Cysteine mutations in the ebolavirus matrix protein VP40 promote phosphatidylserine binding by increasing the flexibility of a lipid-binding loop

Ebolavirus (EBOV) is a negative-sense RNA virus that causes severe hemorrhagic fever in humans. The matrix protein VP40 facilitates viral budding by binding to lipids in the host cell plasma membrane and driving the formation of filamentous, pleomorphic virus particles. The C-terminal domain of VP40 contains two highly-conserved cysteine residues at positions 311 and 314, but their role in the viral life cycle is unknown. We therefore investigated the properties of VP40 mutants in which the conserved cysteine residues were replaced with alanine. The C311A mutation significantly increased the affinity of VP40 for membranes containing phosphatidylserine (PS), resulting in the assembly of longer virus-like particles (VLPs) compared to wild-type VP40. The C314A mutation also increased the affinity of VP40 for membranes containing PS, albeit to a lesser degree than C311A. The double mutant behaved in a similar manner to the individual mutants. Computer modeling revealed that both cysteine residues restrain a loop segment containing lysine residues that interact with the plasma membrane, but Cys311 has the dominant role. Accordingly, the C311A mutation increases the flexibility of this membrane-binding loop, changes the profile of hydrogen bonding within VP40 and therefore binds to PS with greater affinity. This is the first evidence that mutations in VP40 can increase its affinity for biological membranes and modify the length of Ebola VLPs. The Cys311 and Cys314 residues therefore play an important role in dynamic interactions at the plasma membrane by modulating the ability of VP40 to bind PS

Stratospheric influence on the winter North Atlantic storm track in subseasonal reforecasts

Extreme stratospheric polar vortex events, such as sudden stratospheric warmings (SSWs) or extremely strong polar vortex events, can have a significant impact on surface weather in winter. SSWs are most often associated with negative North Atlantic Oscillation (NAO) conditions, cold air outbreaks in the Arctic and a southward-shifted midlatitude storm track in the North Atlantic, while strong polar vortex events tend to be followed by a positive phase of the NAO, relatively warm conditions in the extratropics and a poleward-shifted storm track. Such changes in the storm track position and associated extratropical cyclone frequency over the North Atlantic and Europe can increase the risk of extreme windstorm, flooding or heavy snowfall over populated regions. Skillful predictions of the downward impact of stratospheric polar vortex extremes can therefore improve the predictability of extratropical winter storms on subseasonal timescales. However, there exists a strong inter-event variability in these downward impacts on the tropospheric storm track. Using ECMWF reanalysis data and reforecasts from the Subseasonal to Seasonal (S2S) Prediction Project database, we investigate the stratospheric influence on extratropical cyclones, identified with a cyclone detection algorithm. Following SSWs, there is an equatorward shift in cyclone frequency over the North Atlantic and Europe in reforecasts, and the opposite response is observed after strong polar vortex events, consistent with the response in reanalysis. However, although the response of cyclone frequency following SSWs with a canonical surface impact is typically captured well during weeks 1–4, less than 25 % of the reforecasts manage to capture the response following SSWs with a “non-canonical” impact. This suggests a possible overconfidence in the reforecasts with respect to reanalysis in predicting the canonical response after SSWs, although it only occurs in about two-thirds of the events. The cyclone forecasts following strong polar vortex events are generally more successful. Understanding the role of the stratosphere in subseasonal variability and predictability of storm tracks during winter can provide a key for reliable forecasts of midlatitude storms and their surface impacts.</p

A narrative analysis of career transition themes and outcomes using chaos theory as a guiding metaphor

In a rapidly changing world of work little research exists on mid-career transitions. We investigated these using the open-systems approach of chaos theory as a guiding metaphor and conducted interviews with seven mid-career individuals chosen for their experience of a significant mid-career transition. Four common themes were identified through narrative analysis, where ‘false starts’ to a career were a common experience prior to finding a career ‘fit’. Career transitions, precipitated by a trigger state and/or event such as a period of disillusionment, were an important part of this ‘finding a fit’ process. Overall, career success outcomes were shaped by a combination of chaos elements: chance, unplanned events, and non-linearity of resultant outcomes. We discuss implications for future research and for practice

Plasma Membrane Integrity and Survival of Melanoma Cells After Nanosecond Laser Pulses

