Molecular sensor-based monitoring of toxic algae

Planktonic algae are the most abundant photosynthetic organisms. They are the basis of the marine food web, and changes in phytoplankton communities generally provide an early indication for climate-driven modifications of marine food webs and the whole ecosystem. Harmful algal blooms (HAB) are a subset of planktonic algal species that have negative impacts on humans and aquatic environments. The formation of HABs depends on several multifactorial climate and environmental drivers that influence timing and frequency of these algal blooms. There is some evidence that the occurrence of HAB might become more intense, widespread, frequent and unexpected in future decades due to climate variability. In North ocean regions the global climate change is reflected by changing environmental parameter, such as increasing sea surface temperature, modified water mass stratification and rising freshwater content in the Arctic realm. Within this study the occurrence, abundance and dispersal of toxic algae species is analysed in Nordic seas and the Arctic Ocean, based on molecular detection with a nucleic acid biosensor. Alongside with this it will be assessed whether biosensor systems can serve as an early warning system for human health policies in the field of water resource management

Monitoring seasonal dynamics of selected harmful algae in the North Sea using molecular methods

Microalgae are the major producers of biomass and organic compounds in the aquatic environment. Among them there are toxic species (mainly dinoflagellates) known to have the potential to form Harmful Algal Blooms, the so called HABs. HABs are occurring more often and at new locations. In general, knowledge about biogeographic distribution of harmful algae in the northern hemisphere is limited and patchy. During this project, we will study the seasonal dynamics of marine protists, with special emphasis on toxic algae in the North Sea. Samples will be taken from four geographical distinct locations in the German Bight and from the Orkney Islands. Protist community composition will be assessed by Illumina sequencing and a newly developed fully automated biosensor system. The latter allows for automated sampling and filtration of water samples and automated detection of selected toxic algal species, while the detection is based upon electro chemical quantification of RNA by sandwich hybridization. Here we show first results of calibrating the biosensor for selected toxic algae that are known to occur in the North Sea. Furthermore, we also show preliminary results of the characterization of protist communities from spring to autumn 2016 at four different observation locations in the North Sea via Illumina sequencing

High-resolution monitoring of toxic dinoflagellate species and their biogeographic distribution in the North Atlantic and Polar Seas

Dinoflagellate species of the genus Dinophysis have become target organisms for surveillance and monitoring of microalgae as they may produce potent diarrhetic shellfish toxins and therefore have negative socio-economic impacts. The formation of Dinophysis blooms as well as toxin composition and cellular toxin content depends on several multifactorial climate and environmental drivers and it might be expected that the occurrence of toxic events becomes more intense, widespread, frequent and unexpected in future decades due to climate variability. Conventional methods for the identification of microalgae e.g. microscopy, still have some deficiencies as they are very time-consuming and need special knowledge and experience, especially in case of difficult morphological species distinction. Standard quantification methods also might fail to detect and determine Dinophysis species due to their typically low cell densities and their spatial heterogeneity (=patchiness). Therefore innovative technologies for environmental monitoring of toxic microalgae are needed to prevent humans and aquatic environments from toxic threats and damage. We analysed the occurrence, abundance and dispersal of toxic dinoflagellate species in Nordic seas and the Arctic Ocean. Genetic analyses included a modular composed autonomous rRNA biosensor approach that allows rapid, precise and economically efficient high-resolution quantification and identification of microalgae in aquatic environments. Next generation sequencing (Illumina) was used to get additional information on distributional patterns of the most common dinoflagellate species in the observation area

The pyrrolizidine alkaloid senecionine induces CYP-dependent destruction of sinusoidal endothelial cells and cholestasis in mice

Pyrrolizidine alkaloids (PAs) are widely occurring phytotoxins which can induce severe liver damage in humans and other mammalian species by mechanisms that are not fully understood. Therefore, we investigated the development of PA hepatotoxicity in vivo, using an acutely toxic dose of the PA senecionine in mice, in combination with intravital two-photon microscopy, histology, clinical chemistry, and in vitro experiments with primary mouse hepatocytes and liver sinusoidal endothelial cells (LSECs). We observed pericentral LSEC necrosis together with elevated sinusoidal marker proteins in the serum of senecionine-treated mice and increased sinusoidal platelet aggregation in the damaged tissue regions. In vitro experiments showed no cytotoxicity to freshly isolated LSECs up to 500 µM senecionine. However, metabolic activation of senecionine by preincubation with primary mouse hepatocytes increased the cytotoxicity to cultivated LSECs with an EC50 of approximately 22 µM. The cytochrome P450 (CYP)-dependency of senecionine bioactivation was confirmed in CYP reductase-deficient mice where no PA-induced hepatotoxicity was observed. Therefore, toxic metabolites of senecionine are generated by hepatic CYPs, and may be partially released from hepatocytes leading to destruction of LSECs in the pericentral region of the liver lobules. Analysis of hepatic bile salt transport by intravital two-photon imaging revealed a delayed uptake of a fluorescent bile salt analogue from the hepatic sinusoids into hepatocytes and delayed elimination. This was accompanied by transcriptional deregulation of hepatic bile salt transporters like Abcb11 or Abcc1. In conclusion, senecionine destroys LSECs although the toxic metabolite is formed in a CYP-dependent manner in the adjacent pericentral hepatocytes.</p

