RNA Elements in Open Reading Frames of the Bluetongue Virus Genome Are Essential for Virus Replication

Members of the Reoviridae family are non-enveloped multi-layered viruses with a double stranded RNA genome consisting of 9 to 12 genome segments. Bluetongue virus is the prototype orbivirus (family Reoviridae, genus Orbivirus), causing disease in ruminants, and is spread by Culicoides biting midges. Obviously, several steps in the Reoviridae family replication cycle require virus specific as well as segment specific recognition by viral proteins, but detailed processes in these interactions are still barely understood. Recently, we have shown that expression of NS3 and NS3a proteins encoded by genome segment 10 of bluetongue virus is not essential for virus replication. This gave us the unique opportunity to investigate the role of RNA sequences in the segment 10 open reading frame in virus replication, independent of its protein products. Reverse genetics was used to generate virus mutants with deletions in the open reading frame of segment 10. Although virus with a deletion between both start codons was not viable, deletions throughout the rest of the open reading frame led to the rescue of replicating virus. However, all bluetongue virus deletion mutants without functional protein expression of segment 10 contained inserts of RNA sequences originating from several viral genome segments. Subsequent studies showed that these RNA inserts act as RNA elements, needed for rescue and replication of virus. Functionality of the inserts is orientation-dependent but is independent from the position in segment 10. This study clearly shows that RNA in the open reading frame of Reoviridae members does not only encode proteins, but is also essential for virus replicatio

In search for the sources of plastic marine litter that contaminates the Easter Island Ecoregion

Subtropical gyres are the oceanic regions where plastic litter accumulates over long timescales, exposing surrounding oceanic islands to plastic contamination, with potentially severe consequences on marine life. Islands’ exposure to such contaminants, littered over long distances in marine or terrestrial habitats, is due to the ocean currents that can transport plastic over long ranges. Here, this issue is addressed for the Easter Island ecoregion (EIE). High-resolution ocean circulation models are used with a Lagrangian particle-tracking tool to identify the connectivity patterns of the EIE with industrial fishing areas and coastline regions of the Pacific basin. Connectivity patterns for “virtual” particles either floating (such as buoyant macroplastics) or neutrally-buoyant (smaller microplastics) are investigated. We find that the South American shoreline between 20°S and 40°S, and the fishing zone within international waters off Peru (20°S, 80°W) are associated with the highest probability for debris to reach the EIE, with transit times under 2 years. These regions coincide with the most-densely populated coastal region of Chile and the most-intensely fished region in the South Pacific. The findings offer potential for mitigating plastic contamination reaching the EIE through better upstream waste management. Results also highlight the need for international action plans on this important issue

Profound variation in dihydropyrimidine dehydrogenase activity in human blood cells: major implications for the detection of partly deficient patients

Dihydropyrimidine dehydrogenase (DPD) is responsible for the breakdown of the widely used antineoplastic agent 5-fluorouracil (5FU), thereby limiting the efficacy of the therapy. To identify patients suffering from a complete or partial DPD deficiency, the activity of DPD is usually determined in peripheral blood mononuclear cells (PBM cells). In this study, we demonstrated that the highest activity of DPD was found in monocytes followed by that of lymphocytes, granulocytes and platelets, whereas no significant activity of DPD could be detected in erythrocytes. The activity of DPD in PBM cells proved to be intermediate compared with the DPD activity observed in monocytes and lymphocytes. The mean percentage of monocytes in the PBM cells obtained from cancer patients proved to be significantly higher than that observed in PBM cells obtained from healthy volunteers. Moreover, a profound positive correlation was observed between the DPD activity of PBM cells and the percentage of monocytes, thus introducing a large inter- and intrapatient variability in the activity of DPD and hindering the detection of patients with a partial DPD deficiency. © 1999 Cancer Research Campaig

The influence of learner characteristics on degree and type of participation in a CSCL environment

Computer-Supported Collaborative Learning (CSCL) is often presented as a promising learning method. However, it is also facing some new challenges. Apart from answering the question of whether or not working with CSCL generates satisfying learning outcomes, it is important to determine whether or not all participants profit from collaboration, with the computer as a means of communication. This paper describes the implementation and effects of an experimental program in 5 classes with a total of 120 students in elementary education who, in groups of four, engaged in Knowledge Forum discussion tasks on the subject of healthy eating. The study explores whether or not differences occur in the participation of students who differ in gender, sociocultural background and ability, and whether or not computer skills, computer attitudes, comprehensive reading scores and popularity with classmates are related to student participation. Students’ participation in this CSCL environment appears to be dependent on a number of learner characteristics. Girls contribute more words to the discussions than boys do and are more dependent on their computer skills in this production. Students who are good at comprehensive reading also contribute more words. Popularity among classmates appears to influence the degree of participation further. We also found indications that students with immigrant parents write fewer contributions than those whose parents are not immigrants

