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Brain metastases comprise 40% of all metastatic tumours and breast tumours are among the tumours that most commonly metastasise to the brain, the role that epigenetic gene dysregulation plays in this process is not well understood. We carried out 450 K methylation array analysis to investigate epigenetically dysregulated genes in breast to brain metastases (BBM) compared to normal breast tissues (BN) and primary breast tumours (BP). For this, we referenced 450 K methylation data for BBM tumours prepared in our laboratory with BN and BP from The Cancer Genome Atlas. Experimental validation on our initially identified genes, in an independent cohort of BP and in BBM and their originating primary breast tumours using Combined Bisulphite and Restriction Analysis (CoBRA) and Methylation Specific PCR identified three genes (RP11-713P17.4, MIR124-2, NUS1P3) that are hypermethylated and three genes (MIR3193, CTD-2023M8.1 and MTND6P4) that are hypomethylated in breast to brain metastases. In addition, methylation differences in candidate genes between BBM tumours and originating primary tumours shows dysregulation of DNA methylation occurs either at an early stage of tumour evolution (in the primary tumour) or at a later evolutionary stage (where the epigenetic change is only observed in the brain metastasis). Epigentic changes identified could also be found when analysing tumour free circulating DNA (tfcDNA) in patient’s serum taken during BBM biopsies. Epigenetic dysregulation of RP11-713P17.4, MIR3193, MTND6P4 are early events suggesting a potential use for these genes as prognostic markers.

By shuffling biogeographical distributions, biological invasions can both disrupt long-standing associations between hosts and parasites and establish new ones. This creates natural experiments with which to study the ecology and evolution of host-parasite interactions. In estuaries of the Gulf of Mexico, the white-fingered mud crab (Rhithropanopeus harrisii) is infected by a native parasitic barnacle, Loxothylacus panopaei (Rhizocephala), which manipulates host physiology and behaviour. In the 1960s, L. panopaei was introduced to the Chesapeake Bay and has since expanded along the southeastern Atlantic coast, while host populations in the northeast have so far been spared. We use this system to test the host's transcriptomic response to parasitic infection and investigate how this response varies with the parasite's invasion history, comparing populations representing (i) long-term sympatry between host and parasite, (ii) new associations where the parasite has invaded during the last 60 years and (iii) naïve hosts without prior exposure. A comparison of parasitized and control crabs revealed a core response, with widespread downregulation of transcripts involved in immunity and moulting. The transcriptional response differed between hosts from the parasite's native range and where it is absent, consistent with previous observations of increased susceptibility in populations lacking exposure to the parasite. Crabs from the parasite's introduced range, where prevalence is highest, displayed the most dissimilar response, possibly reflecting immune priming. These results provide molecular evidence for parasitic manipulation of host phenotype and the role of gene regulation in mediating host-parasite interactions.

Misunderstandings regarding the term "false positive" present a significant hurdle to broad adoption of eDNA monitoring methods. Here, we identify three challenges to clear communication of false-positive error between scientists, managers, and the public. The first arises from a failure to distinguish between false-positive eDNA detection at the sample level and false-positive inference of taxa presence at the site level. The second is based on the large proportion of false positives that may occur when true-positive detections are likely to be rare, even when rates of contamination or other error are low. And the third misunderstanding occurs when conventional species detection approaches, often based on direct capture, are used to confirm eDNA approaches without acknowledging or quantifying the conventional approach's detection probability. The solutions to these issues include careful and consistent communication of error definitions, managing expectations of error rates, and providing a balanced discussion not only of alternative sources of species DNA, but also of the detection limitations of conventional methods. We argue that the benefit of addressing these misunderstandings will be increased confidence in the utility of eDNA methods and, ultimately, improved resource management using eDNA approaches. The term false positive is often misused in eDNA research and natural resource management. There are issues of scale of inference, the base rate fallacy, and confirmation errors using conventional methods of detection. We offer a perspective to guide discussions of errors in species detection.

Invasive species surveillance programs can utilize environmental DNA sampling and analysis to provide information on the presence of invasive species. Wider utilization of eDNA techniques for invasive species surveillance may be warranted. This paper covers topics directed towards invasive species managers and eDNA practitioners working at the intersection of eDNA techniques and invasive species surveillance. It provides background information on the utility of eDNA for invasive species management and points to various examples of its use across federal and international programs. It provides information on 1) why an invasive species manager should consider using eDNA, 2) deciding if eDNA can help with the manager's surveillance needs, 3) important components to operational implementation, and 4) a high-level overview of the technical steps necessary for eDNA analysis. The goal of this paper is to assist invasive species managers in deciding if, when, and how to use eDNA for surveillance. If eDNA use is elected, the paper provides guidance on steps to ensure a clear understanding of the strengths and limitation of the methods and how results can be best utilized in the context of invasive species surveillance.

Developing new implant surfaces with anti-adhesion bacterial properties used for medical devices remains a challenge. Here we describe a novel study investigating nanotopography influences on bacterial adhesion on surfaces with controlled interspatial nanopillar distances. The surfaces were coated with proteins (fibrinogen, collagen, serum and saliva) prior to

Ballast water is a leading pathway for the global introduction of aquatic nonindigenous species. Most international ships are expected to install ballast water management systems (BWMS) by 2024 to treat ballast water before release. This study examines if ballast water discharges managed by BWMS are meeting standards for organisms ≥50 μm in minimum dimension (i.e.,10 organisms per m

Incidental detection of species of concern (e.g., invasive species, pathogens, threatened and endangered species) during biodiversity assessments based on high‐throughput DNA sequencing holds significant risks in the absence of rigorous, fit‐for‐purpose data quality and reporting standards. Molecular biodiversity data are predominantly collected for ecological studies and thus are generated to common quality assurance standards. However, the detection of certain species of concern in these data would likely elicit interest from end users working in biosecurity or other surveillance contexts (e.g., pathogen detection in health‐related fields), for which more stringent quality control standards are essential to ensure that data are suitable for informing decision‐making and can withstand legal or political challenges. We suggest here that data quality and reporting criteria are urgently needed to enable clear identification of those studies that may be appropriately applied to surveillance contexts. In the interim, more pointed disclaimers on uncertainties associated with the detection and identification of species of concern may be warranted in published studies. This is not only to ensure the utility of molecular biodiversity data for consumers, but also to protect data generators from uncritical and potentially ill‐advised application of their science in decision‐making.

Developing new implant surfaces with anti-adhesion bacterial properties used for medical devices remains a challenge. Here we describe a novel study investigating nanotopography influences on bacterial adhesion on surfaces with controlled interspatial nanopillars distances. Surfaces were coated with proteins (fibrinogen, collagen, serum and saliva) prior to E. coli-WT adhesion under flow conditions. PiFM provided chemical mapping and showed that proteins adsorbed both between and onto the nanopillars with a preference to areas between the nanopillars. E. coli-WT adhered least to protein-coated areas with low surface nanopillar coverage, most to surfaces coated with saliva, while human serum led to the lowest adhesion. Protein-coated nanostructured surfaces affected the adhesion of E. coli-WT. Abstract text goes here.

