes in overgrazed grassland by investigating, in a pot trial, whether the growth of a geophyte and the size of its USOs are negatively affected by simulated green leaf loss. In a 2 × 2 factorial (clipped vs. unclipped x spring regrowth in the dark vs. light), five replicate plants of Hypoxis hemerocallidea, a common mesic grassland geophyte that resprouts from a corm, were subject to six severe (clipped to 80 mm) defoliations during the growing season and regrown in spring under full or restricted light to measure stored reserve contribution to regrowth. Defoliated plants were resilient to defoliation during the growing season, matching the total biomass production of unclipped plants, though cutting reduced the number of leaves by ¬60% and flowers by almost 85%. Spring regrowth on stored reserves equalled that from reserves plus concurrent photosynthesis, indicating the value of USOs for regrowth. However, there was a marked carry-over effect of previous season defoliation, resulting in a one-third reduction in shoot growth and 40% fewer inflorescence in spring. Crucially, corm mass was more than halved by clipping, which resulted in lower spring biomass and inflorescence production. It was concluded that buried stored reserves are not protected from recurrent defoliation and mechanical damage to aerial plant parts and that continued diminishment of USOs under chronic disturbance by overgrazing or frequent mowing would weaken and likely eventually kill plants, reducing overall forb species richness. Lenient management by infrequent summer mowing or grazing at moderate stocking rates combined with periodic rotational full season resting and dormant-season burning is recommend to maintain the USOs and vigour of grassland geophytes in mesic grassland.

The irregular supply of natural mussel seed for aquaculture has seen an increase in the establishment of hatcheries to produce a reliable source of mussel seed. While many of the factors influencing fertilisation and early survival of hatchery-produced mussel larvae have been explored, little is known about the factors influencing the success of larvae during settlement and development under hatchery conditions. This study examines the effects of feed concentration, airflow rate, rope type and larval stocking density on the growth, settlement and development of Australian blue mussel larvae under hatchery conditions. Manipulation of feeding concentration at the time of settlement (25,000 cell ml−1, 50,000 cell ml−1, 100,000 cell ml−1 and no feed) had little effect on growth, settlement and development of larvae, due to reduced feeding behaviour during this period. In contrast, high airflow rates were found to increase settlement and larval development but did not influence growth. Larvae also show significantly greater settlement rates on more filamentous ropes compared with those on less filamentous rope types. We also detected a strong inverse relationship between larval density and size at settlement, suggesting density-dependent competition may reduce growth under hatchery conditions. These results indicate optimal conditions for settlement and development of mussel larvae in hatcheries may be achieved by decreasing feed concentration during the settlement phase, increasing airflow rates and maintaining moderate densities of larvae. These optimised hatchery conditions have the potential to improve yields while simultaneously reducing production costs.

Cosmopolitan marine pelagic species display variable patterns of population connectivity among the world’s major oceans. While this information is crucial for informing management, information is lacking for many ecologically important species, including apex predators. In this study we examine patterns of genetic structure in the broadnose sevengill shark, Notorynchus cepedianus across its global distribution. We estimate patterns of connectivity among broadnose sevengill shark populations from three major oceanic regions (South Atlantic, Oceania and Eastern Pacific) by contrasting mitochondrial and nuclear DNA haplotype frequencies. We also produced time calibrated Bayesian Inference phylogenetic reconstructions to analyses global phylogeographic patterns and estimate divergence times among distinctive shark lineages. Our results demonstrate significant genetic differentiation among oceanic regions (ΦST = 0.9789, P

Cystic fibrosis (CF) is a rare autosomal-recessive disorder manifested as multisystem organ dysfunction. The cystic fibrosis transmembrane conductance regulator (CFTR) protein functions as an ion transporter on the epithelium of exocrine glands, regulating secretion viscosity. The CFTR gene, encoded on chromosome 7, is required for the production and trafficking of the intact and functional CFTR protein. Literally thousands of human CFTR allelic mutations have been identified, each with varying impact on protein quality and quantity. As a result, individuals harboring CFTR mutations present with a spectrum of symptoms ranging from CF to normal phenotypes. Those with loss of function but without full CF may present with CFTR-related disorders (CFTR-RDs) including male infertility, sinusitis, pancreatitis, atypical asthma and bronchitis. Studies have demonstrated associations between higher rates of CFTR mutations and oligospermia, epididymal obstruction, congenital bilateral absence of the vas deferens (CBAVD), and idiopathic ejaculatory duct obstruction (EDO). Genetic variants are detected in over three-quarters of men with CBAVD, the reproductive abnormality most classically associated with CFTR aberrations. Likewise, nearly all men with clinical CF will have CBAVD. Current guidelines from multiple groups recommend CFTR screening in all men with clinical CF or CBAVD though a consensus on the minimum number of variants for which to test is lacking. CFTR testing is not recommended as routine screening for men with other categories of infertility. While available CFTR panels include 30 to 96 of the most common variants, complete gene sequencing should be considered if there is a high index of suspicion in a high-risk couple (e.g., partner is CFTR mutation carrier). CF treatments to date have largely targeted end-organ complications. Novel CFTR-modulator treatments aim to directly target CFTR protein dysfunction, effectively circumventing downstream complications, and possibly preventing symptoms like vasal atresia at a young age. Future gene therapies may also hold promise in preventing or reversing genetic changes that lead to CF and CFTR-RD.

Context Capture, measurement, genetic sampling, ringing and flagging of shorebirds on their nests are standard techniques that underpin the study and conservation of these species. However, these techniques may reduce hatching success by compromising parental care or nest crypsis, thereby negatively influencing results, study populations and bird welfare. Only a few studies that examine the effect of capture of shorebirds on subsequent egg hatching success are currently available. Aims To compare the hatching success of masked lapwing (Vanellus miles) nests, at which adult capture and associated techniques (ringing, flagging and bleeding) have occurred, with nests at which these did not occur, on the fox-free Phillip Island, Victoria, Australia. Methods Hatching success of masked lapwings was monitored and compared between nests at which adult capture did, and did not, occur. Clutches were aged and age was included in our models to adjust for exposure of clutches to risks such as predators. Key results There was no difference in egg hatching success between nests at which capture occurred and those at which it did not occur: 138 chicks hatched from 178 eggs attended by adults that were captured (77.5% hatched); and 279 chicks hatched from 442 eggs attended by adults that were not captured (63.1%). Conclusion Trapping incubating lapwings using our existing protocols does not compromise egg hatching success, at least where there are no foxes present. Implications Studies of ground-nesting shorebird hatching success in relation to capture can usefully assess for adverse effects of the methods employed. We suggest that further examination of capture of lapwings at the nest be conducted in environments where foxes are present.