Circulating tumor cells (CTCs) photoacoustic detection systems can aid clinical decision-making in the treatment of cancer. Interaction of melanin within melanoma cells with nanosecond laser pulses generates photoacoustic waves that make its detection possible. This study aims at: (1) determining melanoma cell survival after laser pulses of 6 ns at λ = 355 and 532 nm; (2) comparing the potential enhancement in the photoacoustic signal using λ = 355 nm in contrast with λ = 532 nm; (3) determining the critical laser fluence at which melanin begins to leak out from melanoma cells; and (4) developing a time-resolved imaging (TRI) system to study the intracellular interactions and their effect on the plasma membrane integrity. Monolayers of melanoma cells were grown on tissue culture-treated clusters and irradiated with up to 1.0 J/cm2. Surviving cells were stained with trypan blue and counted using a hemacytometer. The phosphate buffered saline absorbance was measured with a nanodrop spectrophotometer to detect melanin leakage from the melanoma cells post-laser irradiation. Photoacoustic signal magnitude was studied at both wavelengths using piezoelectric sensors. TRI with 6 ns resolution was used to image plasma membrane damage. Cell survival decreased proportionally with increasing laser fluence for both wavelengths, although the decrease is more pronounced for 355 nm radiation than for 532 nm. It was found that melanin leaks from cells equally for both wavelengths. No significant difference in photoacoustic signal was found between wavelengths. TRI showed clear damage to plasma membrane due to laser-induced bubble formation

Development and pretesting of an information, education and communication (IEC) focused antenatal care handbook in Pakistan

BACKGROUND: Improvement of maternal morbidity and mortality indicators remains a major challenge for developing countries. Antenatal care is one of the key strategies in maintaining safe motherhood. The objective of this study was to develop and pretest a culturally relevant Antenatal Care Handbook (ANC handbook) utilizing the principles of information, education, and communication (IEC). We developed the ANC handbook after an extensive review of existing literature, available instruments (for keeping track of pregnancy and informing pregnant women), and seeking expert opinion. To pretest the ANC handbook, a cross-sectional approach was adopted, and information was collected from 300 expectant women, 150 women each from the community and from the health facility arm. Trained field workers conducted the pretesting from May 2004 to June 2004. Feedback on messages for pregnant mothers contained in the handbook was also assessed. At the same time, the ANC handbook was reviewed by 25 health care providers (including community health workers, physicians, nurses, and other health staff working at various kinds of health care facilities). Data were analyzed using both quantitative and qualitative methods. FINDINGS: Twenty-three percent of the interviewed women were primigravida, 50% were multigravida and 27% were grandmultipara. The mean age of the women in the community sample was 25.8 SD: 4.9 years and in the hospital sample it was 25.7 SD: 5.2 years. No significant differences were observed between women interviewed at community or health facilities related to their understanding of ANC messages, and the majority of messages were well understood. Similarly, health care providers found all of the instruments useful and workable in the health system. Finally, feedback from pregnant women and health care staff regarding different components of the handbook were incorporated and later verified by them. CONCLUSIONS: Findings of pretesting reveal that a majority of pregnant women have an understanding of the culturally relevant ANC handbook. The handbook was found to be practical by healthcare paraprofessionals and community workers for use in different tiers of the health care system in Pakistan. The ANC handbook can be applied in the health service sector of Pakistan and can be adopted with relevant cultural modifications by countries with a similar context

Self-organization of developing embryo using scale-invariant approach

<p>Abstract</p> <p>Background</p> <p>Self-organization is a fundamental feature of living organisms at all hierarchical levels from molecule to organ. It has also been documented in developing embryos.</p> <p>Methods</p> <p>In this study, a scale-invariant power law (SIPL) method has been used to study self-organization in developing embryos. The SIPL coefficient was calculated using a centro-axial skew symmetrical matrix (CSSM) generated by entering the components of the Cartesian coordinates; for each component, one CSSM was generated. A basic square matrix (BSM) was constructed and the determinant was calculated in order to estimate the SIPL coefficient. This was applied to developing <it>C. elegans </it>during early stages of embryogenesis. The power law property of the method was evaluated using the straight line and Koch curve and the results were consistent with fractal dimensions (fd). Diffusion-limited aggregation (DLA) was used to validate the SIPL method.</p> <p>Results and conclusion</p> <p>The fractal dimensions of both the straight line and Koch curve showed consistency with the SIPL coefficients, which indicated the power law behavior of the SIPL method. The results showed that the ABp sublineage had a higher SIPL coefficient than EMS, indicating that ABp is more organized than EMS. The fd determined using DLA was higher in ABp than in EMS and its value was consistent with type 1 cluster formation, while that in EMS was consistent with type 2.</p