Parasitism can represent a potent agent of selection, and introduced parasites have the potential to substantially alter their new hosts' ecology and evolution. While significant impacts have been reported for parasites that switch to new host species, the effects of macroparasite introduction into naïve populations of host species with which they have evolved remain poorly understood. Here, we investigate how the estuarine white‐fingered mud crab (Rhithropanopeus harrisii) has adapted to parasitism by an introduced rhizocephalan parasite (Loxothylacus panopaei) that castrates its host. While the host crab is native to much of the East and Gulf Coasts of North America, its parasite is native only to the southern end of this range. Fifty years ago, the parasite invaded the mid‐Atlantic, gradually expanding through previously naïve host populations. Thus, different populations of the same host species have experienced different degrees of historical interaction (and thus potential evolutionary response time) with the parasite: long term, short term, and naïve. In nine estuaries across this range, we examined whether and how parasite prevalence and host susceptibility to parasitism differs depending on the length of the host's history with the parasite. In field surveys, we found that the parasite was significantly more prevalent in its introduced range (i.e., short‐term interaction) than in its native range (long‐term interaction), a result that was also supported by a meta‐analysis of prevalence data covering the 50 years since its introduction. In controlled laboratory experiments, host susceptibility to parasitism was significantly higher in naïve hosts than in hosts from the parasite's native range, suggesting that host resistance to parasitism is under selection. These results suggest that differences in host–parasite historical interaction can alter the consequences of parasite introductions in host populations. As anthropogenically driven range shifts continue, disruptions of host–parasite evolutionary relationships may become an increasingly important driver of ecological and evolutionary change.

Interest in developing periphytic diatom and bacterial indicators of nutrient effects continues to grow in support of the assessment and management of stream ecosystems and their watersheds. However, temporal variability could confound relationships between indicators and nutrients, subsequently affecting assessment outcomes. To document how temporal variability affects measures of diatom and bacterial assemblages obtained from DNA metabarcoding, we conducted weekly periphyton and nutrient sampling from July to October 2016 in 25 streams in a 1293 km

Environmental DNA is one of the most promising new tools in the aquatic biodiversity monitoring toolkit, with particular appeal for applications requiring assessment of target taxa at very low population densities. And yet there persists considerable anxiety within the management community regarding the appropriateness of environmental DNA monitoring for certain tasks and the degree to which environmental DNA methods can deliver information relevant to management needs. This brief perspective piece is an attempt to address that anxiety by offering some advice on how end-users might best approach these new technologies. I do not here review recent developments in environmental DNA science, but rather I explore ways in which managers and decision-makers might become more comfortable adopting environmental DNA tools—or choosing not to adopt them, should circumstances so dictate. I attempt to contextualize the central challenges associated with acceptance of environmental DNA detection by contrasting them with traditional “catch-and-look” approaches to biodiversity monitoring. These considerations lead me to recommend the cultivation of four “virtues,” attitudes that can be brought into engagement with environmental DNA surveillance technologies that I hope will increase the likelihood that those engagements will be positive and that the future development and application of environmental DNA tools will further the cause of wise management.

Aim: The introduction of aquatic non-indigenous species (ANS) has become a major driver for global changes in species biogeography. We examined spatial patterns and temporal trends of ANS detections since 1965 to inform conservation policy and management.Location: Global. Methods: We assembled an extensive dataset of first records of detection of ANS (1965–2015) across 49 aquatic ecosystems, including the (a) year of first collection, (b) population status and (c) potential pathway(s) of introduction. Data were analysed at global and regional levels to assess patterns of detection rate, richness and trans-port pathways.Results: An annual mean of 43 (±16 SD) primary detections of ANS occurred—one new detection every 8.4 days for 50 years. The global rate of detections was rel-atively stable during 1965–1995, but increased rapidly after this time, peaking at roughly 66 primary detections per year during 2005–2010 and then declining marginally. Detection rates were variable within and across regions through time. Arthropods, molluscs and fishes were the most frequently reported ANS. Most ANS were likely introduced as stowaways in ships’ ballast water or biofouling, although direct evidence is typically absent.Main conclusions: This synthesis highlights the magnitude of recent ANS detections, yet almost certainly represents an underestimate as many ANS go unreported due to limited search effort and diminishing taxonomic expertise. Temporal rates of detec-tion are also confounded by reporting lags, likely contributing to the lower detection rate observed in recent years. There is a critical need to implement standardized, repeated methods across regions and taxa to improve the quality of global-scale comparisons and sustain core measures over longer time-scales. It will be fundamen-tal to fill in knowledge gaps given that invasion data representing broad regions of the world's oceans are not yet readily available and to maintain knowledge pipelines for adaptive management. Fil: Bailey, Sarah A.. Great Lakes Laboratory For Fisheries And Aquatic Scienc; Canadá Fil: Brown, Lyndsay. Marine Scotland Science; Reino Unido Fil: Campbell, Marnie L.. Deakin University; Australia Fil: Canning-Clode, João. Marine And Environmental Sciences Centre; Portugal Fil: Carlton, James T.. Williams College. Maritime Studies Program; Estados Unidos Fil: Castro, Nuno. Marine And Environmental Sciences Centre; Portugal Fil: Chainho, Paula. Marine And Environmental Sciences Centre; Portugal Fil: Chan, Farrah T.. Great Lakes Institute For Enviromental Research; Canadá Fil: Creed, Joel C.. Universidade do Estado de Rio do Janeiro; Brasil Fil: Curd, Amelia. Ifremer Institut Francais de Recherche Pour L'exploitation de la Mer; Francia Fil: Darling, John. United States Environmental Protection Agency; Estados Unidos Fil: Fofonoff, Paul. Smithsonian Environmental Research Ctr; Estados Unidos Fil: Galil, Bella S.. The Steinhardt Museum Of Natural History; Israel Fil: Hewitt, Chad L.. Murdoch University; Australia Fil: Inglis, Graeme J.. National Institute Of Water & Atmospheric Research Lt; Nueva Zelanda Fil: Keith, Inti. Charles Darwin Foundation; Ecuador Fil: Mandrak, Nicholas E.. University Of Toronto Scarborough; Canadá Fil: Marchini, Agnese. University of Pavia; Italia Fil: McKenzie, Cynthia H.. Fisheries And Ocean Canada. Northwest Atlantic Fisheries Centre; Canadá Fil: Occhipinti‐Ambrogi, Anna. University of Pavia; Italia Fil: Ojaveer, Henn. University of Tartu; Estonia Fil: Pires‐Teixeira, Larissa M.. Universidad de Lisboa; Portugal. Universidade do Estado do Rio de Janeiro; Brasil Fil: Robinson, Tamara B.. Stellenbosch University; Sudáfrica Fil: Ruiz, Gregory M.. United States Environmental Protection Agency; Estados Unidos Fil: Seaward, Kimberley. National Institute of Water & Atmospheric Research Ltd.; Nueva Zelanda Fil: Schwindt, Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; Argentina Fil: Son, Mikhail O.. Institute of Marine Biology; Ucrania Fil: Therriault, Thomas W.. Pacific Biological Station; Canadá Fil: Zhan, Aibin. Chinese Academy of Sciences; República de China