Marked tillers of Heteropogon contortus, Trachypogon spicatus, Tristachya leucothrix and Harpochloa falx were sampled regularly over a two-year period in Highland Sourveld burnt annually in winter and biennially in spring. Parameters recorded were height of shoot apex, tiller mass, number of new lateral tillers and time of flowering. Monitoring of marked tillers before and after each burn enabled the life history of populations of tillers to be followed. In all species investigated, flowering was found to be of minimal importance with most tillers remaining vegetative until death. Shoot apices remained close to the soil surface (

Background Hemolytic disease of the fetus and newborn (HDFN) attributable to anti-M is rare, although case reports implicate anti-M in varying severities of HDFN, including fetal hydrops and intrauterine death. Case description We describe the case of a newborn with HDFN associated with an atypical immunoglobulin (Ig) G anti-M that reacted best at cold temperatures. The maternal antibody detected in pregnancy was not reactive at 37°C, and a direct antiglobulin test (DAT) on red blood cells (RBCs) from the newborn was negative, suggesting an anti-M that should not have been clinically relevant. However, the infant developed hyperbilirubinemia (bilirubin level, 17.6 mg/dL), hemolytic anemia (hemoglobin nadir, 5.5 g/dL), and reticulocytopenia. Laboratory testing demonstrated the presence of an IgG anti-M in maternal and neonatal samples reacting best at 4°C. This passively acquired IgG anti-M provoked hemolytic anemia in the infant and likely suppressed erythropoiesis, resulting in reticulocytopenia with prolonged anemia. He was treated for IgG anti-M HDFN with 10 intravenous Ig infusions and 10 days of oral prednisone followed by a taper. He required seven transfusions with M- RBCs. His hemoglobin level normalized at 3 months of age. Follow-up at 2 years revealed no hematologic or neuro-developmental concerns. Conclusion To our knowledge, this is the second report of HDFN attributable to an IgG anti-M reacting preferentially at cold temperature with no 37°C reactivity. Clinically relevant IgG anti-M may elude standard testing. Early recognition and testing for cold-reacting IgG anti-M should be considered for newborns with hemolysis, a negative DAT, and prolonged anemia.

Three case studies involving two temperate Australian seagrass species – Pondweed (Ruppia tuberosa) and Ribbon Weed (Posidonia australis) – highlight different approaches to their restoration. Seeds and rhizomes were used in three collaborative programmes to promote new approaches to scale up restoration outcomes.

Increasing urbanization and use of urban areas by synanthropic wildlife has increased human and domestic animal exposure to zoonotic diseases and exacerbated epizootics within wildlife populations. Consequently, there is a need to improve wildlife disease surveillance programs to rapidly detect outbreaks and refine inferences regarding spatiotemporal disease dynamics. Multistate occupancy models can address potential shortcomings in surveillance programs by accounting for imperfect detection and the misclassification of disease states. We used these models to explore the relationship between urbanization, slope, and the spatial distribution of sarcoptic mange in coyotes (Canis latrans) inhabiting Fort Irwin, California, USA. We deployed remote cameras across 180 sites within the desert surrounding the populated garrison and classified sites by mange presence or absence depending on whether a symptomatic or asymptomatic coyote was photographed. Coyotes selected flatter sites closer to the urban area with a high probability of use (0.845, 95% credible interval (CRI): 0.728, 0.944); site use decreased as the distance to urban areas increased (standardized $${\widehat{\beta}}$$ = − 1.354, 95% CRI − 2.423, − 0.619). The probability of correctly classifying mange presence at a site also decreased further from the urban area and was probably related to the severity of mange infection. Severely infected coyotes, which were more readily identified as symptomatic, resided closer to the urban area and were most likely dependent on urban resources for survival; urban resources probably contributed to sustaining the disease. Multistate occupancy models represent a flexible framework for estimating the occurrence and spatial extent of observable infectious diseases, which can improve wildlife disease surveillance programs.

Introduction: Patients with sickle cell disease (SCD) have repeated episodes of red blood cell (RBC) sickling and microvascular occlusion that manifest as pain crises, acute chest syndrome, and chronic hemolysis. These clinical sequelae usually increase during pregnancy. Given the racial distribution of SCD, patients with SCD are also more likely to have rarer RBC antigen genotypes than RBC donor populations. We present the management and clinical outcome of a 21-year-old pregnant woman with SCD and an RHD*39 (RhD[S103P], G-negative) variant. Case Presentation: Ms. S is B positive with a reported history of anti-D, anti-C, and anti-E alloantibodies (anti-G testing unknown). Genetic testing revealed both an RHD*39 and homozygous partial RHCE*ceVS.02 genotype. Absorption/elution testing confirmed the presence of anti-G, anti-C, and anti-E alloantibodies but could not definitively determine the presence/absence of an anti-D alloantibody. Ms. S desired to undergo elective pregnancy termination and the need for postprocedural RhD immunoglobulin (RhIG) was posed. Given that only the G antigen site is changed in an RHD*39 genotype and the potential risk of RhIG triggering a hyperhemolytic episode in an SCD patient, RhIG was not administered. There were no procedural complications. Follow-up testing at 10 weeks showed no increase in RBC alloantibody strength. Discussion/Conclusion: Ms. S represents a rare RHD*39 and partial RHCE*ceVS.02 genotype which did not further alloimmunize in the absence of RhIG administration. Her case also highlights the importance of routine anti-G alloantibody testing in women of childbearing age with apparent anti-D and anti-C alloantibodies.