High-resolution monitoring of marine protists based on an observation strategy integrating automated on-board ship filtration and molecular analyses

Information on recent biomass distribution and biogeography of photosynthetic marine protists with adequate temporal and spatial resolution is urgently needed to better understand consequences of environmental change for marine ecosystems. Here we introduce and review a molecular-based observation strategy for high resolution assessment of these protists in space and time. It is the result of extensive technology developments, adaptations and evaluations which are documented in a number of different publications and the results of recently accomplished field testing, which are introduced in this review. The observation strategy is organized at four different levels. At level 1, samples are collected at high spatio-temporal resolution using the remote-controlled automated filtration system AUTOFIM. Resulting samples can either be preserved for later laboratory analyses, or directly subjected to molecular surveillance of key species aboard the ship via an automated biosensor system or quantitative polymerase chain reaction (level 2). Preserved samples are analyzed at the next observational levels in the laboratory (level 3 and 4). This involves at level 3 molecular fingerprinting methods for a quick and reliable overview of differences in protist community composition. Finally, selected samples can be used to generate a detailed analysis of taxonomic protist composition via the latest Next Generation Sequencing Technology (NGS) at level 4. An overall integrated dataset of the results based on the different analyses provides comprehensive information on the diversity and biogeography of protists, including all related size classes. At the same time the cost effort of the observation is optimized in respect to analysis effort and time

Tumor Ulceration Does Not Fully Explain Sex Disparities in Melanoma Survival among Adolescents and Young Adults

Hypertension in kidney transplant recipients (KTRs) is a risk factor for cardiovascular mortality and graft loss. Data on the prevalence of hypertension and uncontrolled hypertension (uHT) in paediatric and young adult KTRs are scarce. Also, it is unknown whether 'transition' (the transfer from paediatric to adult care) influences control of hypertension. We assessed the prevalence of hypertension and uHT among Dutch paediatric and young adult KTRs and analysed the effects of transition. Additionally, we made an inventory of variations in treatment policies in Dutch transplant centres. Cross-sectional and longitudinal national data from living KTRs a parts per thousand currency sign30 years of age (a parts per thousand yen1-year post-transplant, eGFR > 20 mL/min) were extracted from the 'RICH Q' database, which comprises information about all Dutch KTRs <19 years of age, and the Netherlands Organ Transplant Registry database for adult KTRs (a parts per thousand yen18-30 years of age). We used both upper-limit blood pressure (BP) thresholds for treatment according to Kidney Disease: Improving Global Outcomes (KDIGO) guidelines. uHT was defined as a BP above the threshold. A questionnaire on treatment policies was sent to paediatric and adult nephrologists at eight Dutch transplant centres. Hypertension and uHT were more prevalent in young adult KTRs (86.4 and 75.8%) than in paediatric KTRs (62.7 and 38.3%) according to the KDIGO definition. Time after transplantation was comparable between these groups. Longitudinal analysis showed no evidence of effect of transition on systolic BP or prevalence of uHT. Policies vary considerably between and within centres on the definition of hypertension, BP measurement and antihypertensive treatment. Average BP in KTRs increases continuously with age between 6 and 30 years. Young adult KTRs have significantly more uHT than paediatric KTRs according to KDIGO guidelines. Transition does not influence the prevalence of uHT

Cost-Effectiveness of New Cardiac and Vascular Rehabilitation Strategies for Patients with Coronary Artery Disease

Objective: Peripheral arterial disease (PAD) often hinders the cardiac rehabilitation program. The aim of this study was evaluating the relative cost-effectiveness of new rehabilitation strategies which include the diagnosis and treatment of PAD in patients with coronary artery disease (CAD) undergoing cardiac rehabilitation. Data Sources: Best-available evidence was retrieved from literature and combined with primary data from 231 patients. Methods: We developed a Markov decision model to compare the following treatment strategies: 1. cardiac rehabilitation only; 2. ankle-brachial index (ABI) if cardiac rehabilitation fails followed by diagnostic work-up and revascularization for PAD if needed; 3. ABI prior to cardiac rehabilitation followed by diagnostic work-up and revascularization for PAD if needed. Quality-adjusted-life years (QALYs), life-time costs (US $), incremental cost-effectiveness ratios (ICER), and gain in net health benefits (NHB) in QALY equivalents were calculated. A threshold willingness-to-pay of$75 000 was used. Results: ABI if cardiac rehabilitation fails was the most favorable strategy with an ICER of $44 251 per QALY gained and an incremental NHB compared to cardiac rehabilitation only of 0.03 QALYs (95$75 000/QALY. After sensitivity analysis, a combined cardiac and vascular rehabilitation program increased the success rate and would dominate the other two strategies with total lifetime costs of $30 246 a quality-adjusted life expectancy of 3.84 years, and an incremental NHB of 0.06 QALYs (95%CI:−0.24, 0.46) compared to current practice. The results were robust for other different input parameters. Conclusion: ABI measurement if cardiac rehabilitation fails followed by a diagnostic work-up and revascularization for PAD if needed are potentially cost-effective compared to cardiac rehabilitation only