Non-native species pose one of the greatest threats to native biodiversity, and can have severe negative impacts in freshwater ecosystems. Identifying regions of spatial overlap between high freshwater biodiversity and high invasion pressure may thus better inform the prioritization of freshwater conservation efforts. We employ geospatial analysis of species distribution data to investigate the potential threat of non-native species to aquatic animal taxa across the continental United States. We mapped non-native aquatic plant and animal species richness and cumulative invasion pressure to estimate overall negative impact associated with species introductions. These distributions were compared to distributions of native aquatic animal taxa derived from the International Union for the Conservation of Nature (IUCN) database. To identify hotspots of native biodiversity we mapped total species richness, number of threatened and endangered species, and a community index of species rarity calculated at the watershed scale. An overall priority index allowed identification of watersheds experiencing high pressure from non-native species and also exhibiting high native biodiversity conservation value. While priority regions are roughly consistent with previously reported prioritization maps for the US, we also recognize novel priority areas characterized by moderate-to-high native diversity but extremely high invasion pressure. We further compared priority areas with existing conservation protections as well as projected future threats associated with land use change. Our findings suggest that many regions of elevated freshwater biodiversity value are compromised by high invasion pressure, and are poorly safeguarded by existing conservation mechanisms and are likely to experience significant additional stresses in the future.

Ballast water remains a potent vector of non-native aquatic species introductions, despite increased global efforts to reduce risk of ballast water mediated invasions. This is particularly true of intracoastal vessel traffic, whose characteristics may limit the feasibility and efficacy of management through ballast water exchange (BWE). Here we utilize high throughput sequencing (HTS) to assess biological communities associated with ballast water being delivered to Valdez, Alaska from multiple source ports along the Pacific Coast of the United States. Our analyses indicate that BWE has a significant but modest effect on ballast water assemblages. Although overall richness was not reduced with exchange, we detected losses of some common benthic coastal taxa (e.g., decapods, mollusks, bryozoans, cnidaria) and gains of open ocean taxa (e.g., certain copepods, diatoms, and dinoflagellates), including some potentially toxic species. HTS-based metabarcoding identified significantly differentiated biodiversity signatures from individual source ports; this signal persisted, though weakened, in vessels undergoing BWE, indicating incomplete faunal turnover associated with management. Our analysis also enabled identification of taxa that may be of particular concern if established in Alaskan waters. While these results reveal a clear effect of BWE on diversity in intracoastal transit, they also indicate continued introduction risk of non-native and harmful taxa.

Understanding the risks of biological invasion posed by ballast water—whether in the context of compliance testing, routine monitoring, or basic research—is fundamentally an exercise in biodiversity assessment, and as such should take advantage of the best tools available for tackling that problem. The past several decades have seen growing application of genetic methods for the study of biodiversity, driven in large part by dramatic technological advances in nucleic acids analysis. Monitoring approaches based on such methods have the potential to increase dramatically sampling throughput for biodiversity assessments, and to improve on the sensitivity, specificity, and taxonomic accuracy of traditional approaches. The application of targeted detection tools (largely focused on PCR but increasingly incorporating novel probe-based methodologies) has led to a paradigm shift in rare species monitoring, and such tools have already been applied for early detection in the context of ballast water surveillance. Rapid improvements in community profiling approaches based on high throughput sequencing (HTS) could similarly impact broader efforts to catalogue biodiversity present in ballast tanks, and could provide novel opportunities to better understand the risks of biotic exchange posed by ballast water transport—and the effectiveness of attempts to mitigate those risks. These various approaches still face considerable challenges to effective implementation, depending on particular management or research needs. Compliance testing, for instance, remains dependent on accurate quantification of viable target organisms; while tools based on RNA detection show promise in this context, the demands of such testing require considerable additional investment in methods development. In general surveillance and research contexts, both targeted and community-based approaches are still limited by various factors: quantification remains a challenge (especially for taxa in larger size classes), gaps in nucleic acids reference databases are still considerable, uncertainties in taxonomic assignment methods persist, and many applications have not yet matured sufficiently to offer standardized methods capable of meeting rigorous quality assurance standards. Nevertheless, the potential value of these tools, their growing utilization in biodiversity monitoring, and the rapid methodological advances over the past decade all suggest that they should be seriously considered for inclusion in the ballast water surveillance toolkit.

This study presents the first large-scale assessment of cyanobacterial frequency and abundance of surface water near drinking water intakes across the United States. Public water systems serve drinking water to nearly 90% of the United States population. Cyanobacteria and their toxins may degrade the quality of finished drinking water and can lead to negative health consequences. Satellite imagery can serve as a cost-effective and consistent monitoring technique for surface cyanobacterial blooms in source waters and can provide drinking water treatment operators information for managing their systems. This study uses satellite imagery from the European Space Agency's Ocean and Land Colour Instrument (OLCI) spanning June 2016 through April 2020. At 300-m spatial resolution, OLCI imagery can be used to monitor cyanobacteria in 685 drinking water sources across 285 lakes in 44 states, referred to here as resolvable drinking water sources. First, a subset of satellite data was compared to a subset of responses (n = 84) submitted as part of the U.S. Environmental Protection Agency's fourth Unregulated Contaminant Monitoring Rule (UCMR 4). These UCMR 4 qualitative responses included visual observations of algal bloom presence and absence near drinking water intakes from March 2018 through November 2019. Overall agreement between satellite imagery and UCMR 4 qualitative responses was 94% with a Kappa coefficient of 0.70. Next, temporal frequency of cyanobacterial blooms at all resolvable drinking water sources was assessed. In 2019, bloom frequency averaged 2% and peaked at 100%, where 100% indicated a bloom was always present at the source waters when satellite imagery was available. Monthly cyanobacterial abundances were used to assess short-term trends across all resolvable drinking water sources and effect size was computed to provide insight on the number of years of data that must be obtained to increase confidence in an observed change. Generally, 2016 through 2020 was an insufficient time period for confidently observing changes at these source waters; on average, a decade of satellite imagery would be required for observed environmental trends to outweigh variability in the data. However, five source waters did demonstrate a sustained short-term trend, with one increasing in cyanobacterial abundance from June 2016 to April 2020 and four decreasing.