The diverse members of the Enterobacterales are agriculturally and medically relevant species that have continued to undergo taxonomic revision. To assess the current taxonomy of 64 genera of the Enterobacterales , we carried out a phylogenetic analysis using 32 single-copy core proteins. The resulting phylogeny was robust, and shows that eight genera – Biostraticola , Enterobacillus , Gibbsiella , Limnobaculum , Izhakiella , ‘Nissabacter’, Rosenbergiella and Samsonia – are currently assigned to incorrect families. Taxonomic reassignment of these genera was also supported by average amino acid identity comparisons. We propose taxonomic revision of these genera to reflect their phylogenetic position within the Enterobacterales .

Birds are good indicators of ecosystem health. Monitoring populations can provide useful information to inform conservation action. Kaipatiki, Auckland, contains approximately 450 ha of indigenous ...

Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related death in the United States. Despite the high prevalence of Kras mutations in pancreatic cancer patients, murine models expressing the oncogenic mutant Kras (Krasmut) in mature pancreatic cells develop PDAC at a low frequency. Independent of cell of origin, a second genetic hit (loss of tumor suppressor TP53 or PTEN) is important for development of PDAC in mice. We hypothesized ectopic expression and elevated levels of oncogenic mutant Kras would promote PanIN arising in pancreatic ducts. To test our hypothesis, the significance of elevating levels of K-Ras and Ras activity has been explored by expression of a CAG driven LGSL-KrasG12V allele (cKras) in pancreatic ducts, which promotes ectopic Kras expression. We predicted expression of cKras in pancreatic ducts would generate neoplasia and PDAC. To test our hypothesis, we employed tamoxifen dependent CreERT2 mediated recombination. Hnf1b:CreERT2;KrasG12V (cKrasHnf1b/+) mice received 1 (Low), 5 (Mod) or 10 (High) mg per 20 g body weight to recombine cKras in low (cKrasLow), moderate (cKrasMod), and high (cKrasHigh) percentages of pancreatic ducts. Our histologic analysis revealed poorly differentiated aggressive tumors in cKrasHigh mice. cKrasMod mice had grades of Pancreatic Intraepithelial Neoplasia (PanIN), recapitulating early and advanced PanIN observed in human PDAC. Proteomics analysis revealed significant differences in PTEN/AKT and MAPK pathways between wild type, cKrasLow, cKrasMod, and cKrasHigh mice. In conclusion, in this study, we provide evidence that ectopic expression of oncogenic mutant K-Ras in pancreatic ducts generates early and late PanIN as well as PDAC. This Ras rheostat model provides evidence that AKT signaling is an important early driver of invasive ductal derived PDAC., Summary In this study, the authors provide evidence that ectopic expression of oncogenic mutant Kras in pancreatic ducts generates early and late (PanIN) and pancreatic ductal adenocarcinoma (PDAC) . They characterized this Ras rheostat model which reveals elevated Kras mutation frequency and loss of PTEN are important drivers of PanIN and invasive ductal derived PDAC.

A detailed understanding of population genetics in non-native populations helps us to identify drivers of successful introductions. However, separating adaptive change from local signatures of founding populations represents a conceptual and technical difficulty when dealing with invasive populations. The history of introductions, as well as the process of range expansion, can confound interpretation of putative adaption in response to a novel environment. Here, we investigate putative signals of selection in Australian populations of introduced common starlings, Sturnus vulgaris, by examining population wide Single Nucleotide Polymorphisms (SNPs) and identifying SNP outliers associated with environmental variables. We determine that geographic distance plays a strong role in the genetic structure of populations, and that this is most likely strongly influenced by genetic differences in the founding populations, as well as modern day population connectivity. Examining candidate SNPs under putative selection indicated that local adaption has likely occurred, however, strong patterns in genetic variation, likely from founding populations, were visible in SNPs that were strongly associated with environmental variables. When examining putative adaption in invasive populations, we encourage critical interpretation of signatures of selection. Even strongly associated loci and environmental variables, when examined closely, may contain distinct footprints of invasion history or invasion expansion gradients, confounding analysis of the history of selection in these populations.

© 2020 The Authors. Evolutionary Applications published by John Wiley & Sons Ltd Habitat fragmentation imperils the persistence of many functionally important species, with climate change a new threat to local persistence due to climate niche mismatching. Predicting the evolutionary trajectory of species essential to ecosystem function under future climates is challenging but necessary for prioritizing conservation investments. We use a combination of population genetics and niche suitability models to assess the trajectory of a functionally important, but highly fragmented, plant species from south-eastern Australia (Banksia marginata, Proteaceae). We demonstrate significant genetic structuring among, and high level of relatedness within, fragmented remnant populations, highlighting imminent risks of inbreeding. Population simulations, controlling for effective population size (Ne), suggest that many remnant populations will suffer rapid declines in genetic diversity due to drift in the absence of intervention. Simulations were used to demonstrate how inbreeding and drift processes might be suppressed by assisted migration and population mixing approaches that enhance the size and connectivity of remnant populations. These analyses were complemented by niche suitability models that predicted substantial reductions of suitable habitat by 2080; ~30% of the current distribution of the species climate niche overlaps with the projected distribution of the species climate niche in the geographic region by the 2080s. Our study highlights the importance of conserving remnant populations and establishing new populations in areas likely to support B. marginata in the future, and adopting seed sourcing strategies that can help populations overcome the risks of inbreeding and maladaptation. We also argue that ecological replacement of B. marginata using climatically suited plant species might be needed in the future to maintain ecosystem processes where B. marginata cannot persist. We recommend the need for progressive revegetation policies and practices to prevent further deterioration of species such as B. marginata and the ecosystems they support.