Water mass analysis along 22°N in the subtropical North Atlantic for the JC150 cruise (GEOTRACES, GApr08)

This study presents a water mass analysis along the JC150 section in the subtropical North Atlantic, based on hydrographic and nutrient data, by combining an extended optimum multiparameter analysis (OMPA) with a Lagrangian particle tracking experiment (LPTE). This combination, which was proposed for the first time, aided in better constraining the OMPA end-member choice and providing information about their trajectories. It also enabled tracing the water mass origins in surface layers, which cannot be achieved with an OMPA. The surface layers were occupied by a shallow type of Eastern South Atlantic Central Water (ESACW) with traces of the Amazon plume in the west. Western North Atlantic Central Water dominates from 100 to 500 m, while the 13°C ESACW contribution occurs marginally deeper (500–900 m). At approximately 700 m, Antarctic Intermediate Water (AAIW) dominates the west of the Mid-Atlantic Ridge (MAR), while Mediterranean Water dominates the east with a small but non-negligible contribution down to 3500 m. Below AAIW, Upper Circumpolar Deep Water (UCDW) is observed throughout section (900–1250 m). Labrador Sea Water (LSW) is found centered at 1500 m, where the LPTE highlights an eastern LSW route from the eastern North Atlantic to the eastern subtropical Atlantic, which was not previously reported. North East Atlantic Deep Water (encompassing a contribution of Iceland-Scotland Overflow Water) is centered at ~2500 m, while North West Atlantic Bottom Water (NWABW, encompassing a contribution of Denmark Strait Overflow Water) is principally localized in the west of the MAR in the range of 3500–5000 m. NWABW is also present in significant proportions (>25%) in the east of the MAR, suggesting a crossing of the MAR possibly through the Kane fracture zone. This feature has not been investigated so far. Finally, Antarctic Bottom Water is present in deep waters throughout the section, mainly in the west of the MAR. Source waters have been characterized from GEOTRACES sections, which enables estimations of trace elements and isotope transport within water masses in the subtropical North Atlantic

Association of Type 2 Diabetes, According to the Number of Risk Factors Within Target Range, With Structural Brain Abnormalities, Cognitive Performance, and Risk of Dementia

OBJECTIVE: Type 2 diabetes is associated with increased risks of cognitive dysfunction and brain abnormalities. The extent to which risk factor modification can mitigate these risks is unclear. We investigated the associations between incident dementia, cognitive performance, and brain abnormalities among individuals with type 2 diabetes, according to the number of risk factors on target, compared with control subjects without diabetes. RESEARCH DESIGN AND METHODS: Prospective data were from UK Biobank of 87,856 individuals (n = 10,663 diabetes, n = 77,193 control subjects; baseline 2006-2010), with dementia follow-up until February 2018. Individuals with diabetes were categorized according to the number of seven selected risk factors within the guideline-recommended target range (nonsmoking; guideline-recommended levels of glycated hemoglobin, blood pressure, BMI, albuminuria, physical activity, and diet). Outcomes were incident dementia, domain-specific cognitive performance, white matter hyperintensities, and total brain volume. RESULTS: After a mean follow-up of 9.0 years, 147 individuals (1.4%) with diabetes and 412 control subjects (0.5%) had incident dementia. Among individuals with diabetes, excess dementia risk decreased stepwise for a higher number of risk factors on target. Compared with control subjects (incidence rate per 1,000 person-years 0.62 [95% CI 0.56; 0.68]), individuals with diabetes who had five to seven risk factors on target had no significant excess dementia risk (absolute rate difference per 1,000 person-years 0.20 [-0.11; 0.52]; hazard ratio 1.32 [0.89; 1.95]). Similarly, differences in processing speed, executive function, and brain volumes were progressively smaller for a higher number of risk factors on target. These results were replicated in the Maastricht Study. CONCLUSIONS: Among individuals with diabetes, excess dementia risk, lower cognitive performance, and brain abnormalities decreased stepwise for a higher number of risk factors on target