Following decades of ecologic and economic impacts from a growing list of nonindigenous and invasive species, government and management entities are committing to systematic early- detection monitoring (EDM). This has reinvigorated investment in the science underpinning such monitoring, as well as the need to convey that science in practical terms to those tasked with EDM implementation. Using the context of nonindigenous species in the North American Great Lakes, this article summarizes the current scientific tools and knowledge - including limitations, research needs, and likely future developments - relevant to various aspects of planning and conducting comprehensive EDM. We begin with the scope of the effort, contrasting target-species with broad-spectrum monitoring, reviewing information to support prioritization based on species and locations, and exploring the challenge of moving beyond individual surveys towards a coordinated monitoring network. Next, we discuss survey design, including effort to expend and its allocation over space and time. A section on sample collection and analysis overviews the merits of collecting actual organisms versus shed DNA, reviews the capabilities and limitations of identification by morphology, DNA target markers, or DNA barcoding, and examines best practices for sample handling and data verification. We end with a section addressing the analysis of monitoring data, including methods to evaluate survey performance and characterize and communicate uncertainty. Although the body of science supporting EDM implementation is already substantial, research and information needs (many already actively being addressed) include: better data to support risk assessments that guide choice of taxa and locations to monitor; improved understanding of spatiotemporal scales for sample collection; further development of DNA target markers, reference barcodes, genomic workflows, and synergies between DNA-based and morphology-based taxonomy; and tools and information management systems for better evaluating and communicating survey outcomes and uncertainty.

Aim Mapping the geographic distribution of non-native aquatic species is a critically important precursor to understanding the anthropogenic and environmental factors that drive freshwater biological invasions. Such efforts are often limited to local scales and/or to single species, due to the challenges of data acquisition at larger scales. Here, we map the distribution of non-native freshwater species richness across the continental United States and investigate the role of human activity in driving macroscale patterns of aquatic invasion. Location The continental United States. Methods We assembled maps of non-native aquatic species richness by compiling occurrence data on exotic animal and plant species from publicly accessible databases. Using a dasymetric model of human population density and a spatially explicit model of recreational freshwater fishing demand, we analysed the effect of these metrics of human influence on the degree of invasion at the watershed scale, while controlling for spatial and sampling bias. We also assessed the effects that a temporal mismatch between occurrence data (collected since 1815) and cross-sectional predictors (developed using 2010 data) may have on model fit. Results Non-native aquatic species richness exhibits a highly patchy distribution, with hotspots in the Northeast, Great Lakes, Florida, and human population centres on the Pacific coast. These richness patterns are correlated with population density, but are much more strongly predicted by patterns of recreational fishing demand. These relationships are strengthened by temporal matching of datasets and are robust to corrections for sampling effort. Main conclusions Distributions of aquatic non-native species across the continental US are better predicted by freshwater recreational fishing than by human population density. This suggests that observed patterns are driven by a mechanistic link between recreational activity and aquatic non-native species richness and are not merely the outcome of sampling bias associated with human population density.

Nutrient pollution from human activities remains a common problem facing stream ecosystems. Identifying ecological responses to phosphorus and nitrogen can inform decisions affecting the protection and management of streams and their watersheds. Diatoms are particularly useful because they are a highly diverse group of unicellular algae found in nearly all aquatic environments and are sensitive responders to increased nutrient concentrations. Here, we used DNA metabarcoding of stream diatoms as an approach to quantifying effects of total phosphorus (TP) and total nitrogen (TN). Threshold indicator taxa analysis (TITAN) identified operational taxonomic units (OTUs) that increased or decreased along TP and TN gradients along with nutrient concentrations at which assemblages had substantial changes in the occurrences and relative abundances of OTUs. Boosted regression trees showed that relative abundances of gene sequence reads for OTUs identified by TITAN as low P, high P, low N, or high N diatoms had strong relationships with nutrient concentrations, which provided support for potentially using these groups of diatoms as metrics in monitoring programs. Gradient forest analysis provided complementary information by characterizing multi‐taxa assemblage change using multiple predictors and results from random forest models for each OTU. Collectively, these analyses showed that notable changes in diatom assemblage structure and OTUs began around 20 µg TP/L, low P diatoms decreased substantially and community change points occurred from 75 to 150 µg/L, and high P diatoms became increasingly dominant from 150 to 300 µg/L. Diatoms also responded to TN with large decreases in low N diatoms occurring from 280 to 525 µg TN/L and a transition to dominance by high N diatoms from 525–850 µg/L. These diatom responses to TP and TN could be used to inform protection efforts (i.e., anti‐degradation) and management goals (i.e., nutrient reduction) in streams and watersheds. Our results add to the growing support for using diatom metabarcoding in monitoring programs.

Characterizing biodiversity conveyed in ships’ ballast water (BW), a global driver of biological invasions, is critically important for understanding risks posed by this key vector and establishing baselines to evaluate changes associated with BW management. Here we employ high throughput sequence (HTS) metabarcoding of the 18S small subunit rRNA to test for and quantify differences in the accumulation of BW-borne biodiversity among three distinct recipient port systems in the United States. These systems were located on three different coasts (Pacific, Gulf, and Atlantic) and chosen to reflect distinct tradepatterns and source port biogeography. Extensive sampling of BW tanks (n = 116) allowed detailed exploration of molecular diversity accumulation. Our results indicate that saturation of introduced zooplankton diversity may be achieved quickly, with fewer than 25 tanks needed to achieve 95% of the total extrapolated diversity, if sou rce biogeography is relatively limited. However, as predicted, port systems with much broader source geographies require more extensive sampling to estimate diversity, which continues to accumulate after sampling >100 discharges. The ability to identify BW sources using molecular indicators was also found to depend on the breadth of source biogeography and the extent to which sources had been sampled. These findings have implications both for the effort required to fully understand introduced diversity and for projecting risks associated with future changes to maritime traffic that may increase source biogeography for many recipient ports. Our data also suggest that molecular diversity may not decline significantly with BW age, indicating either that some organisms survive longer than recognized in previous studies or that nucleic acids from dead organisms persist in BW tanks. We present evidence for detection of potentially invasive species in arriving BW but discuss important caveats that preclude strong inferences regarding the presence of living representatives of these species in BW tanks., Graphical Abstract