In response to a gap in veterinary care for low-income, under-served populations, the Student Health Outreach for Wellness (SHOW) Clinic in Phoenix, AZ took a One-Health initiative to establish a pilot veterinary care program to serve the veterinary needs of the local homeless pet owners receiving human medical care at the clinic. The study examines the pilot program through the lens of the human-animal bond (HAB) and focuses specifically on the attachment dimension of the bond as measured by the Lexington Attachment to Pets Scale (LAPS). Data was collected from the homeless pet owners, and Rasch modeling was used to investigate the importance of various factors of the owner attachment to pet as described in LAPS items. The results demonstrate the perceptions of the homeless pet owners of ownership. The findings also provide psychometric evidence for revising the LAPS to better serve future research on human-animal relationships. Limitations and possible extensions of the research as related to the HAB and owner attachment to pets are discussed and enhancements proposed for the pilot program to better serve the local homeless people.

B lymphoid development is initiated by the differentiation of hematopoietic stem cells into lineage committed progenitors, ultimately generating mature B cells. This highly regulated process generates clonal immunological diversity via recombination of immunoglobulin V, D and J gene segments. While several transcription factors that control B cell development and V(D)J recombination have been defined, how these processes are initiated and coordinated into a precise regulatory network remains poorly understood. Here, we show that the transcription factor ETS Related Gene (Erg) is essential for early B lymphoid differentiation. Erg initiates a transcriptional network involving the B cell lineage defining genes, Ebf1 and Pax5, which directly promotes expression of key genes involved in V(D)J recombination and formation of the B cell receptor. Complementation of Erg deficiency with a productively rearranged immunoglobulin gene rescued B lineage development, demonstrating that Erg is an essential and stage-specific regulator of the gene regulatory network controlling B lymphopoiesis., B cell development is tightly regulated in a stepwise manner to ensure proper generation of repertoire diversity via somatic gene rearrangements. Here, the authors show that a transcription factor, Erg, functions at the earliest stage to critically control two downstream factors, Ebf1 and Pax5, for modulating this gene rearrangement process.

Modern metabolomic approaches that generate more comprehensive phytochemical profiles than were previously available are providing new opportunities for understanding plant‐animal interactions. Specifically, we can characterize the phytochemical landscape by asking how a larger number of individual compounds affect herbivores and how compounds covary among plants. Here we use the recent colonization of alfalfa (Medicago sativa) by the Melissa blue butterfly (Lycaeides melissa) to investigate the effects of indivdiual compounds and suites of covarying phytochemicals on caterpillar performance. We find that survival, development time, and adult weight are all associated with variation in nutrition and toxicity, including biomolecules associated with plant cell function as well as putative anti‐herbivore action. The plant‐insect interface is complex, with clusters of covarying compounds in many cases encompassing divergent effects on different aspects of caterpillar performance. Individual compounds with the strongest associations are largely specialized metabolites, including alkaloids, phenolic glycosides, and saponins. The saponins are represented in our data by more than 25 individual compounds with beneficial and detrimental effects on L. melissa caterpillars, which highlights the value of metabolomic data as opposed to approaches that rely on total concentrations within broad defensive classes., We use modern metabolomic profiling to dissect the complexity of phytochemistry affecting the development and survival of a specialist caterpillar on a novel host plant. A diversity of effects are uncovered, including positive and negative consequences of variation in metabolites with a range of function. Associations among plant traits are discussed in the context of the evolution of this plant‐insect interaction.

The health of seagrass plants, and thereby the ecosystems they form, is linked to their associated microbial communities. However, the role of the microbiome in holobiont function and health remains poorly understood for most seagrass species and environmental pressures, and there is, therefore, a need to better understand the drivers behind the formation of and external influences on the seagrass microbiome. Using a core microbiome framework, we characterised the leaf microbiomes of 6 estuarine seagrass populations after a precipitation event to explore how the microbiomes vary across different sites and salinities over a regional spatial scale. We found that each estuary had distinct core bacterial community structures (beta-diversity), but shared a more similar fungal core community structure. We hypothesise that the differences in the bacterial members of the microbiomes among estuaries are generally the result of each estuary being influenced by unique watersheds and sources of prokaryotes. In contrast, the similarity in the core fungal communities suggests that the eukaryotic components of the microbiomes are likely under selection or result from similar colonisation pathway(s). We also found that the bacterial taxa driving the differences among estuaries were linked to the salinity of the estuary, likely due to (1) the general epibiotic nature of colonisation (i.e. watershed source and exposure) and (2) members or functional groups within the leaf microbiome assisting seagrasses in coping with the extreme salinities. These results are valuable for linking microbiomes to the resilience of seagrasses living within dynamic estuaries experiencing a range of physicochemical pressures.

Rising ocean temperatures and extreme temperature events have precipitated declines and local extinctions in many marine species globally, but patterns of loss are often uneven across species ranges for reasons that are poorly understood. Knowledge of the extent of local adaptation and gene flow may explain such patterns and help predict future trajectories under scenarios of climate change. We test the extent to which local differentiation in thermal tolerance is influenced by gene flow and local adaptation using a widely distributed intertidal seaweed (Hormosira banksii) from temperate Australia. Population surveys across ~2,000 km of the species range revealed strong genetic structuring at regional and local scales (global F ST = 0.243) reflecting extremely limited gene flow, while common garden experiments (14‐day exposures to 15, 18, 21°C) revealed strong site differences in early development and mortality in response to elevated temperature. Embryos from many sites spanning a longitudinal thermal gradient showed suppressed development and increased mortality to elevated water temperatures, but populations originating from warmer and more variable thermal environments tended to be less susceptible to warming. Notably, there was significant local‐scale variation in the thermal responses of embryos within regions which was corroborated by the finding of small‐scale genetic differences. We expect the observed genetic and phenotypic differentiation to lead to uneven responses to warming sea surface temperatures in this important marine foundation species. The study highlights the challenges of predicting species responses to thermal stress and the importance of management strategies that incorporate evolutionary potential for “climate‐proofing” marine ecosystems.