Interaction of classical swine fever virus with dendritic cells

Functional disruption of dendritic cells (DCs) is an important strategy for viral pathogens to evade host defences. Monocytotropic viruses such as classical swine fever virus (CSFV) could employ such a mechanism, since the virus can suppress immune responses and induce apoptosis without infecting lymphocytes. Here, CSFV was shown to infect and efficiently replicate in monocyte- and in bone marrow-derived DCs. Interestingly, the infected DCs displayed neither modulated MHC nor CD80/86 expression. Stimulation of DCs with IFN-/TNF- or polyinosinic¿polycytidylic acid (pIC) induced phenotypic maturation with increased MHC and CD80/86 expression, both with mock-treated and infected DCs. In addition, the T cell stimulatory capacity of CSFV-infected DCs was maintained both in a polyclonal T cell stimulation and in specific antigen-presentation assays, requiring antigen uptake and processing. Interestingly, similar to macrophages, CSFV did not induce IFN- responses in these DCs and even suppressed pIC-induced IFN- induction. Other cytokines including interleukin (IL)-6, IL-10, IL-12 and TNF- were not modulated. Taken together, these results demonstrated that CSFV can replicate in DCs and control IFN type I responses, without interfering with the immune reactivity. These results are interesting considering that DC infection with RNA viruses usually results in DC activation

Key climate change stressors of marine ecosystems along the path of the east african coastal current

For the countries bordering the tropical Western Indian Ocean (TWIO), living marine resources are vital for food security. However, this region has largely escaped the attention of studies investigating potential impacts of future climate change on the marine environment. Understanding how marine ecosystems in coastal East Africa may respond to various climatic stressors is vital for the development of conservation and other ocean management policies that can help to adapt to climate change impacts on natural and associated human systems. Here, we use a high-resolution (1/4°) ocean model, run under a high emission scenario (RCP 8.5) until the end of the 21st century, to identify key regionally important climate change stressors over the East African Coastal Current (EACC) that flows along the coasts of Kenya and Tanzania. We also discuss these stressors in the context of projections from lower resolution CMIP5 models. Our results indicate that the main drivers of dynamics and the associated ecosystem response in the TWIO are different between the two monsoon seasons. Our high resolution model projects weakening of the Northeast monsoon (December–February) winds and slight strengthening of the Southeast monsoon (May–September) winds throughout the course of the 21st century, consistent with CMIP5 models. The projected shallower mixed layers and weaker upwelling during the Northeast Monsoon considerably reduce the availability of surface nutrients and primary production. Meanwhile, primary production during the Southeast monsoon is projected to be relatively stable until the end of the century. In parallel, a widespread warming of up to 5 °C is projected year-round with extreme events such as marine heatwaves becoming more intense and prolonged, with the first year-long event projected to occur as early as the 2030s. This extreme warming will have significant consequences for both marine ecosystems and the coastal populations dependent on these marine resources. These region-specific stressors highlight the importance of dynamic ocean features such as the upwelling systems associated with key ocean currents. This indicates the need to develop and implement a regional system that monitors the anomalous behaviour of such regionally important features. Additionally, this study draws attention to the importance of investment in decadal prediction methods, including high resolution modelling, that can provide information at time and space scales that are more directly relevant to regional management and policy making

Reduced 5-FU clearance in a patient with low DPD activity due to heterozygosity for a mutant allele of the DPYD gene

5-fluorouracil pharmacokinetics, dihydropyrimidine dehydrogenase-activity and DNA sequence analysis were compared between a patient with extreme 5-fluorouracil induced toxicity and six control patients with normal 5-fluorouracil related symptoms. Patients were treated for colorectal cancer and received chemotherapy consisting of leucovorin 20 mg m−2 plus 5-fluorouracil 425 mg m−2. Blood sampling was carried out on day 1 of the first cycle. The 5-fluorouracil area under the curve0→3h in the index patient was 24.1 mg h l−1 compared to 9.8±3.6 (range 5.4–15.3) mg h l−1 in control patients. The 5-fluorouracil clearance was 520 ml min−1 vs 1293±302 (range 980–1780) ml min−1 in controls. The activity of dihydropyrimidine dehydrogenase in mononuclear cells was lower in the index patient (5.5 nmol mg h−1) compared to the six controls (10.3±1.6, range 8.0–11.7 nmol mg h−1). Sequence analysis of the dihydropyrimidine dehydrogenase gene revealed that the index patient was heterozygous for a IVS14+1G>A point mutation. Our results indicate that the inactivation of one dihydropyrimidine dehydrogenase allele can result in a strong reduction in 5-fluorouracil clearance, causing severe 5-fluorouracil induced toxicity