Cyanobacterial harmful algal blooms are the most common form of harmful algal blooms in freshwater systems throughout the world. However, in situ sampling of cyanobacteria in inland lakes is limited both spatially and temporally. Satellite data has proven to be an effective tool to monitor cyanobacteria in freshwater lakes across the United States. This study uses data from the European Space Agency Envisat MEdium Resolution Imaging Spectrometer and the Sentinel-3 Ocean and Land Color Instrument to provide a national overview of the percentage of lakes experiencing a cyanobacterial bloom on a weekly basis for 2008–2011, 2017, and 2018. A total of 2321 lakes across the contiguous United States were included in the analysis. We examined four different thresholds to define when a waterbody is classified as experiencing a bloom. Across these four thresholds, we explored variability in bloom percentage with changes in seasonality and lake size. As a validation of algorithm performance, we analyzed the agreement between satellite observations and previously established ecological patterns, although data availability in the wintertime limited these comparisons on a year-round basis. Changes in cyanobacterial bloom percentage at the national scale followed the well-known temporal pattern of freshwater blooms. The percentage of lakes experiencing a bloom increased throughout the year, reached a maximum in fall, and decreased through the winter. Wintertime data, particularly in northern regions, were consistently limited due to snow and ice cover. With the exception of the Southeast and South, regional patterns mimicked patterns found at the national scale. The Southeast and South exhibited an unexpected pattern as cyanobacterial bloom percentage reached a maximum in the winter rather than the summer. Lake Jesup in Florida was used as a case study to validate this observed pattern against field observations of chlorophyll a. Results from this research establish a baseline of annual occurrence of cyanobacterial blooms in inland lakes across the United States. In addition, methods presented in this study can be tailored to fit the specific requirements of an individual system or region.

Cyanobacterial harmful algal blooms (cyanoHAB) cause extensive problems in lakes worldwide, including human and ecological health risks, anoxia and fish kills, and taste and odor problems. CyanoHABs are a particular concern in both recreational waters and drinking water sources because of their dense biomass and the risk of exposure to toxins. Successful cyanoHAB assessment using satellites may provide an indicator for human and ecological health protection. In this study, methods were developed to assess the utility of satellite technology for detecting cyanoHAB frequency of occurrence at locations of potential management interest. The European Space Agency's MEdium Resolution Imaging Spectrometer (MERIS) was evaluated to prepare for the equivalent series of Sentinel-3 Ocean and Land Colour Imagers (OLCI) launched in 2016 as part of the Copernicus program. Based on the 2012 National Lakes Assessment site evaluation guidelines and National Hydrography Dataset, the continental United States contains 275,897 lakes and reservoirs >1 ha in area. Results from this study show that 5.6% of waterbodies were resolvable by satellites with 300 m single-pixel resolution and 0.7% of waterbodies were resolvable when a three by three pixel (3 × 3-pixel) array was applied based on minimum Euclidian distance from shore. Satellite data were spatially joined to U.S. public water surface intake (PWSI) locations, where single-pixel resolution resolved 57% of the PWSI locations and a 3 × 3-pixel array resolved 33% of the PWSI locations. Recreational and drinking water sources in Florida and Ohio were ranked from 2008 through 2011 by cyanoHAB frequency above the World Health Organization’s (WHO) high threshold for risk of 100,000 cells mL −1 . The ranking identified waterbodies with values above the WHO high threshold, where Lake Apopka, FL (99.1%) and Grand Lake St. Marys, OH (83%) had the highest observed bloom frequencies per region. The method presented here may indicate locations with high exposure to cyanoHABs and therefore can be used to assist in prioritizing management resources and actions for recreational and drinking water sources.

Cyanobacterial harmful algal blooms are the most common form of harmful algal blooms in freshwater systems throughout the world. However, in situ sampling of cyanobacteria in inland lakes is limited both spatially and temporally. Satellite data has proven to be an effective tool to monitor cyanobacteria in freshwater lakes across the United States. This study uses data from the European Space Agency MEdium Resolution Imaging Spectrometer and the Sentinel-3 Ocean and Land Color Instrument to provide a national overview of the percentage of lakes experiencing a cyanobacterial bloom on a weekly basis for 2008-2011 and 2017. A total of 2,370 lakes across the contiguous United States were included in the analysis. Bloom percentage was calculated for nine United States climate regions to examine regional patterns. Changes in cyanobacterial bloom percentage followed the well-known temporal pattern of freshwater blooms. The percentage of lakes experiencing a bloom increased throughout the year, reached a peak around October, and decreased through the winter. Wintertime data, particularly in the northern latitude regions, was consistently limited due to snow and ice cover. With the exception of the Southeast and South climate regions, regional patterns mimicked patterns found at the national scale. The Southeast and South regions exhibited an unexpected pattern as cyanobacterial bloom percentage peaked in the winter rather than the summer. Several environmental factors and potential satellite limitations can possibly explain these findings. Results from this research can help establish a baseline of annual occurrence of cyanobacterial blooms in inland lakes across the United States.

Understanding the spatial and temporal dynamics underlying the introduction and spread of nonindigenous aquatic species (NAS) can provide important insights into the historical drivers of biological invasions and aid in forecasting future patterns of nonindigenous species arrival and spread. Increasingly, public databases of species observation records are being used to quantify changes in NAS distributions across space and time, and are becoming an important resource for researchers, managers, and policy-makers. Here we use publicly available data to describe trends in NAS introduction and spread across the conterminous United States over more than two centuries of observation records. Available data on first records of NAS reveal significant shifts in dominance of particular introduction patterns over time, both in terms of recipient regions and likely sources. These spatiotemporal trends at the continental scale may be subject to biases associated with regional variation in sampling effort, reporting, and data curation. We therefore also examined two additional metrics, the number of individual records and the spatial coverage of those records, which are likely to be more closely associated with sampling effort. Our results suggest that broad-scale patterns may mask considerable variation across regions, time periods, and even entities contributing to NAS sampling. In some cases, observed temporal shifts in species discovery may be influenced by dramatic fluctuations in the number and spatial extent of individual observations, reflecting the possibility that shifts in sampling effort may obscure underlying rates of NAS introduction.

HK2 has a prominent role in glycolysis, acting as the rate-limiting step in the pathway. HK2 is highly expressed in many cancers, with overexpression associated with drug resistant phenotypes. In parallel, its inhibition has improved the effectiveness of anticancer agents. Evidence indicates that HK2 is epigenetically silenced in normal tissue, and hypomethylation is the driving factor of increased expression in tumours, suggesting HK2 as a potential genetic modifier and therapeutic target. This study determines the role of methylation and its association with expression levels of HK2 in glioblastoma multiforme (GBM) fresh frozen biopsies and patient-derived GBM short-term cultures, by pyrosequencing and quantitative PCR. Anti-proliferative effects of HK2 inhibitors and their mechanism of action was investigated by cytotoxicity assays and FACS analysis. Hypomethylation was seen in all GBM biopsies(n=85) compared to normal brain tissue; average methylation of 4.6% compared to 26% respectively (p< 0.0001). Hypomethylation was also in all culture samples(n=9). An increase in HK2 expression was detected, ranging between 6 to >1000fold change in GBM biopsies(n=50) and 30 to >1000 in cultures(n=10), compared to normal brain tissue. There was a strong correlation between methylation and level of expression (p< 0.0001) as biopsies with HK2 expression greater than the mean fold change additionally displayed hypomethylation levels below mean fold change) was associated with greater sensitivity to BPA. Significantly higher levels of apoptosis were induced after treatment with BPA at ID50 in cultures expressing high HK2 (>mean fold change) (p< 0.0014). This study establishes a strong correlation between hypomethylation and increased expression of HK2. Furthermore, demonstrating the potential of HK2 inhibitors as a novel therapy in a significant subset of GBM.