Transcription start site (TSS) selection is a key step in gene expression and occurs at many promoter positions over a wide range of efficiencies. Here, we develop a massively parallel reporter assay to quantitatively dissect contributions of promoter sequence, NTP substrate levels, and RNA polymerase II (Pol II) activity to TSS selection by "promoter scanning" inSaccharomyces cerevisiae(Pol II MAssively Systematic Transcript End Readout, "Pol II MASTER"). Using Pol II MASTER, we measure the efficiency of Pol II initiation at 1,000,000 individual TSS sequences in a defined promoter context. Pol II MASTER confirms proposed critical qualities ofS. cerevisiaeTSS -8, -1, and +1 positions quantitatively in a controlled promoter context. Pol II MASTER extends quantitative analysis to surrounding sequences and determines that they tune initiation over a wide range of efficiencies. These results enabled the development of a predictive model for initiation efficiency based on sequence. We show that genetic perturbation of Pol II catalytic activity alters initiation efficiency mostly independently of TSS sequence, but selectively modulates preference for initiating nucleotide. Intriguingly, we find that Pol II initiation efficiency is directly sensitive to GTP levels at the first five transcript positions and to CTP and UTP levels at the second position genome wide. These results suggest individual NTP levels can have transcript-specific effects on initiation, representing a cryptic layer of potential regulation at the level of Pol II biochemical properties. The results establish Pol II MASTER as a method for quantitative dissection of transcription initiation in eukaryotes.

The histone acetyltransferase HBO1 (MYST2, KAT7) is indispensable for postgastrulation development, histone H3 lysine 14 acetylation (H3K14Ac), and the expression of embryonic patterning genes. In this study, we report the role of HBO1 in regulating hematopoietic stem cell function in adult hematopoiesis. We used 2 complementary cre-recombinase transgenes to conditionally delete Hbo1 (Mx1-Cre and Rosa26-CreERT2). Hbo1-null mice became moribund due to hematopoietic failure with pancytopenia in the blood and bone marrow 2 to 6 weeks after Hbo1 deletion. Hbo1-deleted bone marrow cells failed to repopulate hemoablated recipients in competitive transplantation experiments. Hbo1 deletion caused a rapid loss of hematopoietic progenitors. The numbers of lineage-restricted progenitors for the erythroid, myeloid, B-, and T-cell lineages were reduced. Loss of HBO1 resulted in an abnormally high rate of recruitment of quiescent hematopoietic stem cells (HSCs) into the cell cycle. Cycling HSCs produced progenitors at the expense of self-renewal, which led to the exhaustion of the HSC pool. Mechanistically, genes important for HSC functions were downregulated in HSC-enriched cell populations after Hbo1 deletion, including genes essential for HSC quiescence and self-renewal, such as Mpl, Tek(Tie-2), Gfi1b, Egr1, Tal1(Scl), Gata2, Erg, Pbx1, Meis1, and Hox9, as well as genes important for multipotent progenitor cells and lineage-specific progenitor cells, such as Gata1. HBO1 was required for H3K14Ac through the genome and particularly at gene loci required for HSC quiescence and self-renewal. Our data indicate that HBO1 promotes the expression of a transcription factor network essential for HSC maintenance and self-renewal in adult hematopoiesis.

Foundational hypotheses addressing plant–insect codiversification and plant defense theory typically assume a macroevolutionary pattern whereby closely related plants have similar chemical profiles. However, numerous studies have documented variation in the degree of phytochemical trait lability, raising the possibility that phytochemical evolution is more nuanced than initially assumed. We utilize proton nuclear magnetic resonance (1H NMR) data, chemical classification, and double digest restriction-site associated DNA sequencing (ddRADseq) to resolve evolutionary relationships and characterize the evolution of secondary chemistry in the Neotropical plant clade Radula (Piper; Piperaceae). Sequencing data substantially improved phylogenetic resolution relative to past studies, and spectroscopic characterization revealed the presence of 35 metabolite classes. Metabolite classes displayed phylogenetic signal, whereas the crude 1H NMR spectra featured little evidence of phylogenetic signal in multivariate tests of chemical resonances. Evolutionary correlations were detected in two pairs of compound classes (flavonoids with chalcones; p-alkenyl phenols with kavalactones), where the gain or loss of a class was dependent on the other’s state. Overall, the evolution of secondary chemistry in Radula is characterized by strong phylogenetic signal of traditional compound classes and weak phylogenetic signal of specialized chemical motifs, consistent with both classic evolutionary hypotheses and recent examinations of phytochemical evolution in young lineages.

Scientists have long been fascinated by the complexity of eukaryotic transcription and the large numbers of proteins involved at each step in the process. In this issue of Cell, Schilbach et al. bring us one important step closer to the goal of a complete understanding of transcription at atomic resolution.

Next-generation sequencing has transformed the fields of ecological and evolutionary genetics by allowing for cost-effective identification of genome-wide variation. Single nucleotide polymorphism (SNP) arrays, or "SNP chips", enable very large numbers of individuals to be consistently genotyped at a selected set of these identified markers, and also offer the advantage of being able to analyse samples of variable DNA quality. We used reduced representation restriction-aided digest sequencing (RAD-seq) of 31 birds of the threatened hihi (Notiomystis cincta; stitchbird) and low-coverage whole genome sequencing (WGS) of 10 of these birds to develop an Affymetrix 50 K SNP chip. We overcame the limitations of having no hihi reference genome and a low quantity of sequence data by separate and pooled de novo assembly of each of the 10 WGS birds. Reads from all individuals were mapped back to these de novo assemblies to identify SNPs. A subset of RAD-seq and WGS SNPs were selected for inclusion on the chip, prioritising SNPs with the highest quality scores whose flanking sequence uniquely aligned to the zebra finch (Taeniopygia guttata) genome. Of the 58,466 SNPs manufactured on the chip, 72% passed filtering metrics and were polymorphic. By genotyping 1,536 hihi on the array, we found that SNPs detected in multiple assemblies were more likely to successfully genotype, representing a cost-effective approach to identify SNPs for genotyping. Here, we demonstrate the utility of the SNP chip by describing the high rates of linkage disequilibrium in the hihi genome, reflecting the history of population bottlenecks in the species.