Molecular sensor based monitoring of harmful algae

Biohazards like harmful microalgae have negative impacts on human population, local environments and economies. Therefore innovative technologies for environmental monitoring and surveillance of harmful microalgae are needed to prevent humans and aquatic environments from toxic threats. In our project we developed a modular composed semi-autonomous approach for rapid, precise and economically efficient monitoring of microalgae in aquatic environments. The approach involves two modules. The automatic and remote-controlled filtration module is a unit for preparation of free nucleic acids from water samples. This includes sampling, filtration and cell lysis, mediated by ultrasound. Subsequently, the free nucleic acids e.g. rRNA can be detected with an automated nucleic acid biosensor, based on a sandwich-hybridization of target DNA with species-specific designed probes. Hybridization is transformed in an electrochemical and measureable signal. Probes, specific for different target organisms can be applied for molecular detection. Thus the nucleic acid biosensor allows the assessment of a broad spectrum of microorganisms including harmful microalgae. Filtration and detection module should be connected to one compact, autonomous system that can be used for different fields of application

Neuropsykiatriska diagnoser och identitet : ungdomars och unga vuxnas upplevelse av att få och leva med en neuropsykiatrisk diagnos.

Neuropsykiatriska diagnoser hos barn och vuxna ökar. Två av de mest förekommande diagnoserna är ADHD och Autismspektrumtillstånd (AST). Båda dessa diagnoser innefattar symptom inom socialt samspel och kognitiv prestation, vilket inverkar på individens vardag, självbild och identitet. Syftet med föreliggande studie var att undersöka individens upplevelse av hur identiteten påverkas av att få och leva med en neuropsykiatrisk diagnos. Elva deltagare i åldersspannet 15 till 30 år rekryterades. Genom semistrukturerade intervjuer och en tematisk analys i två delar; en induktiv och en deduktiv, framkom sammanlagt tolv teman. Resultatet beskriver positiva aspekter med diagnosen såsom identifikation, stöd och förståelse, samt negativa aspekter såsom fördomar, diskriminering och en upplevelse av att vara annorlunda. En teoretisk analys utifrån identitetsteori visade att problemen huvudsakligen uppstod i mötet med omgivningen och att omgivningens syn på och förhållningssätt till neuropsykiatriska funktionsnedsättningar behöver diskuteras för att skapa förändring

Kvinnors upplevelser efter mastektomi vid bröstcancer

Bakgrund: Bröstcancer är den vanligaste cancerdiagnosen hos kvinnor globalt och i Sverige. Diagnosen är den främsta anledningen till en förtidig död bland kvinnor. En vanlig behandling är mastektomi, vilket innebär delvis eller total borttagning av ett eller båda brösten. Kvinnors upplevelser är individuella då de har en egen livsvärld. Syfte: Att beskriva kvinnors upplevelser efter mastektomi vid bröstcancer.  Metod: En kvalitativ litteraturöversikt med induktiv ansats. Analysen har gjorts i fem steg enligt Friberg. Resultatet grundas på 19 kvalitativa artiklar.  Resultat: Resultatet visade en förändrad kroppsbild hos kvinnor som genomgått mastektomi vilket kunde resultera i en försämrad självkänsla. Ett bortopererat bröst orsakade en förändrad syn kring identitet, kvinnlighet och sexualitet. En känsla av normalitet och symmetri blev viktig hos kvinnorna och det kunde bli aktuellt med bröstprotes eller bröstrekonstruktion. Dessa upplevelser påverkade deras liv på olika sätt. Kvinnorna accepterade sin nya situation efter olika lång tid och stöd från omgivningen var betydelsefullt. Kvinnorna hade olika behov och förväntningar på stöd från sjuksköterskor.     Slutsats: Resultatet visar på vad kvinnor kan uppleva efter mastektomi vid bröstcancer. Genom ökad kunskap och förståelse kring kvinnors upplevelser ska sjuksköterskan ge individuellt anpassat stöd utifrån deras livsvärld som kan tillgodose kvinnornas behov