Aberrant cellular metabolism is recognized as a major event in the growth and development of many cancers and the targetting of metabolic defects in tumour cells represents a new therapeutic opportunity. For example, cells that do not express sufficient levels of argininosuccinate synthetase-1 (ASS1) or argininosuccinate lyase (ASL) become auxotrophic for arginine and require exogenous supply. Arginine deprivation using arginine deiminase (ADI-PEG20) is currently under evaluation in clinical trials for adult GBM. In this study, we investigated the arginine biosynthesis pathway in paediatric intracranial ependymoma, comprising 24 fresh frozen biopsies and 17 short-term cell cultures of low passage (

Biological invasions are largely considered to be a "numbers game", wherein the larger the introduction effort, the greater the probability that an introduced population will become established. However, conditions during transport - an early stage of the invasion - can be particularly harsh, thereby greatly reducing the size of a population available to establish in a new region. Some successful non-indigenous species are more tolerant of environmental and anthropogenic stressors than related native species, possibly stemming from selection (ie survival of only pre-adapted individuals for particular environmental conditions) during the invasion process. By reviewing current literature concerning population genetics and consequences of selection on population fitness, we propose that selection acting on transported populations can facilitate local adaptation, which may result in a greater likelihood of invasion than predicted by propagule pressure alone. Specifically, we suggest that detailed surveys should be conducted to determine interactions between molecular mechanisms and demographic factors, given that current management strategies may underestimate invasion risk.

Cyclic nucleotides (cAMP & cGMP) are critical intracellular second messengers involved in the transduction of a diverse array of stimuli and their catabolism is mediated by phosphodiesterases (PDEs). We previously detected focal genomic amplification of PDE1C in >90 glioblastoma multiforme (GBM) cells suggesting a potential as a novel therapeutic target in these cells. In this report, we show that genomic gain of PDE1C was associated with increased expression in low passage GBM-derived cell cultures. We demonstrate that PDE1C is essential in driving cell proliferation, migration and invasion in GBM cultures since silencing of this gene significantly mitigates these functions. We also define the mechanistic basis of this functional effect through whole genome expression analysis by identifying down-stream gene effectors of PDE1C which are involved in cell cycle and cell adhesion regulation. In addition, we also demonstrate that Vinpocetine, a general PDE1 inhibitor, can also attenuate proliferation with no effect on invasion/migration. Up-regulation of at least one of this gene set (IL8, CXCL2, FOSB, NFE2L3, SUB1, SORBS2, WNT5A, and MMP1) in TCGA GBM cohorts is associated with worse outcome and PDE1C silencing down-regulated their expression, thus also indicating potential to influence patient survival. Therefore we conclude that proliferation, migration, and invasion of GBM cells could also be regulated downstream of PDE1C.

The European Union’s Marine Strategy Framework Directive (MSFD) aims to adopt integrated ecosystem management approaches to achieve or maintain “Good Environmental Status” for marine waters, habitats and resources, including mitigation of the negative effects of non-indigenous species (NIS). The Directive further seeks to promote broadly standardized monitoring efforts and assessment of temporal trends in marine ecosystem condition, incorporating metrics describing the distribution and impacts of NIS. Accomplishing these goals will require application of advanced tools for NIS surveillance and risk assessment, particularly given known challenges associated with surveying and monitoring with traditional methods. In the past decade, a host of methods based on nucleic acids (DNA and RNA) analysis have been developed or advanced that promise to dramatically enhance capacity in assessing and managing NIS. However, ensuring that these rapidly evolving approaches remain accessible and responsive to the needs of resource managers remains a challenge. This paper provides recommendations for future development of these genetic tools for assessment and management of NIS in marine systems, within the context of the explicit requirements of the MSFD. Issues considered include technological innovation, methodological standardization, data sharing and collaboration, and the critical importance of shared foundational resources, particularly integrated taxonomic expertise. Though the recommendations offered here are not exhaustive, they provide a basis for future intentional (and international) collaborative development of a genetic toolkit for NIS research, capable of fulfilling the immediate and long term goals of marine ecosystem and resource conservation.

HK2 is highly expressed in several cancers including colon and breast and its role as the rate-limiting step of glycolysis may potentially contribute to tumour growth. Overexpression of HK2 has also been associated with drug resistant phenotypes. Inhibition of HK2 has increased the effectiveness of anticancer agents suggesting HK2 as both a potential genetic modifier and therapeutic target. There is evidence that HK2 is epigenetically silenced in normal tissue and that increased expression in tumours is due to hypomethylation. In this study, methylation status and expression levels of HK2 were determined in 36 glioblastoma multiforme (GBM) fresh frozen biopsies and 9 patient-derived GBM short-term cultures by methylation specific-PCR (MS-PCR) and RT-PCR respectively. More than 90% (n=33) of the GBM biopsies showed some hypomethylation of HK2, including 5 tumour samples that were completely hypomethylated. Total hypermethylation of HK2 was present in only 3 biopsy samples. There was good correlation between methylation status and level of expression; no mRNA transcripts were detected in tumours samples with complete HK2 methylation. There was also complete hypomethylation of HK2 in 4 (44%) of the short-term cultures and the remaining 5 cultures had only partial methlyation. Expression of HK2 was detected in all culture samples. This data demonstrates that hypomethylation of HK2 is a frequent event in GBM compared to normal brain. Furthermore, the high levels of HK2 expression detected in short-term cultures will enable future investigations of HK2 inhibitors as a therapeutic option in GBM.