Background Daratumumab (Dara), an anti-CD38 monoclonal antibody for hematologic malignancies, interferes with routine blood bank testing, specifically affecting the antibody screen and identification panels. In 2016, the AABB recommended performing a baseline phenotype or genotype before a patient (Pt) begins taking anti-CD38 to avoid this interference and potential problems with transfusion. The objective of this study was to assess red blood cell (RBC) utilization and subsequent incidence of alloimmunization to the transfused RBCs in patients receiving Dara. Methods and materials We monitored 244 patients taking Dara to determine their red blood cell transfusions and incidence of clinically significant antibody formation before and following administration of Dara. Poisson generalized estimating equations with log link were used comparing the post-Dara incidence and prevalence to those prior, with significance defined as p Results From September 1, 2015 to December 22, 2018, 244 patients on Dara were identified, of which 145 patients (59.4%) received a red blood cell transfusion. Antibody screens were performed on 97 of the 145 patients at least 2 weeks following RBC transfusion. Four of the total transfused patients (2.8% total, 4.1% patients with follow-up antibody screen testing) formed new clinically significant alloantibodies, which was not significantly different from Asare's hematologic incidence (p = .98/p = .49). Conclusions This study showed our patients on Dara did not form alloantibodies following RBC transfusion at a higher incidence than similar patient populations.

Climate change not only directly impacts marine environments by shifting water temperatures, salinity, pH and dissolved oxygen concentrations, but may also indirectly contribute to the emergence of additional ecosystem stressors, such as infectious diseases, including bivalve disseminated neoplasia. Disseminated neoplasia, a form of cancer found in some bivalves - recently discovered to be transmissible in at least six species - has been shown to impair bivalve health and fitness, with occasional mass outbreaks causing high levels of mortality. As the ability of the host bivalve to respond to disseminated neoplasia, and the survival and transmissibility of disseminated neoplasia both depend on environmental factors, it is crucial to understand the interaction between climate change and disseminated neoplasia epidemiology. Furthermore, with bivalves being species of high ecological and economic importance, there is a rising need for the development of efficient disseminated neoplasia diagnostic tools in order to detect, mitigate and potentially prevent deleterious disseminated neoplasia outbreaks. Therefore, in this study, we reviewed the current knowledge of climate impacted environmental parameters on disseminated neoplasia and identified best practices and methodology for the detection of transmissible disseminated neoplasia in the wild. By exploring the potential effects changing climate has on disseminated neoplasia dynamics, we identified future research directions in order to advance the field. This included using state of the art disease detection methods and taking into account species' ecological niches to understand the dynamic of disseminated neoplasia outbreaks in the wild and to investigate whether disseminated neoplasia is present in freshwater ecosystems. Finally, we provided a comprehensive step-by-step guideline for an evidence-based detection of this disease in marine ecosystems.

Summary Enriched tumor epithelium, tumor-associated stroma, and whole tissue were collected by laser microdissection from thin sections across spatially separated levels of ten high-grade serous ovarian carcinomas (HGSOCs) and analyzed by mass spectrometry, reverse phase protein arrays, and RNA sequencing. Unsupervised analyses of protein abundance data revealed independent clustering of an enriched stroma and enriched tumor epithelium, with whole tumor tissue clustering driven by overall tumor “purity.” Comparing these data to previously defined prognostic HGSOC molecular subtypes revealed protein and transcript expression from tumor epithelium correlated with the differentiated subtype, whereas stromal proteins (and transcripts) correlated with the mesenchymal subtype. Protein and transcript abundance in the tumor epithelium and stroma exhibited decreased correlation in samples collected just hundreds of microns apart. These data reveal substantial tumor microenvironment protein heterogeneity that directly bears on prognostic signatures, biomarker discovery, and cancer pathophysiology and underscore the need to enrich cellular subpopulations for expression profiling., Graphical abstract, Highlights • LMD was used to investigate 3-D molecular heterogeneity in ovarian cancer tissue • Diverse molecular profiles were identified from 3-D spatially separated samples • Molecular heterogeneity impacts HGSOC prognostic sub-type assignment • Proteomic heterogeneity analysis web portal deployed at www.lmdomics.org, Oncology; Cancer systems biology; Proteomics; Transcriptomics

This exploratory pilot study investigated the effects of chronic Δ-9-tetrahydrocannabinol (THC) and cannabidiol (CBD) on cerebral glucose metabolism in people with multiple sclerosis (PwMS). Compared with nonusers, THC users had hypermetabolism of 3 regions (p < 0.039, d >1.17) in left temporal areas, while CBD users had hypometabolism of 5 regions (p < 0.032, d > 1.31) in left temporal areas. This study highlights the need to discriminate between THC and CBD in future cannabis studies. Novelty Chronic THC and CBD use had disparate effects on cerebral glucose metabolism in PwMS.

Genomics and ancient DNA methods have revolutionized many areas of biology, including human evolution. Recently we have seen significant advances in archaeogenetics including the use of large-scale...

Herbivorous insects can defend themselves against pathogens via an immune response, which is influenced by the nutritional quality and phytochemistry of the host plant. However, it is unclear how these aspects of diet interact to influence the insect immune response and what role is played by ingested foliar microbes. We examined dietary protein, phytochemistry, and the caterpillar microbiome to understand variation in immune response of the Melissa blue butterfly, Lycaeides melissa. We also asked if these factors have host plant-specific effects by measuring L. melissa immune response when reared on a recently colonized exotic host plant (Medicago sativa) as compared to the immune response on an ancestral, native host (Astragalus canadensis). L. melissa did not experience immunological benefits directly related to consumption of the novel plant M. sativa. However, we did find negative, direct effects of phytochemical diversity and negative, direct effects of diet-derived microbial diversity on constitutive immune response for caterpillars fed M. sativa, as measured by phenoloxidase activity. Foliar protein did not directly influence the immune response, but did do so indirectly by increasing weight gain. Our results highlight the important effects of host diet on caterpillar physiology and raise the possibility that foliar microbiota, despite being rapidly passed through the gut, can affect the caterpillar immune response.