Recreational and occupational field exposure to freshwater cyanobacteria – a review of anecdotal and case reports, epidemiological studies and the challenges for epidemiologic assessment

Cyanobacteria are common inhabitants of freshwater lakes and reservoirs throughout the world. Under favourable conditions, certain cyanobacteria can dominate the phytoplankton within a waterbody and form nuisance blooms. Case reports and anecdotal references dating from 1949 describe a range of illnesses associated with recreational exposure to cyanobacteria: hay fever-like symptoms, pruritic skin rashes and gastro-intestinal symptoms are most frequently reported. Some papers give convincing descriptions of allergic reactions while others describe more serious acute illnesses, with symptoms such as severe headache, pneumonia, fever, myalgia, vertigo and blistering in the mouth. A coroner in the United States found that a teenage boy died as a result of accidentally ingesting a neurotoxic cyanotoxin from a golf course pond. This death is the first recorded human fatality attributed to recreational exposure to cyanobacteria, although uncertainties surround the forensic identification of the suspected cyanotoxin in this case. We systematically reviewed the literature on recreational exposure to freshwater cyanobacteria. Epidemiological data are limited, with six studies conducted since 1990. Statistically significant increases in symptoms were reported in individuals exposed to cyanobacteria compared to unexposed counterparts in two Australian cohort studies, though minor morbidity appeared to be the main finding. The four other small studies (three from the UK, one Australian) did not report any significant association. However, the potential for serious injury or death remains, as freshwater cyanobacteria under bloom conditions are capable of producing potent toxins that cause specific and severe dysfunction to hepatic or central nervous systems. The exposure route for these toxins is oral, from ingestion of recreational water, and possibly by inhalation. A range of freshwater microbial agents may cause acute conditions that present with features that resemble illnesses attributed to contact with cyanobacteria and, conversely, acute illness resulting from exposure to cyanobacteria or cyanotoxins in recreational waters could be misdiagnosed. Accurately assessing exposure to cyanobacteria in recreational waters is difficult and unreliable at present, as specific biomarkers are unavailable. However, diagnosis of cyanobacteria-related illness should be considered for individuals presenting with acute illness following freshwater contact if a description is given of a waterbody visibly affected by planktonic mass development

Effect of sitagliptin on cardiovascular outcomes in type 2 diabetes

BACKGROUND: Data are lacking on the long-term effect on cardiovascular events of adding sitagliptin, a dipeptidyl peptidase 4 inhibitor, to usual care in patients with type 2 diabetes and cardiovascular disease. METHODS: In this randomized, double-blind study, we assigned 14,671 patients to add either sitagliptin or placebo to their existing therapy. Open-label use of antihyperglycemic therapy was encouraged as required, aimed at reaching individually appropriate glycemic targets in all patients. To determine whether sitagliptin was noninferior to placebo, we used a relative risk of 1.3 as the marginal upper boundary. The primary cardiovascular outcome was a composite of cardiovascular death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for unstable angina. RESULTS: During a median follow-up of 3.0 years, there was a small difference in glycated hemoglobin levels (least-squares mean difference for sitagliptin vs. placebo, -0.29 percentage points; 95% confidence interval [CI], -0.32 to -0.27). Overall, the primary outcome occurred in 839 patients in the sitagliptin group (11.4%; 4.06 per 100 person-years) and 851 patients in the placebo group (11.6%; 4.17 per 100 person-years). Sitagliptin was noninferior to placebo for the primary composite cardiovascular outcome (hazard ratio, 0.98; 95% CI, 0.88 to 1.09; P<0.001). Rates of hospitalization for heart failure did not differ between the two groups (hazard ratio, 1.00; 95% CI, 0.83 to 1.20; P = 0.98). There were no significant between-group differences in rates of acute pancreatitis (P = 0.07) or pancreatic cancer (P = 0.32). CONCLUSIONS: Among patients with type 2 diabetes and established cardiovascular disease, adding sitagliptin to usual care did not appear to increase the risk of major adverse cardiovascular events, hospitalization for heart failure, or other adverse events