Recent years have seen a dramatic rise in the application of genetic methods to understand aquatic biological invasions. In part these methods have been adopted to address fundamental questions in biogeography, evolutionary biology, population ecology, and other fields. But it is also commonly suggested that genetic information has the potential to directly inform the management of aquatic invasions. Here I explore the potential promise of genetic approaches for informing management of aquatic invasive species, the degree to which that promise has been realized in terms of utilization of genetic information by managers and other decision-makers, and the likely limitations to the value of genetic methods (both in principle and in practice) and ways in which these limitations might be overcome. I consider a range of possible applications of genetic tools for management, including molecular detection and identification of cryptic invaders, source tracking and reconstruction of invasion history, and inference of population demographics. Retrospective assessment of the utility of such applications is based on both literature review and solicitation of expert opinion, and suggests that a number of hurdles likely often prevent genetic information from effectively informing decision-making. These include (1) limitations or misunderstandings of the resolution and certainty afforded by genetic analysis; (2) failure to engage decision-makers in problem formulation, research design and research implementation; and (3) complex relationships between basic research and management actions. While some of the obstacles considered are rooted in theoretical and practical limitations of genetic analysis, others are clearly associated with poor communication and insufficient engagement of potential end-users of genetic information. I consider possible avenues for overcoming these obstacles and for improving the applicability of genetic information for supporting management decisions.

International audience; The expanding scale and increasing rate of marine biological invasions have been documented since the early 20th century. Besides their global ecological and economic impacts, non-indigenous species (NIS) also have attracted much attention as opportunities to explore important eco-evolutionary processes such as rapid adaptation, long-distance dispersal and range expansion, and secondary contacts between divergent evolutionary lineages. In this context, genetic tools have been extensively used in the past 20 years. Three important issues appear to have emerged from such studies. First, the study of NIS has revealed unexpected cryptic diversity in what had previously been assumed homogeneous entities. Second, there has been surprisingly little evidence of strong founder events accompanying marine introductions, a pattern possibly driven by large propagule loads. Third, the evolutionary processes leading to successful invasion have been difficult to ascertain due to faint genetic signals. Here we explore the potential of novel tools associated with high-throughput sequencing (HTS) to address these still pressing issues. Dramatic increase in the number of loci accessible via HTS has the potential to radically increase the power of analyses aimed at species delineation, exploring the population genomic consequences of range expansions, and examining evolutionary processes such as admixture, introgression, and adaptation. Nevertheless, the value of this new wealth of genomic data will ultimately depend on the ability to couple it with expanded " traditional " efforts, including exhaustive sampling of marine populations over large geographic scales, integrated taxonomic analyses, and population level exploration of quantitative trait differentiation through common-garden and other laboratory experiments.

HK2 has a prominent role in aerobic glycolysis and has been implicated in many cancer types including GBM, with overexpression associated with drug resistant phenotypes. Previously, we have demonstrated HK2 expression was upregulated between 6 to >1000 fold in GBM biopsy tissue (n=100) and patient derived cell cultures (n=13), compared to normal brain tissue. In the present study we have knocked out HK2 using CRISPR in patient-derived cultures (n=3) and the established cell line U251MG, to determine changes in the rate of cell proliferation and drug sensitivity. Additionally downstream expression changes were investigated via Qiagen profiler arrays, across 84 key genes involved in the regulation and enzymatic pathways of glucose metabolism. A substantial growth rate reduction between 38 to 44% (p

Background: Triple-negative breast cancer (TNBC) has significantly worse prognosis. Acquired chemoresistance remains the major cause of therapeutic failure of TNBC. In clinic, the relapsed TNBC is commonly pan-resistant to various drugs with completely different resistant mechanisms. Investigation of the mechanisms and development of new drugs to target pan-chemoresistance will potentially improve the therapeutic outcomes of TNBC patients. Methods: In this study, 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), combination index (CI)–isobologram, western blot, ALDEFLUOR analysis, clonogenic assay and immunocytochemistry were used. Results: The chemoresistant MDA-MB-231PAC10 cells are highly cross-resistant to paclitaxel (PAC), cisplatin (CDDP), docetaxel and doxorubicin. The MDA-MB-231PAC10 cells are quiescent with significantly longer doubling time (64.9 vs 31.7 h). This may be caused by high expression of p21Waf1. The MDA-MB-231PAC10 cells express high aldehyde dehydrogenase (ALDH) activity and a panel of embryonic stem cell-related proteins, for example, Oct4, Sox2, Nanog and nuclealisation of HIF2α and NF-κBp65. We have previously reported that disulfiram (DS), an antialcoholism drug, targets cancer stem cells (CSCs) and enhances cytotoxicity of anticancer drugs. Disulfiram abolished CSC characters and completely reversed PAC and CDDP resistance in MDA-MB-231PAC10 cells. Conclusion: Cancer stem cells may be responsible for acquired pan-chemoresistance. As a drug used in clinic, DS may be repurposed as a CSC inhibitor to reverse the acquired pan-chemoresistance.

Reconstructing the invasion history of aquatic invasive species can enhance understanding of invasion risks by recognizing areas most susceptible to invasion and forecasting future spread based on past patterns of population expansion. Here we reconstruct the invasion history of the Japanese amphipod Grandidierella japonica Stephensen 1938 combining information from historical collection data with molecular genetic data to better understand post-invasion range expansion and anthropogenic connectivity across the Pacific coast of North America. Compilation of collection data from bays and estuaries of the Pacific North American coast show many new localities have been colonized in the last two decades, moving outward from harbors and bays with high commercial traffic into smaller coastal locations dominated by local recreational traffic. DNA barcode sequence data for G. japonica reveals two distinct clades: one found in San Francisco Bay and sites to the north, and one also found in San Francisco Bay and sites to the south. The two clades differ by an average 7.28 % genetic distance, large enough to consider these invasive amphipods two separate species. Both northern and southern clades exhibit low levels of genetic diversity, suggesting a single introduction event for each. The presence of cryptic diversity within this invasive amphipod highlights the need for more extensive study of the invasive and native populations of aquatic invasive invertebrates to address questions of taxonomy, diversity, and invasion history.

The purpose of this paper is to present a case study focusing on a new technology start-up firm, founded by two graduate students, an engineer and a business major, who met during their university studies. The case is timely, in that only ten percent of new product introductions result in a profitable business. The causes of failure are numerous and include the following: the market may create failure; inadequate funding and capitalization; and competition from established enterprises. Several research studies also point to rising indications of interfunctional conflict in high technology companies. One reason may be that, today, management teams in such companies are typically comprised of greater levels of diversity in age, gender, ethnicity, education, and life experiences, all of which exacerbate conflicts. New venture teams, especially in a technology start-up, may be united because of the product innovation, but they may easily become disconnected and unrealistic when it comes to the management of the enterprise. The presentation and analysis of a conflict management process herein indicates that the way a start-up team manages its conflicts may have a permanent affect on the success of its entrepreneurial venture. Conflict management does not imply terminating conflict, but involves understanding strategies to minimize dysfunction and enhancing constructive effectiveness as a result of conflict. This case exposed the problems that arise due to the differences in the founders’ education, background, experience and understanding of the necessary entrepreneurial mindset for success. By using a model of conflict management that encompasses four negotiation skill sets, including assessment, intervention, resolution and maintenance, their conflicts were resolved quickly and their partnership re-engineered, increasing the chance of their firm’s long-term success.