σV is an extracytoplasmic function (ECF) σ factor that is found exclusively in Firmicutes including Bacillus subtilis and the opportunistic pathogens Clostridioides difficile and Enterococcus faecalis. σV is activated by lysozyme and is required for lysozyme resistance. The activity of σV is normally inhibited by the anti-σ factor RsiV, a transmembrane protein. RsiV acts as a receptor for lysozyme. The binding of lysozyme to RsiV triggers a signal transduction cascade which results in degradation of RsiV and activation of σV . Like the anti-σ factors for several other ECF σ factors, RsiV is degraded by a multistep proteolytic cascade that is regulated at the step of site-1 cleavage. Unlike other anti-σ factors, site-1 cleavage of RsiV is not dependent upon a site-1 protease whose activity is regulated. Instead constitutively active signal peptidase cleaves RsiV at site-1 in a lysozyme-dependent manner. The activation of σV leads to the transcription of genes, which encode proteins required for lysozyme resistance.

Semi-arid to arid areas are prone to droughts which are frequent occurrences in South Africa’s rangelands. Drought can trigger plant species compositional changes in rangelands thus influencing the availability and quality of forage for livestock. The availability and quality of forage drives livestock production, especially among pastoralists who mainly rely on the natural rangeland to sustain their livestock and livelihoods. The aim of this study was to determine the forage selection of cross-bred sheep, Swakara sheep (Ovis aries) and Boer goats (Capra hircus), during the wet season in the summer rainfall Nama Karoo and winter rainfall Succulent Karoo biomes in Namaqualand, South Africa. Since this dryland pastoral system was experiencing drought conditions during the study period of 2016 & 2017, the focus of the study was placed within this context to determine, (1) the availability of forage for small ruminants, (2) the nature of diet selection of different small ruminant groups and how it vary between the two biomes, and (3) the role livestock species, breeds and season play in forage preferences. We established diet selection, preference and dietary overlap by using direct observation of livestock grazing in the veld. Results indicate that availability of forage influence diet selection and preferences of all small ruminant groups. There was a high dietary overlap between mixed bred and Swakara sheep irrespective of the biomes. During 2016 annual herbs made up a large proportion of the sheep’s diets indicating an overdependence to this forage resource when it became available in the rangeland. However, due to the drought spell extending into 2017, annual herbs became less available and therefore altered the forage choices of both sheep groups. Goats were primarily browsers in the Succulent Karoo biome where shrubs dominate, but were considered grazers in the Nama Karoo biome where grasses were dominant. Moreover, all small ruminant groups in this study showed non-selective grazing tendencies which is an important grazing management tool to ensure that rangeland resources are not over exploited. We conclude by arguing that the mixture of different livestock species by pastoralists is an effective diversification strategy for better use of rangelands with herbaceous and woody plants during drought periods.

Characterising adaptive genetic divergence among conspecific populations is often achieved by studying genetic variation across defined environmental gradients. In marine systems this is challenging due to a paucity of information on habitat heterogeneity at local and regional scales and a dependency on sampling regimes that are typically limited to broad longitudinal and latitudinal environmental gradients. As a result, the spatial scales at which selection processes operate and the environmental factors that contribute to genetic adaptation in marine systems are likely to be unclear. In this study we explore patterns of adaptive genetic structuring in a commercially- harvested abalone species (Haliotis rubra) from southeastern Australia, using a panel of genome-wide SNP markers (5,239 SNPs), and a sampling regime informed by marine LiDAR bathymetric imagery and 20-year hindcasted oceanographic models. Despite a lack of overall genetic structure across the sampling distribution, significant genotype associations with heterogeneous habitat features were observed at local and regional spatial scales, including associations with wave energy, ocean current, sea surface temperature, and geology. These findings provide insights into the potential resilience of the species to changing marine climates and the role of migration and selection on recruitment processes, with implications for conservation and fisheries management. This study points to the spatial scales at which selection processes operate in marine systems and highlights the benefits of geospatially-informed sampling regimes for overcoming limitations associated with marine population genomic research.

Access to good-quality forages is one of the major limitations to livestock production in semi-arid pastoral systems. This study aimed to determine whether there are differences in the nutritional quality of diets selected by herded and free-ranging goat and sheep flocks utilising Namaqualand Granite Renosterveld vegetation during the wet and dry seasons. Plant samples collected along the grazing routes of livestock were dried and analysed for their fibre, condensed tannin, total phenolic and mineral nutrient contents. The study showed that a large variety of forages were on offer and livestock groups selected different diets of which some were different to the total diet on offer. In general, significant deficiencies in phosphate, protein and energy in the diets selected by herded and free-ranging goats and sheep were observed in both wet and dry season. The quality of the diets selected by herded and free-ranging livestock was also found to be different from each other, with herded livestock generally selecting more nutrient-dense diets. Herding, therefore, allows livestock to access better-quality forage in the Namaqualand Granite Renosterveld rangeland, where it is generally poor.Keywords: herding, Namaqualand Granite Renosterveld, pastoral livestock, protein deficiencies, small stock nutrition

Understanding and predicting changes in phytoplankton populations requires knowledge of losses due not only to sedimentation and grazing, but also to intrinsic processes (here, collectively termed ���cell death���). Cell death is poorly understood, especially in freshwater phytoplankton, but experiments in culture often suggest involvement of abiotic factors (e.g. temperature, light, nutrients). The occurrence of cell death was examined in a simple, natural environment: a small, well-mixed, temperate, urban pond during a period of phytoplankton growth, from mid-July to mid-November. Abundances of 18 phytoplankton taxa were measured weekly and fluorescence microscopy and staining was used to detect dead cells (using SYTOX which measures loss of membrane integrity) and cells undergoing cell death (using Annexin-V, which measures lipid inversions of membranes, an early signal of cell death). Dead and dying cells occurred in most phytoplankton taxa, but incidence and timing varied considerably, e.g. species like the chlorophyte Ankistrodesmus spiralis showed 20���30% of cells staining with SYTOX and Annexin in late autumn when the population was decreasing, while the dinoflagellate Peridinium sp. showed staining of up to 50% of cells with STYOX throughout the period, and the cyanobacterium Microcystis aeruginosa occasionally showed staining of 100% of cells with SYTOX. Overall, there was some association between cell death staining and growth phase with 10���15% of the total community showing SYTOX and Annexin staining in late autumn, when most populations were declining. Cell death could not be correlated with thresholds or rapid changes in abiotic conditions (e.g. temperature, irradiance) or with indicators of nutrient limitation (e.g. N:P ratios). While abiotic factors have been clearly implicated in cell death within unialgal culture experiments, in natural freshwater ecosystems interactions between biotic factors, such as pathogens or allelopathy, may play greater roles in losses related to cell death and be distinct for different taxa.