Background: Glioblastoma multiforme (GBM) cells are resistant to anticancer drugs. Cancer stem cells (CSCs) are a key mediator of chemoresistance. We have reported that disulfiram (DS), an aldehyde dehydrogenase (ALDH) inhibitor, targets breast CSC-like cells. In this study, the effect of DS and combination of DS and gemcitabine (dFdC) on GBM cells and GBM stem-like cells was investigated. Methods: 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), combination index (CI)-isobologram, western blot, luciferase reporter gene assay, electrophoretic mobility-shift assay and ALDH analysis were used in this study. Results: Disulfiram is cytotoxic in GBM cell lines in a copper (Cu)-dependent manner. Disulfiram/copper enhances the cytotoxicity of dFdC. Combination index-isobologram analysis indicates a synergistic effect between DS/Cu and dFdC. Disulfiram/copper induces reactive oxygen species (ROS), activates JNK and p38 pathways and inhibits nuclear factor-kappa B activity in GBM cell lines. Disulfiram/copper may trigger intrinsic apoptotic pathway via modulation of the Bcl2 family. Disulfiram/copper abolishes stem-like cell population in GBM cell lines. Conclusion: Our findings indicate that the cytotoxicity of DS/Cu and the enhancing effect of DS/Cu on the cytotoxicity of dFdC in GBM stem-like cells may be caused by induction of ROS and inhibition of both ALDH and the NFkB pathway. Both DS and dFdC can traverse the blood–brain barrier. Further study may lead them into GBM chemotherapy.

Understanding the factors contributing to expansion of nonnative populations is a critical step toward accurate risk assessment and effective management of biological invasions. Nevertheless, few studies have attempted explicitly to test hypotheses regarding factors driving invasive spread by seeking correlations between patterns of vector movement and patterns of genetic connectivity. Herein, we describe such an attempt for the invasive tunicate Styela clava in the northeastern Pacific. We utilized microsatellite data to estimate gene flow between samples collected throughout the known range of S. clava in the region, and assessed correlation of these estimates with patterns of intracoastal commercial vessel traffic. Our results suggest that recent shipping patterns have contributed to the contemporary distribution of genetic variation. However, the analysis also indicates that other factors-including a complex invasion history and the influence of other vectors-have partially obscured genetic patterns associated with intracoastal population expansion.

Anthropogenic activities frequently result in both rapidly changing environments and translocation of species from their native ranges (i.e., biological invasions). Empirical studies suggest that many factors associated with these changes can lead to complex genetic patterns, particularly among invasive populations. However, genetic complexities and factors responsible for them remain uncharacterized in many cases. Here, we explore these issues in the vase tunicate Ciona intestinalis (Ascidiacea: Enterogona: Cionidae), a model species complex, of which spA and spB are rapidly spreading worldwide. We intensively sampled 26 sites (N = 873) from both coasts of North America, and performed phylogenetic and population genetics analyses based on one mitochondrial fragment (cytochrome c oxidase subunit 3–NADH dehydrogenase subunit I, COX3-ND1) and eight nuclear microsatellites. Our analyses revealed extremely complex genetic patterns in both species on both coasts. We detected a contrasting pattern based on the mitochondrial marker: two major genetic groups in C. intestinalis spA on the west coast versus no significant geographic structure in C. intestinalis spB on the east coast. For both species, geo-graphically distant populations often showed high microsatellite-based genetic affinities whereas neighboring ones often did not. In addition, mitochondrial and nuclear markers provided largely inconsistent genetic patterns. Multiple factors, including random genetic drift associated with demographic changes, rapid selection due to strong local adaptation, and varying propensity for human-mediated propagule dispersal could be responsible for the observed genetic complexities.

PurposeSocioeconomic stress has been a major influence on organizational development during the past decade, and will continue for the foreseeable future. This treatise aims to focus on The Key and its importance for effective organizational development via leadership responses to communication challenges during the present era. It is a conceptual paper based on an actual case.Design/methodology/approachThe Key is a concept that refers to the thoughts and feelings (based on attitudes) that a leader reflects in communications and interactions with his/her universe, a universe that exists both internally within an organization as well as externally to it. The case focuses on communication challenges encountered by Judith Campbell, executive vice president of Creative Innovations, Ltd, and her interaction with Jonathan Bryant, president of Bryant and Associates, Ltd, an organizational development firm. Communication challenges that had arisen primarily due to socioeconomic stress, were seven in number: tendency to rely on only logical thinking; negative responses to external influences; lack of expectations for positive results; tendency toward reliance on sensory input; actions that ignore an inclusive perspective; failure to value and trust in change; and lack of commitment to interactive relations.FindingsFollowing Bryant's counsel, Campbell chose to address these communication challenges with a training seminar that focused on a paradigm of seven leadership responses related to The Key. These were: Paradoxical thinking; Controlled reflecting; Intentional focusing; Instinctive responding; Inclusive behaving; Purposeful trusting; and Relational being. The training program, as described herein, proved to be very enlightening to the participants and quite effective for the continued organizational development of the firm.Originality/valueThe paper reveals the results of a training program based on seven communication challenges the results of which can be used to aid in continued organizational development of a firm.

PurposeThe purpose of this paper is to explore the issue of crisis management within the Toyota Corporation's series of worldwide recalls for multiple malfunctions on a number of different Toyota brands of vehicles. The analysis relates to the difficulty now faced by Toyota, previously recognized as the world's leading manufacturer of automotive vehicles. The crisis became so great that Toyota corporate leaders even traveled from Japan to testify before a US Congressional Committee hearing.Design/methodology/approachA crisis, typically considered to be a negative issue, can be a positive event in the life of a business firm, such as Toyota, if the management involved seizes the opportunity to make appropriate changes in its operations to facilitate continuing positive growth and development. However, this opportunity was not initially addressed by Toyota in a meaningful way, and the crisis continued to evolve through subsequent stages, bringing a vast array of negative international criticism. The crisis management paradigm that is the focus for this case identifies four stages of a crisis – the preliminary (pre‐) crisis, acute crisis, chronic crisis, and crisis resolution. The present crisis deals with several different malfunctions that were identified, apparently by customers, in various Toyota brands, but publically ignored by Toyota's management. Therefore, the pre‐crisis stage was not appropriately dealt with by Toyota, and the firm was thrust into an acute crisis that has now evolved into a chronic crisis. A brief overview of the historical development of Toyota is presented, and an analysis of the present crisis situation in which the firm found itself is presented in some detail.FindingsIt was concluded that Toyota is now in a very difficult position in the chronic crisis stage due to the failure of its management to facilitate a timely response to the malfunctions of its vehicles.Originality/valueThe paper presents an excellent example of crisis mismanagement by a previously recognized world leader.