While the goal of supplementation programs is to provide positive, population‐level effects for species of conservation concern, these programs can also present an inherent fitness risk when captive‐born individuals are fully integrated into the natural population. In order to evaluate the long‐term effects of a supplementation program and estimate the demographic and phenotypic factors influencing the fitness of a threatened population of Chinook Salmon (Oncorhynchus tshawytscha), we genotyped tissue samples spanning a 19‐year period (1998–2016) to generate pedigrees from adult fish returning to Johnson Creek, Idaho, USA. We expanded upon previous estimates of relative reproductive success (RRS) to include grandparentage analyses and used generalized linear models to determine whether origin (hatchery or natural) or phenotypic traits (timing of arrival to spawning grounds, body length, and age) significantly predicted reproductive success (RS) across multiple years. Our results provide evidence that this supplementation program with 100% natural‐origin broodstock provided a long‐term demographic boost to the population (mean of 4.56 times in the first generation and mean of 2.52 times in the second generation). Overall, when spawning in nature, hatchery‐origin fish demonstrated a trend toward lower RS compared to natural‐origin fish (p 0.05). These results indicate that supplementation programs that reduce the potential for genetic adaptation to captivity can be effective at increasing population abundance while limiting long‐term fitness effects on wild populations.

The question of what determines whether cells are big or small has been the focus of many studies because it is thought that such determinants underpin the coupling of cell growth with cell division. In contrast, what determines the overall pattern of how cell size is distributed within a population of wild type or mutant cells has received little attention. Knowing how cell size varies around a characteristic pattern could shed light on the processes that generate such a pattern and provide a criterion to identify its genetic basis. Here, we show that cell size values of wild typeSaccharomyces cerevisiaecells fit a gamma distribution, in haploid and diploid cells, and under different growth conditions. To identify genes that influence this pattern, we analyzed the cell size distributions of all single-gene deletion strains inSaccharomyces cerevisiae.We found that yeast strains which deviate the most from the gamma distribution are enriched for those lacking gene products functioning in gene expression, especially those in transcription or transcription-linked processes. We also show that cell size is increased in mutants carrying altered activity substitutions in Rpo21p/Rpb1, the largest subunit of RNA polymerase II (Pol II). Lastly, the size distribution of cells carrying extreme altered activity Pol II substitutions deviated from the expected gamma distribution. Our results are consistent with the idea that genetic defects in widely acting transcription factors or Pol II itself compromise both cell size homeostasis and how the size of individual cells is distributed in a population.

Replacement of the exocrine parenchyma by fibrous tissue is a main characteristic of chronic pancreatitis. Understanding the mechanisms of pancreatic fibrogenesis is critical for the development of preventive and therapeutic interventions. Cyclooxygenase-2 (COX-2), a rate-limiting enzyme for prostaglandin synthesis, is expressed in patients with chronic pancreatitis. However, it is unknown whether COX-2 can cause chronic pancreatitis. To investigate the roles of pancreatic acinar COX-2 in fibrogenesis and the development of chronic pancreatitis, COX-2 was ectopically expressed specifically in pancreatic acinar cells in transgenic mice. Histopathological changes and expression levels of several profibrogenic factors related to chronic pancreatitis were evaluated. COX-2 was expressed in the pancreas of the transgenic mice, as detected by Western blot analysis. Immunohistochemical staining showed COX-2 was specifically expressed in pancreatic acinar cells. COX-2 expression led to progressive changes in the pancreas, including pancreas megaly, persistent inflammation, collagen deposition, and acinar-to-ductal metaplasia. Quantitative RT-PCR and immunostaining showed that profibrogenic factors were upregulated and pancreatic stellate cells were activated in the COX-2 transgenic mice. Expression of COX-2 in pancreatic acinar cells is sufficient to induce chronic pancreatitis. Targeting this pathway may be valuable in the prevention of chronic pancreatitis. NEW & NOTEWORTHY COX-2 expression is observed in pancreatic tissues of human chronic pancreatitis. In this study, we showed that COX-2 expression caused the development of chronic pancreatitis in transgenic mice, supporting the idea that COX-2 inhibition may be an effective preventive and therapeutic strategy.

Purpose: Parathyroid hormone-related protein (PTHrP) is required for normal mammary gland development and biology. A PTHLH gene polymorphism is associated with breast cancer risk, and PTHrP promotes growth of osteolytic breast cancer bone metastases. Accordingly, current dogma holds that PTHrP is upregulated in malignant primary breast tumors, but solid evidence for this assumption is missing. Experimental Design: We used quantitative IHC to measure PTHrP in normal and malignant breast epithelia, and correlated PTHrP levels in primary breast cancer with clinical outcome. Results: PTHrP levels were markedly downregulated in malignant compared with normal breast epithelia. Moreover, low levels of nuclear localized PTHrP in cancer cells correlated with unfavorable clinical outcome in a test and a validation cohort of breast cancer treated at different institutions totaling nearly 800 cases. PTHrP mRNA levels in tumors of a third cohort of 737 patients corroborated this association, also after multivariable adjustment for standard clinicopathologic parameters. Breast cancer PTHrP levels correlated strongly with transcription factors Stat5a/b, which are established markers of favorable prognosis and key mediators of prolactin signaling. Prolactin stimulated PTHrP transcript and protein in breast cancer cell lines in vitro and in vivo, effects mediated by Stat5 through the P2 gene promoter, producing transcript AT6 encoding the PTHrP 1-173 isoform. Low levels of AT6, but not two alternative transcripts, correlated with poor clinical outcome. Conclusions: This study overturns the prevailing view that PTHrP is upregulated in primary breast cancers and identifies a direct prolactin–Stat5–PTHrP axis that is progressively lost in more aggressive tumors.